Germplasm Conservation- In Situ, Ex situ and On-Farm;

Short, Medium & Long term Conservation Strategies for

Conservation of Orthodox Seed and Vegetatively propagated

Crops;

Registration of plant genetic sources.

Biodiversity - Definition

The variability among living organisms from allsources including terrestrial, marine, and otheraquatic ecosystems and the ecological complexesof which they are a part; this includes diversitywithin species, between species and of ecosystems.

Biodiversity can be observed at three levels

1) Genetic diversity:-Each member of any animal or plants species

differs widely from other individual in its genetic makeup.

2) Species diversity :-Number of species of plants and animals that are

present in a region constitutes its species diversity.

3) Ecosystem diversity :-There are large variety of different ecosystems on

earth. Each having their own complement of distinctiveinterlinked species based on the differences in the habitat.

Bio-Geographic Zones

Hot Spots

Biodiversity is essential to -

1. To ensure the production of 5F’s

(Food, Fruit, Fiber, Fuel, Fodder)

2. To maintain other ecosystem services

3. To allow adaptation to changing

environmental conditions including-

Climate, Temp., Rain, Weather change

4. To sustain rural peoples' livelihoods

status

Germplasm Conservation

The germplasm has to be maintained in such

a state that there is minimum risk for its loss

and that either it can be prepared for planting

with relative ease; this is called germplasm

conservation.

OR

The management of human use of the

biosphere so that it may yield the sustainable

benefit to present generations, while

maintaining its potential to meet the needs

and aspirations of future generations.

Why Conservation ?

Conservation of plant genetic resources is necessary for foodsecurity and agro-biodiversity. Genetic diversity providesoptions to develop through selection and breeding of newand more productive crops, resistant to biological andenvironmental stresses (Rao, 2004).

Biodiversity provides a valuable source of compounds to themedical, food and crop protection industries. For more food, itwill be necessary to make better use of a broader range ofgenetic diversity across the glob. Many plant species are nowin danger of becoming extinct (Panis and Lambardi, 2005).

Genetically uniform modern varieties are being replaced withhighly diverse local cultivars and landraces of traditionalagro-ecosystems. Deforestation, urbanization, pollution,habitat destruction, fragmentation and degradation, spread ofinvasive alien species, climate change, changing life styles,globalization, market economies, over-grazing and changesin land-use pattern are contributing indirectly to the loss ofdiversity (Pitman and Jorgensen, 2002; Rao, 2004).

Objectives (Long term)

1. To improve the effectiveness of sustainable managementand conservation of biodiversity through adequateconservation, use and handling of genetic resources.

2. To increase the availability of diversified species with poorseed storability for use in breeding programmes (seedsupply).

3. To improve medium and long term conservationtechnologies of genetic resources (genetic conservation).

4. To build and strengthen research capabilities indeveloping countries through training and transfer ofknowledge and technology and the establishment of aninformal network of researchers on agriculture seeds fromdeveloping and developed countries (technology transferand sharing of experience).

1. Scientific aspects: To characterize the seed storagebehaviour (recalcitrant, intermediate or short lived orthodox)of specific, valuable species and to recommend regimes fortheir short and medium term storage.

2. Seed supply aspects: To develop effectivetechnologies/methods for seed collecting, transport, storage,testing and seed health aspects.

3. Genetic conservation aspects: To develop guidelines forgenetic conservation of seeds of crops species (or groups ofspecies).

4. Technology transfer: To produce a publication andpractical guidelines for the handling of crops seed species,dealing with all aspects from ripeness, harvest and storage totesting and sowing. To develop and publish guidelines for

Objectives (Immediate)

Types of Germplasm

1. Land Races

2. Obsolete Varieties

3. Varieties in Cultivation

4. Breeding Lines

5. Special Genetic Stocks

6. Wild form and Wild Relatives

Germplasm Conservation

The Germplasm has to be maintained in such a state thatthere is minimum risk for its loss and that either it can beplanted directly in the field or it can be prepared for planted forplanting with relative ease.

