1. Collection, Evaluation and Documentation of
Germplasm
Submitted by:
Patel Yash N.
Reg. no.: 2020219035
M. Sc. Horticulture (Fruit Sci.)
Submitted to:
Dr. A. K. Pandey
Assistant Professor
Department of Fruit Science

2. India has a rich and varied heritage of biodiversity,
encompassing a wide spectrum of habitats from tropical rainforests
to alpine vegetation and from temperate forests to coastal wetlands.
Out of 18 biodiversity hot spots identified in the world, four
hotspots, i.e. Western Ghats, Western Himalaya, Indo-Burma region,
and Nicobar islands (Sunderland) are in India. It is estimated that
there are 8.7 million species of the world’s biota. Out of them only
1.7 million have been described to date, and their distribution is
highly uneven. India contributes significantly to the biodiversity of
the world by accounting 7.31 % of the global plant diversity from
2.4% of the world’s area.

3. Primary centre Mango, Citrus, Jack Fruit, Bael, Aonla, Ber,
Khejri, Jamun, Tamarind, Phalsa, Lasoda,
Karonda, Wood apple, Pilu, Bilimbi,
Garcinia, Under Utilized Fruits
Secondary
centre
Banana, Pomegranate, Mulberry, Malus,
Pyrus, Prunus, Rubus
India-Centre of Diversity of fruit Crops:
(Source: Singh et al. 2009)

4. COLLECTION OF GERMPLASM

5. REASONS FOR GERMPLASM COLLECTING
• The demand of germplasm is unpredictable and dynamic but
in practice, some prioritization is necessary both at species
level and geographic regions.
• It is in danger of extinction or even erosion;
• A clear need exists for it, as it was expressed by farmers;
• The diversity it represents is missing from or insufficiently
represented in, existing ex-situ germplasm collections;
Definition:
Tapping of genetic diversity from various sources and
assembling at one place is called germplasm collection.

7. EXPLORATION PROGRAMME: STEPS
• Planning
• Making contacts with local research organization
• Gathering equipment and preparation
• Meeting with local researchers in area to be surveyed
• Sorting out of collected samples
• Reporting to the Headquarters
• Preparation & publication of reports
• Distribution/conservation of collected samples

8. AREAS TO BE SURVEYED
• Diversity/Variability rich area
• Under explored area
• Tribal dominated area
• Hot spots
• Threatened habitats
• Farmers’ field

9. PLANNING OF EXPLORATION MISSION
Prioritization of Species and Areas after Gap Analysis:
• The areas to be explored and crops/species to be
collected should be prioritized after thorough gap
analysis based on information from different sources
including database/National Genebank status at
NBPGR.
• The explorer should be well-versed with the nature
and extent of diversity and breeding behaviour of the
crop/species to be collected.

10. • Plan well in advance to facilitate the preparations of
the proposed missions except those to be carried out
under special situations like rescue collecting.
• Visit to herbaria should be made to know the range of
distribution, localities, diversity pattern and period of
collection particularly for wild species.
• Phytosanitary regulations should be followed in case of
transportation of material from foreign country.

11. FINALIZATION OF MISSION
Gathering eco-geographic information:
● Information on topography, climatic conditions, vegetation,
crops in cultivation and their maturity, etc. needs to be gathered
to finalize the tour path of collecting mission.
● Besides, explorers should establish local contacts especially at
grass root level to seek the social, cultural, ethnic and other
information of interest.
● Types of survey:
o Coarse grid survey should be conducted in unexplored areas
to capture the overall variability.
o Fine grid survey should be carried out to build-up more
collections for specific trait(s) known to exist in identified
pockets in previously explored areas.

12. PERIOD OF COLLECTION:
• For seed producing crops and species, exploration should be
undertaken when these are physiologically mature and ready
for harvest.
• In case of species with shattering nature, missions are executed
rather earlier (7-10 days depending on crop/species) before
their maturity. Further, longer duration (2-3 weeks) mission and
repeat visits are suggested for collection of wild species.
• For vegetatively propagated crops/species, the targeted areas
should be surveyed first for identification and marking of elite
types at the time of flowering/fruiting and subsequently the
collections are made at appropriate time.
• Within the country should be of at least 10-15 working days
(excluding journey period) and more than a month when
organised in foreign countries.

