Seed Processing

SEED PROCESSING

Accession: A distinct, uniquely identifiable sample of seeds representing a cultivar, breeding line or a population, which is maintained in storage for conservation and use. Accession number: A unique identifier that is assigned by the curator when an accession is entered into a collection. This number should never be assigned to another accession. Achene: A one-seeded dry indehiscent fruit with the seed attached to the pericarp at only one point. Active collection: A germplasm accession that is used for regeneration, multiplication, distribution, characterization and evaluation. Active collections are maintained in short- to mediumterm storage and usually duplicated in a base collection maintained in medium- to long-term storage. Bar-coding:  ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) A computerized coding system that uses a printed pattern or bars on labels to identify germplasm accessions. Barcodes are read by optically scanning the printed pattern and using a computer program to decode the pattern. Base collection: A collection of germplasm that is conserved in long-term, secure storage and is not used as a routine distribution source. Seed is usually stored at sub-zero temperatures and low moisture content. Base unit: The number of seeds needed to ensure the successful implementation of a procedure such as registration or regeneration. Breeding line: A group of identical pure-breeding diploid or polyploid organisms that are distinguished from other individuals of the same species by a unique phenotype and genotype. Capsule: A dry dehiscent fruit derived from an ovary with two or more carpels that splits partly open at maturity. Characterization: The recording of highly heritable characters that can be easily seen and are expressed in all environments. Collection: A group of germplasm accessions maintained for a specific purpose under defined conditions. 124 Handbooks for Genebanks No. 8 Cultivar: A crop variety produced by scientific breeding or farmerselection methods. Database: An organized set of interrelated data assembled for a specific purpose and held in one or more storage media. Database management  ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) system: A piece of software that controls the organization, storage, retrieval, security and integrity of data in a database—it accepts requests from the application and instructs the operating system to transfer the appropriate data. The major vendors are Oracle, IBM, Microsoft and Sybase. MySQL is a very popular open-source product. Dehiscent fruits: Fruits that open at maturity to shed their seeds (see follicle, capsule). Descriptor: An identifiable and measurable trait, characteristic or attribute observed in an accession that is used to facilitate data classification, storage, retrieval and use. Descriptor list: A collection of all individual descriptors of a particular crop or species. Desiccator: A short glass jar fitted with an air-tight cover and containing a desiccating agent such as silica gel or  ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )calcium chloride, above which the material to be dried is supported on a perforated platform. Distribution: The process of supplying samples of germplasm accessions to breeders and other users. Documentation: The organized collection of records that describe structure, purpose, operation, maintenance, and data requirements. Donor: An institution or individual responsible for donating germplasm. Dormancy: The state in which certain live seeds do not germinate, even under normally suitable conditions. Exploration: The act of searching for genetic diversity in the field. Equilibrium moisture content: The moisture content at which a seed is in equilibrium with the relative humidity of the surrounding air. Evaluation: The recording of those characteristics whose expression is often influenced by environmental factors. Ex situ conservation: The conservation of biological diversity outside its natural habitat—in the case of plant genetic resources, 125 Manual of Seed Handling this may be in seed genebanks, in vitro genebanks or as live collections in field genebanks. Field  collection: A collection of germplasm maintained as living plants—germplasm that would otherwise be difficult to maintain as seed is commonly maintained in field collections. Follicle: A dry, single-celled, many-seeded fruit consisting of a single carpel, dehiscing by the ventral suture. Funicle: A stalk by which an ovule or seed attaches itself to the fruit wall. Genebank: A centre for conserving genetic resources under suitable conditions to prolong their lives. Genetic diversity: The variety of genetic traits that result in differing characteristics. Genetic drift: Changes in the genetic composition of a population when the number of individuals is reduced below the frequency of certain alleles within it. Genetically modified organism (GMO): An organism whose genetic material has been  ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) deliberately altered (see also transgenic plant). Genotype: The genetic constitution of an individual plant or organism. Germplasm: The genetic material that forms the physical basis of heredity and that is transmitted from one generation to the next by germ cells. Germination: The biological process that leads to the development of a seedling from a seed. Radicle emergence is the first visible sign of germination, but may be followed by no further growth or by abnormal development. According to ISTA rules, only seedlings showing normal morphology are considered to have germinated. Germination test: A procedure to determine the percentage of seeds that are capable of germinating under a given set of conditions. Hard seeds: Seeds that fail to imbibe and germinate when placed in a moist medium because they are impermeable to water. Humidification: The process in which the moisture content of very dry seeds is raised by placing them in a humid environment; humidification helps to prevent damage to seeds from rapid uptake of water. Imbibition damage: Damage caused by rapid uptake of water in very dry seeds (see also humidification). 126 Handbooks for Genebanks No. 8 Indehiscent fruit: Fruit that does not open at maturity (see also achene). In vitro collection: A germplasm collection maintained as plant tissue ranging from protoplast and cell suspensions to callus cultures, meristems and embryos. Inventory: A list of samples (and their characteristics) that is stored in a genebank. Isotherm: A graph showing the relationship between seed moisture content and percentage relative humidity (see also sorption isotherms). Laminated aluminium foil packets: Packets constructed of a laminate consisting of an  ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) inner layer of polyethylene, a middle layer of aluminium foil and an outer layer of polyester. Landrace: A crop cultivar that has evolved through many years of farmer-directed selection and that is specifically adapted to local conditions; landraces are usually genetically heterogeneous. Long-term conservation: The storage of germplasm for a long period, such as in base collections and duplicate collections. The period of storage before seeds need to be regenerated varies, but is at least several decades and possibly a century or more. Long-term conservation takes place at sub-zero temperatures. Mass maturity: The stage in development at which seeds attain maximum dry weight. Medium-term conservation: The storage of germplasm in the medium-term such as in active and  ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )working collections; it is generally assumed that little loss of viability will occur for approximately ten years. Medium-term conservation takes place at temperatures between 0°C and 10°C. Micropylar region: The point on a seed that was the orifice (pore) of the ovule. Moisture content (wet-weight basis): The weight of free moisture divided by the weight of water plus dry matter, expressed as a percentage. Monitoring: The periodic checking of accessions for viability and quantity. Monitoring interval: The period of storage between monitoring tests. Most original sample (MOS): A sample of seeds that have undergone the lowest number of regenerations since the material was acquired by the genebank, as recommended for storage as a 127 Manual of Seed Handling base collection. It may be a sub-sample of the original seed lot or a seed sample from the first regeneration cycle if the original seed lot required regeneration before storage. Multiplication: The representative sample of an accession grown to multiply the quantity of conserved material for distribution. 