Ex-situ Conservation - Germplasm conservation is attemptedoutside or away from its natural habitat.

In-situ Conservation – Conservation of germplasm in itsnatural habitat or in area where it grows naturally.

On-farm Conservation One of new approach to in situconservation of genetic resources, focusing on conservingcultivated plant species in farmers' fields. Its nothing but “Thesustainable management of genetic diversity of locallydeveloped traditional crop varieties, with associated wild andweedy species or forms, by farmers within traditionalagricultural, horticultural or agri-silvicultural cultivation systems"(Maxted et al. 1997).

In – Situ / Ex – Situ

In – Situ Ex – Situ

Merits Not only conserve

the genetic diversity

its also allows

evolution to genetic

with climate

Its not allows evolution to

genetic with climate

its allows new

alleles and new

gene combination

would appear with

time

No any new gene

combination due to fix

environment condition

Demerit

s

Its difficult to

establish and very

difficult to maintain

Required costly and well

occupied lab

Very prone to biotic

and abiotic stresses

Types of Ex-situ conservation1. Seed Gene bank

- All gene banks are essentially seed banks

- Seed storage in containers of Glass, Plastic and Tin for 50 to 100 year

Roberts (1973) has classified seeds into two groups for storage purpose ; viz.

Orthodox- Seeds which can be dried to low moisture content (5%) and stored at low temperature without losing their viability for long periods of time. More than 90% of plants spp. Belong to this group.

Recalcitrant- Seeds which show very drastic loss in viabilitywith a decrease in moisture content below 12 - 30% are knownas recalcitrant seeds- cocoa, coconut, mango, tea, coffee,rubber, jackfruit and oil palm seeds.

Such seeds cannot be conserved in seed banks and,therefore require in-situ conservation (on-farm).

Types conservation

Seeds are very convenient for storage

because they occupy smaller space than

whole plants.

In the seed banks, there are three types of

conservation, viz.,

1. Short term (Working

Collections)

2. Medium Term (Active

Collections)

Long Term (Base Collections)

Short term storage: (working collections )

1. Short term storage:

working collections are

stored for short term (>3-

5 years) at 10-150C at

10% Moisture.

The accessions being

actively used in crop

improvement programmes.

These collections are

maintained by the breeders

using them.

Medium term storage: (Active collections )

Medium Term storage: Theaccessions in an active collectionare stored at temperatures below150C (often near 00C), and theseed moisture is kept at 5%.

The storage is for mediumduration, i.e., 10-15 years.

These collections are used forevaluation, multiplication anddistribution of the accessions.

Active collections are usuallymaintained by multiplying theseeds of their own accessions.

But from time to time, basecollection material should be usedfor regeneration of thesecollections.

3. Long term storage: Theseconsist of all the accessionspresent in the germplasm of acrop, which are stored at about -200C with 5% moisture content;they are disturbed only forregeneration.

Germination tests are doneevery 5-10 years.

When the germination of anaccessions falls below, usually,95% of its germination at thestart of storage, the accession isregenerated.

High quality orthodox seeds canmaintain good viability upto 100years.

Long term storage: (Base collections )

Seed Gene Bank

Disadvantages Seeds of recalcitrant

species cannot be stored

in seed banks.

Failure of power supply

may lead to loss of

viability and thereby loss

of germplasm.

It requires periodical

evaluation of seed

viability.

After some time

multiplication is essential

to get new or fresh seeds

for storage.

Advantages• Large number of

germplasm samples

can be conserved in a

very small space.

• Handling of

germplasm is easy

• Germplasm is

conserved under

pathogen and insect

free environment.

2. Field Gene Bank

(Plant gene bank, ex-situ

conservation)

Those plant species that have

recalcitrant seeds or do not

produce seeds readily are

conserved in field gene banks.

In field gene banks, germplasm

is maintained in the form of

plants as a permanent living

collection.

Field gene banks are often

established to maintain working

collections of living plants for

experimental purposes.