13. Team composition: Team consisting of 2-3 members including a
collaborator and need-based local-aid be formed preferably a
botanist/ breeder as leader.
Area and route of exploration: This should be fine-tuned in
consultation with the subject experts of local bodies as soon as the
team reaches to the starting point keeping in view the targeted
species and areas of the proposed mission.
Items and equipments required: As per the nature of the
germplasm to be collected (fruit/ seed/ vegetative propagule/ in
vitro/ live plants) and the area(s) to be explored, the required items
and equipments have been listed.
Domestic quarantine: All precautions including need-based
domestic quarantine should be followed for pest-free collection
and its transportation.

14. TYPES OF COLLECTIONS
Currently three types of collections are held at
most genetic resources centres.
1. Base collection
• The base collection is defined as a set
of accessions, which in terms of
genetic integrity, is as close as
possible to the sample provided
originally, which is preserved for long
term future.
• Seeds can be conserved under long
term (50 to 100 years), at about
-20˚C with 5% moisture content.
• They are disturbed only for
regeneration.

15. 2. Active collection
• An active collection comprises accessions
which are immediately available for
multiplication and distribution for use.
• Accessions are stored for short to medium
periods of time (generally upto 20 years).
• Active collections should be kept at such a
conditions at least 65% germination can be
obtained after for 10 to 20 years.
• Active collections are generally held at
temperatures between 0-10˚C depending
upon the species stored, the prevailing
ambient environment and cost factors.

16. 3. Working collection
• Seeds are stored for 3-5 years
at 5-10˚C and the usually
contain about 10% moisture.
• Such materials are regularly
used in crop improvement
programmes.
• These collections are
maintained by breeders.

17. SAMPLING METHOD
1. Random Sampling
Random sampling is usually carried out by randomly
selecting a starting point at a collection site and taking a single
spike or few pods at every second and third place along a number
(50-60) transacts evenly dispersed in a target sets and bulking them
to form a random sample.
2. Biased sampling
This method is generally followed for sampling rare
phenotype variants or observable spontaneous mutants occurring
in the target population. The attention is given to identify specific
variants for direct utilization.

18. 3. Clustered Sampling
This method is applicable for collection of wild relatives or
weedy species. The collection site is divided into several evenly
spread clusters. Samples are taken independently from each
cluster and then bulked together to form a multiple sample.
4. Coarse Grid Sampling
This technique of sampling is used to collect random bulk
samples from a site by harvesting parts/plants from several spots
of the site. In coarse grid sampling is made at wide interval over
the entire region.
5. Fine grid sampling
This sampling technique is used in interested or intensive
areas of variation identified after the coarse grid survey. Collector
may walk across the site or field twice in cross or zigzag manner
avoiding sampling from borders.

19. COLLECTING WILD RELATIVES OF CROP SPECIES
By wild relatives and related taxa can be classified into primary,
secondary and tertiary gene pools:
1. Primary genepool (GP-1)
Wild species in the primary genepool can produce fertile hybrids with
cultivated types and hence are easy to exploit. These are wild progenitors
closely related to crops.
2. Secondary genepool (GP-2)
The wild species are relatively distantly related and cross compatible
and hence contribute germplasm less easily.
3. Tertiary genepool (GP-3)
Distantly related and unrelated taxa of different genera/species which
can only be used with difficulty for some crops for a limited number of genetic
traits are considered as tertiary gene pool. Hybrids are sterile. The wild
species and the weed races represent the highest level of genetic
heterozygosity and heterogeneity among the different classes of germplasm.

21. COLLECTION TECHNIQUES
Seed propagated, cultivated and wild species
1) Randomly selected sample of 50-100 plants of self pollinated
crop and 100-200 plants of cross pollinated crops.
2) Collect fully and physiologically mature seeds.
3) Collect about 50 seeds (as per the availability) from each
plant and bulk to make a population sample of 2000-4000
seeds in self pollinated and 4000-8000 seeds in cross
pollinated crops.
4) Record passport data and important plant traits for each
sample.
5) Sample as many sites as possible as per the availability of
time.
6) If considerable morphological variation is present make
separate samples of each type.

