Conservation of chickpea genetic resources

Contributors to this page: ICRISAT, Patancheru, India (Hari D Upadhyaya, Shivali Sharma, Cholenahalli L Laxmipathi Gowda, Dintyala Sastry, Sube Singh); NBPGR, New Delhi, India (Shyam Sharma); ICARDA, Aleppo, Syria (Ahmed Amri, Kenneth Street, Natalya Rukhkyan), SARC-RIPP, Piestany, Slovak Republic (Gabriela Antalikova); Institute of Plant Genetic Resources ‘K.Malkov’, Sadovo, Bulgaria (Siyka Stoyanova); Department of Primary Industries, Victoria, Australia (Bob Redden); IPK, Gatersleben, Germany (Andreas Börner).

Importance of chickpea conservation

Archeological chickpea remains found from Syria to Greece dating from the very early days of farming show its important role in the dispersal of farming habits. Like many other crops, it has been subjected to different selection pressures in various ecological and cultural environments and consequently diversified into a range of varieties. It is likely that chickpea has a higher potential to yield more than the yields currently achieved on farmers' fields. This potential can only be fully utilized if a broad range of genetic materials are available to select from or to breed for specific traits. Therefore, breeding for higher and more stable yields and also for enhanced resistance to pests and diseases is one of the immediate objectives of many chickpea programmes.

• A more efficient and effective conservation of the chickpea genepool worldwide, including both cultivated and wild species, is essential and potentially very important for research and crop improvement.
• Some of the wild relatives are potentially important to incorporate resistance to abiotic and biotic stresses.
• Tolerance to drought is a particularly important trait for chickpea that often grows in extremely harsh and dry environments.
• Priority should be given to conservation of chickpea in primary and secondary centres of diversity.
• Urgent priorities identified for chickpea conserved in many genebanks are the need for regeneration, characterization, safety duplication and long-term conservation.
• Cicer genetic resources could be much better utilized if a representative core collection (10% of the entire collection) and a mini core collection (10% of the core or 1% of entire collection) are developed and evaluated extensively for useful traits.
• More data is available at the accession level for economic traits to identify trait-specific germplasm.
• Many small collections contain unique materials that can be extremely important for the genetic improvement of the crop.

Chickpea seed diversity (photo: ICRISAT)	Chickpea diversity of pods (photo: ICRISAT)
Chickpea germplasm diversity in the field (photo: ICRISAT)	A chickpea genebank (photo: ICRISAT)

Major chickpea collections

Chickpea collections include landraces, breeding material and wild species. It is estimated that more than 80 000 accessions are conserved in more than 30 genebanks worldwide. The genebank at ICRISAT Patancheru, India, is one of the largest genebanks holding >20 000 accessions of chickpea from 60 countries. Other major collections (more than 5000 accessions) are held at NBPGR, New Delhi, India; ICARDA Aleppo, Syria; Australian temperate field crops collection, Victoria, Australia; USDA, USA and the Seed and Plant Improvement Institute, Iran.

It was only after 1970 that wild Cicer species were actively collected. Currently there is a reasonable number of wild annual species but still limited availability of perennial species. Less than 1% of the Cicer accessions (conserved in about tengenebanks worldwide) represent wild species. The ICRISAT genebank holds 308 accessions belonging to 18 species of wild Cicer from 14 countries.

References and further reading

Abbo S, Redden RJ, Yadav SS. 2007. Utilization of wild relatives. In: Yadav SS, Redden R, Chen W, Sharma B, editors. Chickpea Breeding & Management. CABI, Wallingford, UK. pp 338-354.

Chickpea [online]. Available from URL: http://www.icrisat.org/crop-chickpea.htm. Date accessed: 29 January 2010.

FAO/IPGRI. 1994. Genebank standards. Food and Agriculture Organization of the United Nations, Rome and International Plant Genetic Resources Institute, Rome. Available in English, Spanish, French and Arabic.

Global Crop Diversity Trust. 2008. Global strategy for the ex situ conservation of chickpea (Cicer L.) [online]. Available from: http://www.croptrust.org/documents/web/CicerStrategy_FINAL_2Dec08.pdf. Date accessed: 25 May 2009.