Seed Processing

Genetically modified organisms (GMOs) Several countries have established regulatory frameworks for importing and handling GMOs. These are largely based on the Cartagena Protocol on Biosafety (CPB) which came into force in September 2003 (see www.biodiv.org/biosafety/default.aspx). National biosafety authorities, in collaboration with phytosanitary authorities, are responsible for issuing import permits, conducting risk assessments and enforcing biosafety guidelines. Researchers wishing to import GMOs must submit an application providing: details of the genetic material to be introduced; accompanying information on research and testing of the GMOs in question; and a plan for safety measures to be followed during ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

introduction. Approval is issued if the national authority determines that the GMO or GMO product poses no risk to the environment, biological diversity or human health. Prior to export of GMOs, authorization from the exporting country’s biosafety authority is also required. No authorization for export will be given if the exporting country bans the GMO or GMO product. 121 Manual of Seed Handling ANNEX II Serological methods for detecting plant pathogens General procedure using antigen-coated plate (ACP)-ELISA 1. Harvest fresh samples of leaf from samples and controls and weigh 0.2 g of each. Grind each sample in 2 ml coating buffer (0.05 M carbonate buffer)+2% w/v polyvinyl pyridine and 0.2% w/v Na2SO3. Transfer the sap to a labelled Eppendorf tube. Spin for five ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