22. 7) Add biased sample if some morphotypes are not included in
random
8) Make herbarium specimens wherever possible.
9) Take photographs of the important variants materials.
10) Write desired field notes
b) Vegetatively propagated crops:
1) Sample each distinct morphotype in the area.
2) Supplement with seed collections wherever possible give
separate number for same plants or seeds from several
plants (bulk samples)
3) The collected materials for grafting with 3-4 days send by
speed post or special messengers

23. No. Crop NAG site* (NAGS) No. of accessions
currently
available
1. Vegetables NBPGR, Pusa Campus, New Delhi 16,139
2. Potato CPRI, Shimla (Himachal Pradesh) 2,375
3. Spices NRC for Spices, Marikunnu, Calicut (Kerala) 2,847
4. Plantation
Crops
CPCRI, Kasaragod (Kerala) 307
5. Medicinal &
Aromatic
Plants
AICRP on Medicinal & Aromatic Plants,
NBPGR, New Delhi
375
6. Fruits (semi-
arid)
All India Coordinated Project (Semi Arid
Fruits), HAU, Hisar (Haryana)
541
7. Fruits
(Subtropical
& Temperate)
NBPGR Regional Station, Phagli, Shimla
(HP)
454
8. Fruits (all) IIHR, Bangalore (Karnataka) 13,118
9. Citrus NRC for Citrus, Nagpur (Maharashtra) 51
10. Mango CIHNP, Lucknow (UP) 587
11. Tuber Crops CTCRI, Sreekariyam, Trivandrum (Kerala) 3,586
Indian Plant Genetic Resources System - Directory of National Active Germplasm Sites (NAGS)

25. EVALUATION OF GERMPLASM

26. Definition:
It deals with the assessing the agronomic potential of an
accession including quality parameters and response to various abiotic
and biotic stresses.
• Evaluation of germplasm resources is necessary to identify the
appropriate germplasm with a target trait for their further utilization.
• Evaluation of germplasm is a multi-disciplinary approach and it should
be done in collaborative mode involving germplasm curator, plant
breeder, physiologist, pathologist, entomologist, biochemist etc.
• Proper plant spacing, fertilizer application, weeding, irrigation, plant
protection measures need to be followed.

27. • The number and row length should be more for cross pollinated
species than those for self-pollinated ones.
• The observations should be recorded on the plants from the
middle row to avoid the border effects.
• After evaluation, the promising accessions should be further
validated through multi-locational, multi-season and multi-year
evaluations.
• Evaluation trails should be carried out in at least three diverse
environments to minimize Genotype x Environment (G x E)
interaction.

28. • Identification of resistance source against a particular
race/strain/biotype within a particular location does not guarantee its
resistance response in other locations may vary depending upon the
agro-meteorological conditions.
Evaluation for biotic stresses:
• Therefore, screening of germplasm for
biotic stresses should be accompanied with
identification race/strain/biotype of the
pest and pathogens existing at that
particular location.
• These four factors i.e. susceptible host,
virulent pathogen, environment and time,
which cause disease called is called
“Disease Quadrangle”.

29. • Under these circumstances, repeated field screening with proper
experimental design followed by stringent screening under artificial
challenged inoculation condition under field and glasshouse are
essential to establish a real resistance or tolerance response of
germplasm.
• For recording of the observations on fungal, bacterial and viral
diseases, standard evaluation system (SES) scale should be
followed.
• For recording of the data on defoliators, percentage infestation in
each accession should be recorded.
• In case of sucking insect pests, the number of insects per unit
plant/leaves/inflorescence should be counted.

30. • Preliminary screening/phenotyping should be done for one year
with large number of accessions under field conditions.
• To narrow down the large number of germplasm accessions,
preliminary screening for abiotic stress should be carried out for
one year with large number of accessions under field conditions
peculiar to the stress under study.
• Evaluation of germplasm for tolerance to different abiotic stresses
is to be carried out under well-defined controlled conditions so
that the optimum stress can be imposed at the desired stage.
Evaluation for abiotic stresses:

31. • Like, evaluation for light and temperature stresses should be carried out
under phytotron facility.
• Evaluation for drought tolerance should be undertaken in drought plots
with rain out shelters and with well defined moisture conditions.
• Microplots with well defined electrical conductivity including specific
salt/ions should be used for evaluating germplasm under saline/alkaline
conditions.
• Standard checks identified for specific abiotic stress should be used for
proper comparison of the germplasm.
Phytotron facility

32. Evaluation for quality parameters:
• Quality evaluation plays an important role for in identification
of value rich germplasm keeping in view of the nutritional
and health security.
• Quality traits like oil, fatty acids, protein, phenols, sugar,
amino acids, vitamins, minerals and anti-nutritional factors
requires specialized equipments.
• Properly labeled and packed material containing complete
experimental details along with passport information should
be made available for quality analysis.