IBPGR, ICRISAT, ICARDA. 1993. Descriptors for Chickpea (Cicer arietinum L.). International Board for Plant Genetic Resources, Rome, Italy; International Crops Research Institute for the Semi-Arid Tropics, Patancheru, India and International Center for Agricultural Research in the Dry Areas, Aleppo, Syria. ISBN 92-9043-137-7. Available here.

ICRISAT Chickpea Germplasm Collection Reports. Genetic Resources Unit, International Crops Research Institute for the Semi-Arid Tropics, Patancheru, A.P. 502 324, India. GRU - Progress Reports – 20 no’s.

ICRISAT Chickpea collection. SINGER [online]. Available from http://www.singer.cgiar.org/index.jsp?page=showkeycount&search=coc=co38. Date accessed: 30 January 2010.

Hanson J. 1985. Procedures for handling seeds in genebanks. Practical manuals for genebanks no. 1. International Board for Plant Genetic Resources. http://www.bioversityinternational.org/index.php?id=19&user_bioversitypublications_pi1[showUid]=2235.

Jambunathan R, Singh U, Subramanian V. 1981. Grain quality of sorghum, pearl millet, pigeonpea and chickpea. In: Proceedings of a workshop held at International Crops Research Institute for the Semi-Arid Tropics, Patancheru-502 324, Andhra Pradesh, India.

Singh KB, Pundir RPS, Robertson LD, van Rheene HA, Singh U, Kelley TJ, Rao PP, Johansen C, Saxena NP. 1997. Chickpea. In: Fuccillo D, Sears L, Stapleton P, editors. Biodiversity in Trust. Cambridge University Press. pp. 100-113.

Rao NK, Hanson J, Dulloo ME, Ghosh K, Nowel D, Larinde M. 2006. Manual of seed handling in genebanks. Handbooks for Genebanks No. 8. Bioversity International, Rome, Italy. Available in English (1.5 MB),  Spanish (1.4 MB) and French (1.9 MB).

Redden B, Furman BJ, Upadhyaya HD, Pundir RPS, Gowda CLL, Coyne C, Enneking D. 2007. Biodiversity Management in Chickpea. In: Yadav SS, Redden R, Chen W, Sharma B, editors. Chickpea Breeding & Management. CABI, Wallingford, UK. pp 355-368.

Upadhyaya HD, Gowda CLL, Sastry DVSSR. 2008. Management of germplasm collections and enhancing their use by mini core and molecular approaches. APEC-ATCWG Workshop. Capacity building for risk management systems on genetic resources. pp. 35-70. http://www.tari.gov.tw/taric/uploads/publication_no135_08.pdf.

Upadhyaya HD, Gowda CLL, Sastry DVSSR. 2008. Plant genetic resources management: collection, characterization, conservation and utilization. Journal of SAT Agricultural Research 6. http://www.icrisat.org/Journal/Volume6/ChickPea_PigeonPea/HD_Upadhyaya.pdf.

Upadhyaya HD, Laxmipathi Gowda CL. 2009. Managing and Enhancing the Use of Germplasm – Strategies and Methodologies. Technical Manual no. 10. Patancheru 502 324, Andhra Pradesh, India: International Crops Research Institute for the Semi-Arid Tropics. 236 pp.

Registration of cultivated chickpea and wild relatives genetic resources

Contributors to this page: ICRISAT, Patancheru, India (Hari D Upadhyaya, Shivali Sharma, Cholenahalli L Laxmipathi Gowda, Dintyala Sastry, Sube Singh); NBPGR, New Delhi, India (Shyam Sharma); ICARDA, Aleppo, Syria (Ahmed Amri, Kenneth Street, Natalya Rukhkyan), SARC-RIPP, Piestany, Slovak Republic (Gabriela Antalikova); Institute of Plant Genetic Resources ‘K.Malkov’, Sadovo, Bulgaria (Siyka Stoyanova); Department of Primary Industries, Victoria, Australia (Bob Redden); IPK, Gatersleben, Germany (Andreas Börner).

Rationale for registration

Registration is done to allow the genebank curator to keep a record of samples held in the genebank and to produce inventory lists for conservation, distribution and other aspects of germplasm management.

• Each sample should have assigned a unique accession number which distinguishes it from all other accessions in the genebank.
• Samples should be registered when first entering the genebank.

