RIRDC · seed post harvest. A seed testing kit should be developed for producers to test the seed...

Understanding and Managing the Causes of Abnormal Seedlings in Lucerne

RIRDCInnovation for rural Australia

RIRDC Publication No. 08/023

08-023 Final Report covers.indd 2 29/05/2008 9:46:25 AM

Understanding and Managing the Causes of Abnormal

Seedlings in Lucerne

by James De Barro

February 2008

RIRDC Publication No 08/023 RIRDC Project No. DEB-4A

ii

© 2008 Rural Industries Research and Development Corporation. All rights reserved. Understanding and Managing the Causes of Abnormal Seedlings in Lucerne ISBN 1 74151 612 9 ISSN 1440-6845 Publication No. 08/023 Project No. DEB-4A The information contained in this publication is intended for general use to assist public knowledge and discussion and to help improve the development of sustainable regions. You must not rely on any information contained in this publication without taking specialist advice relevant to your particular circumstances. While reasonable care has been taken in preparing this publication to ensure that information is true and correct, the Commonwealth of Australia gives no assurance as to the accuracy of any information in this publication. The Commonwealth of Australia, the Rural Industries Research and Development Corporation (RIRDC), the authors or contributors expressly disclaim, to the maximum extent permitted by law, all responsibility and liability to any person, arising directly or indirectly from any act or omission, or for any consequences of any such act or omission, made in reliance on the contents of this publication, whether or not caused by any negligence on the part of the Commonwealth of Australia, RIRDC, the authors or contributors. The Commonwealth of Australia does not necessarily endorse the views in this publication. This publication is copyright. Apart from any use as permitted under the Copyright Act 1968, all other rights are reserved. However, wide dissemination is encouraged. Requests and inquiries concerning reproduction and rights should be addressed to the RIRDC Publications Manager on phone 02 6271 4165.

Researcher Contact Details James De Barro Alpha Group Consulting Pty Ltd P.O. Box 292 KEITH SA 5267 Ph: 08 87551502 Fax: 08 87551 501 Mobile: 0417 946 053 www.thealphagroup.com.au Email: [email protected]

RIRDC Contact Details Rural Industries Research and Development Corporation Level 2, 15 National Circuit BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6271 4100 Fax: 02 6271 4199 Email: [email protected]. Web: http://www.rirdc.gov.au Published in February 2008 by Canprint

http://www.thealphagroup.com.au/

iii

Foreword The Australian lucerne seed industry is estimated to be valued over A$100 million to the Australian economy with approximately 90% of production occurring in a 40 km radius surrounding the township of Keith in South Australia. Whether lucerne seed is used for sowing pasture or sprouting, its value is based fundamentally on market forces and seed quality. The Australian market is significantly export oriented and supply and demand factors are, in general, not manageable from the farm gate. However, seed quality is manageable within the farm gate as well as through the seed cleaning process. Whether seed is produced under a certified scheme or not, there are world standards that apply to differentiate high quality seed from that of lesser quality. The final cleaned seed sample is assessed in accredited International Seed Testing Association (ISTA) laboratories according to international seed testing rules. Seed quality is defined as percentages of normal seedlings, hard seed, dead seed and abnormal seedlings which are present after a set period of germination according to the ISTA guidelines. To meet the highest Australian standard, seed lines must have no less than 85% of the representative seed sample being comprised of a normal seedling and hard seed count. A problem arises when the abnormal seedling presence (and extremely rarely dead seed) exceeds 15% which reduces the seed line below the 85% minimum. Sub-standard seed lines are more difficult to sell, particularly in low demand markets, and can be devalued significantly. Individual producers have received discounts of 12-20% on their seed sales which significantly reduces gross margin profits. This research identified that the principal cause of abnormal seedlings is due to harvest damage with minimal abnormality due to other reasons prior to harvest. In addition, the research also found that certain lucerne seed crops were predisposed to higher risk of harvest damage and hence abnormal seedling production. A high correlation was found between seed damage and abnormal seedling production. The research determined that the process of harvest can be managed to mitigate the percentage of abnormal seedlings and that a simple Ferric Chloride test can be used through the harvesting process to monitor seed damage and hence abnormal seedling creation. These research findings will permit producers to identify high risk paddocks, assisting in managing the seed post harvest. A seed testing kit should be developed for producers to test the seed through harvest, to assist in header management as well as identify problem lines prior to delivery. Producers will be able to make judged decisions regarding their seed marketing by virtue of knowing their seeds likely quality. It is also important that producers are aware that lucerne seeds are living organisms and that different paddocks may require different header management during harvest. This project is funded by De Barro Agricultural Consulting, Waite Analytical Services, Seed Services Australia and industry revenue that is matched by funds provided by the Australian Government. This report, an addition to RIRDC’s diverse range of over 1700 research publications, forms part of our Pasture Seeds R&D sub-program, which aims to facilitate the growth of a profitable and sustainable pasture seeds industry based on a reputation for the reliable supply, domestically and internationally, of a range of pasture species. Most of our publications are available for viewing, downloading or purchasing online through our website: • downloads at www.rirdc.gov.au/fullreports/index.html • purchases at www.rirdc.gov.au/eshop Peter O’Brien Managing Director Rural Industries Research and Development Corporation

http://www.rirdc.gov.au/fullreports/index.html

http://www.rirdc.gov.au/eshop

iv

Acknowledgments The author wishes to acknowledge the co-operation of the following seed producers: Makin Farms D. Hore B. & L. Loller Scotswell Partners N. Densley P.S. & J.H. Meyer Mardango Props R. Densley G. & W. Ryan Bungalally Farms Narkoona Partners K. & R. Sherriff Kermel Pty Ltd Kalimnah Downs R. & T. Wilsdon The Smart Group W. & L. Lehmann P. & A. Davidson W.R. Hunt & Sons H.A & L.M. Densley P. Richardson J. & H. Cozens G. & H. Lehmann A special mention is made to Graham Ramsdale and staff of Tatiara Seeds for their assistance. Special mention is made to staff of Seed Services Australia in particular Heather Lawrie and Annie Dale – especially for the photography of abnormal seedlings. Additionally Seed Services Australia, Waite Analytical Services, Seedmark, Keith Seeds, Alison Graham and Glenn McDonald and the University of Adelaide, RIRDC, Kongal Seeds and staff of De Barro Agricultural Consulting are acknowledged for their contributions.

About the author James De Barro owns and manages De Barro Agricultural Consulting. James has an honours degree in Agricultural Science and a Graduate Diploma of Business and was awarded a Churchill Fellowship in 1999. James is an inaugural member of the Lucerne Australia executive committee. James resides in Keith in South Australia and specialises in consulting to lucerne seed producers and industry regarding all facets of seed production in dryland and irrigated systems. James is responsible for the ongoing research focus of the business that finances several projects. James is involved in industry policy development and promoting the lucerne seed industry through membership of Lucerne Australia.

Don’t find a fault, find a remedy

Henry Ford

v

Contents Foreword.......................................................................................................................................... iii Acknowledgments ............................................................................................................................iv About the author ...............................................................................................................................iv List of Tables and Graphs ............................................................................................................... vii Executive Summary ....................................................................................................................... viii 1. Introduction....................................................................................................................................1 1.1 General Overview.................................................................................................................1

1.1.2 Research area......................................................................................................................1 1.2 Seed quality standards ..........................................................................................................1

1.2.1 Seed production chain ........................................................................................................1 1.2.2 Seed assessment protocol ...................................................................................................2 1.2.3 Seed standards ....................................................................................................................2 1.2.4 Trading of Australia’s lucerne seed....................................................................................2

1.3 Literature review ..................................................................................................................3 1.3.1 Industry funded literature review .......................................................................................3 1.3.2 Summary of review ............................................................................................................3 1.3.3 Photo album of abnormal seedlings ...................................................................................3

2. Objectives ......................................................................................................................................4 2.1 Purpose of research ......................................................................................................................4 3. Methodology..................................................................................................................................5 3.1 Overview......................................................................................................................................5

3.1.1 Sample selection.................................................................................................................5 3.1.2 Sample Collection ..............................................................................................................6 3.1.3 Sample management and preparation.................................................................................6 3.1.4 Sample testing ....................................................................................................................6 3.1.5 Refinement of research over time.......................................................................................6 3.1.6 Development of Ferric Chloride test ..................................................................................7 3.1.7 Assessment of the test ........................................................................................................7

4. Summary of Results.......................................................................................................................8 4.1 Seasonal records ..........................................................................................................................8 4.2 Growing conditions......................................................................................................................8 4.3 Environmental conditions ............................................................................................................8 4.4 Variety, chemical, windrowing, time of harvest factors ..............................................................8 4.5 Seed moisture...............................................................................................................................8 4.6 Handling damage .........................................................................................................................8 4.7 Header type ..................................................................................................................................8 4.8 Nutritional analysis ......................................................................................................................8 4.9 Harvest damage..........................................................................................................................14 4.10 Ferric Chloride assessment ......................................................................................................15 4.11 Ferric Chloride field evaluation ...............................................................................................16

vi

5. Discussion of results ....................................................................................................................17 5.1 Growing conditions....................................................................................................................17 5.2 Environmental conditions ..........................................................................................................17 5.3 Variety, chemical, windrowing, time of harvest factors ............................................................17 5.4 Seed moisture.............................................................................................................................18 5.5 Handling damage .......................................................................................................................18 5.6 Header type ................................................................................................................................18 5.7 Nutritional analysis ....................................................................................................................18 5.8 Harvest damage..........................................................................................................................18 5.9 Ferric Chloride assessment ........................................................................................................19 5.10 Ferric Chloride field evaluation ...............................................................................................21 6. Implications of research...............................................................................................................22 7. Recommendations........................................................................................................................23 8. References....................................................................................................................................24 Appendix 1 Literature Review.........................................................................................................25 Appendix 2: 03/04 seed quality data................................................................................................60 Appendix 3: 03/04 harvest data .......................................................................................................64 Appendix 4: 04/05 seed quality data................................................................................................68 Appendix 5: 04/05 harvest data .......................................................................................................73 Appendix 6: 05/06 seed quality data................................................................................................78 Appendix 6: 05/06 seed quality data................................................................................................78 Appendix 7: 05/06 harvest data .......................................................................................................81 Appendix 7: 05/06 harvest data .......................................................................................................81 Appendix 8: Ferric Chloride test assessment data ...........................................................................84 Appendix 9: Ferric Chloride test header data ..................................................................................84 Appendix 10: Photo album of abnormal seedlings ..........................................................................85

vii

List of Tables and Graphs Table 1: 02/03 pre research commencement lucerne seed nutritional analysis 9 Table 2: 03/04 lucerne seed nutritional analysis 10 Table 3: 04/05 lucerne seed nutritional analysis 11 Graph 1: Average lucerne seed quality through harvest process 03/04 14 Graph 2: Average lucerne seed quality through harvest process 04/05 14 Graph 3: Average lucerne seed quality through harvest process 05/06 15 Table 4: Comparison of Ferric Chloride staining of 05/06 cleaned header samples 15 Graph 4: Lucerne seed staining through harvest process 06/07 16

viii

Executive Summary

The longer you can look back, the further you can look forward Winston Churchill

What is this report about? The report presents a 5 year research project that investigated the cause of abnormal seedlings in lucerne. An in depth literature review focused the areas of research and a methodology was designed to test certain hypotheses. The report outlines the findings of the research and how they can be practically utilised in the production of lucerne seed. Who is report targeted at? Lucerne is the world’s most widely grown and popular perennial legume pasture species. It is grown to some degree in most countries of the world for grazing feed and conserved fodder production such as hay. The main lucerne seed producing countries to supply world demand include Australia and North America with Canada and South America also being suppliers. The Australian lucerne seed industry is estimated to be valued over A$100 million to the Australian economy with approximately 90% of production occurring in a 40 km radius surrounding the township of Keith in South Australia. Other areas of seed production in Australia include the Lachlan valley region of NSW as well as areas around Deniliquin, small areas of western and northern Victoria such as Shepparton and Minimay and the Jamestown, Frances and Naracoorte regions of SA. There are developing areas of lucerne seed production in southern WA as well as Tasmania. The research project and its findings are highly relevant to all seed producers and the wider industry of Australia. Background Whether lucerne seed is used for sowing pasture and hay crops or sprouting its value is based fundamentally on market forces and its quality. The Australian market is significantly export oriented and supply and demand factors are in general not manageable from the farm gate. To a significant degree seed quality is manageable within the farm gate as well as through the seed cleaning process. Lucerne seed is sold on the basis of its purity and germination quality. Purity is in general related to the weed status of the seed sample and germination is specifically the assessment of the seeds potential to germinate and presumably grow into a healthy lucerne plant. Whether seed is produced under a certified scheme or not there are world standards that apply to differentiate high quality seed from that of lesser quality. The final cleaned seed sample is assessed in accredited ISTA laboratories according to international seed testing rules. Seed quality is defined as percentages of normal seedlings, hard seed, dead seed and abnormal seedlings which are present after a set period of germination according to the ISTA guidelines. To meet the highest Australian standard, seed lines must have no less than 85% of the representative seed sample being comprised of a combined normal seedling and hard seed count. A problem arises when the abnormal seedling presence (and extremely rarely dead seed or a combined abnormal seedling - dead seed count) exceeds 15% which reduces the seed line below the 85% minimum. Sub-standard seed lines are more difficult to sell, particularly in low demand markets, and can be devalued significantly. Individual producers have received discounts of 12-20% on their seed sales which significantly reduces gross margin profits. Objective The research aimed to determine why abnormal seedlings occur and if there are practical measures available to reduce their impact on seed quality and the seed producer. Methodology The research assessed many key factors noted in the literature as having potential to cause abnormal seedlings either in lucerne or other legume and coarse grains. Factors such as the growing environment and prevailing weather, pest and diseases, method of preparation for harvest and seed moisture were not found to cause the degree of abnormal seedlings sometimes associated with lucerne seed. The research was conducted over 5 years and over 100 lucerne seed crops. Seed samples of lucerne were taken from the crop and throughout the harvesting and handling process. These samples were tested for nutritional status and

ix

germination percentage. The methodology was relatively simple and organised and permitted the development and evaluation of a ferric Chloride seed test. Key Findings The research identified that the principal cause of abnormal seedlings is damage during the harvest process and that certain lucerne seed crops were predisposed to higher risk of harvest damage and hence abnormal seedling production. A high correlation was found between seed damage and abnormal seedling production and that modifying the header configuration or harvest speed reduces seed damage. Given that the process of harvest can be managed to mitigate the percentage of abnormal seedlings a simple test was applied to permit the header operator to monitor the seed quality during harvest and modify accordingly. The simple 5-15 minute Ferric Chloride test can be used through the harvesting process to monitor seed damage and hence abnormal seedling creation. Implications The key findings of the research should permit producers to identify high risk paddocks and configure and operate the header in a manner that will reduce the potential for seed damage. Each paddock harvested may require header management different to the previous paddock and header operators and producers need to assess crop risks and seed quality prior to and during harvest. Utilisation of the seed test will allow producers to make judged decisions regarding their seed marketing by virtue of predicting the seeds likely quality. A key fundamental to the research and its findings is that all those involved in the production, handling and marketing of lucerne seed need to remind themselves that lucerne seed is a living organism and that how it is treated can have a lasting effect on how it subsequently grows. Recommendations The Ferric Chloride test should be developed into a user friendly kit for header operators to monitor the seed damage through the harvest process. Headers should be operated to suit the crop rather than use standard operating procedures on any type of lucerne seed crop. Producers need to accept that there is a limit on how a header can be operated to reduce damage in high risk crops but that it is also possible to mitigate damage and hence abnormal seedling creation. All current management practices leading up to harvest as well as post harvest are not notably contributing to abnormal seedling creation and no specific changes are deemed necessary.

1

1. Introduction 1.1 General Overview The Australian lucerne seed industry produces in excess of 6,200 tonne of seed per financial year of which over 90% is produced in South Australia. There is at least 8,000 ha of irrigated seed production in the Upper South East of South Australia and over 90% of Australia’s total lucerne seed production is produced in the project’s research region, Keith, South Australia. Other established production areas in Australia include the Jamestown region in the mid north of SA and the Lachlan Valley region in Forbes, NSW. Recent production is occurring around Deniliquin, NSW, Shepparton , Victoria and more recently in Tasmania. The export value of lucerne seed exceeds A$25.0 million and the value of the lucerne seed production industry to the Australian economy is estimated to be over $A100 million. The increasing value of lucerne seed to the Australian pasture seed industry, as well as to the rural economy, defines it as a commodity that requires research designed to preserve seed quality and quantity as well as grower returns. Australian produced lucerne seed is an export commodity that is traded according to global supply and demand factors in accordance to its quality – particularly in low demand periods. This research was required to assist in preserving the quality of Australian produced lucerne seed.

1.1.2 Research area Lucerne seed production in the research region is the district’s key crop. The majority of seed production is irrigated and there is a significant and increasing dryland production base dependent on seasonal rainfall. The research findings are readily transferable to all production areas in Australia as well as overseas.

1.2 Seed quality standards

1.2.1 Seed production chain In general Australian lucerne seed crops are harvested when as close as possible to 100% ripeness. Agronomic management preceding harvest greatly contributes to the attainment of full ripeness prior to harvest. In dry conditions producers will wait until the crop is fully ripe before either spraying with a chemical desiccant (e.g. Reglone) or windrowing in preparation for harvest. There are times when a crop will be prepared prior to 100% ripeness such as if there is impending rainfall or if the seasonal break has occurred and there is a need to harvest the seed crop as soon as possible to mitigate rain damage. The desiccated or windrowed crop is harvested with an open front header or combine harvester. In a desiccated crop the header typically uses a reel or air front to feed the crop into the machine as it moves forward. In windrowed crops the majority of headers use a windrow pickup to feed the crop into the machine although some use an air front. Either conventional or rotary header systems are used, both of which are suitable for harvesting lucerne seed if set up appropriately. In both systems it is important to release the lucerne seed from the pods and separate the seed from the empty pods and chaff. This is achieved by driving the header at an appropriate speed and setting internal fan, rotor or thresher speeds appropriately as well as selecting suitable concave and sieve settings. Australian lucerne seed producers either use their own headers or employ contract harvesters. Through the harvest process the operator will monitor the seed moisture content which needs to be no greater than 12% for storage prior to cleaning. Harvested seed is transferred via auger from the header into either a field bin or truck. If seed is stored in a field bin it is augured into a truck which then delivers the seed to a seed cleaning/processing business. Upon delivery the seed moisture is tested and if it is above 12% it is either dried in a drying plant or it is handled in a manner that permits the seed to dry below 12%. High moisture can be attributed to weed content, green pods, stalk and seed which is often a consequence of inadequate desiccation or harvesting prior to the desiccant or windrow drying down satisfactorily. If the seed is not to be dried it is stored in a

2

silo, seed bin or bulka bag pending cleaning. Movement of seed from the truck or storage silo is via a seed elevator into a container, which is emptied through the cleaning process. If seed is to be dried in a drying plant it is it is moved via an elevator from truck to plant to storage silo. Stored seed is cleaned over a gravity table where the primary seed is separated from weeds, dead seed, chaff and lighter lucerne seed. The gravity table separates seed based on its specific gravity with the primary seed being an operator judged range of heaviest seed which separates from lesser quality seed and waste. Once the seed is cleaned a random sample is assessed for germination quality and seed purity.

1.2.2 Seed assessment protocol Lucerne seed is sold on the basis of its purity and quality. Seed purity and quality is assessed according to the rules and guidelines established by the International Seed Testing Association (ISTA) which has specific rules for lucerne seed and aims to provide uniformity in seed testing across the world. The aim of seed assessment is to qualify the ability of the seed to be sown with the confidence that it will germinate and grow into the required cultivar. Random seed samples of 250 g are taken from all seed lines of up to 10 tonne per line. A sub sample is used for purity and germination testing. The purity test assesses the composition of the seed line differentiating between pure seed from other species of crops or weeds as well as inert matter such as seed coats. The germination test assesses the emergence and development characteristics of lucerne seed under specified standard conditions in accordance to ISTA rules. The seed germination is assessed over 10 days and the results should validate the ability of seeds to grow into plants under satisfactory growing conditions once planted. Significant discussion of the seed testing process is provided in the attached literature review.

1.2.3 Seed standards Once the seed has been tested a certificate of analysis is issued which reports the purity and germination results. Lucerne seed is sold into an international market place and the buyer standards vary from country to country. Typically the market requires seed with 99.9 % purity and no less than 85% germination, comprising normal seedlings and hard seeds. Some markets, such as the sprouting seed market, require seed with 90% germination. The balance of the germination percentage is comprised of abnormal seedlings and dead seeds. It is extremely rare that a seed line will be dominated by the presence of dead seed but it is not uncommon that a seed line will be greatly influenced by the presence of abnormal seedlings. If the abnormal seedlings count exceeds 15% it results in the germination percentage dropping below 85% and this can have marketing complications. Abnormal seedlings are discussed in detail in the attached literature review.

1.2.4 Trading of Australia’s lucerne seed Once the seed has been tested a certificate of analysis is issued which reports the purity and germination results. The producer uses this certificate to validate the seed quality that is for sale. The producer will generally sell seed to a seed trader with very minimal seed being traded between farms. The seed trader will buy the seed on the basis of the certificate of analysis as well as demand factors. Once purchased the seed trader requires a copy of the certificate to validate the seed quality to the market place in which it is to be traded. The majority of the market for Australia’s lucerne seed is off shore and it is for this reason that international seed testing rules and guidelines are critical. If the certificate of analysis states that the germination is below 85% the producer may be penalised in the form of a reduced price per tonne. Many producers grow private varieties under a contractual basis. The contracts define the seed quality required. If the seed fails to meet the requirements it is classed as inferior seed and may suffer a price penalty. The market forces determine the degree of price penalty. If there is an under supply for the demand the penalty is typically less than when supply exceeds demand. The

3

degree of penalty is inconsistent but has been known to be up to 20%. An example of the impact of the price penalty is as follows: A contract price is $4.00/kg clean seed subject to meeting the purity and germination criteria. If abnormal seedlings exceed 15% the seed fails to meet the contract standards. A 20% price reduction provides a farm gate value of $3.20/kg. A 500 kg/ha seed crop meeting the criteria grosses $2000/ha whereas the same crop failing the standards will gross $1600/ha. If cost of production is $1200/ha the failure to meet the germination standards reduces net return by 50%.

1.3 Literature review 1.3.1 Industry funded literature review Prior to the commencement of the research project in August 2003 and with funding support from Seedmark and Keith Seeds, the author commissioned Alison Graham and Glenn McDonald from the University of Adelaide to undertake a literature review into causes of abnormal seedlings in lucerne. The review – ‘Causes of abnormal seedlings in lucerne and some solutions to the problem’ is attached as Appendix 1.

1.3.2 Summary of review The review explains the seed testing process and the interpretations of normal and abnormal seedlings. In addition the review explored causes for abnormal seedlings and possible management options. Key areas of causation such as mechanical damage, crop stage at harvest, mineral deficiency, growing conditions and chemical damage are discussed. The review assisted in forming the direction of the research and the methodology that would be most suited to testing several hypotheses regarding the cause and management of abnormal seedlings in lucerne seed production.

1.3.3 Photo album of abnormal seedlings Seed Services Australia provided for the purpose of this report a compilation of photos taken by their seed camera microscope. The photos catalogue abnormal seedlings classified under ISTA rules for lucerne seed germination analysis. To the authors knowledge this is the best compilation of photos of abnormal lucerne seedlings in any literature available in the world. The photos are compiled in Appendix 10.

4

2. Objectives

2.1 Purpose of research Abnormal seedlings can create problems for the farm gate return of the lucerne seed producer if they exceed a minimum percentage set either by contractual obligations or by buyer requirements. The international lucerne seed industry has debated the principal cause of abnormal seedlings since the early 1960’s with mechanical damage suggested as being the primary cause. Over time and until the present day debate has continued with genetic, weather, chemical and fertility aspects all proposed to be possible causes of abnormal seedling creation in lucerne seed production. The financial losses that can be associated with high levels of abnormal seedling production warranted specific research into the causes of abnormal seedlings and, if possible, to identify the primary factor. Once the primary factor was identified a management tool needed to be developed. Examination of the literature suggested that it would be possible to assess all the listed possible causes and settle upon a principal causal factor. All the proposed causes had an equivalent potential management option and the research would evaluate options for managing the primary cause. The research set itself the aim of determining the primary reason for abnormal seedling presence and, if possible, a management solution.

5

3. Methodology

3.1 Overview The various hypotheses that would be tested in this research required a broad methodology that allowed for multiple assessments and evaluations. There were several key aspects that needed investigation and they outlined below: Pest Damage Environment Header Type Growing Conditions Biotic Factors Time of Harvest Harvest Damage Chemical Damage Nutritional Content Handling Damage Variety Influence Seed Moisture The key factors were all identified in the literature review as possible causes of abnormal seedlings. They have all been recorded in lucerne or other crop species as being a causal agent of abnormal seedlings. The research needed a methodology that could examine lucerne seed samples from selected seed crops with a recorded crop production history and assess the impact of the key factors on the abnormal seedling component within and between samples over the five years of investigation. The designed and implemented methodology needed to be simple and flexible yet robust and accurate enough to evaluate the data and collated information obtained from within the complexities of commercial seed production.

