GENETIC IMPROVEMENT FOR PULP AND WOOD ......Presented By O P Shukla Chief General Manager (Raw...

Presented By

O P Shukla

Chief General Manager (Raw Material)

JK PAPER LIMITED

Unit: CPM, Fort Songadh,

Dist: Tapi, Gujarat, India- 394660

GENETIC IMPROVEMENT FOR PULP AND WOOD

PROPERTIES IN EUCALYPTUS CLONES Khanna N K, Shukla O P, Narkhede S L & Chauhan SKS

Tyre Cement

Agri Genetics V-Belts- Oil Seals & Power Transmission

Systems

Clinical Researc

h

Hospital & Health services

> US $ 4 billion

29 manufacturing plants

Present in over 100 countries

More than 40,000 employees

Paper

Education

Dairy Products

Defence Electronics

JK Group

http://www.jklakshmi.com/default.asp

Core Businesses & Products

9 Plants in India,3 Plants in Mexico

20 Million Tyres per year.

5 plants, 8.4 Million Tons per Year 5 Plants, Power Transmission Items

PAPER TYRE

CEMENT POWER TRANSMISSION

3 Plants in India, 0.60 million TPA

JK Paper – A Profile

JK Paper

JKPM

JK

Paper

CPM

- Capacity : 302,000 TPA

- Employees : 1459

- Estd. In : 1962

- Distance from : 200 km

nearest airport

- Capacity:162000TPA

- Employees : 1180

- Estd. In : 1992

- Distance from : 80 km

nearest airport

JK PAPER

LIMITED

(2 PLANTS) Present Combined Capacity

4,64,000 TPA

Recently acquired

Sirpur Paper Mills,

Kagaznagar, Telangana

state. Total capacity

`100000 TPA

JK PAPER LIMITED

JK Paper Limited Present Capacity , 464000 TPA

Unit: CPM Unit: JKPM

•Capacity: 162000 TPA

•Products: Cut size multi purpose office paper and

security paper, Packaging Board for consumer products

•Capacity: 302000 TPA

•Products: Cut size multi purpose office paper and

security paper

Production of quality planting material – quality clones.

Extension & motivation activities for farm forestry

promotion.

Distribution of clones to farmers at subsidized cost.

States covered by CPM: Gujarat, Maharashtra & Madhya

Pradesh

Services to farmers include financial support through

banks, technical guidance for pulpwood farming,

harvesting & buy back assurance for wood.

Logging & logistics from farmers field.

Focus on Plantation R & D

JKPL: Plantation Initiatives

Slide No.

Agro-forestry Models

•Raw material cost of Brazil (60-65 $ per ADMT- pulp yield 50 to 52 % bleached •Raw material cost of India (100-110 $ per ADMT- Pulp yield 40-41 % bleached

Cost structure of paper

Raw material costs of countries like Brazil, Indonesia are less than India Because- 1. Government has not allotted captive

land for Plantation. 2. Less focus on Research and

Development of Pulpwood plantations.

Ways to reduce cost

of Raw material

To increase fiber yield.

To Decrease

lignin content.

To improve fiber

morphology.

To increase wood density

These 4 factors are responsible

for increasing pulp yield and

increasing pulp production

of the mill.

Present RM Scenario on India

40%

60%

Raw MaterialCost

Other variablecost

1. No captive land for paper industry in India for plantations.

2. More than 85% farmers are small and marginal- Less than 2 Ha land

holding.

3. Tremendous increasing pressure on agriculture land to meet food

demand.

4. Per capita annual paper consumption in India is 13Kg which is very

low over world’s average of 57Kg.

5. The demand for the paper will continuously increase in India which

requires increasing wood supply for paper production.

6. No captive land for paper industry in India for plantations.

7. India wood cost: US $ 100/ADMT to US $110/ADMT, Indonesia (US

$60- US $ 70/ADMT), Brazil (US $.60- 65/ADMT) etc.