Verifying accompanying documentation

Basic documentation is essential to track germplasm material. Health and IPR certificates are very important for entry to the country. The following documents should accompany each consignment of plant germplasm and should be recorded (using a Genebank Information Management System, GIMS):

• Standard Material Transfer Agreement (SMTA) and /or Germplasm Acquisition Agreement (GAA), whose status with regard to conservation and use is clearly defined.
• Phytosanitary certificate from donor.
• Plant import permit.
• GMO-free certificate.
• List of material.
• Seed source.
• Associated passport data, preferably in the Multi-crop Passport Descriptors (MCPD) format, preferably electronically.

Verifying the consignment

Checking the content

• Arrange the material received by the genebank according to alphabetical order of names of variety, or numeric order of number, depending on the identification provided on the packets/bags.
• Check all the packets as per the list provided with the samples.
• Prepare a new list if details are not provided with the samples.
• Request any missing passport information.

Checking the genebank

• Check the passport database file or collector's number and other information to ensure that the sample does not already exist in the genebank.
• If the material is already represented in the genebank, grow the putative duplicate alongside the existing accession to compare the morpho-agronomic characters.
  • If you are convinced that they are duplicates, they can be stored together and maintained with the existing number.

Checking the germplasm

• Verify the health certificates (in the regional Quarantine service, according to Quarantine prescriptions).
• File the official quarantine release papers.
• Open each packet of seeds and inspect their condition by checking for any insect damage, fungal growth, damaged, broken, empty or shriveled seeds which probably would not germinate.
• Visually inspect the seeds to be correctly designated – as desi or kabuli seed, seed colour, unmixed pure sample if a released variety, or possibly mixed if it is a landrace.
• Compare the received seed with the request / import order with identification provided on the imported seed packet/container for veracity, otherwise send to donor/provider for confirmation and to obtain the correct data.
• Check with the photographic record of the accession, if available.
• If seeds are in poor condition, discard the sample and make a note in the files of the action taken and reasons for it. Write to the donors requesting fresh samples.

Assigning numbers

• If the accompanying data is incomplete or seeds are insufficient or of poor quality, assign a temporary number.
• Assign an accession number only if the sample received is unique and sufficient seeds are available.
• If two samples have identical or very similar names, identical seed characteristics, maturity and other morphological and agronomical features, bulk them to make one sample and assign an accession number.

Recording information during registration

The following information must be recorded for each consignment:

• Passport information – the minimum passport data required for collected/donated samples like country of origin, location of collecting site, local name or cultivar name, pedigree information for breeding lines and improved varieties and source of collection (use descriptors for passport data).
• Seed health – seeds should be absolutely free from pathogens and insects (see seed health page for more details).
• Seed quality and quantity – quantity sufficient for at least for four regenerations. Seed viability should not be <85% for cultivated species and <75% for wild species (record the seed status regarding quality and quantity).
• If the seed meets the above standards and does not exist in the genebank, assign accession number following a standard procedure.
• Steps for registration are described in detail in the Genebank Manual of ICRISAT (Upadhyaya and Laxmipathi, 2009).
• Identification of duplicates: if it appears to be a duplicate, assign a new seed lot number under the original accession number (of suspected duplicate). The ‘duplication’ should be verified later (at first growth out and/or during characterization).
• Otherwise, if sample appears ‘original’, assign a new unique accession number (entering data in genebank database).
• Record all other information available on the sample (this will be a permanent record of the source material).
• Enter the genebank number (a unique accession number).
• Regularly do electronic database maintenance.

References and further reading

GIMS (Genebank Information Management System) – a standalone facility of ICRISAT Genebank. International Crops Research Institute for the Semi-Arid Tropics, Patancheru, A.P. 502 324, India. In: Manual of Genebank Operations and Procedures - Technical Manual no. 6. International Crops Research Institute for the Semi-Arid Tropics, Patancheru 502 324, Andhra Pradesh, India: ISBN 92-9066-421-5.

IBPGR, ICRISAT, ICARDA. 1993. Descriptors for Chickpea (Cicer arietinum L.). International Board for Plant Genetic Resources, Rome, Italy; International Crops Research Institute for the Semi-Arid Tropics, Patancheru, India and International Center for Agricultural Research in the Dry Areas, Aleppo, Syria. ISBN 92-9043-137-7. Available here.

