Genebank Management

Ex situ Conservation of Germplasm

Veena Gupta* and Sherry Rachel Jacob *Principal Scientist and Head

Division of Germplasm Conservation ICAR- NBPGR

Biodiversity

PGR

Cultivated PGR

250,000 higher plant species described taxonomically

46% of total biodiversity (115,000 species)

14% of total biodiversity (35,000 species)

Seeds of orthodox species can be conserved Ex situ in Genebanks at -18oC – safest and cheapest method of conservation

• Seed genebanks – unique archives of genetic wealth

• Rich reservoir of potential genes

• Temporal and spatial restrictions in material availability addressed through

genebanks

Ensures the supply of desired germplasm for diverse crop improvement

programmes

Germplasm conserved Ex situ includes-

Parental Lines of Hybrids

Genebank

Genetic Stocks

Wild & Weedy relatives

Improved cultivars

Released Varieties

Land races

Farmers Varieties

•1,750 individual genebanks are reported worldwide

•8 have more than 100,000 accessions

•130 hold more than 10,000 accessions

•The CGIAR established 11 genebanks for conservation and use of PGR .

• CG germplasm is now held ‘in-trust’ for the world community under signed agreements with FAO in 1994

Circles with red indicator represent the CG genebanks.

NETWORK COLLABORATORS

DARE/ ICAR

(The Umbrella Organization)

NBPGR and

its 10

Regional

Stations

59 National Active

Germplasm Sites (NAGS)

comprising ICAR institutes, Project

Directorates, NRCs, AICRIPs,

SAUs

Other National

Stakeholders

International

Collaborators

NBPGR

(The Nodal Institute)

Management of Plant Genetic Resources in India

The NGB located at the headquarters in New Delhi, with the network of 10 regional stations at

different agro-climatic zones and linkages with 59 National Active Germplasm Sites of National

Agricultural Research System is involved in sustainable conservation of PGR.

National Genebank

Network (NBPGR responsible for PGR

activities at national level)

Long-term modules at

-200C

(12)

Medium-term

modules at +4 to

+80C

(6)

In Vitro Genebank at

25+20C (6 culture

rooms)

Cryogenebank at

-180 to -1960C

(6 cryotanks)

NBPGR,

New Delhi

NBPGR, Regional

Stations (10)

Medium-term

modules at

+4 to 80C

(18)

National Active

Germplasm Sites (NAGS)

(59 crop-based ICAR

institutes and SAUs)

Field

Genebanks

Germplasm conservation facilities at ICAR-NBPGR

National Genebank In vitro Genebank

Field Genebank Cryo bank

Short term storage

Storage period: 12–18

months.

Temperature: 18–20 °C

RH: 45–50 %

Storage container type: Cloth

bags, paper bag, glass bottles.

Types of storage in Seed Genebank Medium term storage

• Storage period: 5–10 Years

• Temperature: 0–10 °C

• RH: 35–40 %

• Storage container type: Cloth bag, paper bag, metal can, glass bottle and plastic jars

Long term storage

Storage period: 50–100

Years

Temperature: -18 - -20 °C

RH: No control

Processing of the germplasm at NGB Seed Cleaning

Seed cleaning is the removal of debris, inert material, damaged and infested or infected seeds and seeds of different species (e.g., weeds) to achieve clean and pure samples of seeds of high physiological quality for storage Moisture testing Moisture testing by International Seed Testing Association (ISTA) method Seed viability testing A viable seed is one that is capable of germinating and producing a normal seedling. Tetrazolium Test The tetrazolium test distinguishes between viable and dead tissues of the embryo on the basis of their relative respiration rate in the hydrated state.

Abnormal seedlings in Triticum aestivum L.

Viable Seeds

Non-Viable Seeds

Processing of germplasm at NGB Seed drying Specially designed seed dryers which can maintain 15% RH and 15 °C temperature is used to allow slow and safe drying process at a temperature where minimum loss of seed quality occurs.

Seed Packaging

Specially designed tri-layered aluminum foil packets with following specifications have been found most effective for long term conservation of all types of seeds Outer polyester layer: 12 µm. Middle aluminium foil layer: 12 µm. Inner polyethene layer: 250 gauge

Genebank Information Management System (GBIMS)

No

* Self-pollinated (>2000), Cross-pollinated (>4000) and wild (>500)

** Viability relaxation for wild species in Vegetable/Medicinal/Rare

endangered /forage species (50-70%)

No

Yes

No

No

No

Yes

Yes No

Yes

Are the documents complete?

Verification of Documents

i. Passport Data sheets in case of germplasm

ii. Proposals in case of release varieties

iii. Registration Proforma in case of registered germplasm

Correspond with the Donor

and get more information

Domestic Quarantine (DQ) number allotment and entry in

GHU database and check for the duplicates in the National

Genebank database

Is the sample Unique?

Is the sample with sufficient

quantity*?

Correspond with donor for

more information

Program it for

regeneration/multiplication

Is the seed sample clean?

Remove debris, infested

and broken seeds

Is the seed health satisfactory as

per standards of plant quarantine?