3.1.1 Sample selection Lucerne seed crops were selected during the course of the production season and not prior to the commencement of the season. This was done to allow the flexibility to sample crops that would contribute most to achieving the project’s objectives. Both dryland and irrigated crops were selected in the first two seasons (03/04 and 04/05) on the basis that the research needed to assess crops that were: (i) harvested by different header makes and models (ii) desiccated or windrowed (iii) dry with low plant production or highly productive, vigorous crops (iv) representative of the diversity of varieties produced (v) representative of typical production and handling techniques, management and systems.

Abnormal Seedlings

6

If the sampled crops were irrigated they were to be representative of the methods of irrigation, chiefly border check and centre pivot systems.

3.1.2 Sample Collection Seed samples were collected from the selected crops prior to harvest, from the header, field bin (if used) and truck. The final cleaned sample was collected and managed by both the seed processor and the seed testing laboratory. The pre harvest sample was taken before and after desiccation or before and after windrowing. The random sample was taken across the production area to be representative of the crop.

3.1.3 Sample management and preparation The hand collected samples (‘hand samples’) consisted of a large mass of pods and stalks. The method used to prepare refined seed sample was laborious but needed to be sufficiently gentle so as not to damage the seed. Each sample was hand threshed by rubbing the pods gently between gardening gloved hands over a collection tub. The subsequent mix of seed and offal was gently separated using a combination of light compressed air and a 3 mm sieve. The seed sample was further refined by gentle use of compressed air to blow off dust and small pieces of pods and stems. The refined seed sample as well as the header, field bin and truck samples were cleaned using a scale model gravity table made available to the research project by Tatiara Seeds in Bordertown. The gravity table separated the primary lucerne seed from weeds, offal and secondary lucerne seed. The primary and secondary lucerne seed samples are collected, labeled and stored pending germination analysis.

3.1.4 Sample testing In 03/04 and 04/05, the cleaned seed samples were split into thirds. One third was sent to the Waite Analytical Services (WAS) in Adelaide for nutritional testing, one third was sent to Seed Services Australia (SSA) for germination analysis and the remaining third was stored for reference. In 04/05, the secondary seed samples from the header samples were also sent to SSA. In 05/06, seed samples were only sent to SSA. SSA conducted germination tests in accordance to the ISTA rules and in the same manner as commercial lucerne seed lines are purity and germination tested. In 02/03, prior to the commencement of the research, collected and cleaned header samples were sent to WAS to assist in the development of the research project. WAS is accredited by the Australasian Plant and Soil Analysis Council for analysis of plant tissue. The WAS acid digested the seed and analysed the digest by Inductively Coupled Plasma Atomic Emission Spectroscopy. The elements detected in the seed sample were Aluminium, Boron, Cadmium, Calcium, Cobalt, Copper, Iron, Manganese, Magnesium, Molybdenum, Nickel, Phosphorous, Potassium, Sodium, Sulphur and Zinc. Sub samples from the retained pre harvest and header 05/06 samples were treated with Ferric Chloride as explained later. All the nutritional and germination test results were collated and assessed. In addition, all the key factors were noted and relevant information pertaining to them were recorded as required. These are discussed in the results section.

3.1.5 Refinement of research over time Over the period the research, as results were assessed, refinements were made to paddock selection and treatment of samples. It became apparent that many of the potential key factors were not significant contributors to the existence of abnormal seedlings. By 05/06 and 06/07, the research focused on the harvest process which was highly correlated to the development of abnormal seedlings.

7

3.1.6 Development of Ferric Chloride test As suggested in 3.1.5, the research determined that there existed a strong link between the harvest process and abnormal seedlings. The lucerne seed industry had not openly investigated this relationship even though there were suggestions from production areas in Australia and North America over the past 45 years. Up until the commencement of this research project, the industry had chosen to speculate about the causes of abnormal seedlings and not pursued any investigation into historical data and research. Further research by the author and assistance by staff at Seed Services Australia unearthed a relatively simple seed test to assess the presence of mechanical injury to legume seed. It is unclear of the origin of this test but the imperial measurements indicate it was pre 1966 in Australia or is has North American history. The presentation of the seed test suggests it is pre 1966. The seed test in this research is referred to as the Ferric Chloride test. 100 ml of 42% FeCl3 is diluted in 400 ml of rain water. 100 lucerne seeds are placed in a petri dish and the diluted solution is poured over the seeds to completely cover the seeds. Some seeds float but the majority will not. The staining process stains cracks and chips in the seed coat black. Within 5 minutes, damaged seeds begin to stain and the treatment process is concluded in 15 minutes. The number of stained seeds is expressed as a percentage of the total number of seeds in the test sample. The ferric chloride solution can be strained after use and stored for reuse.

3.1.7 Assessment of the test The Ferric Chloride test was evaluated over the 05/06 header samples to correlate staining percentages and the abnormal seedling percentage in each sample. The testing procedure used was that described in section 3.1.6. In 06/07 three seed crops were tested through the harvest process to monitor abnormal seedling levels. The Ferric Chloride test, as described above, was used on hand and header samples with the header settings being modified in response to the test results through harvest.

8

4. Summary of Results

4.1 Seasonal records The research collected a range of data through its progression and this data is provided in Appendices 2-9. Such data includes variety, location, production method, crop health, germination percentages, moisture content at harvest, desiccation or windrow records, harvest date, header type and settings. This data was used to validate the impact of key factors on the cause of abnormal seedlings and aspects of the data are graphically represented in the following sections.

4.2 Growing conditions RIRDC research projects: Dividing the Droplet (Pub. No. 05/116) and The Invisible Reality of Groundwater Salinity (Pub No. 06/053) as well as DWLBC projects: Minimising salt accession in the South East of South Australia – the Border Designated Area and the Hundred of Stirling (report nos. 2006/19 and 2007/pending) and Volumetric Conversion in the South East of South Australia (DWLBC 2006/30) recorded specific weather factors. The author either designed, conducted, contributed or produced the research in these projects, particularly the collation of weather records through the lucerne seed crop growing seasons. In seasons 00/01 to 06/07, temperature, relative humidity, global radiation, wind speed and rainfall were recorded for the calculation of evapotranspiration (ET) in the assessment of crop water use calculations. This data was used to evaluate the growing conditions over the period of this research period.

4.3 Environmental conditions The environment for lucerne seed production in the research area was consistent from year to year. Soil type and irrigation water quality were annually consistent across the production area and rainfall influenced all crops similarly. Soil type, irrigation water quality, rainfall, pest presence or biotic influences were evaluated on the abnormal seedling count of the hand samples.

4.4 Variety, chemical, windrowing, time of harvest factors

Impact of variety, chemical desiccation, windrowing and time of harvest was observed and recorded on the abnormal seedling count of the hand samples. This data is presented in Appendices 3, 5 and 7.

4.5 Seed moisture Seed moisture was recorded at the time of seed delivery. This is presented in Appendices 2, 4 and 6.

4.6 Handling damage Handling damage was recorded as the change in abnormal seedling presence from the harvest sample through to the field bin and truck sample. This is presented in Appendices 2, 4 and 6.

4.7 Header type Header type (i.e. conventional or rotary systems), model and settings are recorded in Appendices 3, 5 and 7.

4.8 Nutritional analysis Tables 1, 2 and 3 present the lucerne seed nutritional analyses for seasons 02/03, 03/04, and 04/05.

9

Table 1: 02/03 pre research commencement lucerne seed nutritional analysis Date Reported : 8/12/03

Samples Received : 20/11/03Analysis Instrument ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL

Mass (1) Diln. (1) Fe Mn B Cu Mo Co Ni Zn Ca Mg Na K P S Al CdSample No. Abnormal % gm ml mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg

Hallmark Dryland 25 0.597 25 60 13 17 7.9 2.1 < 0.9 3.0 38 1250 2200 53 10700 7200 2900 < 6 < 0.4Siriver Irrigation 12 0.608 25 71 15 16 8.9 2.3 < 0.9 1.1 46 1410 2300 199 11500 7900 3000 < 6 < 0.4

Hunter River Irrigation 7 0.606 25 68 16 19 7.6 1.9 < 0.9 1.8 47 1340 2300 164 12100 7800 3000 < 6 < 0.4Siriver Irrigation 10 0.603 25 71 17 18 8.8 2.1 < 0.9 < 1 48 1570 2300 230 11600 7400 3000 < 6 < 0.4Rippa Dryland 7 0.602 25 80 16 18 11 1.9 < 0.9 3.3 50 1460 1970 87 10800 6800 3100 < 6 < 0.4

Hallmark Dryland 18 0.606 25 66 14 19 7.4 1.7 < 0.9 2.4 46 1340 2200 66 11400 7400 3100 < 6 < 0.4Siriver Irrigation 14 0.603 25 90 15 17 8.5 1.6 1.1 1.9 64 1650 2100 137 11300 7500 3100 < 6 0.47

Hallmark Dryland 13 0.606 25 68 13 18 8.6 2.4 < 0.9 2.7 43 1390 2200 51 11100 7300 3000 < 6 < 0.4FG Dryland 6 0.603 25 79 13 19 8.0 1.3 < 0.9 2.0 49 1690 2000 43 10400 6000 3000 < 6 < 0.4FG Dryland 7 0.603 25 88 18 18 5.1 0.99 < 0.9 4.6 57 1640 2000 31 10700 6600 3100 < 6 < 0.4

Super Siriver Irrigation 5 0.608 25 71 16 14 6.8 3.1 < 0.9 2.2 48 1210 2400 270 11600 7400 3000 < 6 < 0.4Super Siriver Irrigation 8 0.607 25 69 16 14 7.8 3.1 < 0.9 2.1 47 1330 2400 350 11800 7300 3000 < 6 < 0.4

Genesis Dryland 5 0.605 25 67 22 19 8.4 < 0.9 < 0.9 4.5 48 1660 1940 42 10100 5700 2900 < 6 < 0.4Hunter River Irrigation 9 0.607 25 66 18 18 6.3 1.9 < 0.9 2.5 32 1150 2600 440 12000 7200 2900 < 6 < 0.4Super Siriver Irrigation 10 0.605 25 69 14 17 6.9 2.9 < 0.9 1.9 45 1410 2300 590 11600 7400 2900 < 6 < 0.4

Siriver Irrigation 2 0.601 25 70 17 12 5.8 2.2 2.1 2.1 37 1280 2200 310 11300 7500 2800 < 6 < 0.4Siriver Irrigation 6 0.606 25 74 19 13 5.9 2.0 2.4 2.3 38 1580 2300 410 11800 7400 2900 < 6 < 0.4

FG Irrigation 6 0.605 25 76 20 14 6.9 1.8 1.0 3.1 51 1440 2100 99 11000 7100 3000 < 6 < 0.4FG Irrigation 14 0.602 25 76 21 14 6.8 2.0 < 0.9 2.8 51 1660 2100 112 10400 6800 2900 < 6 < 0.4

Hallmark Dryland 2 0.601 25 67 12 13 8.8 2.0 < 0.9 2.5 44 1360 2100 61 10300 7300 2800 < 6 < 0.4 Notes: • All solid samples were analysed on an oven dried basis. • Samples for ICP analysis were digested with nitric acid and finished with hydrochloric acid. This digestion method gives good recovery of all the elements shown

above. ARL Sample solutions were analysed by the ARL Inductively Coupled Plasma Atomic Emission Spectrometry (ICPAES).

• To convert mg/kg to %, divide the value by 10,000. • The limit of determination for the sample is calculated as 10 X the standard deviation of the blank. • Where duplicate analyses are carried out for QA purposes there will be 2 lines in the report for that particular sample. Duplicates give an indication of the

homogeneity of the sample. The first line will give the % variation (%rsd) between the 2 results - we aim for less than 10%, unless there is a problem with the sample e.g. contamination from soil or the method does not recover the total amount of that element as indicated above. Also the closer the results approaches the limit of detection the larger the % variation of the duplicates. The second line of results for the duplicate is the average result of the 2 analyses.

• Symbols: < - Indicates the result is less than the limit of detection of determination of the method

10

Table 2: 03/04 lucerne seed nutritional analysis Date Reported : 31/8/04Samples Received : 13/8/04

Analysis Instrument ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARL ARLMass (1) Diln. (1) Fe Mn B Cu Mo Co Ni Zn Ca Mg Na K P S Al Cd

Sample No. gm ml mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg

1 0.597 25 68 19 14 6.5 2.2 < 0.9 3.8 38 1280 2400 179 11000 7900 2900 < 6 < 0.42 0.6 25 84 16 14 9.3 1.3 < 0.9 2.4 51 1690 1950 111 11100 6400 3000 < 6 < 0.44 0.597 25 65 12 14 8.0 1.9 < 0.9 2.9 29 1350 2100 108 11100 6800 2900 < 6 < 0.46 0.603 25 94 19 15 7.6 2.0 < 0.9 1.8 45 2500 2500 198 11800 8100 3000 26 < 0.48 0.603 25 79 20 14 6.1 2.4 < 0.9 2.3 55 1150 2500 370 11100 7600 3100 < 6 < 0.412 0.597 25 70 19 14 10 2.0 0.92 2.3 35 1280 2600 410 11500 8000 3000 < 6 < 0.414 0.599 25 77 18 15 8.2 < 0.9 < 0.9 2.0 35 1610 2000 89 11500 7100 3100 < 6 < 0.415 0.6 25 82 19 14 8.4 1.6 < 0.9 3.6 40 1430 2000 75 11100 6500 3000 < 6 < 0.417 0.602 25 70 22 13 6.6 2.0 1.5 2.3 55 1150 2500 340 11200 7600 3000 < 6 < 0.419 0.599 25 60 14 14 6.8 1.9 < 0.9 3.5 32 1520 2200 200 12000 7500 2900 < 6 < 0.421 0.597 25 69 15 13 7.8 1.5 < 0.9 2.5 39 1220 2300 97 11900 7200 3000 < 6 < 0.425 0.6 25 72 15 13 9.1 1.7 < 0.9 3.6 45 1200 2300 167 12400 7700 3000 < 6 < 0.427 0.604 25 84 18 13 7.6 2.2 < 0.9 3.2 50 1620 2200 106 12200 7600 3100 < 6 < 0.472 0.603 25 79 20 13 6.2 2.2 < 0.9 2.2 37 1450 2400 230 11800 8100 3000 < 6 < 0.473 0.6 25 82 21 14 8.5 3.8 < 0.9 2.3 50 1660 2500 112 12000 8500 3200 < 6 < 0.477 0.603 25 77 20 13 5.5 1.9 < 0.9 1.2 39 1200 2700 250 12100 8400 3100 < 6 < 0.480 0.599 25 78 15 15 6.8 2.4 < 0.9 2.6 44 1470 2300 111 11300 7000 3100 < 6 < 0.483 0.603 25 80 14 13 7.4 2.0 < 0.9 < 1 57 1470 2200 270 12600 9200 3200 < 6 < 0.488 0.606 25 82 17 13 5.6 3.0 < 0.9 2.4 34 1430 2500 290 12100 7800 3100 < 6 < 0.491 0.599 25 86 19 15 8.6 3.0 < 0.9 2.4 56 1770 2400 750 12600 8300 3300 < 6 < 0.492 0.599 25 80 19 14 7.8 2.3 < 0.9 1.6 52 1450 2500 480 12500 7900 3200 < 6 < 0.496 0.603 25 71 14 14 5.5 2.4 < 0.9 2.7 45 1630 2400 540 12300 7500 3100 < 6 < 0.497 0.604 25 76 21 15 7.5 3.0 < 0.9 1.8 48 1750 2600 220 11400 7900 3200 < 6 < 0.498 0.599 25 74 20 14 6.3 3.3 < 0.9 1.6 47 940 2600 200 11700 8400 3100 < 6 < 0.4102 0.601 25 79 23 13 9.5 1.4 < 0.9 1.9 32 1200 2600 360 11900 7800 3000 < 6 < 0.4

Notes: • All solid samples were analysed on an oven dried basis. • Samples for ICP analysis were digested with nitric acid and finished with hydrochloric acid. This digestion method gives good recovery of all the elements shown

above. ARL Sample solutions were analysed by the Radial ARL Inductively Coupled Plasma Atomic Emission Spectrometry (ICPAES).

• To convert mg/kg to %, divide the value by 10,000. • The limit of determination for the sample is calculated as 10 X the standard deviation of the blank. • Where duplicate analyses are carried out for QA purposes there will be 2 lines in the report for that particular sample. Duplicates give an indication of the

homogeneity of the sample. The first line will give the % variation (%rsd) between the 2 results - we aim for less than 10%, unless there is a problem with the sample e.g. contamination from soil or the method does not recover the total amount of that element as indicated above. Also the closer the results approaches the limit of detection the larger the % variation of the duplicates. The second line of results for the duplicate is the average result of the 2 analyses.


11

Table 3: 04/05 lucerne seed nutritional analysis Date Reported : 26/10/05

Samples Received : 28/9/05Instrument CIROS RCIROS R CIROS R CIROS RCIROS RCIROS RCIROS R CIROS RCIROS RCIROS R CIROS R CIROS RCIROS RCIROS RCIROS RCIROS RCIROS RCIROS R CIROS R CIROS R

Mass Diln. Fe Mn B Cu Mo Co Ni Zn Ca Mg Na K P S Al Ti Cr Cd Pb SeSample No. gm ml mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg

1 0.601 25 79 22 15 11 1.0 0.71 1.2 45 1220 2400 280 11600 7700 2700 2.3 < 0.07 < 0.3 < 0.1 < 1 < 52 0.604 25 64 18 18 9.2 3.5 < 0.3 2.4 41 940 2600 240 11500 8300 2700 2.4 < 0.07 < 0.3 < 0.1 < 1 < 53 0.605 25 70 18 18 7.4 1.9 0.35 3.0 41 1420 2200 166 10900 7400 2800 2.2 < 0.07 < 0.3 < 0.1 < 1 < 54 0.596 25 64 10 18 5.5 2.0 < 0.3 2.3 30 1430 2100 165 10500 6800 2800 1.2 < 0.07 < 0.3 < 0.1 < 1 < 55 0.605 25 71 16 20 5.5 2.1 < 0.3 2.3 37 1040 2200 350 10900 6400 2700 0.58 < 0.07 < 0.3 < 0.1 < 1 < 56 0.606 25 60 17 18 8.1 1.8 0.52 2.0 41 1670 2200 155 11800 7300 2800 0.79 < 0.07 < 0.3 < 0.1 < 1 < 57 0.601 25 66 14 19 9.1 2.2 0.66 3.0 48 1250 2000 440 11100 7700 2800 0.73 < 0.07 < 0.3 < 0.1 < 1 < 58 0.603 25 71 21 23 9.0 1.7 1.1 2.2 46 1200 1980 119 10100 5900 2800 4.5 < 0.07 < 0.3 < 0.1 < 1 < 59 0.596 25 67 15 16 8.5 2.3 2.2 1.7 43 1030 2400 510 10800 7500 2700 1.3 < 0.07 < 0.3 < 0.1 < 1 < 5

10 0.605 25 63 20 15 5.2 2.7 < 0.3 1.2 42 1000 2500 200 11200 7500 2700 0.41 < 0.07 < 0.3 < 0.1 < 1 < 511 0.605 25 72 17 15 6.4 1.3 0.35 1.7 24 1140 2600 350 11600 7900 2600 0.43 < 0.07 < 0.3 < 0.1 < 1 < 512 0.597 25 66 19 21 8.6 2.3 0.74 1.9 46 1060 2400 280 10600 6800 2900 0.75 < 0.07 < 0.3 < 0.1 < 1 < 513 0.605 25 61 15 15 7.5 2.9 0.53 2.6 35 1070 2200 112 10300 6700 2700 1.8 < 0.07 < 0.3 < 0.1 < 1 < 514 0.598 25 70 15 18 6.1 1.8 < 0.3 1.7 43 1130 2500 210 11400 8200 2700 0.87 < 0.07 < 0.3 < 0.1 < 1 < 515 0.601 25 70 20 13 6.7 1.2 0.82 2.4 27 1010 2400 320 10900 6900 2700 0.79 < 0.07 < 0.3 < 0.1 < 1 < 516 0.603 25 69 19 21 6.6 2.7 < 0.3 2.2 45 1360 2200 116 10500 6900 2900 0.64 < 0.07 < 0.3 0.12 < 1 < 517 0.605 25 64 16 20 9.5 1.5 0.48 2.8 40 1040 1960 97 10200 6800 2700 0.50 < 0.07 < 0.3 < 0.1 < 1 < 518 0.6 25 61 18 15 5.2 0.73 0.36 1.9 43 1480 1850 102 10900 6400 2700 2.2 < 0.07 < 0.3 < 0.1 < 1 < 519 0.604 25 61 11 17 8.6 2.2 1.0 2.7 40 1210 2100 51 9600 6500 2600 1.1 < 0.07 < 0.3 < 0.1 < 1 < 520 0.606 25 50 20 16 7.7 1.6 0.34 2.0 39 970 2400 250 11000 7500 2600 1.0 < 0.07 < 0.3 < 0.1 < 1 < 521 0.599 25 73 17 20 11 1.5 1.4 2.2 51 910 2500 560 11500 7800 2800 2.6 < 0.07 < 0.3 < 0.1 < 1 < 522 0.597 25 66 13 17 8.5 2.7 0.45 1.5 41 1480 2100 78 10500 7100 2600 0.86 < 0.07 < 0.3 < 0.1 < 1 6.423 0.606 25 65 15 17 6.0 2.4 < 0.3 1.9 39 1020 2200 123 11200 7900 2700 2.4 < 0.07 < 0.3 < 0.1 < 1 < 524 0.602 25 58 18 18 5.0 6.1 0.57 2.7 38 1380 1920 44 9500 5600 2700 1.0 < 0.07 < 0.3 < 0.1 < 1 < 525 0.605 25 59 15 17 9.2 0.74 0.31 3.2 25 1360 2100 55 10700 6700 2600 0.62 < 0.07 < 0.3 < 0.1 < 1 < 526 0.603 25 72 16 16 7.7 2.3 0.37 2.8 41 1280 2200 136 10200 7000 2600 0.35 < 0.07 < 0.3 < 0.1 < 1 < 527 0.603 25 70 15 15 10 1.8 0.71 2.2 56 1330 2100 126 10700 7000 2800 0.52 < 0.07 < 0.3 < 0.1 < 1 < 528 0.597 25 76 17 15 9.1 1.8 < 0.3 1.3 52 1130 2400 340 10900 7400 2800 1.4 < 0.07 < 0.3 < 0.1 < 1 < 529 0.604 25 61 15 17 5.2 1.9 1.1 2.1 35 1160 2300 320 11700 7600 2800 0.61 < 0.07 < 0.3 < 0.1 < 1 < 530 0.599 25 66 18 20 6.8 0.97 < 0.3 2.3 33 1280 1890 62 9900 6600 2700 0.81 < 0.07 < 0.3 < 0.1 < 1 < 531 0.602 25 73 17 15 7.4 2.4 0.77 3.2 42 1310 2200 167 11000 7700 2900 1.2 < 0.07 < 0.3 < 0.1 < 1 < 532 0.598 25 72 16 14 8.0 2.1 0.61 2.5 52 1340 2200 126 10800 7400 2800 2.6 < 0.07 < 0.3 < 0.1 < 1 < 533 0.601 25 73 18 15 7.6 2.4 1.6 2.9 42 1260 2200 270 11200 7800 2800 2.3 < 0.07 < 0.3 < 0.1 < 1 < 534 0.599 25 61 11 14 5.5 2.1 < 0.3 1.9 40 1360 2100 130 10600 6600 2800 1.6 < 0.07 < 0.3 < 0.1 < 1 < 535 0.602 25 54 16 13 8.3 1.5 0.44 2.2 39 970 2300 320 11000 7300 2700 1.5 < 0.07 < 0.3 < 0.1 < 1 < 536 0.603 25 55 16 16 8.1 1.6 0.40 2.1 38 960 2300 320 10900 7300 2700 0.46 < 0.07 < 0.3 < 0.1 < 1 < 537 0.604 25 57 14 14 7.5 2.3 0.46 3.1 33 1060 2100 82 10700 7200 2700 0.94 < 0.07 < 0.3 < 0.1 < 1 < 538 0.602 25 57 15 15 6.5 2.5 0.31 2.9 34 1080 2100 91 10500 7200 2700 1.7 < 0.07 < 0.3 < 0.1 < 1 < 539 0.601 25 66 15 15 9.1 1.6 0.56 3.5 40 1250 1970 69 10400 7100 2700 0.85 < 0.07 < 0.3 < 0.1 < 1 < 540 0.607 25 76 16 14 6.3 0.63 < 0.3 3.0 29 1480 1880 65 10500 7000 2800 1.8 < 0.07 < 0.3 < 0.1 < 1 < 541 0.597 25 65 16 16 9.7 1.4 0.55 3.2 43 1320 1990 66 10700 7200 2700 1.0 < 0.07 < 0.3 < 0.1 < 1 < 542 0.6 25 57 14 18 6.7 2.3 0.51 2.5 38 1490 1860 30 10000 6100 2800 0.94 < 0.07 < 0.3 < 0.1 < 1 < 543 0.6 25 75 19 13 8.8 2.3 0.50 1.7 49 1140 2400 350 10700 7300 2800 1.6 < 0.07 < 0.3 < 0.1 < 1 < 5

12

Instrument CIROS RCIROS R CIROS R CIROS RCIROS RCIROS RCIROS R CIROS RCIROS RCIROS R CIROS R CIROS RCIROS RCIROS RCIROS RCIROS RCIROS RCIROS R CIROS R CIROS RMass Diln. Fe Mn B Cu Mo Co Ni Zn Ca Mg Na K P S Al Ti Cr Cd Pb Se