FAO/IPGRI. 2001. Multi-Crop Passport Descriptors. FAO and IPGRI, Rome, Italy. Available in English, French and Spanish.

Upadhyaya HD, Laxmipathi Gowda CL. 2009. Managing and Enhancing the Use of Germplasm – Strategies and Methodologies. Technical Manual no. 10. Patancheru 502 324, Andhra Pradesh, India: International Crops Research Institute for the Semi-Arid Tropics. 236 pp.

Sample processing of cultivated chickpea and wild relatives genetic resources

Contributors to this page: ICRISAT, Patancheru, India (Hari D Upadhyaya, Shivali Sharma, Cholenahalli L Laxmipathi Gowda, Dintyala Sastry, Sube Singh); NBPGR, New Delhi, India (Shyam Sharma); ICARDA, Aleppo, Syria (Ahmed Amri, Kenneth Street, Natalya Rukhkyan), SARC-RIPP, Piestany, Slovak Republic (Gabriela Antalikova); Institute of Plant Genetic Resources ‘K.Malkov’, Sadovo, Bulgaria (Siyka Stoyanova); Department of Primary Industries, Victoria, Australia (Bob Redden); IPK, Gatersleben, Germany (Andreas Börner).

 
Seed Cleaning

Cleaning is the removal of physical contamination from the plant material after harvest, before it can be stored. The ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009) recommends:

• Hand threshing and manual cleaning as one seed lot for cultivated type and for wild Cicer species.
• Periodic harvesting, threshing and manual cleaning of seeds.

Visual inspection of seeds

This is a quality control that must be done after harvesting the seeds. ICRISAT's Genebank Manual Procedures (Upadhyaya and Laxmipathi, 2009) recommends:

• Spread seeds on a flat surface and examine for damage/infestation.
• Remove damaged and infected seeds and store healthy seeds.
• Use a light wind blower or remove manually shriveled and damaged seeds.

Special treatments

• No chemical seed treatment is recommended (chemicals induce mutations over long time).
• Fumigation can be carried out, however, to control insects.

Disposal of contaminated material

The Germplasm Exchange and Quarantine of ICRISAT Mandate Crops recommends the following procedures:

• Autoclave and incinerate contaminated material.
• Contaminants should be placed in secure containers and buried deep (to assure that pests and disease risks associated with contaminated seeds are neutralized).

Inspection and certification

• Visually observe the samples and compare them with original seed and database information.
• Perform the purity analysis (following the ISTA Standards).

Seed drying

Methods

Most genebanks follow FAO/IPGRI Standards as well as the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009):

Controlled environment facility for germplasm seed drying at ICRISAT (photo: ICRISAT)

• Dehumidified drying (temperature 15-20ºC and RH 15%) and silica gel drying.

IPK (Gatersleben, Germany) uses procedures based on requirements of the IPK quality management system (DIN EN ISO 9001:2000):

• Computer controlled drying chamber.

Drying time

According to procedures of the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009):

• Depends on size of samples, initial moisture content and method of drying – usually one to four weeks.

Moisture content before drying

According to procedures of FAO/IPGRI Standards, as well as the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009):

• About 8-12% (based on practical experience).

Moisture content for storage

• 8-10% for active collection and 5-7% for base collections.

Critical moisture content

The equilibrium relationship between seed moisture content and RH for chickpea storage is described in the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009).

• It can vary between 3 and 7%.

Recording information during seed cleaning and drying

The following information should be recorded for each processing step:

• Accession identifier; National identity; Alternative identity (unique identification of the accession).
• Reference to seed source (to trace the origin of the sample).
• Seed moisture at harvest (% of moisture content of samples collected during harvesting.
• Threshing method employed.
• Drying method employed.
• Plot yield (g) (harvested seed yield).
• Seed samples moisture % after drying.
• Purity (%).
• Seed quantity (g) (cleaned seed yield).
• 100-seed weight (g) (weight of 100 randomly selected seeds).
• Germination % (seed germination from a standard test).
• Date of storage (date, month, year as dd/mm/yyyy).
• *Photographic record of seeds for each accession (100 seeds in petri dish with ID label).
• Remarks (any other information).

* This could be restricted to being done once, during characterization and does not need to be repeated at every regeneration cycle.

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Determination of seed moisture content

Methods

According to the ISTA Standards:

• Use a high, constant temperature oven-drying.