minutes at 10 000 rpm. 2. Label the microtitre plate and load the samples, 100 µl per well. Cover the plate with parafilm and incubate at 4ºC overnight. Weigh 4 g of the healthy sample and grind in 10 ml phosphate-buffered saline solution with Tween (PBST), which acts as a blocking solution. It is made by dissolving 80.0 g NaCl, 2.0 g KH2PO4, 11.0 g Na2HPO4 and 2.0 g KCl in 2000 ml distilled water, adjusting the pH to 7.4 and adding 5.0 ml Tween 20 to make up to 10 l. Weigh the samples and dissolve. Filter through cotton wool and make the filtrate up to 80 ml. Use this to make dilutions of the antiserum. Allow to absorb at 4°C overnight. 3. Make a 0.1% solution of bovine serum albumen in PBST. 4. Rinse the plate in a gentle stream of tap water and wash three times with PBST. 5. Add PBST to all wells of the plate, 150 µl per well. Cover the plate with parafilm and incubate for 30 minutes at room temperature. 6. Completely empty the blocking solution from the plate. Without washing, add the diluted antiserum; use 100 µl per well. Cover the plate with parafilm and incubate three to four hours at room temperature. 7. Make the dilution of the alkaline phosphatase enzyme conjugate in PBST (1/1000 made fresh). 8. Wash the plate three times with PBST, then add the diluted conjugate according to the pattern on the loading diagram, starting with the most diluted. ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

Use 100 µl per well. Cover the plate with parafilm and incubate at 4°C overnight. 9. Wash the plate with PBST three times. 10. Dissolve the substrate tablet (p-NPP, Sigma) in substrate buffer (1mg/ml) (diethanolamine 10%, pH 9.8, stored in a refrigerator). 11. Add the dissolved substrate to all wells, 150 µl per well. Incubate at room temperature for 30 minutes. Evaluate and score the colour intensity of each well with the ELISA reader. 122 Handbooks for Genebanks No. 8 General procedure using tissue-blot immunoassay (TBIA) on nitrocellulose membranes 1. Collect tissues (leaves, ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

petioles, stems, etc.). 2. For thin tissues such as leaves, roll them into a tight core. For batch samples, bind them together with parafilm. 3. Cut a piece of nitrocellulose membrane (NCM), cut the top left corner and mark a grid with in pen. 4. Hold tissues in one hand and cut with a new razor blade in a steady motion with the other hand to obtain a single-cut surface. 5. Press the newly cut surface onto on grid square of the NCM with a firm but gentle force. Document what material has been blotted into which grid on a prepared form. Continue to blot leaves until all grids are full. 6. Wash the NCM three times with PBST at five-minute intervals. 7. Dilute the antiserum in healthy sap in PBST (dilution 1/500– 1/2000) and allow to absorb at room temperature for two hours or 4ºC overnight. 8. Block the NCM in 2 µg/ml polyvinyl alcohol in PBST and incubate one minute at room temperature. 9. Wash as in step 6. 10. Add diluted antiserum and incubate for one hour at room temperature. 11. Wash as in step 6. 12. Add anti-rabbit conjugate (dilution 1/1000–1/5000 in conjugate buffer) and incubate for one hour at room temperature. 13. Wash as in step 6. 14. Add substrate solution ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

 (NBT/BCIP). 15. To stop the reaction, wash with de-ionized water. 123 Manual of Seed Handling ANNEX III Glossary of terms Absolute humidity: The amount of water vapour present in a unit volume of air, usually expressed in kilograms per cubic meter. 