Sample No. gm ml mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg44 0.601 25 83 17 14 6.8 < 0.5 < 0.3 2.4 27 1560 1930 77 10400 7000 2700 1.3 < 0.07 < 0.3 < 0.1 < 1 < 545 0.603 25 83 18 15 9.1 1.4 0.36 2.8 38 1320 1970 64 10300 6400 2800 1.8 < 0.07 < 0.3 < 0.1 < 1 < 546 0.602 25 71 15 16 7.2 2.4 0.49 2.7 50 1630 2300 150 10900 7700 2800 2.7 < 0.07 < 0.3 < 0.1 < 1 < 547 0.6 25 55 11 18 5.4 2.4 0.31 2.3 34 1350 2100 155 10400 6300 2700 2.0 < 0.07 < 0.3 < 0.1 < 1 < 548 0.601 25 52 14 14 7.4 1.7 0.33 3.2 28 1020 2200 108 10800 7100 2600 1.7 < 0.07 < 0.3 < 0.1 < 1 < 549 0.599 25 60 12 14 8.2 3.0 0.90 2.9 42 1280 2000 63 9900 6300 2700 1.8 < 0.07 < 0.3 < 0.1 < 1 < 550 0.597 25 65 16 15 7.3 1.8 0.99 2.0 44 1010 2400 350 11100 7700 2800 0.44 < 0.07 < 0.3 < 0.1 < 1 < 551 0.601 25 64 16 14 7.4 1.7 1.0 2.1 44 1000 2400 380 11100 7800 2800 0.27 < 0.07 < 0.3 < 0.1 < 1 < 552 0.601 25 63 15 16 5.3 1.8 0.37 2.6 41 2500 2300 250 11100 7400 2600 1.3 < 0.07 < 0.3 < 0.1 < 1 < 553 0.599 25 64 17 17 9.9 0.86 0.40 2.6 32 1330 2000 98 10200 6400 2700 1.1 < 0.07 < 0.3 < 0.1 < 1 < 554 0.601 25 59 19 14 6.7 0.78 0.34 2.1 43 1400 1900 86 10600 6600 2800 2.5 < 0.07 < 0.3 < 0.1 < 1 < 555 0.604 25 63 20 14 7.5 0.79 0.35 2.0 46 1540 2000 140 10800 6700 2900 3.0 < 0.07 < 0.3 < 0.1 < 1 < 556 0.599 25 53 16 13 8.8 1.6 0.51 2.1 42 990 2400 380 11200 7400 2800 0.34 < 0.07 < 0.3 < 0.1 < 1 < 557 0.602 25 55 14 13 7.9 1.8 0.39 3.2 28 1080 2300 125 10800 7200 2700 0.88 < 0.07 < 0.3 < 0.1 < 1 < 558 0.604 25 72 12 13 9.1 2.1 0.55 2.0 50 1290 2000 80 10500 7000 2800 1.7 < 0.07 < 0.3 < 0.1 < 1 < 559 0.598 25 67 15 13 5.8 2.1 0.50 3.0 39 1960 2300 240 11400 7700 2700 0.95 < 0.07 < 0.3 < 0.1 < 1 < 560 0.602 25 65 18 14 9.8 0.86 0.39 2.6 31 1380 2100 89 10300 6700 2700 0.63 < 0.07 < 0.3 < 0.1 < 1 < 561 0.608 25 60 18 12 7.1 1.1 0.67 2.7 41 1370 1870 82 9700 5900 2700 1.8 < 0.07 < 0.3 < 0.1 < 1 < 562 0.607 25 71 18 13 8.9 2.2 0.53 1.8 50 1100 2300 320 10900 7600 2800 1.2 < 0.07 < 0.3 < 0.1 < 1 < 563 0.601 25 71 15 13 8.6 2.1 1.8 1.4 52 1140 2400 390 10900 7400 2800 1.1 < 0.07 < 0.3 < 0.1 < 1 < 564 0.597 25 64 14 12 7.2 2.5 2.1 2.2 38 1090 2400 340 10900 7400 2700 1.3 < 0.07 < 0.3 < 0.1 < 1 < 565 0.603 25 73 15 14 9.0 2.0 0.48 2.5 48 1310 2200 380 11100 7700 2800 0.88 < 0.07 < 0.3 < 0.1 < 1 < 566 0.6 25 71 14 16 8.7 2.1 0.36 2.2 48 1260 2100 380 10900 7900 2800 1.2 < 0.07 < 0.3 < 0.1 < 1 < 567 0.599 25 74 15 16 8.7 2.1 0.49 2.6 47 1290 2200 390 11000 7600 2800 0.88 < 0.07 < 0.3 < 0.1 < 1 < 568 0.604 25 66 18 17 7.6 2.9 < 0.3 2.0 42 1030 2600 250 11500 8200 2800 1.3 < 0.07 < 0.3 < 0.1 < 1 < 569 0.603 25 61 18 14 8.4 2.8 < 0.3 2.1 42 1020 2600 250 11000 7900 2700 1.3 < 0.07 < 0.3 < 0.1 < 1 < 570 0.606 25 65 18 17 8.0 2.8 < 0.3 1.9 39 1080 2600 270 11400 7900 2700 1.5 < 0.07 < 0.3 < 0.1 < 1 < 571 0.603 25 56 12 15 7.8 2.2 0.56 2.7 30 1200 2200 91 10800 7500 2800 0.39 < 0.07 < 0.3 < 0.1 < 1 < 572 0.604 25 64 14 16 6.0 1.5 0.37 1.9 42 1090 2500 240 11300 8100 2800 1.5 < 0.07 < 0.3 < 0.1 < 1 < 573 0.603 25 57 12 14 8.2 2.0 0.42 3.0 29 1310 2200 85 10700 7400 2800 0.57 < 0.07 < 0.3 < 0.1 < 1 < 574 0.604 25 72 18 13 8.1 2.2 1.1 2.5 35 1010 2400 320 11200 8000 2800 0.59 < 0.07 < 0.3 < 0.1 < 1 < 575 0.603 25 71 18 14 6.3 2.2 0.58 2.7 40 1080 2400 270 11000 7500 2900 1.7 < 0.07 < 0.3 < 0.1 < 1 < 576 0.596 25 72 17 14 6.8 2.1 0.33 1.9 40 1070 2300 400 11200 7300 2800 1.2 < 0.07 < 0.3 < 0.1 < 1 < 577 0.598 25 64 17 16 6.0 1.5 < 0.3 1.8 31 1100 2500 340 11500 7700 2800 1.0 < 0.07 < 0.3 < 0.1 < 1 < 578 0.605 25 70 17 17 6.7 2.1 < 0.3 1.6 40 1080 2300 450 11600 7300 2800 1.1 < 0.07 < 0.3 < 0.1 < 1 < 579 0.602 25 67 21 14 9.7 1.9 0.94 2.3 46 1540 2300 183 11700 7500 3000 1.6 < 0.07 < 0.3 < 0.1 < 1 < 580 0.605 25 70 16 13 6.8 2.2 < 0.3 1.7 40 1040 2300 450 11300 7300 2800 0.78 < 0.07 < 0.3 < 0.1 < 1 < 581 0.605 25 63 13 12 5.0 1.7 < 0.3 2.0 45 1130 2400 220 10800 7300 2800 0.99 < 0.07 < 0.3 < 0.1 < 1 < 582 0.606 25 63 20 14 6.2 2.1 < 0.3 1.2 41 960 2500 210 10900 7600 2700 1.5 < 0.07 < 0.3 < 0.1 < 1 < 583 0.597 25 67 22 12 6.2 1.3 0.34 2.0 32 1110 2300 210 11100 7200 2800 1.6 < 0.07 < 0.3 < 0.1 < 1 < 584 0.603 25 69 22 15 7.2 1.3 0.36 2.2 33 1160 2300 210 11300 7300 2800 1.1 < 0.07 < 0.3 < 0.1 < 1 < 585 0.6 25 81 16 13 7.6 2.1 0.41 2.7 39 1190 2200 198 10900 7400 2900 1.2 < 0.07 < 0.3 < 0.1 < 1 < 586 0.606 25 67 21 13 8.6 1.9 0.92 2.1 42 1530 2200 148 11600 7700 2900 1.2 < 0.07 < 0.3 < 0.1 < 1 < 587 0.6 25 67 18 13 6.9 1.4 < 0.3 1.7 33 1120 2500 330 11100 7500 2800 5.1 < 0.07 < 0.3 < 0.1 < 1 < 5

13

Instrument CIROS RCIROS R CIROS R CIROS RCIROS RCIROS RCIROS R CIROS RCIROS RCIROS R CIROS R CIROS RCIROS RCIROS RCIROS RCIROS RCIROS RCIROS R CIROS R CIROS RMass Diln. Fe Mn B Cu Mo Co Ni Zn Ca Mg Na K P S Al Ti Cr Cd Pb Se

Sample No. gm ml mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg88 0.607 25 79 17 13 7.5 2.3 0.45 2.6 37 1240 2200 230 10900 7300 2900 1.2 < 0.07 < 0.3 < 0.1 < 1 < 589 0.598 25 71 17 13 7.1 2.0 < 0.3 1.6 41 1010 2300 550 11300 7400 2800 0.77 < 0.07 < 0.3 < 0.1 < 1 < 590 0.603 25 66 21 13 6.3 2.2 < 0.3 1.4 43 1050 2500 240 11100 7500 2800 2.5 < 0.07 < 0.3 < 0.1 < 1 < 591 0.603 25 73 17 14 7.0 2.0 < 0.3 1.7 40 1150 2400 540 11900 7400 2800 0.99 < 0.07 < 0.3 < 0.1 < 1 < 592 0.602 25 70 17 14 6.8 2.0 < 0.3 1.6 40 1170 2300 610 11400 7300 2800 1.6 < 0.07 < 0.3 < 0.1 < 1 < 593 0.607 25 65 20 17 7.7 1.2 0.71 2.9 42 1430 1910 87 10600 6300 2800 2.2 < 0.07 < 0.3 < 0.1 < 1 < 594 0.605 25 64 16 14 9.0 1.5 0.57 3.3 40 1230 2000 96 10600 7200 2700 1.1 < 0.07 < 0.3 < 0.1 < 1 < 595 0.605 25 61 11 14 5.5 2.3 < 0.3 2.0 41 1330 2100 133 10600 6500 2900 1.9 < 0.07 < 0.3 < 0.1 < 1 < 596 0.603 25 74 17 15 9.3 2.1 0.45 1.6 53 1120 2400 310 11200 7700 2900 1.7 < 0.07 < 0.3 < 0.1 < 1 < 597 0.604 25 66 15 15 5.5 2.0 0.43 2.9 43 1100 2300 280 11500 7500 2800 1.1 < 0.07 < 0.3 < 0.1 < 1 < 598 0.6 25 64 16 13 7.4 1.8 1.0 2.2 44 1030 2400 380 11200 7800 2800 0.34 < 0.07 < 0.3 < 0.1 < 1 < 599 0.601 25 76 15 14 8.6 2.1 0.45 2.6 49 1350 2200 450 11400 7800 2900 2.2 < 0.07 < 0.3 < 0.1 < 1 < 5

100 0.601 25 71 18 14 6.7 2.3 0.53 2.7 39 1080 2400 340 11200 7600 2900 1.1 < 0.07 < 0.3 < 0.1 < 1 < 5101 0.599 25 70 15 17 8.8 2.2 1.9 1.5 54 1130 2400 440 11300 7500 2900 1.4 < 0.07 < 0.3 < 0.1 < 1 < 5102 0.603 25 64 18 14 5.9 1.4 < 0.3 1.6 31 1150 2500 360 11200 7300 2700 1.3 < 0.07 < 0.3 < 0.1 < 1 < 5103 0.597 25 72 23 16 10 2.0 1.1 2.5 49 1580 2300 191 11900 7600 3000 1.0 < 0.07 < 0.3 < 0.1 < 1 < 5104 0.596 25 60 14 13 5.0 1.5 < 0.3 2.0 46 1110 2500 270 11000 7400 2900 1.1 < 0.07 < 0.3 < 0.1 < 1 < 5105 0.601 25 74 18 13 7.7 2.3 1.4 3.0 43 1280 2300 310 11400 7800 2900 2.3 < 0.07 < 0.3 < 0.1 < 1 < 5106 0.606 25 71 16 13 7.6 2.5 0.46 2.8 51 1310 2300 157 11300 7700 2900 1.9 < 0.07 < 0.3 < 0.1 < 1 < 5107 0.596 25 65 21 13 6.6 2.2 < 0.3 1.3 43 1030 2600 260 11500 7900 2800 1.8 < 0.07 < 0.3 < 0.1 < 1 < 5

Notes: • All samples were analysed on an OVEN DRIED basis, with extra dilution if required. • Samples for ICP analysis were digested with nitric acid and finished with hydrochloric acid.

LIQUID Samples - This digestion method gives good recovery of all the elements shown in the table above. SOLID Samples - This digestion method gives good recovery of all the elements shown in the table above, except for those indicated below. For nitric/hydrochloric acid digests:

- The aluminium (Al) values are ONLY indicative, as a nitric/hydrochloric acid digest MAY not give a total recovery. - The chromium (Cr) and titanium (Ti) values are ONLY indicative, as a nitric/hydrochloric acid digest does NOT give a total recovery. However these elements (Cr & Ti) MAY be used as a potential indicator for soil/grinder contamination.

• Sample solutions were analysed by either:

• To convert mg/kg to %, divide the value by 10,000. • The limit of determination for the sample is calculated as 10 X the standard deviation of the blank. • Where duplicate analyses are carried out for QA purposes there will be 2 lines in the report for that particular sample. Duplicates give an indication of the homogeneity of the sample. The

first line will give the % variation (%rsd) between the 2 results - we aim for less than 10%, unless there is a problem with the sample e.g. contamination from soil or the method does not recover the total amount of that element as indicated above. Also the closer the results approaches the limit of detection the larger the % variation of the duplicates. The second line of results for the duplicate is the average result of the 2 analyses.


ARL Radial ARL Inductively Coupled Plasma Atomic Emission Spectrometry (ICPAES)

CIROS A Axial CIROS Inductively Coupled Plasma Atomic Emission CIROS R Radial CIROS Inductively Coupled Plasma Atomic Emission Spectrometry (ICPAES)

14

4.9 Harvest damage Graphs 1, 2 and 3 present the average lucerne seed quality through the harvest process for seasons 03/04, 04/05 and 05/06. It is important to note that for logistical reasons the final seed sample was analysed 2-3 months earlier than the research (hand, header, truck, field bin) samples. As a result alterations in relative percentages such as hard seed are a consequence of cleaning rather than hard seed breakdown.

Graph 1: Average lucerne seed quality through harvest process 03/04

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

Hand Header Truck Final

Type of sample

Qua

lity

%

NormalHardAbnormalDead


0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

Hand Good Hand Dry HeaderGood

HeaderGood 2nds

Header Dry Field BinGood

TruckGood

Truck Dry Final Good Final Dry

Type of sample

Qua

lity

%


Note: ‘2nds’ refers to assessment made in this year of the germination percentages of seed that was separated from the primary seed off the gravity table.

15


0

5

10

15

20

2530

35

4045

5055

60

65

70

75

80

8590

HandGood

HandDry

HeaderGood

HeaderDry

FieldBin

Good

FieldBin Dry

TruckGood

TruckDry

FinalGood

FinalDry

TrialHandGood

TrialHandDry

TrialHeaderGood

TrialHeader

Dry

Type of sample

Qua

lity

%


4.10 Ferric Chloride assessment Table 4 presents the assessment of the ferric Chloride test. The test was conducted on the 05/06 cleaned header samples in accordance to the test procedure outlined in section 3.1.6. Each sample test was conducted three times and the stained percentage averaged. Table 4: Comparison of Ferric Chloride staining of 05/06 cleaned header samples

Sample No. Abnormal % Total Seeds Stained %

24 16 100 14 19 6 100 4 2 20 100 18 57 11 100 11 49 10 100 9 62 12 100 11 64 5 100 3 45 14 100 11 53 21 100 19 58 30 100 31 59 40 100 44 60 24 100 22 37 16 100 14 30 10 100 9 54 16 100 16 27 15 100 13 55 19 100 18 35 8 100 6 36 12 100 11 22 18 100 18

16

4.11 Ferric Chloride field evaluation Graph 4 presents the staining data of the Ferric Chloride seed test where 3 seed crops were assessed in 06/07. Appendices 8 and 9 present the field data from the 3 seed crops.

Graph 4: Lucerne seed staining through harvest process 06/07

0

5

10

15

20

25

Hand Header first Header second Final

Type of sample

% s

tain

ed

Seed Crop 1Seed Crop 2Seed Crop 3

17

5. Discussion of results

5.1 Growing conditions The growing conditions were consistent across the time period of this research. Average growing period temperature over the 7 years ranged from 16.89 – 20.86oC and the average relative humidity ranged from 54.12 – 63.74%. In 00/01 and 06/07, the average temperature was the highest being 20.8oC and in these same seasons the relative humidity was the lowest at 54.12% in 06/07 and 55.27% in 00/01. This data reflects the hot dry summers that occur in the research region and that from season to season there are only subtle variances. Wind speeds were consistent over the growing seasons with a constant average of 10.6 km/hr. Of all the seasons, 06/07 had the most days with winds above 40 km/hr. Crop production periods ranged from 88-112 days in 00/01 and 89-119 days in 06/07 to 120-136 days in 01/02. It is known that the growing period is influenced by the weather and that 00/01 and 06/07 were dry and warmer seasons compared to 01/02, which was the coolest of the 7 years. For all the 7 years the ET for lucerne seed crop production ranged from 3.49 – 4.19 ML/ha which was represented by 35 seed crops of mixed varieties. Weather contributes to the quantity of seed production from lucerne crops, especially dryland lucerne seed crops. Lack of rainfall can create a dry, thinner crop on some soil types and this can predispose the crop to increased harvest damage. The region’s net seed production remained consistent over the research period with 04/05 being lower producing than 03/04 and 05/06. In general the weather patterns over the lucerne seed production period are consistent and have no obvious significant bearing on abnormal seedling production as evidenced by the seed quality of all the hand samples (refer section 5.8).

5.2 Environmental conditions The soil type of lucerne seed production paddocks in the research region varies from deep white sand to shallow loamy sand and limestone all over a sand/lime and marl subsoil formation. Irrigated crops in the research project were irrigated by either flood or centre pivot irrigation systems. Irrigation water quality ranged from 2000 – 8000 ppm of total dissolved salts. Equivalent rainfall occurred over the irrigated and dryland seed crops. All seed crops used in the research project were managed under expert agronomical advice available to producers in the region. Crops were sprayed for pest and disease management under integrated pest management strategies based on sound economic thresholds. None of the crops were affected by pest or diseases to a degree that impacted on maximum seed yield potential. Soil type, irrigation water quality, method of irrigation, rainfall, pest and biotic factors have no obvious significant bearing on abnormal seedling production as evidenced by the seed quality of all the hand samples (refer section 5.8).

5.3 Variety, chemical, windrowing, time of harvest factors

Whether the crop was chemically desiccated using a registered desiccant such as Reglone (or an unregistered product such as Sprayseed) or was windrowed, it had no influence on the degree of abnormal seedlings in the hand or final samples. The presence of abnormal seedlings was not increased either by desiccating or by the process of windrowing. It is not known whether desiccation of a crop significantly before full ripeness would increase the abnormal seedling presence but early desiccation is not practiced. It is proposed that this process would reduce yield but not significantly increase abnormal seedling presence, at least not in a hand sample. No variety was less at risk of creating abnormal seedlings and the period of harvest which varied from mid February to early May had no apparent significant impact on the degree of abnormal seedlings in the germination test. The actual time of day of harvest may contribute indirectly to abnormal seedling creation as it is directly relevant to header configuration during the harvest period. Header settings differ on hot harvest days to cool harvest days and as discussed in section 5.8 header configuration can be critical in abnormal seedling creation.

18

5.4 Seed moisture The significant majority of crops used in the research had seed moisture levels less than 12% at point of harvest. In the few instances when seed moisture was above 12% the seed was dried and this had no impact on abnormal seedling presence. If the seed were not dried and stored incorrectly it may have contributed to a rise in the abnormal seedling presence but this is unfounded from this research and the opportunity for this to occur is well outside standard best practice for lucerne seed production. The research determined that seed moisture at harvest had no obvious significant bearing on abnormal seedling production.

5.5 Handling damage

No significant increases in abnormal seedling presence can be attributed to handling damage from header through to final delivery (Graphs 1-3). The use of field bins is not a common practice and not enough data was able to be collected to closely evaluate this extra handling practice. It is suggested it would have a similar result to that of handling from header to truck. If any producer handles the seed harshly it could result in increased abnormal seedling presence. An example of this from the data is in Appendix 6, code E where the handling process from header to field bin greatly increased the abnormal seedling count that remained into the final germination test. It is unknown what actually happened in this case and if in fact there was a sampling error from the header. Given this is the only case of its kind in the research it is treated as an outlier and that, in general, the current process for handing seed from header to processing plant has no significant impact on the overall seed quality.

5.6 Header type Data presented in Appendices 3, 5 and 7 outlines the various header types and settings used during harvest. Regardless of the make, model or whether the header used a conventional or rotary threshing system the header had a direct relationship to the creation of abnormal seedlings. This is discussed in section 5.8.

5.7 Nutritional analysis Seed samples were tested from first and second grade seed, from dryland and irrigated crops, and from all varieties. 20 elements were evaluated in 152 samples over three successive harvests. Tables 1, 2 and 3 when referenced to Appendices 2, 4 and 6 clearly show that there is no relationship between an abnormal seedling and seed nutrition.

5.8 Harvest damage The F-Test for variance differences was undertaken on selected data ranges and no statistically significant differences at the 95% level were detected (e.g. 03/04; Hand vs. Header, F=0.14 < F 0.975 (40,27) = 1.99). On this basis the data as presented in Appendices 2, 4 and 6 were statistically evaluated using a 2 sample, two tailed t-Test assuming equal variances. In Graphs 2 and 3, the terminology of the sample type is used to distinguish between ‘Good’ being typical crops at the point of desiccation or windrowing and ‘Dry’ being crops rated as having dried down too early in the pod filling and ripening period to the point where yield may be compromised. Graph 1 shows there were significantly more abnormal seedlings in the header sample than the hand sample (t=6.89, t0.975 (67) = 1.99) and significantly more abnormal seedlings in the final seed sample than the hand sample (t=5.78, t0.975 (64) = 1.99). There was no significant difference between the ‘hand good’ and ‘hand dry’ sample (t=1.43, t0.975 (28) = 2.05) (Graph 2). In 04/05 there were significantly more abnormal seedlings created by the header in dry crops than in good crops (t=1.75, t0.975 (42) = 2.01) and although in 05/06 this was not significantly different (t=1.55, t0.975 (16) = 2.11) the trend was very strong. There existed no significant differences in

19

years 03/04, 04/05 or 05/06 between header and truck samples or the level of abnormal seedlings in these samples compared to the final seed sample. There were significantly more abnormal seedlings in the header ‘good’ or ‘dry’ samples than the combined hand sample (t=7.37, t0.975 (69) = 1.99) and significantly more abnormal seedlings in the final seed sample than the combined hand sample (t=7.18, t0.975 (57) = 2.00) (Graph 2). There was no significant difference in 04/05 between the final ‘good’ and ‘dry’ samples. A similar result was recorded in 05/06 and presented in Graph 3. There was no significant difference between the ‘hand good’ and ‘hand dry’ sample (t=0.71, t0.975 (20) = 2.09) and there was no significant difference between the final ‘good’ and ‘dry’ samples (t=1.29, t0.975 (20) = 2.09). There were significantly more abnormal seedlings in the header samples than the hand samples ((t=4.95, t0.975 (38) = 2.02) and in the final samples compared to the hand samples (t=7.22, t0.975 (42) = 2.01). In 05/06, and represented in Graph 3, a small trial plot header was used to harvest replicated plots in a bay of flood irrigated lucerne that had been under irrigated with the final irrigation. The irrigation event did not water the entire bay leaving a reasonable area to dry off dramatically before desiccation. Prior to desiccation hand samples were taken from the well irrigated (‘good’) and under irrigated (‘dry’) areas. The small plot header harvested two 40 m2 plots in each treatment. There was no significant difference detected in the abnormal seedling count of the hand samples but there was a significant rise in the presence of abnormal seedlings in both treatments in the header samples. The presence of abnormal seedlings was 12.5% higher in the ‘dry’ plots compared to the ‘good’ plots, which had 20% more abnormal seedlings than the hand samples. This highlighted the impact a header can have on seed quality.

5.9 Ferric Chloride assessment

From table 4 it is clear that there is a very high correlation between the abnormal seedling percentage and the percentage of seed stained in the Ferric Chloride test. This result establishes the validity of the testing procedure to identify damaged seed. The result also crystallises the statistical evidence of the relationship between seed damage caused by headers and abnormal seedling existence.

Figure 1: Lucerne seed sample after 1 minute in Ferric Chloride solution Figure 1 is a photo of lucerne seed in a petri dish soon after addition of Ferric Chloride. Figure 2 is the same seed after 15 minutes in solution. It is evident that there is a presence of black stained seed which highlights damaged seed coat.

20

Figure 2: Lucerne seed sample after 15 minutes in Ferric Chloride solution Figure 3 presents a close up image of the seed in solution depicting, more clearly, the black stained seed. It is evident to see the cracks in the seed coat of some seeds.