Yes

Send it to concerned crop curator for

viability testing

Is viability** >85%

Sent back to the

donor for

multiplication

Assign National ID (PGEC Division) and document information in database and

conserve the sample in the Genebank

Fumigated samples

Germplasm Acquisition

Fumigate or discard the

sample under intimation to

donor

Initial

Germination

(%)

Monitoring interval (years)

Active collection (4 °C) Base collection (-20 °C )

Non-oil

rich

seeds

Oil rich

seeds

Non-oil rich

seeds

Oil rich

seeds

<85 5 3 10 5

85-95 8 5 15 8

>95 10 8 20 10

Monitoring of the conserved germplasm

Crop Group Present status

Cereals 161897

Millets 58443

Forages 6930

Pseudo-cereals 7295

Legumes 65675

Oilseeds 58574

Fibre crops 15586

Vegetables 26077

Fruits & Nuts 273

Medicinal & Aromatic plants & Narcotics 7947

Ornamental 653

Spices and Condiments 3074

Agroforestry 1642

Duplicate safety Samples (Lentil, Pigeonpea) 10235

Trial Material (Wheat, Barley) 10771

Total 435072

List of genebank s as per ranking

Accns Species

USDA 584449 15720

ICAR-NBPGR 435072 1900

ICGR-CAAS China 415232 360

NI Vavilov All-Russian Scientific Research Institute of Plant Industry (VIR)

325,000 2532

NARO, JAPAN h

224000 -

Republic of Korea National Agrobiodiversity centre

201,889 1617

IPK, Germany 151002 2933

Plant Gene Resources of Canada

110,444 1036

Embrapa Genetic Resources, Brazil

102000 500

MSB, Kew 80,000 37600

Global Genebanks Vs Indian National Genebank

Thrust areas for future

•Gaps in explored and

conserved accessions

•CWRs

•Repatriation

•Alternate site – Zero

energy conservation

20

National Permafrost Facility

GUIDELINES FOR SENDING GERMPLASM FOR LTS CONSERVATION

• Only mature and clean seeds free of any insect pests and fungi and weed seeds Undersized, shriveled and immature seeds

• 2,000 seeds for self pollinated crops and 4000 seeds number for cross pollinated crops

• packed in moisture permeable pouches • correctly and neatly labeled. If possible typed or computer generated

labeling should be done. • Seeds should not be treated with any chemical • five major type of data needed for any accession meant for conservation

viz., Passport data, Management data, Characterization data, Evaluation data and Mode of reproduction.

S.No. Collector No.

IC. No. Crop Name Botanical Name

Vernacular Name

Biological status Type of

material

Date of

collection

Source

Frequency

S.No.

Coll. No.

Sample Type Sampling Method

Habitat

Site of Collection Latitude

Longitude

Altitude

Remarks

Village Mand

al District State

PASSPORT DATA FORM

Collector’s Name and Address:

Collaborating Institute: Name of Scientist(s) and Address:

Area surveyed:

Biological Status Descriptor State Definitions

Sl.

No.

Descripto

r

Definition Letter

code

1 Wild All species other than

cultivated of PGR Value/

potential value.

WL

1a

Crop Wild

Relative

All wild species that are

related to cultivated

species and part of the

gene pools from which

genetic introgression into

cultivated species has

occurred.

WL_CWR

2 Weedy Weedy forms of cultivated

species occurring in

companionship with some

other cultivated species.

WD

3 Landrace/

Traditional

cultivar

All cultigens in farmer’s field

with/without specific names

frequently associated with unique

traits and geographical region

identified by farmers

LR_TC

3a Landrace All cultigens in farmer’s field

frequently associated with unique

traits and names identified by farmers.

LR

3b Traditional

cultivars/

Primitive

cultivars/

folk variety

All heterogenous cultigens cultivated

by farmers with specific vernacular

names frequently associated either

with unique traits or geographical

region often with some specific traits.

TC

3c Farmers

variety

Farmers variety registered under the

PPV&FR Act viz-

‘Varieties which have been

traditionally cultivated and evolved by

farmers in their fields or those that are

wild relatives or land races of a variety

about which farmers possess common

knowledge.’

FV

4 Breeding/

Research

material

Semi-finished products or

segregating material generated

out of breeding program to meet

specific breeding objectives.

BR

4a Hybrid It is cross breed developed by

crossing of two inbred lines,

varieties, species or genera.

BR_H

4b In-breds Homogenous offspring of cross -

pollinated species obtained as a

result of repeated forced self

pollination

BR_I

4c Segregating

population

Genetically diverse progeny from

crosses between parents that

differ for one or more traits under

genetic control.

BR_SG

4d Breeding

lines

Lines developed in breeding

programmes with valuable trait/

gene recombination.

BR_BL

4e Parental

lines of

hybrids

Parents of developed hybrids. BR_PL

5 Genetic

stock

Trait and gene

specific germplasm,

experimentally

developed or

identified through

scientific

interventions (e.g.

sources of resistance,

cytogenetic stock ,

mutants, mapping

population, MS lines

GS

5a Registered

germplasm

Germplasm registered

for unique trait(s)

with ICAR-NBPGR.

GS_RG

6 Elite line Crop varieties under

advanced varietal trial

but not yet released/

notified. Selection

from population from

the coordinated trial

(AVT I & II line).

EL

7 Improved

cultivar

Improved cultigens of common knowledge that are in commercial

cultivation (extant varieties, and varieties released by institution/

organization/ State).

IC

7a Released

variety

Varieties released and notified by the Central Sub-Committee on Crop

Standards, Notification and Release of Varieties of Agricultural and

Horticultural Crops.

IC_RC

7b Released

Hybrid

Hybrids released and notified by the Central Sub-Committee on crop

standards, Notification and Release of Varieties of Agricultural and

Horticultural Crops.

IC_RH

7c State

released

Varieties released by the State Committee of Agricultural and Horticultural

Crops.

IC_SR

7d Institute

released

Varieties released by the Institute Committee of Agricultural and

Horticultural Crops.

IC_IR

7e Exotic

varieties

Exotic varieties introduced through the process of germplasm exchange IC_EX

8 Others Doubtful germplasm with unknown biological status. OT

Thank You

IRC, 2016