Seed Processing

International Code of Conduct for Plant Germplasm Collecting and Transfer The International Code, adopted by the FAO Conference in 1993, provides a general framework for collecting and transfer of germplasm. It sets out the minimum responsibilities of collectors and curators regarding germplasm collecting and transfer. Although a voluntary instrument, the code is compatible with the CBD and serves as a reference for countries to establish their own regulations for germplasm collection and exchange (www.fao.org/ag/agp/agps/ pgr/icc/icce.htm). Germplasm Acquisition Agreements Article 15 of CBD stipulates that access to ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

genetic resources shall be on mutually agreed terms and subject to prior informed consent. Prior informed consent means that the providing country can grant or refuse access to germplasm following a request from the applicant. Access is on mutually agreed terms when both the supplier and recipient agree. This usually implies a contractual arrangement executed on a bilateral basis, which often takes the form of germplasm acquisition agreement (GAA) setting out the terms on which the genetic material is acquired and transferred. 119 Manual of Seed Handling International Treaty and Multilateral System for Access and Benefit Sharing In 2001, the FAO Conference adopted the International Treaty on PGRFA, recognizing that: (i) agriculture in all countries depends largely on PGRFA that originated elsewhere; (ii) future advances in crop improvement require continued access ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

to a wide genetic base without major restrictions; and (iii) a purely bilateral approach to access and benefit-sharing is not well suited to genetic resources of major food crops. The Treaty creates a Multilateral System for Access and Benefit Sharing covering 64 major crops and forages, and provides for facilitated access to genetic resources in the Multilateral System. The contracting parties are obliged to provide access for the purposes of food and agriculture research, breeding, and training when: • they are requested to do so by another party, a legal entity under the jurisdiction of a party, or by an international institute that has signed an agreement with the governing body; and • PGRFA have been acquired under these terms. By the terms of the Treaty, countries agree that prior informed consent is not required for access to a defined category of PGRFA, but that a set of mutually agreed-upon terms will apply. The standard material transfer agreement (SMTA) enables access to plant genetic resources and establishes benefit-sharing based on royalties levied on ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

commercial products that use material obtained through the multilateral system. For more details, see www.fao.org/ag/cgrfa/itpgr.htm. CITES Permits The Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) is an international agreement between national governments that helps member countries control and monitor protected plant and animal populations. CITES regulates trade and exchange through permits and certificates. When importing samples of a species that is listed in the appendixes of CITES, a CITES import permit from the importing country and a CITES export permit from the country of origin must be obtained. The species covered by CITES are listed in three appendixes according to the degree of protection they ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

need. 1. Appendix I lists species that are the most endangered. 2. Appendix II lists species that are not currently threatened but face extinction unless trade is closely controlled. 3. Appendix III is a list of species for which the cooperation of other countries is needed to prevent unsustainable or illegal exploitation. Seeds of Appendix II plants and seeds of artificially propagated hybrids of Appendix I plants are exempt from CITES controls. 120 Handbooks for Genebanks No. 8 Plants grown from exempt seeds are protected, however, and require CITES permits for import and export. The CITES-( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

listed species database, including the three appendixes and national focal points for permits and certificates, is available 

Seed Processing

 INOVAMER COMPANY HAS THE BEST IN SEED PROCESSING

• Harvest individual plants within an accession when there are differences in flowering and maturity between plants. • Mix an equal proportion of seeds from different mother plants to avoid maternal effects. • Bags holding harvested seeds or heads should be made of porous material enabling good air circulation for drying. • Options for harvesting depend on the crop: • Harvest plants individually, preferably by hand. If machineharvesting, use purpose-built machinery because commercial machinery cannot be cleaned adequately between regeneration plots. • Harvest infructescences individually by hand. • Initiate seed drying immediately after harvesting to prevent seed deterioration. • If seeds cannot be processed quickly, they should be placed in a temporary holding area in a controlled environment such as an air-conditioned room. ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )Documentation Regeneration is performed as a result of information generated by seed monitoring.