Figure 3: Close up of lucerne seed sample in Ferric Chloride solution Figures 4, 5 and 6 present lucerne seeds before and after staining and show the presence of cracks in the seed coat that not evident to the naked eye and are difficult to discern under a seed microscope in the absence of staining. The staining procedure highlights the damage in black which is easily viewed by the naked eye.

21

Crack highlighted black

Crack highlighted black

Figure 4 Figure 5 Figure 6 Figure 4: A single lucerne seed prior to staining.

Figure 5: The seed on the left is unstained and the seed on the right is stained black across a crack on its tip.

Figure 6: A stained lucerne seed with a faint crack on its bottom tip.

5.10 Ferric Chloride field evaluation

Three lucerne seed crop seed samples were assessed by the Ferric Chloride seed test in 06/07. All samples had a low moisture content (<10%). It is evident in Graph 4 that in sites 2 and 3 there was no significant increase in the stained percentage of seeds in the header sample compared to the hand sample. This indicated the strong likelihood that the final seed sample would not have a high level of abnormal seedlings, i.e. it would be under 15%. After cleaning the primary line was tested by Seed Services Australia under the standard ISTA guidelines as previously mentioned. The abnormal seedling percentage of the crops tested was, as predicted, under 15%. It is important in the testing of header samples to appreciate that the level stained is likely to be higher than that of the final sample as there is percentage of stained material in the header sample that will be removed in the cleaning process (e.g. broken seed). The interesting observation was site 1. The initial header sample indicated a 20% increase in the presence of damaged seed between the hand sample and the header sample (Graph 4). The crop was typical of many older irrigated or dryland crops with sufficient plant population for profitable seed production but variable plant production/m2. This impacted on the volume of straw that feeds into the header as it harvests the crop. As suggested in the literature review, when reduced plant material feeds into the header there is the likelihood of more seed damage due to the physical impact of the machine components. If this crop were harvested without review of the settings it is predicted that the abnormal seed percentage would have rendered the final seed lot as failing to fulfil the industry’s minimal standards. The seed test identified the problem and the header’s speed was increased from 7 km/hr to 12 km/hr with no change to any of the other settings (Appendix 9). This increase in speed was ‘foreign territory’ to the contract harvester whom over the years had been confronted with high abnormal seedling percentages in client’s crops. The increase in header speed resulted in a reduction of the seed test stained damaged seed from 23% to 9.0% (Graph 4). The final seed sample, after cleaning, had an abnormal seedling count of 7.0%. The impact of the 15 minute seed test was that the harvested seed exceeded the minimum standards and that it achieved the highest market price possible.

22

6. Implications of research The process of harvesting lucerne seed presents the most significant threat to seed quality. Other factors such as genetics and the environment can influence seed quality minimally in the context of optimum seed quality for marketing. This is evidenced by the low degree of abnormal seedlings in hand samples. The current standard of post harvest seed handing does not affect seed quality significantly. Seed cleaning refines the harvested sample but does not cause abnormal seedlings. Harvest damaged seed (e.g. cracked seed coat) that is of the same specific gravity as an undamaged seed cannot be separated over the gravity table. Apart from removing most weed seeds and offal such as pods, seed cleaning will remove lighter seed (whether damaged or not – this seed is the secondary seed) and a very high proportion of broken seed and husks. The resultant seed sample is generally cleaned to 99.9% purity according to seed analysis results. Harvest damaged seed in the primary sample appears, to the naked eye, to be no different, and until germinated, the presence and degree of abnormal seedlings mostly created by header damage is impossible to quantify. Whilst the research identified that the use of a header is highly correlated to increasing the abnormal seedling presence above a ‘background’ level prior to harvest, there is no option of not using a header to harvest lucerne seed. Consequently, how the header is used is critical. Having stated this there is only so much a header and its operator can do to reduce seed damage and the creation of abnormal seedlings. When a lucerne seed crop has a low plant population the volume of plant mass that feeds into the header front is lower than a crop with a greater plant population harvested at the same ground speed. Header drivers need to drive at a speed that balances the intake of crop into the header with the ability of the separation mechanisms to adequately separate seed from sticks and pods. If the mass passing through the separation mechanisms is too fast, lucerne seed will not separate out into the header box but will fall out the back of the header with the offal. Consequently, the ground speed of the header is limited to a point where too much quality seed is being wasted. Some header operators do not concern themselves with the ground speed of the header but drive according to the grain loss monitor. For example, they keep their header traveling at a speed just below where the grain loss monitor records too much seed passing out the back of the header. Given the relationship between speed and grain loss there are situations in thin crops, whether they are dry or not prior to desiccation or windrowing, that the risk of header damage is high because of the lack of mass feeding into the header concaves and across the thresher. A good flow of crop through the concave permits a softer process of breaking the pods. Due to the need to give lucerne pods a reasonable ‘crush’ to free the seed, the concaves often tend to be closed, especially in tougher (cooler/damper) harvest conditions. A thinner crop has a higher risk of damage through the concave and thresher as the buffer of numerous pods is not available and seed has more opportunity to bounce around and suffer impact damage. This research highlighted that there are certain lucerne seed crops that are at a higher risk of abnormal seedling creation. The highest risk crops are thin and dry with thin good crops and dry dense crops also being at high risk. A header operator should be aware of these risks and set up and operate the header to suit the risks. Frequent seed inspections and the use of the Ferric Chloride test is a tool that can be used, as best as possible, to mitigate the creation of abnormal seedlings. The findings of this research can be applied to all lucerne seed crops grown anywhere in the world. Provided there is consistency and suitability of the prevailing climate and environment, and that crop and seed handling management is of a comparable standard to this research region, the assessment of key factors by this research is predicted to yield similar results. Furthermore, this research is adaptable to all lucerne producing areas in Australia. This research has provided the RIRDC Pasture Seeds Program, and levy paying, certified lucerne seed producers, a number of highly effective and useful outcomes.

23

7. Recommendations The following are recommendations from the research findings: (1) Develop the Ferric Chloride test into a user friendly kit for header operators to monitor the seed

damage through harvest process.

(2) Headers should be operated to suit the crop rather than use standard operating procedures on any type of lucerne seed crop.

(3) Accept that there is a limit on how a header can be operated to reduce damage in high risk crops but that it is also possible to mitigate damage and hence abnormal seedling creation.

(4) All current management practices leading up to and after harvest are not notably contributing to abnormal seedling creation and no specific changes are deemed necessary.

24

8. References 1: De Barro J.E. (2005) ‘Dividing the Droplet: A water balance study for lucerne seed production resourced by an underground aquifer’. RIRDC publication 05/116. 2: De Barro J.E. (2006) ‘The Invisible reality of groundwater salinity: monitoring salt accessions in irrigated lucerne seed production.’ RIRDC publication 06/053. 3: DWLBC (2006) ‘Minimising salt accession in the South East of South Australia – the Border Designated Area and the Hundred of Stirling’. Reports 2006/19 and 2007/pending. 4: DWLBC (2006) ‘Volumetric conversion in the South East of South Australia’. Report DWLBC 2006/30.

25

Appendix 1 Literature Review

Causes of Abnormal Seedlings in Lucerne and

Some Solutions to the Problem:

a review of the literature

FACULTY OF SCIENCES DEPARTMENT OF PLANT SCIENCE November 2002

26

TABLE OF CONTENTS

ACKNOWLEGEMENTS ..................................................................................................................29 EXECUTIVE SUMMARY.................................................................................................................30 1. Introduction..................................................................................................................................32 1.1 Lucerne ......................................................................................................................................32 1.2 Lucerne seed industry ................................................................................................................32

1.2.1 Seed testing and certification......................................................................................33 1.2.2 The problem of abnormals..........................................................................................33

2. Seed testing ..................................................................................................................................35 2.1 The germination test ..................................................................................................................35

2.1.1 Normal seedlings..............................................................................................................35 2.1.2 Ungerminated seeds .........................................................................................................36 2.1.3 Abnormal seedlings..........................................................................................................37

2.2 Normal seedlings of lucerne ......................................................................................................37 2.2.1 Root ..................................................................................................................................38 2.2.2 Hypocotyl .........................................................................................................................38 2.2.3 Shoot apex........................................................................................................................38 2.2.4 Cotyledons........................................................................................................................38 2.2.5 Secondary infection..........................................................................................................38

2.3 Abnormal seedlings of lucerne ..................................................................................................39 2.3.1 Damaged seedlings...........................................................................................................39 2.3.2 Deformed seedlings..........................................................................................................40 2.3.3 Decayed seedlings ............................................................................................................41

3. Causes of abnormal seedlings......................................................................................................42 3.1 Harvesting operations and conditions........................................................................................42

3.1.1 Mechanical injury.............................................................................................................42 3.1.2 Inaccurate timing of harvest .............................................................................................45

3.2 Mineral deficiency of the parent plant .......................................................................................46 3.2.1 Manganese........................................................................................................................46 3.2.2 Calcium ............................................................................................................................46 3.2.3 Potassium .........................................................................................................................47 3.2.4 Boron................................................................................................................................47

3.3 Poor growing conditions............................................................................................................48 3.3.1 High temperatures during ripening...................................................................................48 3.3.2 Chemical treatments .........................................................................................................48 3.3.3 Biotic injury .....................................................................................................................49

3.4 Unsuitable storage conditions ....................................................................................................49 4. Managing abnormal seedling production.....................................................................................50 4.1 Reduced injury at harvest ..........................................................................................................50

4.1.1 Harvester configuration....................................................................................................50 4.1.2 Plant breeding...................................................................................................................50

4.2 Harvesting date ..........................................................................................................................51 4.3 Mineral nutrition of the seed crop..............................................................................................51

27

4.3.1 Adequate fertiliser strategies ............................................................................................51 4.3.2 Seed nutrient treatments ...................................................................................................52

4.4 Use of chemicals ........................................................................................................................52 4.5 Crop protection ..........................................................................................................................53 4.6 Storage conditions......................................................................................................................53 5. Conclusions..................................................................................................................................54 6. References....................................................................................................................................55

28

LIST OF FIGURES Figure 1 Structure of the lucerne seed, showing the location of essential features ..................32

Figure 2 Examples of abnormal lucerne seedlings, classified as damaged...............................39

Figure 3 Examples of abnormal lucerne seedlings, classified as deformed..............................40

Figure 4 An example of an abnormal lucerne seedling, classified as decayed .........................41

LIST OF TABLES Table 1 Percentage of abnormal amaranth seedlings produced after sowing primed (-1.25 MPa, 10 days, 15°C) or non-primed, combine-harvested and threshed or hand-harvested and threshed seed in a greenhouse. Adapted from Pill et al. (1994)........................................43

Table 2 Percentage of abnormal lucerne seedlings produced following classification of mechanical injury to the seed coat. Adapted from Cobb and Jones (1960).............................43

Table 3 Percentage of abnormal lupin seedlings from seed samples harvested at different thresher speeds and moisture contents. Adapted from Hawthorne (1982) ..............................44

Table 4 Percentage of abnormal seedlings produced from parent plants supplied with various rates of calcium and potassium ....................................................................................47

Table 5 Percentage of abnormal sorghum seedlings from parent plants treated with various rates of glyphosate at different times after full bloom. Adapted from Baur et al. (1977)........................................................................................................................................48

29

ACKNOWLEGEMENTS

The photograph of a field of lucerne growing in the South East of South Australia was

kindly provided by Dr. Trevor Garnett, South Australian Research and Development

Institute (SARDI) Lucerne Breeding Group, Adelaide, South Australia.

30

EXECUTIVE SUMMARY

The Australian lucerne seed industry produces over 5000 tonnes of seed per year, with an export value of over $13.5 million. Over 85% of this seed is produced near the town of Keith, in South Australia’s Upper South East. Lucerne seed production is a highly specialised, high value farming enterprise. Every effort is made by growers to produce a superior product that will meet the standards set down by the Australian Seeds Committee, and thus attain certification. However, if a sample is found to contain more than 15% abnormal seedlings during the germination test, it is downgraded, and the grower will receive a lower price. Abnormal seedlings are those broken, weak and/or malformed seedlings that do not have the capacity to develop into a normal plant when grown under favourable field conditions. On average 10% of lucerne seed samples tested in South Australia contain more than 15% abnormals, making this a significant problem within the Australian lucerne seed industry. This review has identified several potential causes of abnormal seedlings in lucerne, and a number of recommendations for future research are suggested. POSSIBLE CAUSES OF ABNORMAL SEEDLINGS

Mechanical injury

Mechanical damage is any sort of breakage to the seed, and is usually caused by rough treatment during harvesting and associated operations. Lucerne is particularly susceptible to mechanical breakage of the embryo parts, due mainly to the orientation of the embryo within the mature seed. Since most mechanical injuries are related to the impact of the seed in the revolving drum of the harvester, the percentage of abnormal seedlings may be reduced by maintaining a constant amount of material within the harvester, and by reducing the speed of the cylinder. There is some evidence from other crops and pastures that genetic variation for resistance to seed injury exists, but little is known of this with respect to lucerne.

Harvest date

The timing of harvest of a lucerne seed crop is critical. Premature harvesting of immature seeds has been shown to correlate with a high incidence of abnormal seedling production in seed crops, while a delay in harvesting will result in an increased number of mechanical injuries, since very dry, brittle seeds are more susceptible to fracturing during handling operations. An optimum moisture content for lucerne seed at harvest is approximately 13%, when seeds are dry enough to prevent the release of destructive enzymes upon impact, and yet not so dry that fracturing will occur.

Mineral deficiency

Seeds of several crop plants deficient in manganese, calcium, potassium and/or boron have been shown to produce a much higher percentage of abnormal seedlings than their nutrient sufficient counterparts. Furthermore, soils of South Australia’s Upper South East are deficient in a number of trace elements, including manganese. Calcium and potassium have also been found deficient in the grain of cereal crops from this area. This suggests that nutritional deficiencies may be a significant cause of abnormal seedling production within a lucerne seed crop.

Poor growing conditions High temperatures during seed development and maturation, particularly during the drying down time from physiological maturity to harvest maturity, have been shown to increase the percentage of abnormal seedlings in a number of crop and pasture species, including lucerne. Biotic injuries too, such as infection by fungi or bacteria and insect attack, may lead to the production of abnormal

31

seedlings in a lucerne seed crop, although this problem can be limited by correct application of pest control measures. Overuse of chemicals, such as fungicides and pre-harvest desiccants, also has the potential to cause abnormal seedlings within a seed crop; however it is unknown whether this is a significant problem among lucerne seed producers in southern Australia. RECOMMENDATIONS FOR FUTURE RESEARCH This review of the literature has identified a number of research areas with the potential to clarify the causes of abnormal seedling production in lucerne seed crops in southern Australia. We suggest that further investigations into the following areas are warranted:

• identification of the genetic variation that exists among lucerne cultivars for resistance to mechanical injury, with a view to breeding lucerne seed with this resistance in the future

• determination of the magnitude of mineral nutrient deficiency as a cause of abnormal seedlings in lucerne

• clarification of the role of chemicals in giving rise to abnormal seedlings in lucerne seed crops of southern Australia.

We believe this research is necessary in order to elucidate the causes of abnormal seedlings from lucerne seed, and represents the way forward for a reduction in the percentage of abnormal seedlings in the crops of Australian lucerne seed producers.

32

1. Introduction 1.1 Lucerne Lucerne (Medicago sativa L.) is a deep-rooted perennial pasture legume that is sown on approximately 1.8 million hectares throughout Australia (SARDI 2001). Lucerne is adapted to a wide range of climatic conditions and will grow on a range of soil types, however it is best suited to deep, well-drained soils of medium to light texture with a good supply of calcium. It does not do well on heavy clays or cracking soils. Seedlings are sensitive to water logging and salinity. Lucerne is a dicotyledonous plant. The seed (Figure 1) is composed predominantly of two cotyledons, or seed leaves, which are pulled above the soil surface during germination by the growth and elongation of the hypocotyl. Once exposed to sunlight, the cotyledons act as true leaves, and begin to photosynthesise. The seed also contains the radicle, or primary root, and the epicotyl, or shoot growing point, in which the first true leaves are present (Bass et al. 1988). A layer of endosperm lies between the cotyledons and the testa, or seed coat (Gunn 1972).

Figure 1 Structure of the lucerne seed, showing the location of essential features (A) External features. (B) Internal features, longitudinal section. (C) Internal features, transverse section. From Gunn (1972). c = cotyledon, en = endosperm, ep = epicotyl, h = hilum, l = lens, m = micropyle, r = radicle, s = seed coat.

Externally a lucerne seed has several visible structures: a centrally located hilum, where the seed was attached to the seed pod; a lens, which is a weak point in the seed coat and is visible as a minute bump; and a micropyle, which is a remnant of the small opening that the pollen tube grew through during the process of fertilisation (Bass et al. 1988). The colour of the seed coat is usually either yellow or olive green to brown, rarely white or black (Gunn 1972).

1.2 Lucerne seed industry The Australian lucerne seed industry produces over 5000 tonnes of seed per year, with an export value of over $13.5 million (De Barro 2001). In 1998/99 there were 298 certified producers of lucerne seed in South Australia, and together these growers produce over 90% of Australia’s total lucerne seed production (Hassall and Associates 2001). Lucerne seed production is a highly specialised, high value farm enterprise. Approximately two-thirds of South Australia’s lucerne seed is grown under irrigation (Teague Australia 2000). However

33

specialised production involves expensive inputs and valuable land, and therefore lucerne seed production is also a high cost and high risk farming venture. In a survey of 40 pasture seed producers across New South Wales, Victoria, South Australia and Western Australia, growers identified a number of items as important contributors to the cost of their production, including fuel, labour, chemicals, repairs and maintenance, seed cleaning and seed testing and certification costs. In addition, this survey also found that lucerne seed production, on a per hectare basis, incurred the greatest production costs of all pasture seeds surveyed (Hassall and Associates 2001). Such an intensive high input crop has an inherent requirement for high successive yields and grower returns. However seed yields of lucerne are known to vary considerably with crop management and seasonal conditions (Morthorpe 1986). Experienced producers are able to limit this variation and associated risk by judicious irrigation management (aimed at optimising the balance between vegetative and reproductive growth), pollination management (optimal number and placement of hives), crop protection (number and timing of measures to control weeds, pests and diseases) and harvest management (optimising both weather conditions and maturity of crop for timing of harvest) (Morthorpe 1986).

1.2.1 Seed testing and certification The greatest return for the lucerne seed grower comes from the production of certified seed. In Australia, the Australian Seeds Committee has set seed purity and seed germination standards, and a crop will not be certified if the minimum requirements for either of these parameters are not met during seed analysis (Hassall and Associates 2001).

The purity analysis gives an indication of the physical quality of the seed lot by defining the composition of the sample, including the identity and legislative status of any foreign seeds within the sample, and the identity of inert particles such as stones or soil within the sample. The grower, therefore, has some control over the purity of his/her seed crop (Primary and Industries and Resources SA 1999).

The germination analysis determines the maximum germination potential of a seed lot and thus gives an estimation of its field planting value. The results of a germination test provide data on the relative proportions of the following groups within the seed lot:

(1) Normal seedlings: those which can be expected to develop into healthy and productive plants in the field;

(2) Abnormal seedlings: those with defects, such as lack of a functional root or shoot system, and are therefore not expected to develop into productive plants; and

(3) Hard and dead seed, and those which have taken in water but are slow to develop, known as fresh, ungerminated seed (Primary Industries and Resources SA 1999).

1.2.2 The problem of abnormals Since the results for hard seed are incorporated into the positive results of a germination analysis, it is predominantly the percentage of abnormal seedlings that determines the quality of each particular sample. If the test results indicate that a sample contains greater than 15% abnormal seedlings then that sample is considered to be of a lower standard, the seed lot is downgraded and the grower will receive a lower price.

Germination test results from Primary Industries and Resources SA (PIRSA) Seed Services’ laboratory over the past seven years indicate that approximately 10% of lucerne seed samples have

34

over 15% abnormal seedlings. The losses associated with this devalued seed over this time are estimated to be in excess of $1 million in export earnings, and an equivalent loss to seed producers (J. De Barro, pers. comm.). Failure of a seed lot to contain 85% normal seedlings reduces a producer’s harvest advance (in many cases to nil) and significantly slows the sale of his/her seed. Consequently cash flow is restricted and with the need to continue financing future production, as well as debt repayments, the grower’s financial status is reduced and the ultimate value of his/her seed is further reduced by losses in interest payments on debts owed in the period awaiting seed payments (J. De Barro, pers. comm.).

Although a high percentage of abnormal seedlings within lucerne seed lots is a frequent occurrence, relatively little information is available as to the causes of this phenomenon. As a result, a grower has little control over the ways in which the crop can be managed in order to avoid a high percentage of abnormals within the seed lot.

Anecdotal evidence appears to rule out the prevailing environmental conditions during growth as a cause of abnormal seedling production, since abnormals appear to occur at random, and do not seem to be related to season or location. Neither does the production of abnormal seedlings appear to be a genetic trait, since the problem does not appear to occur more often in one variety or another, but occurs in all varieties (J. De Barro, pers. comm.). Other evidence suggests that the production of abnormals could be the result of harvester damage, or even crop nutrition. This review of the literature is an attempt to bring together what is known about the production of abnormal seedlings in crop plants, to discuss some of the possible causes of abnormality, both generally and more specifically in lucerne, and finally to recommend some priority areas for future research, so that this problem of abnormal seedlings may be kept to a minimum within the Australian lucerne seed industry.

35

2. Seed testing One of the greatest hazards in agriculture is sowing seed that does not have the capacity to produce an abundant crop of the required cultivar. Seed testing is used to minimise this risk by assessing the quality of seed before it is sown. Seed quality is a concept made up of different attributes and is measured by specific tests including purity, germination, tetrazolium staining, moisture content, sprouting and vigour. These attributes are of interest to various segments of the industry; producers, processors, merchants, farmers, certification authorities, and agencies responsible for seed control. In all cases the ultimate objective is to determine the value of seed for planting (International Seed Testing Association 1985a). Uniformity in seed testing across international boundaries is ensured by organisations such as the International Seed Testing Association (ISTA).

Two of the most important seed tests for all crops are the purity test and the germination test. The purity test defines the composition of a particular seed lot and is based on the physical determination of the components present. Results indicate percentages by weight of: (1) pure seed, (2) other crop seed, (3) weed seed, and (4) inert matter (Copeland and McDonald 1995). The germination test examines the emergence and development of a seedling and indicates whether or not it is able to develop into a satisfactory plant under favourable conditions in soil. This is discussed at length below.

2.1 The germination test A laboratory germination test should indicate the percentage of pure seeds that will produce seedlings capable of continued development under standardised conditions of substrate moisture supply and temperature (Wellington 1966). The germination percentage of a seed lot is therefore the proportion of individual seeds capable of producing normal plants (Justice and Bass 1979). The ultimate objective of a germination test is to gain information about the field planting value of a seed lot and to provide results that can be used to compare the value of different seed lots. The testing instructions established by the ISTA include the germination media required, the temperature for germination, and the duration of the test period for a particular species (Copeland and McDonald 1995).

The rules and conditions for lucerne germination, as set by the ISTA, are a paper substrate (the seeds are either placed on top of a piece of blotting paper or between two blotters), a constant temperature of 20°C, and a duration of 10 days with a first count after 4 days to remove sufficiently well developed seedlings from the substrate to avoid over-crowding. It is also recommended that lucerne be given a pre-chilling treatment to break dormancy (International Seed Testing Association 1985b).

Germination in a laboratory test is defined as the emergence and development of those essential structures from the seed embryo which indicate the seedling’s ability to grow into a normal plant under favourable field conditions. The conditions for laboratory germination must not only be precise enough to initiate the seed’s growth, but must also favour seedling development, within a limited period of time, to a stage in which all essential structures can be evaluated. The results give an indication of the numbers of normal seedlings, ungerminated seeds and abnormal seedlings that will be found within a seed lot (Van Geffen 1986).

2.1.1 Normal seedlings Normal seedlings show the capacity for continued development into satisfactory plants when grown in good quality soil and under favourable conditions of moisture, temperature and light. To be classified as ‘normal’ a seedling must conform to one of the following categories:

[1] Intact seedlings: seedlings with all their essential structures well developed, complete, in proportion and healthy. Depending on the species being tested these seedlings show a specific

36

combination of the following essential structures:

(a) A well developed root system, to ensure adequate uptake of water and nutrients, and to anchor the plant in the soil,

(b) A well developed shoot axis, capable of transporting water and organic materials, and of maintaining the green leaves of the plant in the appropriate position for photosynthesis,

(c) One cotyledon for seedlings of monocotyledons and two cotyledons for seedlings of dicotyledons,

(d) Green and expanding primary leaves,

(e) A terminal bud or shoot apex, the development of which varies with the species being tested.

[2] Seedlings with slight defects: seedlings that show limited damage to their essential structures (such as only one cotyledon in dicotyledons, if there is no evidence of damage to the shoot apex or surrounding tissue), but otherwise show satisfactory and balanced development comparable to the intact seedlings in the same test.

[3] Seedlings with secondary infection: seedlings that would have conformed to [1] or [2] above, but which have been affected by fungi or bacteria from a source other than the parent seed (Wellington 1966, International Seed Testing Association 1985 a,b).

2.1.2 Ungerminated seeds Ungerminated seeds are those which have not germinated by the end of the test period when tested under the specific conditions for that species, as classified below:

[1] Hard seeds: seeds which remain hard at the end of the test period because the seed coat does not allow them to imbibe water. Hardseededness is a form of dormancy which is common in many species of the Leguminosae, but may also occur in other families. The percentage of hard seeds is reported as part of the total percentage germination.