They are used as source of

germplasm for species such as

coconut, rubber, mango,

The conservation of germplasm in field genebank

involves the collecting of materials and planting in the

orchard or field in another location. Field genebank has

traditionally been used for perennial plants, including:

Species producing recalcitrant seeds;

Species producing little or no seeds;

Species that are preferably stored as clonal material;

Species that have a long life cycle to generate breeding

and/or planting material.

Field genebanks are commonly used for such species

as cocoa, rubber, coconut, coffee, sugarcane, banana,

tuber crops, tropical and temperate fruits, vegetatively

propagated crops (e.g. wild onion and garlic) and forage

grasses (e.g. sterile hybrids or shy seed producers).

Many important varieties of field, horticultural and forestry

species are either difficult or impossible to conserve as

seeds (i.e. no seeds are formed or if formed, the seeds are

recalcitrant) or reproduce vegetatively. Hence they are

conserved in field genebanks(FGB).

FGBs provide easy and ready access to conserved material

for research as well as for use. For a number of plant

species, the alternative methods have not been fully

developed so that they can be effectively used (Ramanatha

Rao and Riley 1995; Ramanatha Rao et al. 1998).

It is one of the options of a complementary strategy for the

conservation of germplasm of many plant species. At the

same time, efforts to develop and refine other methods,

such as in vitro conservation and on – farm conservation,

must continue (Ramanatha Rao et al. 1998).

There is a possibility that a few well-managed gardens can

emphasise on conservation of certain groups of species as

Field Gene Bank

Disadvantages Field gene banks cannot

cover the entire genetic

diversity of a species. It can

cover only a fraction of the

full range of diversity of a

species.

The germplasm in field gene

banks is exposed to

pathogens and insects and

some-times is damaged by

natural disasters such as

bush fires, cyclones, floods,

etc.

Maintenance of germplasm

in the field gene banks is

Advantages It provides

opportunities for

continuous

evaluation for

various economic

characters.

It can be directly

utilized in the

breeding

programme.

Field Gene Banks in INDIA

Location/ Centre Plant Species Holdings

Issapur, NewDelhi Low chilling and minor fruits 305

Shimla HP Temperate fruits species, species of Rosaceae 800

CAZRI, JodhpurArid zone multipurpose trees, Jojoba, Jatropha,

Acacias350

Thrissur KRBanana, Jackfruit, Pepper, Root and

Rhizomatous crops (8 Perennial ssp.)539

Shilong, MEGABanana, Guava, Ornamentals, Citrus, Passion

Fruit71

Himalayan Zone Fruits and Herbal ssp. 74

Lakhnow, UPTamarind, Jamun, Bael, Jackfruit and other

Medicinal plant ssp.400

Bhowali Medicinal plant ssp. 261

CRYOPRESERVATION

Cryopreservation involves storageof plant material at low temp. ( -196 °C), in liquid nitrogen ornitrogen vapor ( -154 to -196 °C).

At this temperature the celldivision and metabolic processesstop.

Thus plant material can be storedfor longer period withoutalteration.

Cryopreservation of those speciesthat can easily be regeneratedinto whole plants is a promisingoption for the safe, long-termstorage of germplasm.

Cryopreservation requires limitedspace, involves very littlemaintenance and is considered tobe a cost-effective option.

MERISTEM GENE BANKS

For conservation of meristemcultures, meristem or shoot tipbanks are established. Germplasm of asexually

propagated species can beconserved in the form ofmeristem.

Widely used for conservation andpropagation of horticulturalspecies.

In vitro method can be used intwo ways.

1. storage of tissues under slowgrowth conditions.

2. long-term conservation of

POLLEN STORAGE/POLLEN GENE

BANKS

Pollen storage was mainly developedas a tool for controlled pollination ofasynchronous flowering plantsespecially fruit free species.

Advantage The relatively small quantity of the specimen

required for a single accession.

Exchange of germplasm through pollenpossesses fewer quarantine problemscompared with seed or other propagules.

Disadvantage Pollen storage alone cannot conserve the

cytoplasmic genetic diversity of a species.

There is need to assess the potentialdrawbacks of excluding maternal genes andfeasibility of ovule storage and in-vitrofertilization techniques.