As regeneration methods vary according to 115 Manual of Seed Handling species, the types of descriptors used to record information also vary. The following descriptors will help in documenting the data: • Regeneration site • Collaborator (where applicable) • Plot reference • Sowing date • Germination in the field • Number of plants established • Days from sowing to flowering • Breeding system • Pollination control method used • Harvest date • Number of plants harvested • Quantity of seeds harvested Further reading Ellis, R.H., Hong, T.D. and Roberts, E.H. 1985. Handbook of seed technology for genebanks. Volume 1. Principles and Methodology. Handbooks for Genebanks. No. 2, IBPGR, Rome, Italy. FAO/IPGRI. 1994. Genebank Standards. FAO and IPGRI, Rome, Italy. Sackville Hamilton, N.R. and Chorlton, K.H. 1997. Regeneration of accessions in seed collections: A decision guide. (J. Engels, ed.). Handbook for Genebanks No. 5. IPGRI, Rome, Italy. Thormann, I., Metz, T. and ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

Engels, J.M. 2004. IPGRI species compendium, Version 1.0, December 2004. IPGRI, Rome, Italy. www.bioversityinternational.org/Themes/Genebanks/Species_ Compendium/default.asp 116 Handbooks for Genebanks No. 8 Table 8.4. Reproductive behaviour and pollination control mechanisms for regeneration of important crops. Crop Species Pollination behaviour (crossing rate) Pollination mechanism Method of regeneration Alfalfa (lucerne) Medicago sativa Mainly CP (84-94%) Insects (tripping) Isolation; screened cages with pollinators Amaranth Amaranthus spp. CP Wind Isolation; bagging Barley Hordeum vulgare SP Black gram Vigna mungo SP Bottle guard Lagenaria siceraria CP; monoecious Insects Bagging and hand-pollination Brown mustard Brassica juncea ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

Mainly SP (4-14% cross-pollination) Insects Bagging Buck wheat Fagopyrum esculentum CP; self-incompatible Wind Bagging and hand-pollination Buffel grass Cenchrus ciliaris CP Wind Isolation; bagging Cabbage Brassica oleracea var. capitata CP Insects Screened cages with pollinators Carrot Daucus carota CP; protandrous Insects Screened cages with pollinators Castor bean Ricinus communis CP; monoecious Wind Bagging and hand pollination Cauliflower Brassica oleracea var. botrytis Mainly CP Insects Bagging Chickpea Cicer arietinum SP Chicory Cichorium intybus CP; strongly self-incompatible Insects Spatial isolation; Bagging; insect-proof cages Common bean Phaseolus vulgaris Mainly SP; cross-pollination 8-20% Insects Isolation; insect-proof cages; bagging

Cotton Gossypium spp. Mainly SP; natural outcrossing 10-50% Insects Bagging; insect-proof cages Cowpea Vigna unguiculata Mainly SP Crotalaria Crotalaria juncea Mainly SP Cucumber Cucumis sativus CP; monoecious Insects Bagging and hand pollination Eggplant Solanum melongena Partially SP; natural outcrossing up to 48% Insects Isolation; bagging Faba bean Vicia faba Mainly SP; 4-8% outcrossing Insects Isolation; bagging Finger millet Eleusine corocana SP Foxtail millet Setaria italica SP Grass pea Lathyrus sativus SP; significant levels of CP can occur Bagging Hyacinth bean Lablab purpureus Lentil Lens culinaris SP Lettuce Lactuca sativa Mainly SP; natural outcrossing 1-6% Insects Bagging; insect-proof cages Lima bean Phaseolus lunatus Mainly SP (up to 18% natural crossing) Insects Isolation Linseed (flax) Linum usitatissimum Normally SP; natural crossing up to 12% Insects Isolation; bagging Lupin Lupinus spp. Mainly SP; some CP can occur Insects Isolation; insect-proof cages or bagging 117 Manual of Seed Handling Crop Species Pollination behaviour (crossing rate) Pollination mechanism Method of regeneration Maize Zea mays CP; monoecious Wind Bagging ear and hand pollination with pollen pool Mung bean Vigna radiata SP Oat Avena sativa SP Okra Abelmoschus esculentus Partially SP; natural outcrossing 4-19% Insects Isolation; insect-proof cages or bagging Onion Allium cepa Mainly CP; protandrous Insects Screened cages with pollinators Pea Pisum sativum Mainly SP Peanut Arachis hypogaea SP Pearl millet Pennisetum glaucum Mainly CP;

 ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) protogynous Wind Bagging; hand-cross with pollen pool Pepper, chilli Capsicum annuum Often CP; heterostyly Insects Bagging Pigeonpea Cajanus cajan Normally SP; natural outcrossing 5-40% Insects Isolation; bagging; insectproof cages Potato Solanum tuberosum Mainly CP Insects Isolation; bagging Pumpkin Cucurbita moschata CP; monoecious Insects Bagging and hand pollination Radish Raphanus sativus CP; strongly self-incompatible Insects Screened cages with pollinators Red clover Trifolium pratense CP; strongly self-incompatible Insects Screened cages with pollinators Rice Oryza sativa SP Rye Secale cereale CP; strongly self-incompatible Wind Bagging and hand-pollination with pollen pool Rye grass Lolium perenne CP Wind Bagging Safflower Carthamus tinctorius SP Sesame Sesamum indicum Mainly SP; cross pollination up to 5% Insects Sorghum Sorghum bicolor Mainly SP; cross-pollination up to 1-50% Wind Isolation; bagging Soya bean Glycine max SP Spinach Spinacea oleracea CP; dioecious Wind Spatial isolation Strawberry Fragaria ananassa Mainly CP Insects Insect-proof cages Sugar beet Beta vulgaris CP; self-incompatible Wind, Insects Spatial isolation, Screened cages with pollinators Sunflower Helianthus annuus Partially CP; protandrous Insects Bagging and hand pollination Sweet clover Melilotus albus SP Tobacco Nicotiana tabacum SP Tomato Lycopersicon esculentum Normally SP; some species selfincompatible with moderate to high CP Triticale Triticosecale CP Wind Isolation; bagging Vetch Vicia sativa SP Watermelon Citrullus lanatus CP; ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) monoecious Insects Bagging and hand pollination Wheat Triticum aestivum SP SP= Self-pollinating; CP= Cross-pollinating 118 Handbooks for Genebanks No. 8 ANNEX I International policies and frameworks influencing access to and exchange of germplasm Germplasm assemblage and distribution essentially involve movement of seeds across locations or regions. In assembling germplasm, genebanks acquire or import material from germplasm collectors or other suppliers from within and outside the country. Distribution involves exporting seed samples to users worldwide. In addition to the phytosanitary regulations described in Chapters 2 and 7, the following international policies, frameworks and agreements influence the access and exchange of germplasm. Convention on Biological Diversity (CBD) The Convention on Biological Diversity (CBD), which came into force in December 1993, provided a legal framework for the conservation and sustainable use of plant genetic resources. Prior to the CBD, genetic resources were considered to be the common heritage of humankind and were freely available for use without restrictions. The CBD affirmed national ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )sovereignty over genetic resources; in Article 15, it provided guidelines for access and use, including fair and equitable sharing of benefits from resource utilization (www.biodiv.org/convention/articles.asp). 