[2] Fresh seeds: seeds which are neither hard nor have germinated, but remain clean, firm and apparently viable at the end of the test period. Fresh seeds result mainly from physiological dormancy. They may be able to imbibe water under the test conditions but further development is blocked.

[3] Dead seeds: seeds which are neither hard nor fresh, nor have produced any part of a seedling at the end of the test period. These seeds are usually soft and discoloured, and are frequently mouldy.

[4] Other categories: under some circumstances ungerminated seeds may be further classified according to the following categories:

(a) Empty seeds: these contain only some residual tissue, or may be completely empty.

(b) Embryoless seeds: these seeds contain fresh endosperm or gametophytic tissue, but no embryonic cavity or embryo.

(c) Insect damaged seeds: seeds which contain insect larvae, or show other evidence of insect attack affecting the ability of the seed to germinate (International Seed Testing Association 1985 a,b).

37

2.1.3 Abnormal seedlings Abnormal seedlings do not have the capacity to develop into a normal plant when grown in good quality soil under favourable conditions of moisture, temperature and light. One or more of the essential structures of the seedling is irreparably defective, and it is therefore of little agricultural value. One or a combination of the following defects in a seedling renders it abnormal:

(1) The primary root is stunted, stubby, retarded, missing, broken, split from the tip, constricted, spindly, trapped in the seed coat, glassy, exhibits negative geotropism or is decayed as a result of primary infection.

(2) The hypocotyl, epicotyl or mesocotyl is short and thick, deeply cracked or broken, split right through, missing, constricted, tightly twisted, bent over, forming a loop or spiral, spindly, glassy or decayed as a result of primary infection.

(3) The cotyledons are swollen or curled, deformed, broken or otherwise damaged, separate or missing, discoloured, necrotic, glassy or decayed as a result of primary infection.

(4) The primary leaves are deformed, damaged, missing, discoloured, necrotic, less than ¼ of normal size or decayed as a result of primary infection.

(5) The terminal bud and surrounding tissue is deformed, damaged, missing or decayed as a result of primary infection.

(6) The seedling as a whole is deformed, fractured, yellow or white, spindly, glassy, has the cotyledons emerging before the root, has two cotyledons fused together, has a persisting endosperm collar or is decayed as a result of primary infection (International Seed Testing Association 1985 a,b).

Abnormal seedlings can also be classified into one of the following three categories:

[1] Damaged seedlings: seedlings with physical damage to the tissues of the embryo so that one or more of their essential structures are missing or so badly and irreparably damaged that balanced development cannot be expected to occur.

[2] Deformed seedlings: seedlings with an irregular pattern of growth giving rise to either weak or unbalanced development, or essential structures which are misshapen or out of proportion.

[3] Decayed seedlings: seedlings with tissues that have undergone major physiological or pathological breakdown. The essential structures of these seedlings are so diseased or decayed as a result of primary infection (from the parent seed) that normal development is prevented. These seeds have been infected by fungi or bacteria, which may be either pathogenic or saprophytic. An intact embryo is immune to saprophytic organisms, but these may gain entry if injury causes the death of any part of the embryo. A seedling is regarded as decayed only if it is clear that the decaying organism is liable to spread (Thomson 1979, International Seed Testing Association 1985 a).

2.2 Normal seedlings of lucerne The ISTA has established a number of guidelines with which to identify the essential structures (roots, hypocotyl, shoot apex, cotyledons) that a perfectly normal lucerne seedling should possess by the end of a germination test (United States Department of Agriculture 1952, Wellington 1970). These guidelines for the evaluation of a normal lucerne seedling are outlined below.

38

2.2.1 Root A normal lucerne seedling must possess a well developed root system, which includes the primary root. The primary root should be long and slender, usually with root hairs. A seedling can be classified as normal if it has roots that are slightly stubby, provided the seedling is otherwise normal. A seedling with short splits on the roots can also be considered normal, provided the split does not extend into the central conducting tissues of the hypocotyl, that root hairs are present, and that the seedling is normal in other respects. Seedlings must not be classified as normal if the primary root fails to develop, or is stunted, even if the hypocotyl elongates and one or more secondary roots develop.

2.2.2 Hypocotyl A normal lucerne seedling must also possess a long, well-developed, undamaged hypocotyl. If the hypocotyl has superficial damage, such as slight cracks or breaks, the seedling may be classified as normal, provided this damage does not extend into the conducting tissues. Seedlings must not be classified as normal if there is evidence of damage to the hypocotyl in the form of a constriction, grainy lesion, or open split which appears likely to interfere with the conducting tissues. Seedlings must not be classed as normal if the hypocotyl is damaged or decayed to an extent which would prevent it from functioning normally, particularly if this damage occurs at the point of attachment to the cotyledons.

2.2.3 Shoot apex The shoot apex of a normal lucerne seedling must take the form of an intact, undamaged epicotyl and plumular bud, although this will not be visible at the end of the test period. It may be assumed that the epicotyl and the plumular bud will develop normally when there is no evidence that the surrounding tissues are damaged or decayed.

2.2.4 Cotyledons To be considered normal a lucerne seedling must possess two attached, undamaged cotyledons, which have opened. However a seedling with one complete cotyledon and no evidence of damage to the epicotyl may also be classified as normal, provided development of the other essential structures is normal. A seedling may also be classified as normal when one or both of the cotyledons are partially damaged or decayed, provided that at least half of the total area appears capable of functioning normally, there is no evidence of damage to the shoot apex, and the other essential structures are normal. If the point of attachment of the cotyledons to the hypocotyl cannot be seen at the end of the test period, the seed coat should be peeled back to determine whether a break or decay has occurred, or whether there is any damage to the epicotyl and plumular bud.

2.2.5 Secondary infection A seedling that is seriously decayed by fungi or bacteria may only be classified as normal when it is clearly evident that the parent seed was not the source of infection. All essential structures must also have been present. It should be established that the initial infection was remote from the point of contact between the seedling and its seed coat, and was due to proximity to other seeds or seedlings during the test. The spread of infection by Ascochyta spp. is particularly rapid during tests on lucerne, and special precautions, such as extra wide spacing of individual seeds, may be necessary to prevent secondary infection.

39

2.3 Abnormal seedlings of lucerne The ISTA has also established a series of guidelines that describe the appearance of abnormal lucerne seedlings, in order that a seed analyst may accurately classify a seedling as abnormal (United States Department of Agriculture 1952, Wellington 1970). These descriptions fall into the three general categories of damaged, deformed or decayed seedlings, and are outlined below.

2.3.1 Damaged seedlings Lucerne seedlings are classified as abnormal when one or more of the essential structures fail to develop normally because of damage to the embryo. Such damaged seedlings include those with no primary root (Figure 2b), and those with the primary root split longitudinally into two parts. Damaged seedlings are also abnormal if there is a constriction, grainy lesion or open split in the hypocotyl, which appears likely to interfere with the conducting tissues (Figure 2c). Other damaged seedlings classified as abnormal include those with no cotyledons attached, or with only one cotyledon attached and damage to the epicotyl, and seedlings with more than half the total area of both cotyledons broken off or not capable of functioning normally.

Figure 2 Examples of abnormal lucerne seedlings, classified as damaged

(A) Normal seedling. (B) No primary root. (C) Hypocotyl with constriction, grainy lesion, or split likely to interfere with the conducting tissues. Arrows indicate the area of damaged tissue. Adapted from Wellington (1970).

40

2.3.2 Deformed seedlings Deformed lucerne seedlings are classified as abnormal when their development as a whole is out of proportion compared with that of a normal seedling germinated at the same time. Included in this category are seedlings that are short and weak (Figure 3b), or have a spindly or watery appearance, provided this is not the result of excess moisture in the substrate. Deformed seedlings are also classified as abnormal when the primary root is short and stubby, and this is usually associated with a short and thickened hypocotyl (Figure 3c). Seedlings are abnormal if the hypocotyl is twisted, or watery and translucent in appearance. In addition, seedlings with two enlarged cotyledons and a very short radicle are classed as abnormal.

(A)

Figure 3 Examples of abnormal lucerne seedlings, classified as deformed

(A) Normal seedling. (B) Short and weak seedling. (C) Short and stunted primary root, with short, thick hypocotyl. Arrows indicate the areas of deformed tissue. Adapted from Wellington (1970).

41

2.3.3 Decayed seedlings This category includes those lucerne seedlings with any of their essential structures so badly decayed that normal development would be prevented, provided the decay did not spread to the seedling from an adjacent seed or seedling. Decayed seedlings are those with more than half of the total area of the cotyledons decayed (Figure 4b), often caused by infection with Ascochyta spp., and those with decay of the hypocotyl or primary root.

Seedlings with any decay or discolouration of the cotyledons at the point of attachment to the hypocotyl are also classified as abnormal, since the shoot apex is likely to be affected.

(A)

Figure 4 An example of an abnormal lucerne seedling, classified as decayed (A) Normal seedling. (B) Decayed cotyledons. Arrows indicate the area of decayed tissue. Adapted from Wellington (1970).

42

3. Causes of abnormal seedlings The establishment of a normal seedling has been found to depend on the physical environment during germination, the physiological condition of the embryo, and the availability of stored nutrients within the seed. For the purposes of this review, a seedling’s physical environment during germination can be disregarded, since here we are concerned with the reasons for the development of abnormal seedlings within the standard conditions of a germination test, prior to planting in the field. Justice (1972) has identified a number of causes of abnormal seedlings in crop plants. Those that contribute to the deterioration of an embryo’s physiological condition include mechanical injury, chemical treatments, biotic injury (including infection by pathogenic organisms and insect attack), and declining vitality as a result of unfavourable storage conditions. Justice (1972) also identifies mineral deficiency in the soil on which the seed is produced as a cause of abnormal seedling development.

3.1 Harvesting operations and conditions

3.1.1 Mechanical injury Mechanical injury is any sort of breakage to the seed, and is usually caused by rough treatment during the harvesting, threshing, handling and processing operations (Justice 1972). The actual amount of damage depends on how well or poorly the equipment was adjusted, and whether or not the correct equipment was employed in all operations. The greater the impact on the seed, the greater the level of injury, and the lower the germination percentage (Friedman 1985). While breaks and fractures are the predominant types of mechanical injury sustained by dry seeds, seeds with sufficient moisture for toughness may also suffer injuries, normally in the form of bruising (Moore 1972). A mechanical injury may be externally visible, such as a crack in the seed coat, or it may be internal and may not be discovered until the seed has germinated.

3.1.1.1 GENERAL MECHANICAL DAMAGE Borthwick (1932) was among the first to describe the various types of seed injuries that can be sustained by mechanical damage. In a study of machine-threshed lima beans, he found that practically every part of the embryo was susceptible to thresher injury. These injuries included detachment of the cotyledons, loss of the radicle, and breaks or splits in the hypocotyl. Borthwick (1932) also noted that injuries to the radicle and hypocotyl were completely absent in hand-shelled beans, while injuries to the cotyledons and plumule were also less frequent than in machine-threshed beans. Similarly Pill and colleagues (1994) showed that combine-harvesting resulted in 7.8% abnormal amaranth seedlings, while hand-harvesting produced only 2.8% abnormals. An osmotic priming treatment, used to invigorate seeds and permit pregerminative physiological and biochemical activities, had no significant effect on abnormal seedling percentage (Table 1). This phenomenon of hand harvested and threshed seeds sustaining a much smaller degree of mechanical damage than machine harvested and threshed seeds has been described in several crop species, including maize (Tatum and Zuber 1943), lucerne (Cobb and Jones 1960, Popescu and Craiu 1996), green beans (Clark and Kline 1965), soybean (Green et al. 1966, Prakobboon 1982), kidney beans (Wilson and McDonald 1992) and lupins (Dracup et al. 2001).

43

Table 1 Percentage of abnormal amaranth seedlings produced after sowing primed (-1.25 MPa, 10 days, 15°C) or non-primed, combine-harvested and threshed or hand-harvested and threshed seed in a greenhouse. Adapted from Pill et al. (1994)

Some species are more susceptible to mechanical injury of than others, mainly because of the size and location of the embryo within the seed. Lucerne has been found to be particularly susceptible to mechanical breakage of the embryo parts, along with the large-seeded pulse crops (beans, peas, soybeans), clovers and some of the cereals (Colbry et al. 1961). Mechanical damage results in abnormal seedlings with injuries to the cotyledons, so that the total cotyledon area is reduced to less than half; fractures of the hypocotyl or epicotyl; and injuries to the plumular bud. Essential structures may have open splits or constrictions likely to affect the conducting tissues. Root damage is also found, and although the seedling may develop adventitious roots to compensate for the loss of the radicle, it is unlikely to produce a normal plant in a crop (MacKay 1972).

Cracking of the seed coat is another important type of mechanical injury sustained during the harvesting and handling procedures. Cracked seed coats not only allow rapid entry of oxygen and moisture, which will cause seed deterioration during storage, but also provide an entry point for fungi and bacteria which can result in decayed seedlings. Moreover, a cracked seed coat often indicates that the seed has suffered serious injury to one or more of its essential structures and may therefore give rise to a damaged seedling (Cobb and Jones 1960). This is particularly evident in legumes, since any cracks in the seed coat of these plants will be in contact with the embryo, either over the embryonic axis, or over the cotyledons, and will thus cause damage to living tissue. In a visual examination of lucerne seed coats, Cobb and Jones (1960) found that the percentage of abnormal seedlings increased with the degree of injury to the testa. Seeds with an intact seed coat produced an average of 11% abnormal seedlings, those with a slight seed coat fracture produced 36% abnormals, while those with cracked or broken seed coats gave rise to over 60% abnormal seedlings (Table 2).

Table 2 Percentage of abnormal lucerne seedlings produced following classification of mechanical injury to the seed coat. Adapted from Cobb and Jones (1960)

Harvesting and threshing injuries are clearly related to the speed of the revolving drum in the harvester. Hawthorne (1982), working with lupins in the South East of South Australia, showed that an increase in thresher speed significantly increased the percentage of abnormal lupin seedlings from

44

31% to 65%, but only at a low seed moisture content of <11.7% (Table 3). Thresher speed did not influence abnormal seedling percentage at higher seed moisture contents. Other researchers have demonstrated similar results in lupins (Blanchard 1990, Dracup et al. 2001), and a number of other crop plants, including soybean (Green et al. 1966), navy beans (Singh and Linvill 1977), field peas (Blanchard 1990) and amaranth (Krishnan et al. 1994). Popescu and Craiu (1996) also found that the effect of mechanical harvesting on injury varied depending on the hour of the day, with the percentage of damaged lucerne seed being 12% at 12.30 pm and 26.5% at 5.45 pm. Again, this is most likely due to the variations in moisture content of the seed at different times of the day.

Table 3 Percentage of abnormal lupin seedlings from seed samples harvested at different thresher speeds and moisture contents. Adapted from Hawthorne (1982)

* Abnormal percentage includes ungerminated seeds in these results.

3.1.1.2 MECHANICAL DAMAGE IN LUCERNE Cobb and Jones (1960) were among the first to discover that mechanical damage is a major cause of germination failure in lucerne seed. Factors that contribute to the degree of damage sustained by the seed include resistance to removal from the pod, seed size and shape, and seed moisture content (Bass et al. 1988). During development of the embryo in lucerne, and other Medicago and Trifolium species, two lobes first become differentiated as the cotyledons, and then the hypocotyl elongates and becomes curved so that it lies parallel to the cotyledons when the seed is mature, and not between them as in the large-seeded legumes. In this position it is more exposed to mechanical injury, and breaks at the point of attachment of the cotyledons to the hypocotyl, with accompanying injury to the epicotyl, are very common in lucerne seeds that have been mechanically damaged (United States Department of Agriculture 1952, Cobb and Jones 1960, Wellington 1970).

A simple test for recognising mechanically damaged lucerne seeds was developed by Cobb and Jones (1960), based on the visual examination of the seed coat. Using a binocular microscope at 10x magnification, these researchers were able to examine the seed coats of 400 newly harvested lucerne seeds in less than 30 minutes, and this information was then used to estimate the germination percentage. This test has been routinely used on lucerne seed lots certified in California (Bass et al. 1988). French and colleagues (1962) further developed this method with the use of a stain, indoxyl acetate, to aid in the identification of seed coat fractures in lucerne and other light coloured legume seeds. In many cases this removed the need for seed magnification, since only cracked seeds were stained, and the stained area became much larger than the seed coat crack.

While embryo injuries are usually accompanied by broken seed coats, not all mechanical damage to lucerne seed can be observed by examination of the seed coat. Internal injuries, including undeveloped and broken cotyledons, radicles and shoot apices, in lucerne seed showing no external damage were found by Hruskova (1992) to be caused by mechanical injury, and the conclusion was made that more careful seed handling is necessary in order to reduce internal mechanical damage.

45

3.1.2 Inaccurate timing of harvest Seed germination percentage has been found to be markedly influenced by harvest date, and hence by seed maturity and seed moisture content (Nichols et al. 1978). Seed moisture content is a critical factor in seed deterioration (Bass et al. 1988). The moisture content of seeds at time of harvest is determined principally by existing weather conditions. Failure to have a crop in the correct condition at harvest may lead to increased mechanical damage of the seed, which reflects on the quality of seed placed on the market (RIRDC 1997). Mechanical injury has been found to be related directly to seed moisture content. Seeds with an optimum level of moisture at harvest, approximately 14% depending on the crop, are dry enough to prevent cell rupturing and release of destructive, hydrolytic enzymes upon impaction, and yet are not dry and brittle enough to promote fracturing (Moore 1972).

3.1.2.1 PREMATURE HARVESTING Harvesting of immature, high moisture content seeds has been shown to correlate with a low germination percentage and a high incidence of abnormal seedling production in a number of crops, including pulses (Nichols et al. 1978) and vegetables (Spurr et al. 2002). If a crop is prematurely harvested, when nutrients are still being transferred from the plant to the seeds, the seeds are likely to have a high moisture content which will make them liable to heat rapidly, unless they are artificially dried, leading to the production of abnormal seedlings when germination occurs (Wellington 1966).

3.1.2.2 DELAYED HARVESTING Although mechanical harvesting at a high moisture content does cause injury, predominantly bruising, to seeds, it does not cause as much damage as harvesting at a low moisture content. Very dry and brittle seeds are more susceptible to fracturing of their essential seed parts, resulting in more split seeds, cracked seedcoats and damaged embryos. This was reported by Tatum and Zuber (1943) in maize, and later in a number of other crop species, including soybean (Green et al. 1966, Prakobboon 1982), navy beans (Wijandi and Copeland 1974), lupins (Blanchard 1990, Dracup et al. 2001), field peas (Blanchard 1990) and kidney beans (Wilson and McDonald 1992). Most of these researchers claim an optimal moisture level for harvesting to be 12-14%, with physical damage likely to increase significantly as moisture decreases below this level. However often a suitable moisture content for seeds during storage must also be taken into account. The recommended maximum moisture content for lucerne seed in storage is 10% (Morthorpe 1995), and therefore a compromise must be achieved. A high moisture content will reduce the longevity of seeds in storage, particularly if bruising has occurred, and it may be necessary to reduce the moisture content of high-moisture seeds soon after harvest.

46

The drying of seed artificially after harvest in order to reduce its moisture content to a safe level for storage may affect germination if the temperature is too high. In barley, high temperature during drying has produced deformed seedlings in which the elongation of the seminal roots was restricted or prevented, while in others the plumule did not elongate and failed to emerge from its covering layers (MacKay 1972).

3.2 Mineral deficiency of the parent plant Deficiencies of certain mineral elements in the soil on which seeds are produced have been found to generate seeds that germinate poorly, and produce abnormal, weakened seedlings, even if the seeds are germinated in complete nutrient media. Moreover soils that are deficient in one or more macro- or micronutrients are known to occur throughout the southern Australian lucerne seed producing districts. This is discussed at length in section 4.3.1.1 of this review. Although specific evidence for lucerne seed crops is not available, seeds of a number of crop plants deficient in manganese, calcium, potassium or boron have been shown to produce a much higher percentage of abnormal seedlings than their nutrient sufficient counterparts, and many of these abnormalities are characteristic of the particular nutrient deficiency.

3.2.1 Manganese Seed manganese deficiency in peas and beans is characterised by the appearance of a sunken, brown, and slightly pithy area in the centre of the flat surfaces of the cotyledons, and sometimes browning of the plumules. This condition is known as ‘marsh spot’ and although shoots may develop in the axils of the cotyledons, usually the plumular bud is destroyed and the resultant seedlings do not produce normal plants (Reynolds 1955, Justice 1972).

A low manganese seed content in lupins has been found to produce weak, chlorotic, wilted seedlings. Other abnormalities include cotyledons that appear above the soil surface but with no further growth, and upside down emergence, with radicles above the soil surface (Longnecker et al. 1996).

3.2.2 Calcium Low seed calcium concentrations are also associated with reduced seed germination and an increase in the production of abnormal seedlings. Harrington (1960) was one of the first to demonstrate this fact, when he showed that calcium deficiency reduced germination, decreased the percentage of normal seeds produced, and shortened the storage life of seeds of three vegetable crops, pepper, carrot and lettuce. The author concluded that calcium deficiency is more harmful for seed germination than deficiencies of nitrogen, phosphorus or potassium. Frost and Kretchman (1989) showed that cucumber seeds produced in a hydroponic solution containing low calcium

(40 mg Ca l 1) produced a significantly higher percentage of abnormal seedlings (58% vs 4%) than

those produced with adequate calcium (160 mg Ca l-1

). Similarly, Keiser and Mullen (1993) found that a decrease in the percentage of abnormal soybean seedlings from 25.5% to 9.3% coincided with an increase in calcium added to the nutrient solution from 0 mM to 2.5 mM (Table 4). Abnormal seedlings produced from soybean seed deficient in calcium showed undeveloped or absent primary roots, cracked or decayed hypocotyls, decayed epicotyls and/or absence of the terminal bud.

47

Table 4 Percentage of abnormal seedlings produced from parent plants supplied with various rates of calcium and potassium

3.2.3 Potassium Results obtained by Harrington (1960), in pepper and carrot seeds, demonstrate that potassium deficiency also results in a low germination percentage, and further, that seed from potassium deficient plants deteriorates faster in storage than seed from control plants supplied with adequate

nutrients. These results were supported by Ison (1980), who reported that 50 kg potassium ha-1

supplied to a potassium deficient sandy loam soil decreased the percentage of abnormal seedlings produced by french bean seeds from 43% down to 5% (Table 4).

3.2.4 Boron Abnormal seedlings are also produced as a result of low seed boron concentrations. Bell et al. (1989) found abnormalities including the absence of the shoot apex, epicotyl, one or both cotyledons and/or the primary root in black gram seedlings produced from seed containing low levels of boron.

The authors conclude that a boron concentration of 6 mg kg-1

is critical for the production of normal seedlings. Abnormal seedlings from B deficient seed have also been observed in peas (Leggatt 1948), turnip rape (Saarela 1985) and soybean (Rerkasem et al. 1997).

Rerkasem et al. (1997) suggest that soybean seeds with a low concentration of boron (7-10 mg kg-1

) have permanently damaged embryos, which prevents their germination or produces defective seedlings. At slightly higher boron concentrations, embryos are not permanently damaged, but require a higher level of boron in the soil in order to produce normal seedlings, than those seeds with higher concentrations of seed boron.

Germination percentage also appears to be influenced by the ratio of boron to calcium in the seed. Keiser and Mullen (1993) found that normal seedling percentage was negatively correlated to the B:Ca ratio in soybean seeds. As the seed B:Ca ratio increased, the percentage of normal seedlings decreased linearly, and the percentage of abnormal seedlings increased.

48

3.3 Poor growing conditions

3.3.1 High temperatures during ripening A low germination percentage in certain crops has also been attributed to high temperatures during seed development and maturation, particularly during the drying down period from physiological maturity to harvest maturity (Dotzenko et al. 1967). Controlled environment experiments with lucerne have shown that seeds produced under a high temperature regime have a reduced germination percentage (Dotzenko et al. 1967, Walter and Jensen 1970), and it has been suggested that this may explain the differences in abnormal seedling production from year to year, or in the same year, from location to location (Dotzenko et al. 1967).

In the US, soybean seed produced from later planting dates that reach maturity after hot, dry weather, generally has a higher germination percentage and field emergence than seed that matures during hot, dry growing conditions (Amable 1976). High temperatures, high relative humidity, and precipitation will speed field deterioration of seed, and this has been reported in a number of crops, including pulses (Amable 1976), annual pastures (Jansen and Ison 1994) and vegetables (Spurr et al. 2002). Furthermore, seed exposed to weathering in the field is more susceptible to mechanical damage, and therefore to the associated production of abnormal seedlings, as outlined above.

3.3.2 Chemical treatments Seeds that have been incorrectly exposed to chemicals, particularly those that have been over-treated with toxic fungicides, commonly produce abnormal seedlings. The symptoms include stunting and thickening of the roots and hypocotyls. In some observed cases the treatment had permanently killed certain of the essential seedling organs (Colbry et al. 1961). Seeds that have been treated or accidentally subjected to certain chemical pesticides, such as 2,4-D and phenol compounds, may also produce abnormal seedlings (Justice 1972). The herbicide glyphosate, applied as a pre-harvest desiccant to aid harvest, has been shown by Baur and colleagues (1977) to significantly increase the number of abnormal seedlings from the seed of sorghum, from 2% (nil glyphosate applied) up to

49% (4.48 kg ha-1

glyphosate) (Table 5). Bennett and Shaw (2000) also found certain varieties of soybean to be susceptible to seed injury by glyphosate. Although there is no direct evidence that chemicals are responsible for abnormal seedlings in lucerne, these results from other crops suggest that a lucerne seed crop may also be at risk from the over-use of chemicals.