33. DOCUMANTATION OF GERMPLASM

34. Germplasm conservation, in its various stages, includes a
range of activities for which information is required or from
which information is derived. This may refer to species, their
sites of origin, or activities or stages of conservation. The action
of recording, organizing, and analyzing conservation data is
known as documentation.
WHAT IS DOCUMENTAION?

35. OBJECTIVES
• Understand what documentation of plant genetic
resources (PGRs) signifies, and its importance in the
routine management and scientific use of a germplasm
bank
• Define the stages of constructing a documentation
system
• Document the most common operational procedures of
a germplasm bank

36. Stages for the establishment of a
documentation system:
• Obtaining information on the bank’s needs.
• Defining documentation objectives.
• Analyzing procedures.
• Identifying significant descriptors.
• Developing data formats and recording forms.
• Developing documentation procedures and implementing
the system.

37. Documenting common procedures
The most common procedures in germplasm banks
include activities are:
• Registering samples (data of accessions)
• collection
• cleaning
• drying
• viability testing
• storage
• Characterization and evaluation
• regeneration
• distribution

39. Accession: An accession is a group of related plant material from
a single species which is collected at one time from a specific
location. Each accession is an attempt to capture the diversity
present in a given population of plants.
Accession Number: A unique number assigned to each accession.

40. MULTI CROP PASSPORT DISCRIPTORS:
• Institute code (INSTCODE)
• Accession number (ACCENUMB)
• Collecting number (COLLNUMB)
• Collecting institute code (COLLCODE)
• Genus (GENUS)
• Species (SPECIES)
• Species authority (SPAUTHOR)
• Subtaxa (SUBTAXA)
• Subtaxa authority (SUBTAUTHOR)
• Common crop name (CROPNAME)
• Accession name (ACCENAME)
• Acquisition date [YYYYMMDD] (ACQDATE)
• Country of origin (ORIGCTY)
• Location of collecting site (COLLSITE)
• Latitude of collecting site (LATITUDE)

41. • Longitude of collecting site (LONGITUDE)
• Elevation of collecting site (masl) (ELEVATION)
• Collecting date of sample [YYYYMMDD] (COLLDATE)
• Breeding institute code (BREDCODE)Biological status of
accession (SAMPSTAT)
• Ancestral data (ANCEST)
• Collecting/acquisition source (COLLSRC)
• Donor institute code (DONORCODE)
• Donor accession number (DONORNUMB)
• Other identification (numbers) associated with the accession
(OTHERNUMB)
• Location of safety duplicates (DUPLSITE)
• Type of germplasm storage (STORAGE)
• Remarks (REMARKS)
Continue…

43. Registering samples
The registration of samples consists of assigning each
sample (or accession) a unique identification number and
recording data received with the samples, including those known
to be descriptors of the accession. The data recorded would
include:
• Accession number
• Other numbers associated with the accession (e.g., code numbers for
collectors and donors)
• Scientific name (genus, species, subtaxa, and authorities)
• Common name(s) of the cultivated species
• Cultivar name(s)
• Date of acquisition of sample (incorporation into the germplasm bank)
• Date of last regeneration

44. Collection data
Collection data are also known as passport data and refer to the data
reported when the sample was first collected. These collection data or
descriptors can be numerous, depending on the degree of detail in which
information is needed. FAO and IPGRI have jointly prepared a list of passport
descriptors of many crops to provide uniform coding systems for common
passport descriptors of various crops (FAO and IPGRI 2001).
The passport data most commonly documented on registering samples include:
• Collection date
• Collector’s name, number, and institute
• Country and province or state of collection
• Locality, latitude, longitude, and altitude of collection site
• Origin of sample (e.g., household garden, market, or farm)
• State of sample (e.g., wild, landrace, or advanced cultivar)
• Number of sampled plants

