1.6. GENETIC RESOURCES “Genetic resources or germplasm of a crop species may be describes as the sum total of the genetic information present in the species and its wild relatives. The genetic information comprises all the alleles of all the g«-ies present in the given crop species and its related species. The germplasm, in practical terms, consists of the following materials: (i) varieties in cultivation, (i1) obsolete or old varieties, (iii) breeding lines, (iv) special genetic stocks, (v) land races or local varieties, (vi) wild forms and (vii) wild relatives. A wild form is the wild species from which the given crop has been directly derived by domestication. A wild relative is one of the other species that are related to the crop species by descent during the evolution. The first four categories of the materials broadly constitute the adapted or elite germplasm, while the remaining materials comprise the unadapted germplasm. The germplasm originating in the country under consideration are called indigenous germplasm, while those from another country are referred to as exotic germplasm. The germplasm is thus the chief source of desirable alleles and genes that are used to construct the improved varieties through hybridization. The germplasm accessions that easily hybridize with the crop species and yield fertile hybrids are grouped into the primary gene pool (GP1). Generally, GP1 germplasm comprises the strains of the concerned crop species and is the most extensively used. The secondary gene pool (GP2) accessions present difficulties in hybridization with the crop species, and the hybrids are partially fertile. GP2 serves as a valuable source of useful genes and alleles. The hybridization between members of the tertiary gene pool (GP3) with gene transfers into the crop species generally requires special techniques and great amount of work.The genetic variability present in the crops and their relatives is consistently declining due to various human activities; this is termed as genetic erosion. The variability in the cultivated germplasm is declining due to widespread adoption of the improved varieties. It is ironical that the improved product derived from the genetic variability represented in the germplasm is adversely affecting the genetic variability itself. The wild forms and the wild relatives of the crop species are threatened by extension of cultivation and grazing into the forests, developmental activities like railways, roads, industrial areas, hydroelectric projects etc., and even introduction of some species that invade the natural habitats. In view of the above, it is necessary to conserve the germplasm of crop species to ensure their availability in the future. The wild forms and wild relatives can be conserved in the areas where they occur by protecting them from human interference; this is called in situ germplasm conservation. The protected areas are known as gene sanctuaries or biosphere reserves. They are best established in the centres of origin of crop species. In India, 14 biosphere reserves have been established. The germplasms present in gene sanctuaries continue to evolve in response to the natural forces. Still it is generally very difficult to protect these areas from human interference. At the same time, the cultivated germplasm cannot be conserved in gene sanctuaries. The cultivated germplasm has to be collected and conserved as germplasm collections, often called gene banks. This is known as ex-situ germplasm conservation, Even the wild forms and wild relatives of crops collected and conserved in gene banks. The germplasm can be conserved in one of the following five ways: (i) stored as seeds, (ii) grown in the fields as plants, (iii) maintained as slow growing shoot cultures, (iv) kept frozen in liquid nitrogen at - 196°C as cell and organ cultures or even embryos, and (v) stored as DNA fragments as cosmid clones or phage lysates. Storage as seed is used wherever it is feasible. This is because seed storage is easy, safe, and is amenable to rapid use after storage. Most crop species produce orthodox seed, which can be dried to 5% or lower moisture content. These seeds (with 5% moisture) are stored for long periods (20-100 years depending on species) at -20°C; this constitutes base collection. The seeds in an active collection are stored at a temperature close to 0°C for 10-15 years. The working collection is maintained at ~15°C for up to 5 years. Government of Norway has established a Global Gene Vault at Svalbard, near North Pole in 2008. In India, Defense Research and Development Organization (DRDO) has set up similar Gene Vault at Chang La in Ladakh in 2009. The genetic resources of the asexually propagated species are maintained plants in the field, as slow-growth shoot cultures or as cryopreserved cells, tissues and organs including embryos. The genetic material from the extinct or nearly extinct species can be stored as cosmid clones or phage lysates (Table 1.3). The large gene banks having germplasm from all over the