Table 5 Percentage of abnormal sorghum seedlings from parent plants treated with various rates of glyphosate at different times after full bloom. Adapted from Baur et al. (1977). Seed moisture content (%) Abnormal seedlings (%)

Glyphosate applied (kg ha-1) 0 1.12 2.24 4.48

35 2 24 54 49

27 4 10 18 47

20 2 4 4 6 15 2 5 10 17

49

3.3.3 Biotic injury 3.3.3.1 INFECTION WITH PATHOGENIC ORGANISMS Pathogenic organisms in the field may infect seed during its formation and ripening, and seedlings grown from it may be attacked during germination tests (MacKay 1972, Wu and Cheng 1990). These infected seeds usually initiate growth, but one or more of the essential seedling structures may frequently be damaged or destroyed by the fungi or bacteria. As discussed above, seedlings infected with Ascochyta spp. may show decay or discolouration of the cotyledons, and if this affects the area adjacent to the shoot apex, or covers more than one half of the total cotyledon area, the seedling is considered abnormal. Since the manifestations of disease on the seedlings are largely dependent on environmental conditions during the test period, germination results may be erratic unless the conditions are carefully controlled (Justice 1972). Lucerne seed frequently presents problems in this respect, as does seed of the large-seeded legumes, sweet clover, cereals, cotton, rhubarb, celery, radish and flax.

3.3.3.2 INSECT DAMAGE Insect infestation is common in seeds such as vetch, field peas, cowpea, some clovers and lucerne (Colbry et al. 1961). Seeds that have been infested with insects may produce damaged seedlings that lack an essential part or structure, or the seedling may be severely stunted or weakened. Weevils, for example, have been found responsible for abnormal seedling production in sorghum, field peas and cowpeas (Justice 1972). In lucerne in South Australia the principal insect problem is lucerne seed wasp (Bruchophagus roddi) damage (De Barro 2001), but it is unknown whether this insect is directly involved in the production of abnormal seedlings.

3.4 Unsuitable storage conditions Germination capacity declines as seed ages during storage, but complete death is usually preceded by the production of abnormal seedlings whose development is weak or unbalanced because the loss of vital functions does not occur simultaneously in the different tissues (MacKay and Flood 1969, MacKay 1972). Seeds that have been subjected to unfavourable storage conditions have been found to produce a number of abnormal seedlings in the same way; some tissues remain physiologically alive, even though the embryo as a whole has lost the ability to grow into a normal plant (Thomson 1979). Typical symptoms include the absence or stunting of one or more essential seedling structures, such as restricted root and shoot development (Justice 1972). In the Leguminosae and Cruciferae an additional symptom of declining seed viability is the breakdown of hypocotyl tissue, which then gives a glassy or watery appearance (Haferkamp et al. 1953, MacKay 1972). Furthermore, under high moisture storage conditions, decay of some essential seedling structures may be frequent (MacKay 1972).

As discussed above, the effect of mechanical damage on the germination percentage of seeds can be quite serious. However the delayed effects of injury to seeds can also cause problems (Moore 1972). During storage the injured areas act as sites for infection, and this results in accelerated aging and a shortened duration of viability (MacKay and Tonkin 1967). Injured areas may also promote rapid weakening and early death of surrounding normal tissues. An initial injury may be non-critical, but if it is located on or near an essential part of the embryo structure, a viable seed may readily become non-viable with only a small amount of additional deterioration. Injuries located in the vicinity of the embryonic axis usually bring about a more rapid loss of viability in storage than injuries of a similar size located in less important areas of the seed (Moore 1972).

50

4. Managing abnormal seedling production

4.1 Reduced injury at harvest

4.1.1 Harvester configuration A standard combine harvester is used to pick up a windrowed lucerne seed crop and thresh the seed from the pod. The equipment must be carefully adjusted to separate the very small seed from the large amount of plant material without damaging it. Mueller (1998) suggests that most lucerne seed injury during harvesting is due to either impact in the cylinder of the combine because of small loads, or because of excessive cylinder speeds. It has been found that when small amounts of material pass through the cylinder seed damage tends to be greater than when the load is heavy, although losses of seed increase with heavy loads. Increasing or reducing the speed of the harvester as it moves through heavy and light areas of the paddock can balance loads, however. By varying the speed of the harvester, the amount of straw fed into the machine can remain fairly constant.

Excessive cylinder speeds can also lead to unnecessary seed injury, therefore a balance between damaged seed and incomplete threshing must be achieved (Mueller 1998). Reducing cylinder speed will reduce damage to the seed, but if speeds are too low seeds will not be removed from the pod during threshing. Lower speeds can also leave the straw in better condition, so that the seed can be separated and cleaned more efficiently with less loss over the rear of the machine.

Mueller (1998) suggests that a valuable estimate of damage can be obtained in a short time by checking 4 to 10 seed samples. One damaged seed, identified using a hand lens, in a sample of 100 seeds indicates 3-5% damage. Two or three damaged seeds indicate 5-10% damage. If the injury is greater than 5% the cylinder speed should be adjusted downward until the damage index falls below 5%. Seed samples should be tested throughout the harvest period to determine if changed conditions of weather, crop or combine during harvest have altered the damage estimate.

Every lost or damaged seed occurring during harvest is removed from the grower’s profit. It is therefore essential that proper threshing methods are carried out, at a correct moisture content, in order to avoid damage and the production of abnormal seedlings, and maintain high seed quality.

4.1.2 Plant breeding There is some evidence that certain species have lines and cultivars that are more susceptible to mechanical damage during harvest and handling than others. Soybeans, for example, have been found to show highly significant varietal differences for resistance to mechanical damage (Costa et al. 1987, Vieira et al. 1994, Carbonell and Krzyzanowsky 1995). Genetic variation for resistance to mechanical damage has also been demonstrated in snap beans, and a number of mechanisms responsible for the greater resistance of some lines have been suggested, including tighter adherence of the seed coat to the cotyledons and resistance of the cotyledons to cracking (Dickson and Boettger 1976) and better embryonic axis protection by the cotyledons (Bay et al. 1995). More recently Peltonen-Sainio et al. (2001) have demonstrated differences between naked oat cultivars in susceptibility to mechanical damage, and they suggest groat hardness and protrusion of the embryo as two potential methods for screening naked oat lines for injury resistance.

Anecdotal evidence suggests that abnormal seedlings in lucerne seem to occur equally in all varieties, and there has been little or no research into genetic variation among lucerne cultivars for resistance to seed mechanical damage. However the results from other crops suggest that this may be another method of decreasing the number of abnormal seedlings produced in lucerne seed crops, and an investigation into the genetic variation for resistance to seed injury that exists among

51

Australian and overseas lucerne cultivars is warranted.

4.2 Harvesting date While little can be done to prevent abnormal seedlings occurring as a result of unfavourable growing conditions, such as high temperatures during maturation, choice of harvest date is somewhat more manageable by the grower. Kowithayakorn and Hill (1982) propose that lucerne seed should not be harvested until maturity at 40 days after pollination, at which stage maximum seed dry weight (physiological maturity) has been attained. At this stage approximately 50% of the pods may be yellow and the remainder green (Morthorpe 1995). Husman (1999) goes further to suggest that the harvesting of a lucerne seed crop should begin when seed moisture content is 13% or less. If moisture content is above 13%, there is risk of heating, seed damage and combine losses. Premature harvesting is therefore detrimental to the quality of a seed crop, but likewise a delay in harvesting is also potentially harmful and may also give rise to abnormal seedlings. As discussed above, mechanical injuries to seed increase significantly as seed moisture content falls below 12%. Clearly timing of harvest is one of the most critical management factors that influence seed quality, as a grower must attempt to maximise seed weight, germinability and vigour, while minimising seed moisture content in order to optimise the quality and quantity of his/her seed lot.

4.3 Mineral nutrition of the seed crop

4.3.1 Adequate fertiliser strategies The detrimental effects of seed nutrient deficiencies in the germination of abnormal seedlings may be corrected by growing the seed under adequate nutrient conditions. There is little information in the literature concerning the production of abnormal seedlings in lucerne by seeds deficient in mineral nutrients, but the evidence provided by a range of other crops suggests that deficiencies of manganese, calcium, potassium and/or boron in the seed can give rise to abnormal seedlings with characteristic deformities following germination.

4.3.1.1 INFERTILE SOILS OF THE UPPER SOUTH EAST Mineral deficiencies occur in crops and pastures on a range of soils across southern Australia, including South Australia’s Upper South East, where 85% of Australia’s total lucerne seed production takes place. Soils in this area are highly infertile, and include dune ridges of calcareous sands, extensive areas of leached polsolized sands, and solonetz soils consisting of surface sand overlying a clay subsoil rich in sodium. Soils such as the Laffer sand and allied types are also common throughout the Upper South East (Tiver 1955). Once known as the Ninety Mile Desert of heath and scrub (Riceman 1948), nutrient deficiencies can still occur in this area, despite the widespread use of fertilisers since the 1950’s. In a survey of nutrient concentrations in barley grains from each Hundred of South Australia covering the decade 1983-1993, Reuter (1995) found low levels of several nutrients in the grain from the Upper South East, including manganese, calcium and potassium. Thus the potential exists for deficiencies of these nutrients to occur in lucerne seed crops grown in this area, although there is no published data to confirm this at this stage.

4.3.1.2 MANGANESE DEFICIENCY Reuter (1995) recorded low manganese concentrations (< 11 mg kg

-1) in barley grain from the Upper

South East in all four years surveyed (1983, 1989, 1992 and 1993), and a number of crops have been shown to respond to manganese applications in this area of South Australia, including subterranean clover, oats and peas (Tiver 1955), lupins (Hannam and Riggs 1985), safflower (Lewis and McFarlane 1986), wheat (Mackareth 1996) and faba beans (Paull 2002). Furthermore, the amelioration of manganese deficiency can often be difficult in areas such as this, since manganese is quickly rendered unavailable on these highly calcareous soils, often within a week of application, and uptake is further decreased in the cool, wet winter months (Tiver 1955, McDonald et al. 2001).

52

4.3.1.3 POTASSIUM DEFICIENCY Low potassium concentrations in the barley grain from the Upper South East were recorded by Reuter (1995) in three of the four years surveyed (1983, 1989 and 1993). In some Hundreds this concentration was less than 0.4% potassium. Potassium deficiency in lucerne has been reported in northern New South Wales (Grewal and Williams 1998), and in the western United States, where it is described in irrigated crops growing on alkaline calcareous soils (James 1988). Furthermore, this author and colleagues suggest that fertiliser effects on low potassium soils are short lived, and that moderate rates of potassium fertiliser should be applied on a yearly basis, in order to correct this problem (James et al. 1995).

4.3.1.4 CALCIUM DEFICIENCY Reuter (1995) also found generally low levels of calcium in the barley grain harvested from the cropping areas throughout South Australia, despite the relative abundance of calcareous soils in the State. The author suggests that this may be due to the relative immobility of calcium in plants. Calcium deficiency in lucerne has been reported in Europe (Petkov 1979), as well as in northern New South Wales (Grewal and Williams 1998).

4.3.1.5 SUMMARY It can be seen from the above that deficiencies of the nutrients likely to give rise to abnormal seedlings have been shown to occur in lucerne crops, and further, that it is likely that these deficiencies will occur on the sandy soils of South Australia’s Upper South East. Further studies are required to clarify the role of these and other nutrients in the germination of seeds and seedling establishment, but at present the evidence suggests that lucerne seed producers need to employ optimum fertiliser tactics for their crops, thus to ensure that inadequate nutrition is not responsible for abnormal seedling production within their seed lots.

4.3.2 Seed nutrient treatments It should be noted that a number studies have shown the possibility of alleviating some of these detrimental effects of seed nutrient deficiencies. Application of calcium nitrate to the germination medium was shown to restore viability to calcium deficient seed in snap bean (Clark and Kline 1965) and soybean (Smiciklas et al. 1989), while adding boron to the germination medium was shown to prevent abnormal seedlings when the boron concentration in the seed is marginal in pea (Leggatt 1948) and soybean (Rerkasem et al. 1997). It may therefore be possible to treat seed of low nutrient contents with the appropriate nutrient (particularly calcium and boron) before sowing in order to limit abnormal seedling generation (Rerkasem et al. 1997). These results are of little use to the seed producer however, since it is the percentage of abnormal seedlings produced under the constant conditions of the germination test that will determine the certification of his/her seed lot, not whether the adverse effects of low seed nutrient content can be mitigated when sown for the next crop.

4.4 Use of chemicals Although the evidence in the literature is somewhat limited, it appears that some chemicals applied to crops (particularly overexposure of certain chemicals) can give rise to abnormal seedlings from the seed of the treated parent plant. It is acknowledged that herbicides and pesticides are an essential part of lucerne seed production, but the judicious use of some chemicals would seem prudent in the light of the evidence available. Clearly additional research is necessary to further elucidate the effect of chemicals on lucerne seed crops, and to determine the likelihood of these chemicals generating abnormal seedlings in the next generation.

53

4.5 Crop protection As outlined above, certain insect and pathogenic organisms may attack a lucerne seed crop, and may be responsible for the generation of abnormal seedlings from the seed. It is likely, however, that most growers would have an integrated pest management program in place for the control of these pests, since not only do they give rise to abnormal seedlings, but they usually also bring about a more immediate and visible effect, a reduction in yield. Therefore there is possibly little more that a grower can do in order to eliminate abnormal seedlings produced in this way, beyond the usual crop protection strategies of crop monitoring and correct identification of, and control measures for, the target species.

4.6 Storage conditions Legume seeds in general store well, and lucerne is no exception (Wilton et al. 1978, Roberts 1999). Providing the seed is of good initial quality and stored under recommended conditions (approximately 3-7% moisture content for legume seeds, and at less than 18°C), seed viability can be maintained for many years, and the production of abnormal seedlings will remain at an acceptable percentage. If seed is of low initial viability however, or if inferior storage conditions are used, viability may drop sooner to unsatisfactory levels (Roberts 1999). Abnormal seedlings produced during storage are of little concern to the lucerne seed producer, however, since it is the initial quality of his/her seed lot that is assessed for certification requirements.

54

5. Conclusions This literature review has identified a number of causes of abnormal seedlings within lucerne seed lots, and has proposed several ways in which a grower may be able to limit the production of these abnormals, and thus maintain an acceptable standard for certification at the germination test. Possible areas of future research into the phenomenon of abnormal seedling production have been highlighted.

1 Mechanical injury: Lucerne has been found to be a particularly susceptible crop to mechanical damage, which can occur at any time during the harvesting and handling operations. Mechanical injuries are largely related to the impact of the seed in the revolving drum of the harvester, and can be reduced by maintaining a constant amount of material within the harvester, and by reducing cylinder speed. This origin of abnormal seedlings can explain why one grower may experience a high level of abnormals within his/her seed lot, while his neighbour may not. Further research is necessary to determine the potential for breeding lucerne seed with resistance to mechanical injury.

2 Harvest date: The timing of harvest of a lucerne seed crop is critical. Lucerne seed should be harvested at a optimal seed moisture content that is dry enough to prevent embryo injury from bruising upon impaction, and yet not dry and brittle enough to aggravate mechanical damage by fracturing.

3 Mineral deficiencies: Soils deficient in certain mineral nutrients, particularly manganese, calcium, potassium and boron, can give rise to abnormal seedlings from the seeds produced on them. Manganese deficiency is common on the highly alkaline soils of South Australia’s Upper South East, while calcium and potassium deficiencies have also been reported on the soils of this area. Although further research is necessary to clarify the magnitude of this problem in lucerne, a grower can ensure that nutrient deficiency is not responsible for abnormal seedlings produced within his/her lucerne seed crop by maintaining an adequate fertiliser regime.

4 Chemical treatments: Overuse of some chemicals has been found to produce abnormal seedlings in the next generation of several crop species. Further research is required to determine if this is a significant problem among the lucerne seed producers of southern Australia.

5 Biotic injury: Pests and diseases are responsible for a limited number of abnormal seedlings within seed lots. Grower vigilance and attention to conventional pest management practices may limit this source of embryo injury.

It can be seen from this review of the literature that a producer of lucerne seed has a number of options available with which to limit the number of abnormal seedlings in his/her crop, thereby improving his/her chances of attaining seed certification for the seed lot. Future research may further increase these options available to growers, which in turn will reduce the overall abnormal seedling percentage within the industry from the current 10%, and thus allow the Australian lucerne seed industry to maintain and expand its existing markets.

55

6. References Amable, R. (1976). The effects of harvest date and storage conditions on soybean seed and quality.

In: Doku, E.V. (Ed.) Proceedings of the Joint University of Ghana-Council for Scientific and Industrial Research Symposium on Grain Legumes in Ghana. University of Ghana : Legon, Ghana. pp. 109-118.

Bass, L.N., Gunn, C.R., Hesterman, O.B. and Roos, E.E. (1988). Seed physiology, seedling performance and seed sprouting. In: Hanson, A.A., Barnes, D.K. and Hill, R.R. (Eds) Alfalfa and alfalfa improvement. American Society of Agronomy, Crop Science Society of America and Soil Science Society of America : Madison, Wisconsin. pp. 961983.

Baur, J.R., Miller, F.R. and Bovey, R.W. (1977). Effect of preharvest dessication with glyphosate on grain sorghum seed. Agron. J. 69: 1015-1018.

Bay, A.P.M., Taylor, A.G. and Paine, D.H. (1995). Mechanical damage resistance of snap bean (Phaseolus vulgaris L.) seeds: protection of the embryonic axis by the cotyledons. Plant Var. Seeds. 8: 151-159.

Bell, R.W., McLay, L., Plaskett, D., Dell, B. and Loneragan, J.F. (1989). Germination and vigour of black gram (Vigna mungo (L.) Hepper) seed from plants grown with and without boron. Aust. J. Agric. Res. 40: 273-279.

Bennett, A.C. and Shaw, D.R. (2000). Effect of preharvest dessicants on Group IV Glycine max seed viability. Weed Sci. 48: 426-430.

Blanchard, E.D. (1990). The effect of mechanical damage on the seed viability of lupin and field pea, grain legume seeds. In: Agricultural engineering conference, Toowoomba, 11-14 November 1990. The Institution of Engineers, Australia : Barton, A.C.T. pp. 4649.

Borthwick, H.A. (1932). Thresher injury in baby lima beans. J. Agric. Res. 44: 503-510.

Carbonell, S.A.M. and Krzyzanowsky, F.C. (1995). The pendulum test for screening soybean genotypes for seeds resistant to mechanical damage. Seed Sci. Technol. 23: 331339.

Clark, B.E. and Kline, D.B. (1965). Effects of water temperature, seed moisture content, mechanical injury and calcium nitrate solutions on the germination of snap bean seeds in laboratory germination tests. Proc. Assoc. Off. Seed Anal. 55: 110-120.

Cobb, R.D. and Jones, L.G. (1960). Germination of alfalfa (Medicago sativa L.) as related to mechanical damage of seed. Proc. Assoc. Off. Seed Anal. 50: 101-108.

Colbry, V.L., Swofford, T.F. and Moore, R.P. (1961). Tests for germination in the laboratory. In: Seeds: The yearbook of agriculture 1961. United States Department of Agriculture : Washington D.C. pp. 433-443.

Copeland, L.O. and McDonald, M.B. (1995). Principles of seed science and technology. 3rd

ed. Chapman and Hall : New York.

Costa, A.V., Kueneman, E.A. and Monteiro, P.M.F.O. (1987). Varietal differences in soybeans for resistance to physical damage of seed. Soybean Genet. Newsl. 14: 73-76.

De Barro, J. (2001). Evaluating and managing lucerne seed wasp in lucerne seed crops. Rural Industries Research and Development Corporation : Barton, A.C.T.

Dickson, M.H. and Boettger, M.A. (1976). Factors associated with resistance to mechanical damage in snap beans (Phaseolus vulgaris L.). J. Amer. Soc. Hort. Sci. 101: 541-544.

Dotzenko, A.D., Cooper, C.S., Dobrenz, A.K., Laude, H.M., Massengale, M.A. and Feltner, K.C. (1967). Temperature stress on growth and seed characteristics of grasses and legumes. Colo. Agric. Exp. Stn. Tech. Bull. 97: 1-27.

56

Dracup, M., Thomson, R. and Galwey, N. (2001). Restricted branching narrow-leafed lupin. 3. Poor establishment. Aust. J. Exp. Agric. 41: 781-786.

French, R.C., Thompson, J.A. and Kingsolver, C.H. (1962). Indoxyl acetate as an indicator of cracked seed coats of white beans and other light colored legume seeds. J. Amer. Soc. Hort. Sci. 80: 377-386.

Friedman, M. (1985). A contribution to lucerne seed resistance to mechanical injury. In: Third international conference on physical properties of agricultural materials. VSZ : Prague, Czechoslovakia. pp. 213-217.

Frost, D.J. and Kretchman, D.W. (1989). Calcium deficiency reduces cucumber fruit and seed quality. J. Amer. Soc. Hort. Sci. 114: 552-556.

Grewal, H.S. and Williams, R. (1998). Nutrient deficiencies in lucerne pastures under rotation. [Online, accessed 8 November 2002] URL: http://www.dpi.gov.au/extra/pdf/fsi/life_dec1998.pdf

Green, D.E., Cavanah, L.E. and Pinnell, E.L. (1966). Effect of seed moisture content, field weathering and combine cylinder speed on soybean seed quality. Crop Sci. 6: 7-10.

Gunn, C.R. (1972). Seed characteristics. In: Hanson, A.A. (Ed.) Alfalfa science and technology. American Society of Agronomy : Madison, Wisconsin. pp. 677-687.

Haferkamp, M.E., Smith, L. and Nilan, R.A. (1953). Studies on aged seeds I. Relation of age of seed to germination and longevity. Agron. J. 45: 434-437.

Hannam, R.J. and Riggs, J.L. (1985). The effect of manganese applied from an aircraft as a low-volume foliar spray in preventing manganese deficiency in Lupinus angustifolius. L. Fert. Res. 6: 146-156.

Harrington, J.F. (1960). Germination of seeds from carrot, lettuce, and pepper plants grown under severe nutrient deficiencies. Hilgardia 30: 219-235.

Hassall and Associates Pty Ltd (2001). A study of the costs of production of lucerne, medic and clover seeds in Australia. Rural Industries Research and Development Corporation : Barton, A.C.T.

Hawthorne, W.A. (1982). Survey observations on lupin seed quality. In: Agronomy Australia 1982: Proceedings of the Second Australian Agronomy Conference, Riverina College of Advanced Education, Wagga Wagga, July 1982. Australian Society of Agronomy : Parkville, Victoria. pp. 227.

Hruskova, H. (1992). Internal seed damage and germination in lucerne. Rostl. Vyroba.

38: 73-80,

Husman, S.H. (1999). Growing alfalfa for seed in Arizona. [Online, accessed 2 September 2002] URL: http://ag.arizona.edu/pubs/crops/az1129.pdf

International Seed Testing Association (2002). What is ISTA. [Online, accessed 21 October 2002] URL: http://www.seedtest.org

International Seed Testing Association (1985a). International Rules for Seed Testing Rules 1985. Seed Sci. Technol. 13: 299-355.

International Seed Testing Association (1985b). International Rules for Seed Testing Annexes 1985. Seed Sci. Technol. 13: 356-513.

Ison, R. (1980). The effect of potassium on french bean seed yield and seed vigour. In:

Pathways to productivity: Proceedings of the Australian Agronomy Conference, Queensland Agricultural College, Lawes, April 1980. Consolidated Fertilizers Ltd : Brisbane. pp. 303.

James, D.W. (1988). Leaf margin chlorosis in alfalfa: a potassium deficiency symptom associated

57

with high concentration of sodium in the leaf. Soil Sci. 145: 374-380.

James, D.W., Tindall, T.A., Hurst, C.J. and Hussein, A.N. (1995). Alfalfa cultivar responses to phosphorus and potassium deficiency: biomass. J. Plant Nutr. 18: 24312445.

Jansen, P.I. and Ison, R.L. (1994). Hydration-dehydration and subsequent storage effects on seed of the self-regenerating annuals Trifolium balansae and T. resupinatum. Seed Sci. Technol. 22: 435-447.

Justice, O.L. (1972). Essentials of seed testing. In: Kozlowski, T.T. (Ed.) Seed biology. Volume III: Insects and seed collection, storage, testing and certification. Academic Press : New York. pp. 301-370.

Justice, O.L. (1979). Principles and practices of seed storage. Castle House Publications : Tunbridge Wells, Kent, U.K.

Keiser, J.R. and Mullen, R.E. (1993). Calcium and relative humidity effects on soybean seed nutrition and seed quality. Crop Sci. 33: 1345-1349.

Kowithayakorn, L. and Hill, M.J. (1982). A study of lucerne seed development and some aspects of hard seed content. Seed Sci. Technol. 10: 179-186.

Krishnan, P., Evans, T.A. and Pill, W.G. (1994). Threshing cylinder speed affects germination of Amaranthus cruentus L. seeds. HortSci. 29: 652-654.

Leggatt, C.W. (1948). Germination of boron deficient peas. Sci. Agr. 28: 131-139.

Lewis, D.C. and McFarlane, J.D. (1986). Effect of foliar applied manganese on the growth of safflower (Carthamus tinctorius L.) and diagnosis of manganese deficiency by plant tissue and seed analysis. Aust. J. Agric. Res. 37: 567-572.