Seed Processing

 INOVAMER COMPANY HAS THE BEST IN SEED PROCESSING

• Harvest individual plants within an accession when there are differences in flowering and maturity between plants. • Mix an equal proportion of seeds from different mother plants to avoid maternal effects. • Bags holding harvested seeds or heads should be made of porous material enabling good air circulation for drying. • Options for harvesting depend on the crop: • Harvest plants individually, preferably by hand. If machineharvesting, use purpose-built machinery because commercial machinery cannot be cleaned adequately between regeneration plots. • Harvest infructescences individually by hand. • Initiate seed drying immediately after harvesting to prevent seed deterioration. • If seeds cannot be processed quickly, they should be placed in a temporary holding area in a controlled environment such as an air-conditioned room. ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )Documentation Regeneration is performed as a result of information generated by seed monitoring. As regeneration methods vary according to 115 Manual of Seed Handling species, the types of descriptors used to record information also vary. The following descriptors will help in documenting the data:

 • Regeneration site • Collaborator (where applicable) • Plot reference • Sowing date • Germination in the field • Number of plants established • Days from sowing to flowering • Breeding system • Pollination control method used • Harvest date • Number of plants harvested • Quantity of seeds harvested Further reading Ellis, R.H., Hong, T.D. and Roberts, E.H. 1985. Handbook of seed technology for genebanks. Volume 1. Principles and Methodology. Handbooks for Genebanks. No. 2, IBPGR, Rome, Italy. FAO/IPGRI. 1994. Genebank Standards. FAO and IPGRI, Rome, Italy. Sackville Hamilton, N.R. and Chorlton, K.H. 1997. Regeneration of accessions in seed collections: A decision guide. (J. Engels, ed.). Handbook for Genebanks No. 5. IPGRI, Rome, Italy. Thormann, I., Metz, T. and ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

Engels, J.M. 2004. IPGRI species compendium, Version 1.0, December 2004. IPGRI, Rome, Italy. www.bioversityinternational.org/Themes/Genebanks/Species_ Compendium/default.asp 116 Handbooks for Genebanks No. 8 Table 8.4. Reproductive behaviour and pollination control mechanisms for regeneration of important crops. Crop Species Pollination behaviour (crossing rate) Pollination mechanism Method of regeneration Alfalfa (lucerne) Medicago sativa Mainly CP (84-94%) Insects (tripping) Isolation; screened cages with pollinators Amaranth Amaranthus spp. CP Wind Isolation; bagging Barley Hordeum vulgare SP Black gram Vigna mungo SP Bottle guard Lagenaria siceraria CP; monoecious Insects Bagging and hand-pollination Brown mustard Brassica juncea ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )

Mainly SP (4-14% cross-pollination) Insects Bagging Buck wheat Fagopyrum esculentum CP; self-incompatible Wind Bagging and hand-pollination Buffel grass Cenchrus ciliaris CP Wind Isolation; bagging Cabbage Brassica oleracea var. capitata CP Insects Screened cages with pollinators Carrot Daucus carota CP; protandrous Insects Screened cages with pollinators Castor bean Ricinus communis CP; monoecious Wind Bagging and hand pollination Cauliflower Brassica oleracea var. botrytis Mainly CP Insects Bagging Chickpea Cicer arietinum SP Chicory Cichorium intybus CP; strongly self-incompatible Insects Spatial isolation; Bagging; insect-proof cages Common bean Phaseolus vulgaris Mainly SP; cross-pollination 8-20% Insects Isolation; insect-proof cages; bagging