Sampling frequency

This is important to cover the diversity in the accessions especially the seed traits.

• 5% of total number of seed samples/accessions.
• Or a minimum of ten accessions.

Sample size

According to the ISTA Standards:

• Use two replicates of 1.5-2.0 g of seeds.

Grinding

According to the ISTA Standards:

• It is recommended to coarse grind the samples.

Oven drying temperature

According to the ISTA Standards:

• 130±2ºC for one hour.

Others

According to the ISTA Standards:

• Use two replicates per sample.

Recording information during determination of seed moisture content

The following information should be recorded for each step:

• Accession number (unique identification of the accession).
• Replication number.
• W1 [weight of empty dish (g)].
• W2 [weight of dish + fresh sample (g)].
• W3 [weight of dish + dried sample (g)].
• Drying temperature.
• Time and date.
• (W2-W3)/(W2-W1) x100 [% of moisture content (on wet basis) of the replication].
• Mean moisture content (%) of the replications.
• Additionally, data field names are mentioned in Handling Seed in Genebanks (Hanson 1985) and Manual of Seed Handling in Genebanks (Rao et al., 2006).

References and further reading

Chakrabarty SK, Anitha K, Girish AG, Sarath BB, Prasada RRDVJ, Varaprasad KS, Khetarpal RK, Thakur RP. 2005. Germplasm Exchange and Quarantine of ICRISAT Mandate Crops. Information Bulletin No. 69. Rajendranagar 500 030, Andhra Pradesh, India: National Bureau of Plant Genetic Resources; and Patancheru 502 324, Andhra Pradesh, India: International Crops Research Institute for the Semi-Arid Tropics. 80 pp. ISBN 92-9066-481-9.

FAO/IPGRI. 1994. Genebank standards. Food and Agriculture Organization of the United Nations, Rome and International Plant Genetic Resources Institute, Rome. Available in English, Spanish, French and Arabic.

Hanson J. 1985. Procedures for handling seeds in genebanks. Practical manuals for genebanks no. 1. International Board for Plant Genetic Resources, Rome. http://www.bioversityinternational.org/index.php?id=19&user_bioversitypublications_pi1[showUid]=2235.

ISTA. 1993. International rules for seed testing. International Seed Testing Association. Bassersdorf, Switzerland. Seed Science and Technology 21 (Supplement):1-288.

ISTA. 2003. International rules for seed testing. International Seed Testing Association. Bassersdorf, Switzerland.

ISTA. 2005. International rules for seed testing. International Seed Testing Association. Bassersdorf, Switzerland.

Rao NK, Hanson J, Dulloo ME, Ghosh K, Nowel D, Larinde M. 2006. Manual of seed handling in genebanks. Handbooks for Genebanks No. 8. Bioversity International, Rome, Italy. Available in English (1.5 MB), Spanish (1.4 MB) and French (1.9 MB).

Stoyanova SD. 1992. Drying chickpea seeds for long-term preservation in genebanks. PGR-Newsletter 90:19-27.

Upadhyaya HD, Laxmipathi Gowda CL. 2009. Managing and Enhancing the Use of Germplasm – Strategies and Methodologies. Technical Manual no. 10. Patancheru 502 324, Andhra Pradesh, India: International Crops Research Institute for the Semi-Arid Tropics. 236 pp.

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Viability of cultivated chickpea and wild relatives genetic resources

Contributors to this page: ICRISAT, Patancheru, India (Hari D Upadhyaya, Shivali Sharma, Cholenahalli L Laxmipathi Gowda, Dintyala Sastry, Sube Singh); NBPGR, New Delhi, India (Shyam Sharma); ICARDA, Aleppo, Syria (Ahmed Amri, Kenneth Street, Natalya Rukhkyan), SARC-RIPP, Piestany, Slovak Republic (Gabriela Antalikova); Institute of Plant Genetic Resources ‘K.Malkov’, Sadovo, Bulgaria (Siyka Stoyanova); Department of Primary Industries, Victoria, Australia (Bob Redden); IPK, Gatersleben, Germany (Andreas Börner).

Viability testing

Laboratory methods

Describes the various recommended options, to test the viability and quality of chickpea seeds. The percentage of germination of the stored seeds of the accessions determines when regeneration of the accessions should take place and if the accession can be distributed to the users.