Longnecker, N., Crosbie, J., Davies, F. and Robson, A. (1996). Low manganese concentration and decreased emergence of Lupinus angustifolius. Crop Sci. 36: 355-361.

MacKay, D.B. (1972). The measurement of viability. In: Roberts, E.H. (Ed.) Viability of seeds. Chapman and Hall : London. pp. 172-208.

MacKay, D.B. and Flood, R.J. (1969). Investigations in crop seed longevity. III. The viability of grass and clover seed stored in permeable and impermeable containers. J. Natl. Inst. Agric. Bot. 11: 521-546.

MacKay, D.B. and Tonkin, J.H.B. (1967). Investigations in crop seed longevity. I. An analysis of long-term experiments, with special reference to the influence of species, cultivar, provenance and season. J. Natl. Inst. Agric. Bot. 11: 209-225.

Mackerath, T. (1996). Independent evaluation of LIG-ZINC and MANGANESE in beans. [Online, accessed 5 November 2002] URL: http://www.sjbagnutri.com.au/ligzinkmnbatrials.htm

McDonald, G.K., Graham, R.D., Lloyd, J., Lewis, J., Lonergan, P. and Khabas-Saberi, H. (2001). Breeding for improved zinc and manganese efficiency in wheat and barley. Proc. 10th Aust. Agron. Conf. Hobart. www.regional.org.au/au/asa/2001/

Moore, R.P. (1972). Effects of mechanical injuries on viability. In: Roberts, E.H. (Ed.) Viability of seeds. Chapman and Hall : London. pp. 94-113.

Morthorpe, K. (1995). Developments in lucerne seed production. In: Lowe, K.F. (Ed.)

Australian lucerne breeding and agronomy workshop: held at Yanco Agricultural Institute, Yanco, NSW, 7-11 December 1986. Department of Primary Industries, Queensland : Brisbane. pp. 99-105.

Mueller, S. (1998). Seed crop notes: August 1998. University of California : Fresno, California.

58

Nichols, M.A., Scott, D.J., Warrington, I.J. and Green , L.M. (1978). Pre-harvest treatment effects on some quality criteria of pea seeds. Acta Hortic. 83: 113-124.

Paull, J. (2002). Faba bean breeding. [Online, accessed 5 November 2002] URL: http://www.adelaide.edu.au/sciences/plant/research/breeding_groups/jp.html

Peltonen-Sainio, P., Muurinen, S., Vilppu, M., Rajala, A., Gates, F. and Kirkkari, A.M. (2001). J. Agric. Sci. (Camb.) 137: 147-156.

Petkov, V. (1979). The physiology of calcium nutrition of lucerne. IV. Effect of calcium deficiency on formation of dry matter and reproductive organs, its absorption and accumulation by plants. Rasteniev’d Nauki. 16: 3-10.

Pill, W.G., Evans, T.A. and Krishnan, P. (1994). Priming improves germination and emergence of combine-harvested Amaranthus cruentus L. seeds. HortSci. 29: 655-658.

Popescu, S. and Craiu, D. (1996). Mechanical damage, the cause of decreased seed germination in alfalfa. Prob. Agrofitoteh. Teor. Apl. 18: 193-199.

Prakobboon, N. (1982). A study of abnormal seedling development in soybean as affected by threshing injury. Seed Sci. Technol. 10: 495-500.

Primary Industries and Resources SA (1999). Seed Analytical Services. [Online, accessed 2 September 2002] URL: http://www.pir.sa.gov.au

Rerkasem, B., Bell, R.W., Lodkaew, S. and Loneragan, J.F. (1997). Relationship of seed boron concentration to germination and growth of soybean (Glycine max). Nutrient Cycling in Agroecosystems. 48: 217-223.

Reuter, D.J. (1995). Survey of nutrient levels in barley grains: a potential indicator of soil fertility status. Grains Research and Development Corporation, Unpublished Report CSO135S.

Reynolds, J.D. (1955). Marsh spot of peas: a review of present knowledge. J. Sci. Food Agric. 6: 725-734.

Riceman, D.S. (1948). Mineral deficiency in plants on the soils of the ninety-mile plain in South Australia. J. Coun. Scient. Ind. Res. (Aust.) 21: 229-235.

RIRDC (1997). Compendium of research projects completed between January 1996 and June 1997: Pasture seeds. [Online, accessed 13 September 2002] URL:

http://www.rirdc.gov.au/pub/97comp/pasture.html

Roberts, E.H. (1999). The influence of storage conditions on seed viability. In: Bennett,

S.J. and Cocks, P.S. (Eds) Genetic Resources of Mediterranean Pasture and Forage Legumes. Kluwer Academic Publishers : Dordrecht, The Netherlands. pp. 132-140.

Saarela, I. (1985). Plant-available boron in soils and the boron requirement of spring oilseed rapes. Annls. Agric. Fenniae. 24: 183-265.

SARDI (2001). Lucerne. [Online, accessed 16 October 2002] URL: http://www.sardi.sa.gov.au

Singh, B. and Linvill, D.E. (1977). Determining the effect of pod and grain moisture content on threshing loss and damage of navy beans. Trans Am. Soc. Agric. Engrs. 20: 226-227.

Smiciklas, K.D., Mullen, R.E., Carlson, R.E. and Knapp, A.D. (1989). Drought-induced stress effect on soybean seed calcium and quality. Crop Sci. 29: 1519-1523.

Spurr, C.J., Fulton, D.A., Brown, P.H. and Clark, R.J. (2002). Changes in seed yield and quality with maturity in onion (Allium cepa L., cv. ‘Early Cream Gold’). J. Agron. Crop Sci. 188: 275-280.

Tatum, L.A. and Zuber, M.S. (1943). Germination of maize under adverse conditions. J. Amer. Soc. Agron. 35: 48-59.

59

Teague Australia (2000). Production statistics – seed. [Online, accessed 11 October 2002] URL: http://www.tjt.com.au

Thomson, J.R. (1979). An introduction to seed technology. Leonard Hill : Glasgow.

Tiver, N.S. Deficiencies in South Australian soils. J. Dep. Agric. S. Aust. 59: 100-113.

United States Department of Agriculture (1952). Manual for testing agricultural and vegetable seeds: Agriculture handbook No. 30. United States Department of Agriculture : Washington D.C.

Van Geffen, A. (1986). An introduction to germination testing. In: Srivastava, J.P. and Simarski, L.T. (Eds). Seed production technology. International Centre for Agricultural Research in Dry Areas : Aleppo, Syria. pp. 160-183.

Vieira, C.P., Vieira, R.D. and Paschoalick, J.H.N. (1994). Effects of mechanical damage during soybean seed processing on physiological seed quality and storage potential. Seed Sci. Technol. 22: 581-589.

Walter, L.E. and Jensen, E.H. (1970). Effect of environment during seed production on seedling vigor of two alfalfa varieties. Crop Sci. 10: 635-638.

Wellington, P.S. (1966). Seed production and testing. J. R. Agric. Soc. Engl. 127: 164186.

Wellington, P.S. (1970). Handbook for seedling evaluation. Proc. Int. Seed Test. Assoc. 35: 449-597.

Wijandi, S. and Copeland, L.O. (1974). Effect of origin, moisture content, maturity, and mechanical damage on seed and seedling vigor of bean. Agron. J. 66: 546-548.

Wilson, D.O. and McDonald, M.B. (1992). Mechanical damage in bean (Phaseolus vulgaris L.) seed in mechanized and non-mechanized threshing systems. Seed Sci. Technol. 20: 571-582.

Wilton, A.C., Townsend, C.E., Lorenz, R.J. and Rogler, G.A. (1978). Longevity of alfalfa seed. Crop Sci. 18: 1091-1093.

Wu, W.S. and Cheng, K.C. (1990). Relationships between seed health, seed vigor and the performance of sorghum in the field. Seed Sci. Technol. 18: 713-720.

60

Appendix 2: 03/04 seed quality data Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture %

83 A VENUS Keith I Final Normal 1 50 11 39 0 67 A VENUS Keith I Hand Normal 1 51 1 43 4 10 A VENUS Keith I Header Normal 1 52 7 41 0 1 A VENUS Keith I Truck Normal 1 47 12 41 0

51 B FG Keith D Final Normal 1 88 5 7 0 59 B FG Keith D Hand Normal 1 89 2 8 1 3 B FG Keith D Header Normal 1 84 4 12 0 2 B FG Keith D Truck Normal 1 87 6 5 0

119 C FG Keith I Final Normal 1 62 5 32 0 <10.0 60 C FG Keith I Hand Normal 1 67 3 26 4 11 C FG Keith I Header Normal 1 51 5 43 1 8 C FG Keith I Truck Normal 1 66 10 24 0

113 D FG Laffer D Final Normal 1 73 12 15 0 7.1 53 D FG Laffer D Hand Normal 1 86 3 11 0 57 D FG Laffer D Hand Normal 1 84 4 12 0 63 D FG Laffer D Hand Normal 1 72 6 22 0 16 D FG Laffer D Header Normal 1 81 8 10 1 15 D FG Laffer D Truck Normal 1 82 7 11 0 107 E HALL Laffer D Final Normal 1 86 8 6 0 6.3 29 E HALL Laffer D Hand Normal 1 83 2 14 1 30 E HALL Laffer D Hand Normal 1 88 1 11 0 49 E HALL Laffer D Hand Normal 1 76 3 21 0 65 E HALL Laffer D Hand Normal 1 86 1 13 0 5 E HALL Laffer D Header Normal 1 82 7 11 0

18 E HALL Laffer D Header Normal 1 82 11 7 0 4 E HALL Laffer D Truck Normal 1 82 7 11 0

19 E HALL Laffer D Truck Normal 1 82 8 10 0 75 F HR Keith I Final Normal 1 50 5 44 0 58 F HR Keith I Hand Normal 1 40 3 56 1

61

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 12 F HR Keith I Header Normal 1 44 7 48 1 13 F HR Keith I Truck Normal 1 47 5 47 1 69 G FG Keith I Final Normal 1 49 9 41 1 66 G FG Keith I Hand Normal 1 32 2 62 4 9 G FG Keith I Header Normal 1 44 11 44 1

72 G FG Keith I Truck Normal 1 45 8 46 1 118 H S/SIR Keith I Final Normal 1 86 6 7 0 <10.0 54 H S/SIR Keith I Hand Normal 1 41 2 48 9 95 H S/SIR Keith I Hand Normal 1 36 5 57 1 102 H S/SIR Keith I Header Normal 1 43 7 50 0 103 H S/SIR Keith I Truck Normal 1 49 5 46 0 44 I FG Laffer I Field Bin Normal 1 63 7 29 1 108 I FG Laffer I Final Normal 1 52 29 19 0 37 I FG Laffer I Hand Normal 1 64 2 34 0 98 I FG Laffer I Header Normal 1 68 14 18 0 76 I FG Laffer I Truck Normal 1 61 15 21 3 114 J FDALE Laffer D Final Normal 1 87 7 6 0 9.1 48 J FDALE Laffer D Hand Normal 1 92 3 4 1 52 J FDALE Laffer D Hand Normal 1 88 5 7 0 80 J FDALE Laffer D Header Normal 1 80 12 8 0 78 J FDALE Laffer D Truck Normal 1 79 12 9 0 115 K SIR Laffer I Final Normal 1 67 8 25 0 10.1 71 K SIR Laffer I Hand Normal 1 65 8 27 0 82 K SIR Laffer I Hand Normal 1 63 10 27 0 77 K SIR Laffer I Header Normal 1 61 11 27 1 121 L SIRO Brimbago D Final Normal 1 88 3 9 0 13.5 32 L SIRO Brimbago D Hand Normal 1 82 4 12 2 61 L SIRO Brimbago D Hand Normal 1 75 2 22 0 26 L SIRO Brimbago D Header Normal 1 82 7 11 0 8.5 27 L SIRO Brimbago D Truck Normal 1 86 3 11 0 116 M AUR Keith I Final Normal 1 65 7 27 0 9.4 36 M AUR Keith I Hand Normal 1 37 7 45 11

62

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 62 M AUR Keith I Hand Normal 1 50 7 38 5 81 M AUR Keith I Header Normal 1 53 14 29 4 86 M AUR Keith I Truck Normal 1 60 7 33 0 90 N FG Tintinara I Field Bin Normal 1 80 6 13 1 122 N FG Tintinara I Final Normal 1 73 9 17 0 8.5 39 N FG Tintinara I Hand Normal 1 96 1 3 0 91 N FG Tintinara I Header Normal 1 83 8 9 0 94 N FG Tintinara I Truck Normal 1 80 10 10 0 35 O S/CUF Coombe I Final Normal 1 46 6 48 0 43 O S/CUF Coombe I Hand Normal 1 28 5 51 16 73 O S/CUF Coombe I Header Normal 1 42 4 54 0 79 O S/CUF Coombe I Truck Normal 1 42 11 46 1 109 P FDALE Coombe I Final Normal 1 75 15 7 3 13.5 41 P FDALE Coombe I Hand Normal 1 33 2 56 8 96 P FDALE Coombe I Header Normal 1 45 6 38 11 70 P FDALE Coombe I Truck Normal 1 41 4 55 0 110 Q AUR Laffer D Final Normal 1 67 28 5 0 <10.0 50 Q AUR Laffer D Hand Normal 1 74 4 22 0 68 Q AUR Laffer D Hand Normal 1 79 6 13 2 20 Q AUR Laffer D Header Normal 1 82 5 13 0 7.0 21 Q AUR Laffer D Truck Normal 1 83 6 11 0 111 R FG Laffer D Final Normal 1 64 33 3 0 < 7.0 28 R FG Laffer D Hand Normal 1 61 2 37 0 31 R FG Laffer D Hand Normal 1 79 5 16 0 24 R FG Laffer D Header Normal 1 75 6 18 0 25 R FG Laffer D Truck Normal 1 78 3 19 0 117 S FG Brimbago I Final Normal 1 44 3 53 0 11.0 45 S FG Brimbago I Hand Normal 1 40 2 53 5 100 S FG Brimbago I Header Normal 1 39 7 54 0 97 S FG Brimbago I Truck Normal 1 41 8 51 0 120 T HR Keith I Final Normal 1 72 8 20 0 <10.0 46 T HR Keith I Hand Normal 1 36 3 59 2

63

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 55 T HR Keith I Hand Normal 1 31 3 64 2 64 T HR Keith I Hand Normal 1 37 1 61 1 7 T HR Mt Monster I Header Normal 1 60 10 30 0 6 T HR Mt Monster I Truck Normal 1 56 5 39 0

104 U S/SIR Coombe I Final Normal 1 85 8 7 0 7.0 104 U S/SIR Coombe I Final Normal 1 85 8 7 0 7.0 40 U S/SIR Coombe I Hand Normal 1 36 6 50 8 89 U S/SIR Coombe I Header Normal 1 42 6 52 0 93 U S/SIR Coombe I Header Normal 1 41 9 49 1 101 U S/SIR Coombe I Header Normal 1 42 8 50 0 87 U S/SIR Coombe I Truck Normal 1 43 7 50 0 92 U S/SIR Coombe I Truck Normal 1 47 8 45 0 99 U S/SIR Coombe I Truck Normal 1 41 8 51 0 105 V AQUA Coombe D Final Normal 1 80 12 8 0 <7.0 34 V AQUA Coombe D Hand Normal 1 91 1 9 0 56 V AQUA Coombe D Hand Normal 1 81 4 14 1 23 V AQUA Coombe D Header Normal 1 84 6 10 0 14 V AQUA Coombe D Truck Normal 1 83 10 6 1 106 W SIRO Coombe I Final Normal 1 74 8 18 0 8.1 42 W SIRO Coombe I Hand Normal 1 30 7 46 17 47 W SIRO Coombe I Hand Normal 1 24 3 59 14 74 W SIRO Coombe I Header Normal 1 49 9 39 3 88 W SIRO Coombe I Header Normal 1 35 5 58 2 84 W SIRO Coombe I Truck Normal 1 39 7 53 1 85 W SIRO Coombe I Truck Normal 1 43 9 46 2 112 X AUR Keith I Final Normal 1 81 13 6 0 8.8 33 X AUR Keith I Hand Normal 1 92 1 6 1 38 X AUR Keith I Hand Normal 1 89 1 10 0 17 X AUR Keith I Header Normal 1 90 3 7 0 22 X AUR Keith I Truck Normal 1 88 3 9 0

64

Appendix 3: 03/04 harvest data

Sample No. Code Variety Harvest

Date Header Concave Rotar Thresher Fan Speed Front Comments

1 A VENUS

10 A VENUS 3/04/2004 CASE 2388 0 (closed) 940 760 5.5 p/up Windrow Harvest

67 A VENUS Pre Windrow Sample: 30/3/04 83 A VENUS 8 B FG 11 B FG 14/04/2004 INT 1460 1 830 570 2 7 Desiccated Harvest 60 B FG Post desiccation sample: 14/4/04 2 C FG 3 C FG 2/03/2004 INT 1460 1 830 540 5 7 m Harvested in mid afternoon. Desiccated 51 C FG 59 C FG Pre desiccation sample:L12 17/2/04 15 D FG 16 D FG 20/02/2004 NH 8080 5-7 (open) 600 7 p/up Mid afternoon windrow harvest 53 D FG Pre windrow sample: 16/2/04 57 D FG Post harvest sample/30 mm rain: 11/3/04 63 D FG Windrow, day of harvest 4 E HALL 5 E HALL 4/03/2004 NH 8080 3-7 (open) 590 7.5 p/up Windrow, day of harvest; hot/windy 18 E HALL 8/03/2004 NH 8080 2 (closed) 630 9 7.5 m 19 E HALL 29 E HALL Post desiccation sample: day of harvest 30 E HALL Pre Desiccation sample 49 E HALL Windrow sample, day of harvest 65 E HALL Pre windrow sample 12 F HR 5/04/2004 JD1075 7-10 mm 900 850 4 6 m Mid afternoon harvest. Desiccated 13 F HR 58 F HR Post desiccation sample, 5/4/04

65



75 F HR 9 G FG 19/04/2004 JD1075 7-10 mm 900 850 4 6 m Mid afternoon harvest. Desiccated sample 66 G FG Post Desiccation sample: 19/4/04 69 G FG 72 G FG 54 H S/SIR Pre desiccation sample: 13/4/04 95 H S/SIR 21/4/04: Desiccated sample

102 H S/SIR 21/04/2004 JD9650

CTS 0 850 825 3 103 H S/SIR 37 I FG Pre windrow sample 44 I FG 76 I FG 98 I FG 13/04/2004 NH 8080 1-2 (open) 600 4 p/up Windrow harvest 48 J FDALE Pre rain and pre desiccation sample 52 J FDALE Post rain and desiccation sample 78 J FDALE 80 J FDALE 17/02/2004 NH 8080 1-2 (open) 600 4 p/up 71 K SIR Windrow, pre rain sample 77 K SIR 13/04/2004 NH 8080 1-2 (open) 600 4 p/up Windrow harvest 82 K SIR Windrow, post rain sample

26 L SIRO 11/02/2004 JD 9650

CTS 6 900 6 7.5 Desiccated 27 L SIRO 32 L SIRO Pre Desiccation sample 61 L SIRO Post desiccation sample 36 M AUR Pre Harvest, Post windrow sample 62 M AUR Pre windrow sample: 13/4/04 81 M AUR 21/04/2004 JD9610 0 900 900 86 M AUR 39 N FG Post windrow, pre harvest sample 90 N FG

66



91 N FG JD1075 3-6 mm 900 6 Windrow sample: harvest 14:30 94 N FG 20 O AUR 10/02/2004 JD 9660 0 740 900 7 7.5 m Desiccated sample 21 O AUR 50 O AUR Desiccated. 30/1/04 68 O AUR Post desiccation, day of harvest; 13:00 24 P FG 3/03/2004 JD 9660 0 740 900 7 7.5 Desiccated 25 P FG 28 P FG Pre desiccation sample 31 P FG Post desication sample 35 Q S/CUF 43 Q S/CUF Windrow sample 73 Q S/CUF 15/02/2004 JD 9660 0 760 920 3 7.5 m Windrow harvest 79 Q S/CUF 41 R FDALE Windrowed 70 R FDALE 96 R FDALE 4/05/2004 JD9660 0 750 920 5 Windrowed, 30/4/04 45 S FG 97 S FG Desiccated sample

100 S FG 6/05/2004 JD9610 0 840 860 4.5 Desiccated crop 6 T HR

7 T HR 12/04/2004 CASE 2388 0.5 1150 750 5 9 m Desiccated crop

46 T HR Desiccated. C-probe rings - 18/5/04 55 T HR Pre desiccation, 31/3/04 64 T HR Post desiccation, pre harvest 40 U S/SIR Post desiccation 87 U S/SIR Header 2

89 U S/SIR 22/04/2004 CASE 2388 0 1020 780 Windrowed, Header 2, AFX rotor

92 U S/SIR Header 3

93 U S/SIR 22/04/2004 CASE 2388 0 1020 780 Header 1

67



99 U S/SIR

101 U S/SIR 22/04/2004 CASE 2388 0 1020 780 Standard rotor

104 U S/SIR 4% of the C1 seed had a 19% abs level. 104 U S/SIR 14% of line had 19% abs. 42 V SIRO Windrowed 47 V SIRO Windrow, day of harvest

74 V SIRO 10/05/2004 CASE 2388 0 1000 790 Windrowed, Header 2

84 V SIRO Header 1 85 V SIRO Header 2

88 V SIRO 10/05/2004 CASE 2388 0 950 790 Windrowed, Header 1, AFX rotor

106 V SIRO 14 W AQUA

23 W AQUA 4/03/2004 CASE 2388 0 1000 790 9 Desiccated crop

34 W AQUA Post Des./Pre Harvest sample 56 W AQUA Pre desiccation sample: 24/2/04

105 W AQUA

17 X AUR 14/03/2004 CASE 2166 0 (closed) 880 780 5 p/up

22 X AUR 33 X AUR Pre windrow and rain sample 38 X AUR Pre harvest sample. 13.5 mm on 8/3/04

68

Appendix 4: 04/05 seed quality data

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 133 A VENUS Keith I Final Normal 1 78 6 16 0 29 A VENUS Keith I Hand Normal 1 87 2 11 0 52 A VENUS Keith I Header Normal 1 85 7 7 1 97 A VENUS Keith I Header Normal 2 85 5 10 0 59 A VENUS Keith I Truck Normal 1 87 5 8 0

108 AA S/SIR Coombe I Final Normal 1 85 5 10 0 10.0 26 AA S/SIR Coombe I Hand Normal 1 54 5 37 0 78 AA S/SIR Coombe I Header Normal 1 68 5 27 0 89 AA S/SIR Coombe I Header Normal 1 75 4 21 0 91 AA S/SIR Coombe I Header Normal 2 73 9 18 0 92 AA S/SIR Coombe I Header Normal 2 73 8 19 0 76 AA S/SIR Coombe I Truck Normal 1 71 6 22 1 80 AA S/SIR Coombe I Truck Normal 1 78 3 19 0

125 B SIR Keith D Final Dry 1 64 20 16 0 9.0 4 B SIR Keith D Hand Dry 1 81 4 15 0 34 B SIR Keith D Header Dry 1 80 10 10 0 94 B SIR Keith D Header Dry 2 78 12 9 1 47 B SIR Keith D Truck Dry 1 75 15 10 0

131 BB FG Coombe D Final Normal 1 77 7 16 0 <10.0 30 BB FG Coombe D Hand Normal 1 81 3 15 1 44 BB FG Coombe D Header Normal 1 77 6 17 0 40 BB FG Coombe D Truck Normal 1 80 7 13 0 36 C SIR Keith I Field Bin Normal 1 87 6 6 0

114 C SIR Keith I Final Normal 1 86 7 7 0 8.8 20 C SIR Keith I Hand Normal 1 80 4 16 0 56 C SIR Keith I Header Normal 1 87 5 8 0 35 C SIR Keith I Truck Normal 1 86 8 6 0

109 CC 9B78 Coombe I Final Normal 1 75 10 15 0 5 CC 9B78 Coombe I Hand Normal 1 68 4 26 2

69

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 88 CC 9B78 Coombe I Truck Normal 1 70 6 23 1

112 D SIR Keith I Final Normal 1 84 11 5 0 <10.0 14 D SIR Keith I Hand Normal 1 82 5 11 2 81 D SIR Keith I Header Normal 1 82 9 9 0

104 D SIR Keith I Header Normal 2 83 10 7 0 72 D SIR Keith I Truck Normal 1 76 10 14 0

117 DD AUR Keith D Final Normal 1 74 14 12 0 10.5 8 DD AUR Keith D Hand Normal 1 97 3 0 0 61 DD AUR Keith D Header Normal 1 87 10 3 0 93 DD AUR Keith D Header Normal 2 80 17 3 0

113 E AUR Keith D Final Dry 1 74 9 17 0 12.5 18 E AUR Keith D Hand Dry 1 93 4 2 1 55 E AUR Keith D Header Dry 1 82 14 4 0 54 E AUR Keith D Truck Dry 1 78 17 5 0

118 F HALL Laffer D Final Normal 1 65 21 13 0 6.3 22 F HALL Laffer D Hand Normal 1 74 3 23 0 71 F HALL Laffer D Header Normal 1 89 7 4 0 73 F HALL Laffer D Truck Normal 1 86 10 3 1

127 G AQUA Keith I Final Normal 1 71 22 7 0 12 G AQUA Keith I Hand Normal 1 60 3 37 0 75 G AQUA Keith I Header Normal 1 64 6 30 0