world are recognized as world collections (Table 1.4). In India, the germplasm aremaintained as base collections at the National Bureau of Plant Genetic Resource (NBPGR), New Delhi. In 2006, the NBPGR had 339,194 accessions of germplasm of different crop species. Table 1.3: A summary of the various S approaches for germplasm co conservation Accessions ined as Growing plants in mplasm m of wild ‘stablished in centres of conservation reserves protected forms and wild origin/diversity; difficult to from human relatives. maintain interference Ex situ Plants, seeds, shoot Cultivated Established in places with conservation cultures ete. in gene germplasm, wild appropriate facilities banks forms, and wild relatives (i) Seed Seeds All species producing The most preferred approach banks orthodox* seeds a, Base Seeds with 5% The ultimate source Disturbed only for collection moisture at -20°C of active collections regeneration** and germination tests b. Active Seeds with 5% Used for evaluation, Periodically replaced by collection moisture at close multiplication and accessions from the base 10 0°C distribution collection ¢. Working Seeds with 10% Used in breeding Maintained by the breeders collection moisture at~ 15°C programmes. (ii) Field Plant maintained in Asexually propagated Requires large areas and banks the field species considerable resources (iii) Slow- Slow growing shoot Asexually propagated A substitute for field banks; growth cultures species Tequires tissue culture facility; cultures in use for some species (iv) Cryopre- Cells, tissues, Asexually propagated Requires considerable servation organs, e.g., embryos, species expertise and sophistication; kept at -196°C in in use for some species liquid nitrogen (v) DNA. DNA fragments as Extinct or nearly Highly sophisticated; needs banks, cosmid clones or extinct species; a considerable resources phage lysates source of desired DNA sequences *Seeds that can be dried to 5% or lower moisture level without any adverse effect on their viability. **Growing the stored seeds in the field, and using the freshly harvested seeds for further storage.Table 1.4: World collections of germplasm. The various international crop research institutes, in addition, maintain large collections of germplasm of the concerned crops World collection located at Coll Ethiopia, Africa Groundnut Bambey, Senegal Potato Cambridge, UK Sugarcane Canal Point, Florida, USA’ Sweet potato New Zealand Small grain crops Beltsville, USA. All crops Institute of Plant Industry, Russia All crops Royal Botanic Gardens, Kew, England All crops Svalbard Global Seed Vault*, near North pole * Popularly known as doomsday vault, It has more than 230,000 permafrost zone near North Pole to safeguard against natural and human-mediated disasters. The germplasm stored in gene banks has to be collected by exploration or introduction (collection of germplasm). All germplasm is obtained by exploration, i.c., trips for collection of germplasm. Explorations generally cover the areas of centres of origin/diversity, and in areas where the diverse genotypes of the crops are expected to be found. Centres of origin are the areas where the crop species are believed to have originated. These areas show great plant diversity, and often termed as centres of diversity. Initially, only eight centres of origin were proposed, but the total number is now 11 (China, Hindustan, Central Asia, Asia Minor, Mediterranean, Abyssinia, Central America, Peru, Brazil- Paraguay, Chile and USA). Similarly, 12 megacentres of plant diversity have been recognized; these generally overlap the centres of origin. Within each! centre, there are relatively much smaller areas that show very high variability; 4 these areas are called microcentres. Microcentres are the first to be explored for germplasm collection. y The germplasm accessions have to be evaluated for the various traits of interest to enable their judicious utilization (evaluation). This multidisciplinary activity is the most critical and expensive step in germplasm conservation. Each germplasm accession is given a unique identification number, and all the data available for it are stored in a suitable format that facilitates its retrieval when Tequired. This information is published by the gene banks as catalogues (cataloguing). The germplasm accessions are stored, their germination is Periodically assessed and the accessions are regenerated when germination Teaches a critical level. Regeneration consists of growing the accessions in the field and using their fresh seeds for further storage. The germplasm accessionsare multiplied and distributed to the research workers as per their research needs. The germplasm accessions can be used for selection of suitable lines for direct release as new varieties, as parents in hybridization or in other numerous types of investigation (utilization). In India, large number (nearly 200) varieties have been developed through selection in germplasm accessions; over 50 varieties have been developed in rice alone.