Type of test

When sufficient seed is available:

• Use a standard viability test (between paper, BP), following the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009).

When the seed sample is small:

• Use a sequential viability test (following the FAO/IPGRI standards, 1994 and ISTA standards).

Number of seeds and replicates

• Use 50-100 seeds in 2-4 replications (following the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009); FAO/IPGRI standards, 1994; Normative ISTA and AOSA; ‘Methodics examinations seed and plant, of State variety trials’. Bratislava. 1999. Slovak Republic).

Or

• 10 seeds in 2-4 replications (following the FAO/IPGRI standards, 1994).

Pre-treatment

Follow the recommendations of the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009):

• Apply a dry seed treatment with fungicide at 3 g/kg seed.
• Scarify the seed coat for hard seeds, especially wild Cicer species.

To avoid seed damage resulting from rapid moistening by liquid water in germination substrate, and to improve germination:

• Re-humidify (90% RH) seeds stored in dry and cool environments (-18^oC).
• Pre-chill the re-humidified seeds (10^oC for 48 hours) followed by germination according to ISTA standards.

Media

Following the recommendations of the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009):

• Standard paper towel is best and cost effective.
• Alternatively, germination boxes with artificial soil (turf, vermiculate, etc.) using tap water may be used.

Temperature

According to the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009) and the ISTA Standards:

• 20-25ºC.

Light

According to the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009):

• Not required.

Duration of test

According to the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009) and the ISTA Standards:

• 5-10 days after incubation.

Interpretation of results

According to the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009) and the ISTA Standards (2003, 2005), Association of Official Seed Analysts (AOSA) Standards (2005):

• Scarify hard and ungerminated seeds and evaluate 14 days after incubation.
• If the mean viability percentage of the accession is below 85% (75% for wild species), repeat the test.
• Calculate the mean viability percentage from the results of the two tests and use this as the overall test result.
• If the results are far below the germination standard, the accession should be sent to regeneration.

Monitoring intervals

According to the ICRISAT Genebank Manual (Upadhyaya and Laxmipathi, 2009):

For the active collection (4ºC and 20-30% RH)

• Germination less than 85% - 5 years.
• Germination 85-95% - 8 years.
• Germination 95% or more - 10 years.

For the base collection (-20ºC)

• Germination less than 85% - 10 years.
• Germination 85-95% - 15 years.
• Germination 95% and more - 20 years.

Other option

According to Ellis' and Robert's predictive model with log parameters for initial viability, seed storage temperature and moisture content were used to monitor viability. This was more efficient than routine calendar interval monitoring of viability of collections such as every 5–10 years:

• As per predicted longevity: 20-30 years after initial regeneration and viability testing.

Recording information during viability testing

The following information must be recorded for each testing step:

• Accession number (unique identification of the accession).
• Taxonomic identification (name).
• Date of storage (date when sample was stored in cold rooms).
• Date of testing (date when sampled was tested for viability).
• Substrate (type of paper used as substratum).
• Temperature [incubation temperature (ºC)].
• Incubation time [incubation period (days)].
• Replications / No. of seeds (number of replications and number of seeds tested).
• Date (date of testing).
• Pre-treatment.
• Days (days from the test date).
• Total germinated [number of seeds germinated in each category (normal, abnormal, non-germinated, hard seeds)].
• Abnormal (number of abnormal seedlings).
• Hard/dormant (number of hard/dormant seeds).
• Dead (number of dead seeds).
• Germination % (counted on normal seedlings only).
  • Seedlings should be judged as normal according to specific criteria - use ISTA Standards (2003, 2005), Association of Official Seed Analysts (AOSA) Standards, (2005).

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Routine monitoring

Describes the recommended monitoring methods to assure minimum viability and quantity of seeds in storage.

• Routine inventory management of active collection and base collection using a Genebank Information Management System (GIMS).
• Aim viability of germplasm seed stock at 85% or more, based upon seed age and predicted viability from storage trial (to keep viable seed of all accessions available for distribution, and that seed stock is sufficient for both regeneration and distribution requirements).
• Monitor when seed quantity in active storage falls below 100 g.
• Monitoring – five years in the active collection, ten years in the base collection (recommended in the FAO/IPGRI standards, 1994).
• Use the computerized stock control system to monitor the quality and quantity of seed accessions in the cold store.