100 G AQUA Keith I Header Normal 2 74 6 20 0 74 G AQUA Keith I Truck Normal 1 58 3 39 0

110 H SIR Keith I Final Normal 1 62 7 31 0 3 H SIR Keith I Hand Normal 1 56 1 41 2 33 H SIR Keith I Header Normal 1 75 7 18 0

105 H SIR Keith I Header Normal 2 78 8 14 0 31 H SIR Keith I Truck Normal 1 74 6 19 1

130 I AuR Brimbago I Final Normal 1 83 4 13 0 9 I AUR Brimbago I Hand Normal 1 81 2 17 0 63 I AUR Brimbago I Header Normal 1 74 8 18 0

101 I AUR Brimbago I Header Normal 2 75 13 12 0

70

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 64 I AUR Brimbago I Truck Normal 1 92 3 5 0

121 J FG Laffer I Final Normal 1 66 28 6 0 12.0 10 J FG Laffer I Hand Normal 1 57 3 38 1 90 J FG Laffer I Header Normal 1 94 5 1 0

107 J FG Laffer I Header Normal 2 86 10 3 1 82 J FG Laffer I Truck Normal 1 91 6 3 0 68 K FDALE Laffer I Field Bin Normal 1 80 7 13 0

132 K FDALE Laffer I Final Normal 1 77 8 15 0 <10.0 2 K FDALE Laffer I Hand Normal 1 74 3 20 3 69 K FDALE Laffer I Header Normal 1 74 9 17 0 70 K FDALE Laffer I Truck Normal 1 75 8 17 0

119 L FDALE Laffer D Final Normal 1 78 15 7 0 9.0 25 L FDALE Laffer D Hand Normal 1 80 2 18 0 60 L FDALE Laffer D Header Normal 1 79 7 14 0 53 L FDALE Laffer I Truck Normal 1 77 8 15 0

120 M FDALE Laffer D Final Normal 1 78 15 7 0 9.0 13 M FDALE Laffer D Hand Normal 1 79 3 18 0 48 M FDALE Laffer D Header Normal 1 80 10 10 0 57 M FDALE Laffer D Truck Normal 1 83 7 10 0

111 N SIR Brimbago I Final Normal 1 68 7 25 0 11.4 27 N SIR Brimbago I Hand Normal 1 67 5 28 0 46 N SIR Brimbago I Header Normal 1 75 19 6 0

106 N SIR Brimbago I Header Normal 2 76 15 9 0 32 N SIR Brimbago I Truck Normal 1 76 13 11 0

134 O SIROSAL Brimbago D Final Normal 1 84 7 9 0 19 O SIROSAL Brimbago D Hand Normal 1 85 3 12 0 58 O SIROSAL Brimbago D Header Normal 1 89 6 5 0 49 O SIROSAL Brimbago D Truck Normal 1 89 6 5 0

115 P AUR Keith I Final Normal 1 79 17 4 0 15 P AUR Keith I Hand Normal 1 56 1 38 5 83 P AUR Keith I Header Normal 1 70 3 26 1 84 P AUR Keith I Truck Normal 1 71 8 21 0

71

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 116 Q HR Keith I Final Normal 1 88 8 4 0 11 Q HR Keith I Hand Normal 1 55 3 40 2 77 Q HR Keith I Header Normal 1 60 3 37 0

102 Q HR Keith I Header Normal 2 72 5 23 0 87 Q HR Keith I Truck Normal 1 65 3 31 1 67 R FG Tintinara I Field Bin Normal 1 85 7 7 1

124 R FG Tintinara I Final Normal 1 77 14 9 0 10.0 7 R FG Tintinara I Hand Normal 1 95 2 2 1 65 R FG Tintinara I Header Normal 1 83 11 6 0 99 R FG Tintinara I Header Normal 2 79 11 8 2 66 R FG Tintinara I Truck Normal 1 84 11 5 0 23 S S/CUF Laffer D Hand Normal 1 91 3 5 1 37 S S/CUF Laffer D Header Normal 1 81 7 12 0 38 S S/CUF Laffer D Truck Normal 1 87 5 8 0

123 T S/CUF Laffer D Final Normal 1 77 16 7 0 9.1 17 T S/CUF Laffer D Hand Normal 1 82 2 16 0 39 T S/CUF Laffer D Header Normal 1 82 10 8 0 95 T S/CUF Laffer D Header Normal 2 76 17 7 0 41 T S/CUF Laffer D Truck Normal 1 84 7 9 0

126 U SIR Keith I Final Normal 1 81 10 9 0 28 U SIR Keith I Hand Normal 1 79 2 19 0 62 U SIR Keith I Header Normal 1 82 4 14 0 96 U SIR Keith I Header Normal 2 73 7 20 0 43 U SIR Keith I Truck Normal 1 68 6 26 0

122 V SIR Keith I Final Normal 1 84 7 9 0 7.5 21 V SIR Keith I Hand Normal 1 91 3 5 1 51 V SIR Keith I Header Normal 1 86 5 9 0 98 V SIR Keith I Header Normal 2 84 7 9 0 50 V SIR Keith I Truck Normal 1 87 3 10 0 86 W AUR Brimbago I Field Bin Normal 1 62 5 33 0

136 W AUR Brimbago I Final Normal 1 75 6 19 0 8.0 6 W AUR Brimbago I Hand Normal 1 54 3 43 0

72

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 79 W AUR Brimbago I Header Normal 1 63 7 30 0

103 W AUR Brimbago I Header Normal 2 68 6 25 1 85 W AUR Brimbago I Truck Normal 1 64 9 27 0

135 X FG Brimbago I Final Normal 1 77 6 17 0 11.5 16 X FG Brimbago I Hand Normal 1 63 2 32 3

129 Y UQL1 Brimbago D Final Normal 1 63 30 7 0 24 Y UQL1 Brimbago D Hand Normal 1 76 1 23 0 42 Y UQL1 Brimbago D Truck Normal 1 83 3 14 0

128 Z HR Mt Monster I Final Normal 1 67 23 10 0 9.0 1 Z HR Mt Monster I Hand Normal 1 90 4 6 0

73

Appendix 5: 04/05 harvest data Sample No. Code Variety Harvest Date Header Concave Rotar Thresher Fan Speed Front Comments

133 A VENUS 29 A VENUS Windrowed 52 A VENUS 13/03/2005 CASE2388 0 920 750 5 p/up 97 A VENUS 59 A VENUS

125 B SIR 4 B SIR Post Desiccation 21/2/05 34 B SIR 24/02/2005 JD9600 0 940 9 9 m 94 B SIR 47 B SIR 20 C SIR Windrowed 13/2/05. 56 C SIR 17/02/2005 CASE 2388 0.5-1.0 900 750 p/up 36 C SIR

114 C SIR 35 C SIR

112 D SIR 14 D SIR Windrowed 9/4/05 81 D SIR 13/04/2005 CASE 2388 0.5-1.0 900 750 p/up

104 D SIR 72 D SIR

113 E AUR 18 E AUR Desiccated 7/2/05. Dry crop. 55 E AUR 14/02/2005 CASE 2388 0.5-1.0 900 750 9 m 54 E AUR

118 F HALL 22 F HALL Windrowed14/2/05 71 F HALL 18/02/2005 CASE 2388 0.5-1.0 900 750 p/up

74

Sample No. Code Variety Harvest Date Header Concave Rotar Thresher Fan Speed Front Comments 73 F HALL

127 G AQUA 12 G AQUA Desiccated 13/4/05 75 G AQUA 17/04/2005 JD1075 7-10 mm 900 900 6 m

100 G AQUA 74 G AQUA

110 H SIR 3 H SIR Post Desiccation 11/4/05 33 H SIR

105 H SIR 15/04/2005 JD9600 3-6 mm 950 750 5.5 6 31 H SIR

130 I AUR 9 I AUR Desiccated April 05 63 I AUR

101 I AUR 27/04/2005 Claas 64 I AUR

121 J FG 10 J FG Windrowed 18/4/05 90 J FG 22/04/2005 CASE 2388 0.4 980 750 6.5 p/up Header 12

107 J FG 82 J FG 68 K FDALE Augered 3 times

132 K FDALE 2 K FDALE Windrowed 18/4/05 69 K FDALE 22/04/2005 CASE2388 0.4 900 740 6 p/up 70 K FDALE

119 L FDALE 25 L FDALE Desiccated 21/2/05. 60 L FDALE 28/02/2005 CASE 2388 0.1 960 5 9 m Desiccated 53 L FDALE

75

Sample No. Code Variety Harvest Date Header Concave Rotar Thresher Fan Speed Front Comments 120 M FDALE 13 M FDALE Desiccated 21/2/05 48 M FDALE 28/02/2005 CASE 2388 0.1 960 5 9 m 57 M FDALE

111 N SIR 27 N SIR Windrowed 11/4/05 46 N SIR 15/04/2005 CASE 2388 0.2 970 8 p/up

106 N SIR 32 N SIR

134 O SIROSAL 19 O SIROSAL Windrowed 10/2/05. Dry crop 58 O SIROSAL 14/02/2005 JD9706CTS 5 800 5 p/up 49 O SIROSAL

115 P AUR 15 P AUR Desiccated 21/4/05 83 P AUR 84 P AUR 28/04/2005 JD9610 0 900 900 5.5 7.5

116 Q HR 11 Q HR Desiccated 21/4/05 77 Q HR

102 Q HR 87 Q HR 29/04/2005 JD9610 880 890 6 7.5 67 R FG

124 R FG 7 R FG 15/04/2005 JD1075 0 950 6 p/up Windrowed 65 R FG 99 R FG 66 R FG 23 S S/CUF Desiccated 8/2/05 37 S S/CUF 14/02/2005 JD9660 0 700 8 7.5

76

Sample No. Code Variety Harvest Date Header Concave Rotar Thresher Fan Speed Front Comments 38 S S/CUF

123 T S/CUF 17 T S/CUF Desiccated 8/2/05 39 T S/CUF 15/02/2005 JD9660 0 700 7 7.5 95 T S/CUF 41 T S/CUF

126 U SIR 28 U SIR Windrowed 6/4/05 62 U SIR 10/04/2005 NHTX66 8 mm 910 8 p/up 96 U SIR 43 U SIR

122 V SIR 21 V SIR Windrowed 2/2/05. 51 V SIR 6/02/2005 JD9400 0 800 p/up 98 V SIR 50 V SIR 86 W AUR

136 W AUR 6 W AUR Desiccated 21/4/05 79 W AUR 28/04/2005 JD9610 1 700 5 7.5

103 W AUR 85 W AUR

135 X FG 16 X FG 30/04/2005 JD9610 1 700 5 7.5 Desiccated 21/4/05

129 Y UQL1 14/02/2005 CASE 2388 0 850 780 5.1 9 m 24 Y UQL1 Windrowed 10/2/05. 42 Y UQL1

128 Z HR 25/03/2005 CASE 2388 0.4 850 800 5.1 p/up 1 Z HR Windrowed 21/3/05

108 AA S/SIR 4% of the C1 seed had a 19%

77

Sample No. Code Variety Harvest Date Header Concave Rotar Thresher Fan Speed Front Comments 26 AA S/SIR abnormals level. 78 AA S/SIR 20/03/2005 CASE2388 0 950 770 5.4 p/up Windrowed March 05. 89 AA S/SIR 91 AA S/SIR 20/03/2005 CASE 2388 0 950 800 5.4 p/up Header 11 92 AA S/SIR 76 AA S/SIR 80 AA S/SIR

131 BB FG 30 BB FG Pre Desiccation 8/2/05 44 BB FG 22/02/2005 CASE2388 0 990 780 8 9 m 40 BB FG

109 CC 9B78 5 CC 9B78 17/04/2005 CASE 2388 0 950 780 5.4 p/up Windrowed April 05 88 CC 9B78

117 DD AUR 8 DD AUR Desiccated 8/2/05 61 DD AUR 15/02/2005 CASE 2166 0 880 780 5 p/up 93 DD AUR

78

Appendix 6: 05/06 seed quality data Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture %

71 A SIR Brimbago I Final Dry 1 74 20 6 0 16 A SIR Brimbago I Hand Dry 1 77 6 17 0 24 A SIR Brimbago I Header Dry 1 76 16 8 0 7.0 43 A SIR Brimbago I Truck Dry 1 81 12 7 0 61 B WL525 Brimbago D Final Dry 1 79 16 5 0 70 B WL525 Brimbago D Final Dry 1 74 18 8 0 14 B WL525 Kongal D Hand Dry 1 84 9 7 0 50 B WL525 Brimbago D Truck Dry 1 77 17 6 0 <7.0 68 C SIR Keith D Final Dry 1 73 25 2 0 <7.0 12 C SIR Keith D Hand Dry 1 84 9 7 0 21 D SIR Keith I Field Bin Normal 1 86 11 3 0 <10.0 79 D SIR Keith I Final Normal 1 77 10 13 0 5 D SIR Keith I Hand Normal 1 78 8 14 0 39 E AUR Keith D Field Bin Dry 1 75 19 6 0 78 E AUR Keith D Final Dry 1 77 22 1 0 17 E AUR Keith D Hand Dry 1 92 2 6 0 19 E AUR Keith D Header Dry 1 87 6 6 0 <7.0 56 F HALL Laffer D Field Bin Dry 1 73 23 4 0 44 F HALL Laffer D Field Bin Dry 1 83 8 9 0 69 F HALL Laffer D Final Dry 1 57 21 22 0 13 F HALL Laffer D Hand Normal 1 80 7 13 0 40 F HALL Laffer D Truck Dry 1 78 16 6 0 9.1 23 G S.CUF Keith I Field Bin Dry 1 77 15 8 0 75 G S.CUF Keith I Final Dry 1 78 17 5 0 32 G S.CUF Keith I Hand Dry 1 81 5 14 0 2 G S.CUF Keith I Header Dry 1 71 20 9 0 10.5 76 H S.SIR Keith I Final Dry 1 83 10 7 0

79

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 31 H S.SIR Keith I Hand Dry 1 56 2 42 0 57 H S.SIR Keith I Header Dry 1 59 11 30 0 >7 & < 12 48 H S.SIR Keith I Truck Dry 1 67 9 23 0 77 I SIR Keith I Final Normal 1 73 12 15 0 9 I SIR Keith I Hand Normal 1 57 4 38 0 49 I SIR Keith I Header Normal 1 58 10 32 0 10.0 47 I SIR Keith I Truck Normal 1 52 10 38 0 46 I SIR Keith I Truck Normal 2 58 9 33 0 73 J SIR Brimbago I Final Dry 1 67 14 19 0 74 J SIR Brimbago I Final Dry 1 66 8 26 0 3 J SIR Brimbago I Hand Normal 1 69 8 23 0 62 J SIR Brimbago I Header Normal 1 79 12 9 0 10.0 64 J SIR Brimbago I Header Normal 1 83 5 12 0 10.0 63 J SIR Brimbago I Truck Normal 1 74 16 10 0 65 J SIR Brimbago I Truck Normal 1 79 7 14 0 45 K AUR Laffer D Final Dry 1 77 13 10 0 >7 & <12 4 K AUR Laffer D Hand Dry 1 90 5 5 0 45 K AUR Laffer D Header Dry 1 85 14 1 0 52 K AUR Laffer D Truck Dry 1 86 14 0 0 72 M AUR Mundulla I Final Dry 1 71 20 8 0 7 M AUR Mundulla I Hand Dry 1 70 11 18 0 37 M AUR Mundulla I Header Dry 1 71 16 13 0 <7 33 M AUR Mundulla I Truck Dry 1 66 15 18 1 84 N AUR Brimbago I Final Normal 1 78 17 5 0 1 N SIR Brimbago I Hand Dry 1 40 8 50 0 6 N SIR Brimbago I Hand Normal 1 37 3 60 0 54 N SIR Brimbago I Header Dry 1 61 16 23 0 >7 & <12 30 N SIR Brimbago I Header Normal 1 62 10 28 0 >7 & <12 85 O HR Brimbago I Final Dry 1 76 6 18 0 29 O HR Brimbago I Hand Dry 1 61 3 35 0 15.7

80

Sample No. Code Variety Location Type Process Status Grade Normal % Abnormal % Hard % Dead % Moisture % 28 O HR Brimbago I Truck Dry 1 75 5 20 0 66 P SIR Brimbago I Final Normal 1 85 13 2 0 67 P SIR Brimbago I Final Normal 1 75 21 4 0 8-13.5 11 P SIR Brimbago I Hand Normal 1 54 7 36 0 82 Q FG Coombe D Final Normal 1 77 13 10 0 20 Q FG Coombe D Hand Normal 1 80 12 8 0 27 Q FG Coombe D Header Normal 1 69 15 16 0 < 7 55 Q FG Coombe D Header Normal 1 66 19 15 0 < 7 26 Q FG Coombe D Truck Normal 1 66 16 18 0 83 R FG Coombe I Final Normal 1 86 9 5 0 15 R FG9B78 Coombe I Hand Dry 1 65 4 31 0 18 R FG9B78 Coombe I Hand Normal 1 66 5 28 0 36 R FG Coombe I Header Dry 1 68 12 20 0 >7 & <12 35 R FG Coombe I Header Normal 1 70 8 22 0 >7 & <12 34 R FG Coombe I Truck Dry 1 70 10 20 0 38 R FG Coombe I Truck Normal 1 72 8 20 0 81 S FG Coombe D Final Normal 1 75 15 10 0 8 S FG Coombe D Hand Normal 1 93 3 4 0 22 S FG Coombe D Header Normal 1 72 18 9 0 <7 25 S FG Coombe D Truck Normal 1 74 16 10 0

81

Appendix 7: 05/06 harvest data

Sample No. Code Harvest Date Variety Header Concave Rotar Thresher Fan Speed Front Comments 71 A SIR 16 A SIR Very dry post flowering 24 A 17/03/2006 SIR JD9600 3 mm 880 880 4.5 p/up 31 degrees. 7.5 m windrow width 43 A SIR 61 B WL525 70 B WL525

14 B WL525 Dry finish to flowering and ripening period

50 B 14/02/2006 WL525 JD9600 3 mm 880 800 12.5 9 m

68 C 1/02/2006 SIR JD9600 Closed

(1mm/0) 940 900 9 9 m 12 C SIR Dry all way through production 21 D 3/03/2006 SIR Case 2388 0.5 900 750 10 9 m 79 D SIR 5 D SIR

39 E AUR 78 E AUR 17 E AUR Dry all way through production 19 E 1/02/2006 AUR Case 2388 0.5 900 750 12 9 m 56 F HALL 44 F HALL 69 F HALL 13 F HALL 40 F 8/02/2006 HALL Case 2388 0.5 900 750 10 9 m 23 G S.CUF 75 G S.CUF 32 G S.CUF 2 G 14/03/2006 S.CUF JD1075 7-10 mm 900 900 5.5 6 m Very dry post flowering

76 H S.SIR 31 H S.SIR

82

Sample No. Code Harvest Date Variety Header Concave Rotar Thresher Fan Speed Front Comments 57 H 27/03/2006 S.SIR JD9600 6mm/3mm 950 750 5.5 22 degrees 48 H S.SIR 77 I SIR 9 I SIR

49 I 3/04/2006 SIR JD9600 5mm/3mm 950 750 5.5 23 degrees 46 I SIR 47 I SIR 73 J SIR 74 J SIR 3 J SIR

62 J 31/03/2006 SIR Case 2388 0.5 1000 7 p/up 7.5 m windrow/Chaffy sample

64 J 31/03/2006 SIR JD 9760

STS 9 940 5.5 p/up 7.5 m windrow 63 J SIR 65 J SIR 45 K 2/02/2006 AUR JD 9660 0 700 900 8.5 7.5 m 4 K AUR Dry all way through production

45 K AUR 52 K AUR 80 L CIALFA67 JD 9660 0 740 900 4 7.5 m 10 L CIALFA67 51 L CIALFA67

53 L 4/04/2006 CIALFA67

SARDI small plot harvester 746 kg/ha

58 L 4/04/2006 CIALFA67


59 L 4/04/2006 CIALFA67


60 L 4/04/2006 CIALFA67


72 M AUR

83

Sample No. Code Harvest Date Variety Header Concave Rotar Thresher Fan Speed Front Comments 7 M AUR Very dry post flowering

37 M 29/03/2006 AUR Gleaner

R72 Closed

1/16":1/8" 990 4 5.5 9 m 33 M AUR 84 N AUR 1 N SIR Very dry finish through ripening 6 N SIR

30 N 11/04/2006 SIR JD9610 0 810 800 6 9 m Use grain loss monitor to control speed 54 N 11/04/2006 SIR JD9610 0 810 800 6 9 m Use grain loss monitor to control speed 28 O HR 85 O HR Dried immediately - high moisture due to 29 O HR wireweed contamination 66 P SIR Small line 925 kg 67 P 27/03/2006 SIR Case 2388 0.5 1000 800 7 9 m One delivery dried (13.5%). 11 P SIR 82 Q FG 20 Q FG 27 Q 8/02/2006 FG Case 2388 0.5 950 750 12 p/up 6.3 m windrow width. 30 degrees 55 Q 8/02/2006 FG Case 2388 0.5 950 750 12 p/up 6.3 m windrow width. 30 degrees 26 Q FG 83 R FG 15 R FG9B78 18 R FG9B78 35 R 26/03/2006 FG Case 2388 0.5 950 740 12 p/up 6.3 m windrow width. 24 degrees 36 R 26/03/2006 FG Case 2388 0.5 950 750 12 p/up 6.3 m windrow width. 30 degrees 34 R FG 38 R FG 81 S FG 8 S FG

22 S 6/02/2006 FG Case 2388 0.5 950 750 13 p/up 6.3 m windrow width. 24 degrees 25 S FG

84

Appendix 8: Ferric Chloride test assessment data

Test Code Variety Location Type Process Status Harvest Date

Total Seeds Stained Moisture

(%) Comments

1 1 SIR Keith I Hand Normal 2/02/2007 100% 3.0%

2 1 SIR Keith I Header Normal 2/02/2007 100% 23.0% 7% Smallish seed and dull coloured

3 1 SIR Keith I Header Normal 2/02/2007 100% 9.0% 7%

4 1 SIR Keith I Final Normal 100% 7.0%

5 2 SIR Brimbago I Hand Normal 13/03/2007 100% 2.5% 7%

6 2 SIR Brimbago I Header Normal 13/03/2007 100% 3.0% 7% Excellent seed colour, size and quality

7 2 SIR Brimbago I Final Normal 100% 5.0%

8 3 S/AUR Mt Monster I Hand Normal 27/03/2007 100% 3.3% 7%

9 3 S/AUR Mt Monster I Header Normal 27/03/2007 100% 3.8% 7% Smallish seed but good colour

10 3 S/AUR Mt Monster I Final Normal 100% 6.0%

Appendix 9: Ferric Chloride test header data Code Variety Type Process Status Grade Header Concave Rotar Thresher Fan Speed Front

1 SIR I Header Normal 1 Case 2388 0.5 (closed) 900 890 7 9m 1 SIR I Header Normal 1 Case 2388 0.5 (closed) 900 890 12 9m 2 SIR I Header Normal 1 JD9600 1/16 (closed) 800-900 830 Windrow 3 S/AUR I Header Normal 1 Case 2388 1.8 (open) 820 790 5.1 9m

85

Appendix 10: Photo album of abnormal seedlings

1) Cotyledon only 2) Cotyledon only

3) Cotyledon only 4) Dead seed

5) Decayed and deformed seed 6) Decayed root tip

86

7) Decayed insufficient root 8) Deformed

9) Deformed 10) Deformed

11) Deformed 12) Deformed

87

13) Detached cotyledon: insufficient 14) Detached cotyledon: roots root. trapped

15) Detached cotyledon: root trapped 16) Detached cotyledon: stunted root

17) Fractured seedling: Decayed root 18) Insufficient decayed roots

88

19) Insufficient hypospindly root 20) Loop in hypodecayed root

21) Multiple decayed roots 22) No root

23) Spindly decayed root 24) Stunted decayed root

89

25) Stunted root 26) Stunted seedling

27) Trapped root 28) Trapped and deformed root

29) Trapped and deformed root 30) Yellow detached cotyledon and

trapped roots

Understanding and Managing the Causes of Abnormal Seedlings in Lucerne

The report presents a 5 year research project that investigated the cause of abnormal seedlings in lucerne. An in depth literature review focused the areas of research and a methodology was designed to test certain hypotheses. The report outlines the findings of the research and how they can be practically utilised in the production of lucerne seed.

The Rural Industries Research and Development Corporation (RIRDC) manages and funds priority research and translates results into practical outcomes for industry.

Our business is about new products and services and better ways of producing them.

Most of the information we produce can be downloaded for free from our website: www.rirdc.gov.au.

RIRDC books can be purchased by phoning 02 6271 4160 or online at: www.rirdc.gov.au/eshop.

RIRDC Publication No. 08/023

Contact RIRDC:Level 2

15 National CircuitBarton ACT 2600

PO Box 4776Kingston ACT 2604

Ph: 02 6271 4100Fax: 02 6271 7199

Email: [email protected]: www.rirdc.gov.auwww.rirdc.gov.au/eshop

This publication can be viewed at our web-site—www.rirdc.gov.au. All RIRDC books can be purchased from:.

RIRDCInnovation for rural Australia

08-023 Final Report covers.indd 1 29/05/2008 9:46:20 AM

RIRDC · seed post harvest. A seed testing kit should be developed for producers to test the seed...

Documents

Transcript of RIRDC · seed post harvest. A seed testing kit should be developed for producers to test the seed...