Cotton Gossypium spp. Mainly SP; natural outcrossing 10-50% Insects Bagging; insect-proof cages Cowpea Vigna unguiculata Mainly SP Crotalaria Crotalaria juncea Mainly SP Cucumber Cucumis sativus CP; monoecious Insects Bagging and hand pollination Eggplant Solanum melongena Partially SP; natural outcrossing up to 48% Insects Isolation; bagging Faba bean Vicia faba Mainly SP; 4-8% outcrossing Insects Isolation; bagging Finger millet Eleusine corocana SP Foxtail millet Setaria italica SP Grass pea Lathyrus sativus SP; significant levels of CP can occur Bagging Hyacinth bean Lablab purpureus Lentil Lens culinaris SP Lettuce Lactuca sativa Mainly SP; natural outcrossing 1-6% Insects Bagging; insect-proof cages Lima bean Phaseolus lunatus Mainly SP (up to 18% natural crossing) Insects Isolation Linseed (flax) Linum usitatissimum Normally SP; natural crossing up to 12% Insects Isolation; bagging Lupin Lupinus spp. Mainly SP; some CP can occur Insects Isolation; insect-proof cages or bagging 117 Manual of Seed Handling Crop Species Pollination behaviour (crossing rate) Pollination mechanism Method of regeneration Maize Zea mays CP; monoecious Wind Bagging ear and hand pollination with pollen pool Mung bean Vigna radiata SP Oat Avena sativa SP Okra Abelmoschus esculentus Partially SP; natural outcrossing 4-19% Insects Isolation; insect-proof cages or bagging Onion Allium cepa Mainly CP; protandrous Insects Screened cages with pollinators Pea Pisum sativum Mainly SP Peanut Arachis hypogaea SP Pearl millet Pennisetum glaucum Mainly CP; ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) protogynous Wind Bagging; hand-cross with pollen pool Pepper, chilli Capsicum annuum Often CP; heterostyly Insects Bagging Pigeonpea Cajanus cajan Normally SP; natural outcrossing 5-40% Insects Isolation; bagging; insectproof cages Potato Solanum tuberosum Mainly CP Insects Isolation; bagging Pumpkin Cucurbita moschata CP; monoecious Insects Bagging and hand pollination Radish Raphanus sativus CP; strongly self-incompatible Insects Screened cages with pollinators Red clover Trifolium pratense CP; strongly self-incompatible Insects Screened cages with pollinators Rice Oryza sativa SP Rye Secale cereale CP; strongly self-incompatible Wind Bagging and hand-pollination with pollen pool Rye grass Lolium perenne CP Wind Bagging Safflower Carthamus tinctorius SP Sesame Sesamum indicum Mainly SP; cross pollination up to 5% Insects Sorghum Sorghum bicolor Mainly SP; cross-pollination up to 1-50% Wind Isolation; bagging Soya bean Glycine max SP Spinach Spinacea oleracea CP; dioecious Wind Spatial isolation Strawberry Fragaria ananassa Mainly CP Insects Insect-proof cages Sugar beet Beta vulgaris CP; self-incompatible Wind, Insects Spatial isolation, Screened cages with pollinators Sunflower Helianthus annuus Partially CP; protandrous Insects Bagging and hand pollination Sweet clover Melilotus albus SP Tobacco Nicotiana tabacum SP Tomato Lycopersicon esculentum Normally SP; some species selfincompatible with moderate to high CP Triticale Triticosecale CP Wind Isolation; bagging Vetch Vicia sativa SP Watermelon Citrullus lanatus CP; ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  ) monoecious Insects Bagging and hand pollination Wheat Triticum aestivum SP SP= Self-pollinating; CP= Cross-pollinating 118 Handbooks for Genebanks No. 8 ANNEX I International policies and frameworks influencing access to and exchange of germplasm Germplasm assemblage and distribution essentially involve movement of seeds across locations or regions. In assembling germplasm, genebanks acquire or import material from germplasm collectors or other suppliers from within and outside the country. Distribution involves exporting seed samples to users worldwide. In addition to the phytosanitary regulations described in Chapters 2 and 7, the following international policies, frameworks and agreements influence the access and exchange of germplasm. 

Convention on Biological Diversity (CBD) The Convention on Biological Diversity (CBD), which came into force in December 1993, provided a legal framework for the conservation and sustainable use of plant genetic resources. Prior to the CBD, genetic resources were considered to be the common heritage of humankind and were freely available for use without restrictions. The CBD affirmed national ( Seed Processing, Seed Cleaning, Stone separator, Combo Cleaner, Grading Machine, Gravity Separator  )sovereignty over genetic resources; in Article 15, it provided guidelines for access and use, including fair and equitable sharing of benefits from resource utilization (www.biodiv.org/convention/articles.asp).