Monitoring frequency

Recommends the minimum quantity and minimum viability of seeds below which they need to be regenerated.

Critical quantity

To be detected by data on stocks in the Genebank Information Management System (GIMS) (according to FAO/IPGRI standards, 1994; ISTA Standards).

• For the active collection: critical quantity less than 100 g.
• For the base collection: When the number of seeds falls below the number required for at least three cycles of regeneration.
• For wild Cicer: 300 seeds.

Critical germination level

To be detected by data on stocks in the Genebank Information Management System (GIMS) (according to FAO/IPGRI standards, 1994; ISTA Standards).

• For the active collection: critical germination level less than 75%.
• For the base collection: when viability falls below 85% of the initial value.
• For wild Cicer: critical germination level: less than 75%.

Recording information during storage monitoring

The following information should be recorded for each step:

• Accession identifier/number (unique identifier of the accession).
• Season of crop harvest (mm/yyyy).
• Site of regeneration (place where the samples were regenerated).
• Container (type of container used for storage).
• No. of containers (total number of containers used for storing the sample).
• Date of storage [date on which the sample was stored in the genebank (dd/mm/yyyy)].
• Location in genebank (exact location where the sample is placed in the genebank).
• Seed quantity (g) (quantity of seeds currently available in storage).
• Germination (%) (percentage of seed germination resulting from the recent germination test conducted).
• Date of germination testing [date on which seeds were tested for germination (dd/mm/yyyy)].
• Remarks (any significant observation).
• Active or base storage.
• Age of seed [track age of seed from last regeneration date, or original if seed is from outside sources mostly with unknown storage conditions. This is an indirect guide to viability risk (aim applying predictive model].
• Passport information (follow the Multi-Crop Passport Descriptors, MCPD - Mandatory fields for passport information).
• Seed moisture content (% wet basis).
• Date when accession was sent for regeneration (date, person) [date accessions were sent for regeneration in the field (dd/mm/yyyy)].
• Date of receiving newly regenerated seeds of the accession (date, seed germination, others). According to accepted protocols for seed handling in genebanks.
• Number of regenerations (frequent regeneration can affect germplasm identity).
• Date of next test (one year: if germination % is close to critical value; five years: when it is above 85%).

References and further reading

AOSA (Association of Official Seed Analysts). 2005. In: Rules for Testing Seeds (Capashew Ed.), 4-0, 4-11. Las Cruces, NM. pp.113.

Bratislava. 1999. Methodics examinations seed and plant, of State variety trials. Slovak Republic.

GIMS (Genebank Information Management System) – a standalone facility of ICRISAT Genebank. International Crops Research Institute for the Semi-Arid Tropics, Patancheru, A.P. 502 324, India. In: Manual of Genebank Operations and Procedures - Technical Manual no. 6. International Crops Research Institute for the Semi-Arid Tropics, Patancheru 502 324, Andhra Pradesh, India: ISBN 92-9066-421-5.

FAO/IPGRI. 1994. Genebank standards. Food and Agriculture Organization of the United Nations, Rome and International Plant Genetic Resources Institute, Rome. Available in English, Spanish, French and Arabic.

FAO/IPGRI. 2001. Multi-Crop Passport Descriptors. FAO and IPGRI, Rome, Italy. Available in English, French and Spanish.

ISTA. 1993. International rules for seed testing. International Seed Testing Association. Bassersdorf, Switzerland. Seed Science and Technology 21 (Supplement):1-288.

ISTA. 2003. International rules for seed testing. International Seed Testing Association. Bassersdorf, Switzerland.

ISTA. 2005. International rules for seed testing. International Seed Testing Association. Bassersdorf, Switzerland.

Rao NK, Hanson J, Dulloo ME, Ghosh K, Nowel D, Larinde M. 2006. Manual of seed handling in genebanks. Handbooks for Genebanks No. 8. Bioversity International, Rome, Italy. Available in English (1.5 MB), Spanish (1.4 MB) and French (1.9 MB).

Upadhyaya HD, Laxmipathi Gowda CL. 2009. Managing and Enhancing the Use of Germplasm – Strategies and Methodologies. Technical Manual no. 10. Patancheru 502 324, Andhra Pradesh, India: International Crops Research Institute for the Semi-Arid Tropics. 236 pp.

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