8. Scale of paper production operation is lower in India.

9. Dumping of paper from china & other countries.

Present Status of Indian Paper Sector

Objectives of the Mutation R & D

1. Enhancement of wood productivity

2. Short rotation of crop cycle

3. Improvement in Pulp qualities & productivity.

4. Improvement wood densities.

5. Reduction of lignin contents

Mutations are heritable changes in the phenotypes of organisms. These

changes are the results of chemical changes at the level of genes. Such

changes are capable of bringing about new and heritable character variations

in plants and such variations can be selected and used for the establishment

of plant varieties with new characters.

Tree improvement techniques efficiently used at JKPL:

1. CPTs selection.

2. Breeding & hybridization.

3. Physical Mutation through Gamma rays irradiation.

4. Chemical mutation (EMS)

5. Provenance trials & testing.

6. Clonal seed orchards etc.

Mutation

1. We have applied 5 doses Gamma rays (Co-60) at different -2 frequencies on 10 high

productive commercial eucalyptus clones in Gamma Chamber Model-Mitsubishi GO T1000

at Nuclear Research Laboratory Indian Agriculture Research Institute, Pusa New delhi –

India.

2. 10 Eucalyptus clones selected to induce mutation by irradiation through Gamma rays.

3. The clones are mentioned as under with species.

S. No. Cultivar number Derived from different crosses & selections

1. CPM EC- 1 Eucalyptus camaldulensis

2. CPM EC-2 Eucalyptus urophylla

3. CPM EC-3 Eucalyptus tereticornis X E. camaldulensis

4. CPM EC-4 Eucalyptus urophylla X E. grandis

5. CPM EC-5 Eucalyptus camaldulensis

6. CPM EC-6 Eucalyptus camaldulensis

7. CPM EC-7 Eucalyptus camaldulensis X E. tereticornis

8. CPM EC-8 Eucalyptus camaldulensis X E. deglupta

9. CPM EC-9 Eucalyptus camaldulensis X E. urophylla

10 CPM EC-10 Eucalyptus camaldulensis X E. tereticornis

Mutation in Eucalyptus clones

Gamma rays irradiated Eucalyptus

clonal plants

1. Progeny trials planted in RBD in Jan 2017.

2. Semi irrigated conditions.

3. The spacing of the trial is 3 Meters X 1.5 Meters i.e.

2222 plants per Ha.

4. The soil of the plot is light black alkaline soils.

5. We have recorded growth data on quarterly basis.

Progeny Trials

Field trial of Gamma rays irradiated Eucalyptus clones at R& D

experimental Plot at CPM – Age years

Dose CPM

EC-1

CPM

EC-2

CPM

EC-3

CPM

EC-4

CPM

EC-5

CPM

EC-6

CPM

EC-7

CPM

EC-8

CPM

EC-9

CPM

EC-10 Mean VAR SD

Ht in

Mtrs

T1 10.93 13.13 12.9 14 11.45 11.75 12.06 11.4 13.71 12.2 12.35 0.968 1.037 1.38

T2 9.6 12.17 11.75 13 10.4 10.28 12 11.25 12.2 10.39 11.30 1.072 1.091 0.33

T3 7.58 11.6 9.33 10.6 9.75 8.5 7.58 8.08 8.58 6.9 8.85 1.948 1.471 -2.12

TC 10.3 11.94 10.33 11.8 10.85 10.55 11.3 11.25 11.25 10.15 10.97 0.362 0.634 Control

Average Height increment on age of 3.2 years in meters per Tree

VAR: Variance.

SD: Standard Deviation

Dose CPM

EC-1

CPM

EC-2

CPM EC-

3

CPM

EC-4

CPM

EC-5

CPM

EC-6

CPM

EC-7

CPM

EC-8

CPM

EC-9

CPM

EC-10 Mean VAR SD

T1 0.21 0.32 0.25 0.29 0.27 0.28 0.28 0.32 0.37 0.33 0.26 0.002 0.044

T2 0.19 0.28 0.23 0.24 0.24 0.23 0.26 0.27 0.30 0.27 0.22 0.001 0.031

T3 0.17 0.28 0.19 0.18 0.23 0.19 0.19 0.22 0.20 0.17 0.18 0.001 0.034

TC 0.22 0.31 0.21 0.28 0.27 0.22 0.26 0.29 0.28 0.28 0.23 0.001 0.033

Average Girth increment on age of 3.2 years in meters per tree

VAR: Variance.