In addition effective sample techniques tocover a population or gene pool are needed.

Tissue culture conservation

DNA STORAGE / DNA GENE BANKS

Storage of DNA is in principle,simple to carry out and widelyapplicable in the lab.

Genetic engineering has brokendown the crossability barriers.

Transgenic plants incorporatinggenes from virus, bacteria, fungiand even mice are reality now.

Such efforts have lead to storage oftotal genomic information ofgermplasm in the form of DNAlibraries.

However, strategies andprocedures have to be developedon how to use the material storedin the form of DNA.

Therefore, the role and value of thismethod for PGR conservation is not

Home Gardens Home gardens conservation is similar to on-

farm conservation but the scale is much

smaller.

Home gardens tend to contain a wide

spectrum of species, such as vegetables,

fruits, medicinal plants and species.

Home gardens, as a single unit has very little

value in terms of conservation, but a

community of them in a given area may

contribute significantly to the conservation

and direct use of genetic diversity.

Most of such diversity could be somewhat

unique/rare, as the people tend to grow

unique materials in their gardens and also

under utilized or undomesticated species.

However, the system is vulnerable to change

in management practices.

Home gardens are also known as testing

grounds for farmers for some of the wild

and semi-wild species.

Botanical Gardens

Botanical gardens aims atmaintaining essential ecologicalprocesses and life support systemspreserve genetic diversity and ensuresustainable utilization of species andecosystem.

The role of most Botanical gardens inconserving intra species diversity islimited because most of theseconserve only a few accessions perspecies or taxon.

However, these play a greater role inpublic awareness and education.

Botanical gardens are mainly used todisplay a great number of differentand exotic species.

There is a possibility that a few wellmanaged gardens lay emphasis onconservation of certain group of

Herbal Gardens

Herbal gardensresemble botanicalgardens except thatthese maintainmedicinal and aromaticplants.

Herbal gardens aregetting moreimportance these daysbecause medicinal andaromatic plant group ismost threatened due totheir over-exploitationin natural conditions.

On-farm

conservation On-farm conservation which the CBD defines as “A

form of in situ conservation in the place where thedomesticated or cultivated species have developedtheir distinctive properties.”

There is an urgent need to also pay attention to themany economically important wild species that areneither on-farm nor in protected areas. Thepopulations of many of these wild species are underheavy pressure due to over-exploitation, habitatdegradation and invasive species and agriculturalbiodiversity on farms and in forests.

Their effective in situ conservation will be difficult toaccomplish and therefore presents a huge challenge toconservationists.

To maintenance of domesticates such as landraces or

local crop varieties in farmers’ fields, often referred to

as ‘on-farm’ conservation (Maxted et al. 2002), ‘in

agro’ or ‘inter situ’ (Blixt 1994).

Trees may also be transplanted from native habitats

and managed within an in situ on-farm system using

traditional sylvicultural techniques. The material is

effectively managed within traditional farming systems

by local farmers.

Conservation of crop plants and primitive

cultivars/landraces only in on-farm.

strengthening the scientific basis of on-farm

conservation and management of local landraces,

horticultural crops and wild fruit species.

NBPGR

New

Delhi

Svalbard Seed Bank, Norway

To preserve gene diversity of major food crops,

international Institutions have established a series of green

gene banks, which store samples of genetic material of

various strains of each plant species.

Svalbard Seed Bank is meant as a sort of safety net, a

reserve of last resort and the vault functions like a genetic

safety deposit box. It stores duplicate specimens from gene

banks worldwide and while the Svalbard seed bank owns

the building, the individual depositor owns the contents of

his or her box and the access to individual specimens is

• The Svalbard Global Seed Vault on

February 26, 2008 with the

construction of the vault financed

entirely by Norwegian Government.

The operational cost is currently

shared by Norway and the Global

Crop Diversity Trust.

The seed bank is

located in an old

copper mine on remote

northern island of

Spitsbergen, Norway.