45. SEED CLEANING
The seeds to be conserved in a germplasm bank should
be, as far as possible, clean and free of broken seeds, residues,
or infested or infected seeds. To save time, some banks do not
document this procedure; others consider that such data have
little practical or scientific value. Some descriptors suggested for
this procedure are:
• Accession number
• Date of procedure
• Method used
• Total number of seeds
• Empty seeds (%)
• Operator (name of person who carried out the test)

46. SEED DRYING
In a germplasm bank, orthodox or intermediate seeds are dried to
reduce their moisture content to acceptable levels without affecting their
viability. The initial moisture content is first determined. If this is very high,
then the seeds are dried, using a suitable method, to reduce moisture
content to the desired level. Once seeds are dried, some banks determine the
total weight of the dried seeds and the 100- (Seed index) or 1000-seed weight
(Test Weight), depending on their size.
The most commonly used descriptors for seed drying are:
• Accession number
• Initial moisture content
• Drying method
• Date of measurement
• Final moisture content
• Total dry weight of seeds
• 1000-seed weight
• 100-seed weight (for large seeds)

47. SEED VIABILITY
Germination under laboratory conditions is defined as the
emergence and development of those essential structures that indicate,
for the class of seed being analyzed, the seed’s ability to become a
normal plant under favourable conditions. The results of this test
indicate the percentage of live seeds of an accession that can produce
plants under appropriate conditions.
• Accession number
• Lot reference (any date, code, or number
that uniquely identifies the accession’s
regeneration or multiplication cycle)
• Collection type (e.g., whether base or active
collection)
• Reference for method used (e.g., absorbent
tissue or tetrazolium test)
• Viability (%)
• Operator (name of person who carried out
the test)

48. STORAGE
Once the seeds have been dried and cleaned, and their
percentage of viability recorded, they are stored in cold rooms (or
under normal conditions, according to case). The following data or
descriptors are recorded:
• Accession number
• Lot reference
• Collection type
• Location in cold room
• Total quantity of seeds stored per
accession
• 1000-seed weight
• 100-seed weight (for large seeds)
• Minimum quantity permitted for
seeds (this parameter helps determine
when more seeds should be produced
or multiplied)

49. GERMPLASM CHARACTERIZATION
AND EVALUATION
Germplasm characterization refers to the recording of
highly inheritable descriptors (or data) that are readily seen and
are expressed in all environments. They mostly include:
• Accession number
• Plant descriptors (morphological characterization)
• Susceptibility to abiotic stress (evaluation)
• Susceptibility to biotic stress (evaluation)
• Biochemical markers (molecular characterization)
• Molecular markers (molecular characterization)
• Cytological characteristics

50. REGENERATION OR MULTIPLICATION
Regeneration or multiplication is carried out in response to
data obtained through the seed monitoring control or during the
growth cycle of the vegetative propagated species, which is
conducted at given intervals of time to test the viability of each
accession in storage and ascertain the quantity of seeds it has.
The principal data or descriptors used to record regeneration or
multiplication are:
• Accession number
• Lot reference
• Collection type
• Regeneration site
• Plot reference (of field,
furrow, and plot number)
• Planting date
• Planting density
• Germination in the field (%)
• Established plants (no.)
• Days from planting to
flowering (no.)
• Harvest date
• Cultural practices

51. GERMPLASM DISTRIBUTION:
Linked to the information mentioned above on storage,
information on the distribution of germplasm that the bank
carries out should also be recorded. For example, some banks
continually distribute germplasm for improvement programmes
or for exchange with other banks. In these cases, to maintain
efficient control over the bank’s holdings of materials, a record
must be kept of the materials being distributed. Typical
descriptors that should be considered are:
• Accession number
• Lot reference
• Date of exit of material
• Quantity of seed sent
• Data on receiver
• Plant health certificate number (if applicable)

52. REFERENCES
• FAO; IPGRI. 1994. Genebank standards. Rome. 15 p.
Also available at http://www.ipgri.cgiar.org/
• Konopka J; Hanson J. (1985) Information handling systems for
genebank management. In Konopka J; Hanson J, eds. Proc.
Workshop held at the Nordic Genebank, Alnarp, Sweden, 21-
23 Nov 1984. IBPGR, Rome. pp 21-28
• Stalker HT; Chapman C. (1989) Scientific management of
germplasm: characterization, evaluation and enhancement.
IBPGR, Rome. 194 p