SD: Standard Deviation

Dose CPM

EC-1

CPM

EC-2

CPM

EC-3

CPM

EC-4

CPM

EC-5

CPM

EC-6

CPM

EC-7

CPM

EC-8

CPM

EC-9

CPM

EC-10 Mean VAR SD

T1 0.022 0.059 0.035 0.051 0.037 0.040 0.042 0.050 0.081 0.060 0.04 0.000271 0.016

T2 0.015 0.042 0.028 0.034 0.027 0.023 0.037 0.036 0.047 0.034 0.03 0.000087 0.009

T3 0.009 0.040 0.014 0.016 0.022 0.013 0.013 0.017 0.014 0.018 0.02 0.000081 0.009

TC 0.021 0.049 0.019 0.040 0.034 0.023 0.034 0.041 0.037 0.036 0.03 0.000090 0.009

Average Volume increment on age of 3.2 years in CMT per tree

Clone wise, treatment wise volume increment per

tree in M3 on 3.2 years of age.

TC

T1

T2

T3

0,022

0,059

0,035

0,051

0,037 0,040 0,042

0,050

0,081

0,060

0,015

0,042

0,028

0,034

0,027 0,023

0,037 0,036

0,047

0,034

0,009

0,040

0,014 0,016

0,022

0,013 0,013 0,017

0,014

0,008

0,021

0,049

0,019

0,040

0,034

0,023

0,034

0,041

0,037 0,036

0,000

0,010

0,020

0,030

0,040

0,050

0,060

0,070

0,080

0,090

T1 T2 T3 TC

0,016 0,021

0,026 0,033

0,042 0,050

0,063

0,075 0.081

0,007

0,016 0,019 0,020 0,026 0,027

0,039 0,043 0,047

0,000 0,002 0,004 0,005 0,006 0,008 0,014 0,017

0,021

0,005 0,010 0,013 0,016 0,018 0,020

0,031 0,034 0,037

0,000

0,010

0,020

0,030

0,040

0,050

0,060

0,070

0,080

0,090

jan/18 mar/18 mai/18 jul/18 set/18 nov/18 jan/19 mar/19 mai/19 jul/19 set/19 nov/19 jan/20 mar/20

jan/18 abr/18 jul/18 out/18 jan/19 abr/19 jul/19 nov/19 mar/20

T1 0,016 0,021 0,026 0,033 0,042 0,050 0,063 0,075 0,081

T2 0,007 0,016 0,019 0,020 0,026 0,027 0,039 0,043 0,047

T3 0,000 0,002 0,004 0,005 0,006 0,008 0,014 0,017 0,021

TC 0,005 0,010 0,013 0,016 0,018 0,020 0,031 0,034 0,037

Month wise & treatment wise MAI volume increment per tree in M3

T1

T2

TC

T3

Slide No.

CPM EC- 9 (TC)3.2 year old CPM EC- 9 (T1- Mutant)3.2 year old

Control Vs Mutant for CPM EC- 9

1. The increment readings shown in progeny trials indicated that

there is a positive or induce growth in all lower mutation

frequencies i.e. T1, T2, over TC (Control).

2. The higher doses gamma rays irradiation i.e. T4 & T5 doses of all

10 varieties of clones were died after 1 month of planting.

3. The highest volume increment observed in T1 treatment over all

treatments.

4. We have selected 2 clones namely CPM EC-9 (Eucalyptus

camaldulensis X E. urophylla) & CPM EC-4 (Eucalyptus urophylla X

E. grandis) for DNA fingerprinting at T1 treatment at JK

Agrigenetics Lab, at Hyderabad, India to study effect of mutation

changes.

5. The DNA fingerprinting- PCR with some indicative primers are

presented in slide no. 33-35.

Result & discussions

1. We have observed lot of changes in mutated vs control clonal

plantations especially volume & growth increment.

2. DNA fingerprinting study at JK Agrigenetics Ltd,

Biotechnology Lab, Hyderabad in 2 clones CPM EC-9 and CPM

EC-4.

3. 72 nos of primers were used for fingerprinting the mutated and

control samples.

4. The PCR product was electrophoresed on 3.5% agarose gel

electrophoresis and then the imaging was done in DNA imager.