The facility currently has a capacity to conserve 4.5

million seed samples. With approximately 1.5 million

distinct seed samples of agricultural crops thought to

exist, the Svalbard Seed Bank can store roughly

three of each sample. Under the current temperature

conditions in the vault (temperatures similar to those

in a kitchen freezer) the seed samples can remain

viable to begin new crops for anywhere from 2000 to

20,000 years.

On farm conservation On farm conservation involves the

maintenance of traditional crop cultivars(land races) by farmers within traditionalagricultural system.

It is based on the recognition that farmershave improved and grown genetic diversity.This process will still continue among manyfarmers in spite of social economic andtechnical changes. Farmers should beencouraged to continue their land races byagricultural development policies thatenhance incentives to continue to maintainland races.

Given the role of farmers on farmconservation, meeting development goalssuch as increased farm income is critical.

In a recent conclusion study by NBPGR withIRRI, Philippines and GKV, Raipur in Basterarea of MP suggested that on farmconservation of rice genetic resources is acompliment to ex-situ conservation in tribalarea of Baster Plateau which can bemotivated for maintaining crop diversityprovided it is a viable option and forstrengthening the farmer's assess to

conservation

Conservation Vegetativaly Propagated

Crops

Conservation of

vegetativaly

propagated

crops

Ex-situ as well

In-situ and on-

farm condition

Conservation Vegetativaly propagated

crops

Registration of Plant

Germplasm

1. Plant Germplasm Registration Committee

2. Nodal Agency

3. Application Form

4. Eligibility Criteria for Registration

5. Germplasm Ineligible for Registration

6. Screening of Application(s) and their

Consideration by the PGRC

7. Validity of Registration

8. Publication of Registered Germplasm

9. Conservation, Maintenance and Sustainable

Utilization of Registered Germplasm

10. De-registration

Procedure For Submission Of

Proposal / Germplasm Material

1. Submission of Application and

Germplasm

2. Guidelines for Submitting the Orthodox

Seed Germplasm

3. Guidelines for Submission of

Recalcitrant / Intermediate Seed

Germplasm

4. Guidelines for Submission of

Propagules

National Active Germplasm Sites

In India, Germplasm Maintains and

Exchange

For Agriculture and Horticulture Crops –

NBPGR, New Delhi (1976)

For Mediational and Aromatics Plants

(Botanical Plants) – BSI, Kolkata, WB

(1890)

For Forest Trees – FRI, Dehradun, UKD

References Maxted, N., J.G. Hawkes, B.V. Ford-Lloyd and J.T. Williams.

1997. A practical model for in situ genetic conservation

complementary conservation strategies. Pp. 339-367 in Plant

Genetic Conservation: The In Situ Approach (N. Maxted, B.V.

Ford-Lloyd and J.G. Hawkes, eds.). Chapman and Hall, London.

Harlan, J.R. 1975. Crops and Man. First Edition. American

Society of Agronomy and Crop Science Society of America,

Madison, Wisconsin.

HD Upadhyaya, CLL Gowda and DVSSR Sastry, 2008. Plant

genetic resources management: collection, characterization,

conservation and utilization, An Open Access Journal published

by ICRISAT, INDIA.

http://lloydkahnongoing.blogspot.in/2011/12/svalbardseedbank.h

tml

GUIDELINES for REGISTRATION OF PLANT GERMPLASM

(revised, 2014)

NBPGR (ICAR) Pusa Campus, New Delhi-110 012

Brown, A.H.D. 1999. The genetic structure of crop

landraces and the challenge to conserve them in situ

on farms. In: Brush, S.B. (ed.), Genes in the Field:

Conserving plant diversity on farms. Lewis Publishers,

Boca Raton, FL, USA. pp. 29–48.

Maxted, N., Guarino, L., Myer, L. and Chiwona, E.A.

2002. Towards a methodology for on-farm

conservation of plant genetic resources. Genetic

Resources and Crop Evolution 49(1):31–46.

Mohd Said Saad and V. Ramanatha Rao (2001).

Establishment and management of field genebank . A

Training Manual, IPGRI-APO, Serdang

References