DNA Fingerprinting Study

Result & discussions- Conti---

First set of 24 markers

M 1 2 3 4 5 6 7 8 9 10 11 12 M

A B C D A B C D A B C D A B C D A B C D A B C D A B C D A B C D A B C D A B C D A B C D A B C D

M 13 14 15 16 17 18 19 20 21 22 23 24 M


“M” denotes for standard molecular weight marker. A: CPM EC-9 (control), B: CPM EC-9

(mutated), C: CPM EC-4 (control) D: CPM EC-4(mutated) No-1 to 24 are SSR markers


Second set of 24 markers

M” denotes for standard molecular weight marker. A: CPM EC-9 (control), B: CPM EC-9

(mutated), C: CPM EC-4 (control) D: CPM EC-4 (mutated) No-1 to 24 are SSR markers

M 1 2 3 4 5 6 7 8 9 10 11 12 M


M 13 14 15 16 17 18 19 20 21 22 23 24 M



CPM EC-9 (TC- control) Vs CPM EC-9 (T1- mutated)

1. DNA fingerprints with 72 primers reveals that no differences found between CPM EC-

9 (control) and mutated samples at genome level through SSR markers.

2. This indicates that the real mutation can be identified only through whole genome

sequencing/ SNP (single nucleotide polymorphisms) genotyping study.

3. Progeny trial clearly shows the physiological changes in mutant clone as increase in

volume over its control which is replicated in all progeny trials across geo-climatic

conditions

CPM EC-4 (TC control) Vs CPM EC-4 (T1 mutated)

1. 26 markers out of 72 primers showed the polymorphic difference in the fingerprints

between CPM EC-4 control and mutated samples.

2. The results show that the diversity between these two samples found to be

approximately 40% variations at genome level.

3. Progeny trial clearly shows the physiological changes in mutant clone as increase in

volume over its control which is replicated in all progeny trials across geo-climatic

conditions

Observations from DNA Fingerprinting


Slide No.

Conclusions

1. Low frequencies of gamma rays irradiation i.e. T1 & T2 are showings higher

growth over its control in progeny trials.

2. Clone CPM EC-9 (EC X EU) @ T1 treatment are showing 100 % survival & 105 %

higher growth improvement in progeny trials.

3. Productivity/Ha/Year for CPM EC-9 (T1) was observed: 50 M3/Ha/Year & CPM EC-9

(TC - Control) is 32 M3/Ha/Year at 3.2 year rotation. This can be grown in semi arid

& alkaline conditions.

4. Bulk density improved from 205 KG/ M3 to 233 KG/ M3. The pulp production

capacity can be increased up to 13 % with higher bulk resulting in reducing

variable cost.

5. Similarly rotation age can be reduced to 2-3 years which is critical & important in

country like India where farm forestry is only option for wood based industries &

small land holding.

6. Gamma rays irradiation/ Mutation breeding can play important role in tree

improvement in clonal forestry in short term as well as long term.

Slide No.

Conclusions- Conti----

7. Based on progeny trails at various locations we have released mutant clone

JKSC Jumbo at commercial level for plantations.

8. The said clone is high productive, disease resistant (like resistant to

cylindrocladium in nursery & plantations, gall insect resistant), drought

tolerant & showing lot of plasticity in its performance.

9. Similar wood qualities & productivity improvement observed in other clones

(like CPM EC-4, CPM EC-8 (EC X ED), CPM EC-2 (CPM U 283 turbo) etc of the

trials which are under study & large scale progeny trials.

10.The mutation experimental for further improvement is going on at JKPL for

further improvement.

11.Gamma rays mutation is a cost effective & efficient method of tree

improvement & can be utilized efficiently in advance R & D.

Commercial clonal nursery of

Eucalyptus mutant -JKSC-JUMBO

Commercial plantation of JKSC-

JUMBO- Age 2 months

Commercial Plantation of JKSC-

JUMBO- Age 5 months

Slide No.

Mutant of JKSC-Jumbo Eucalyptus clones

planted at Commercial plantation plot – Age

3 month old

GENETIC IMPROVEMENT FOR PULP AND WOOD ......Presented By O P Shukla Chief General Manager (Raw...

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