Seed conservation - A global approach

Seed conservation is an important activity and a strategy to save, preserve, safeguard and conserve plant biological resources mostly in the form of seeds both at national and international level. Many organizations, agencies and institutes are involved in conservation realizing the importance of rare and endangered plant species in very existence of mankind now and in future. There are two broad approaches namely in situ conservation and ex situ conservation. Little effort is done to brief some of the techniques in seed conservation here in this presentation.

1. THANK YOU WELCOME

2. Seed conservation : Global approach V V Gadad PGS14AGR6337 Ph.D Scholar Seed Science & Technology UAS Dharwad

3. Flow of presentation Introduction Conservation-global concern In- situ conservation Ex- situ conservation International organisations for seed conservation The ultimate seed vault Wrap up!

4. Biodiversity Degree of variations of life forms The totality of genes, species and ecosystems of region It can be defined as the variability among and between the living organisms and species of surrounding ecosystems and ecological complexes of their life support.

5. Genetic resources Animal genetic resources Plant genetic resources Microbial genetic resources Marine genetic resource

6. One third of the global plant species are threatened in different level. -International Union of Conservation of Nature (IUCN) Major threat to rapid loss and extinction of genetic diversity- Habitat destruction Pollution Climate change Invasion of exotic species Human population pressure Ever increasing agricultural pressure and practices and Life style change ( Opdam and Wascher , 2004 )

7. Conservation –Global challenge Global population approaching towards 9.1 billion in 2050 –need of 70% increase in the food production ( Godfray et al ., 2010) Undertaking of effective as well as productive agricultural land uses has raised global challenge of conserving biodiversity. ( Tscharntke et al ., 2012) Conserve plant genetic diversity- essential for food security -FAO Hence, International Treaty on Plant Genetic Resources for Food and Agriculture came into exist in 2001 to recognize farmers contribution to the diversity of crops, establish mechanism to access plant genetic materials and share benefits of developing genetic materials (FAO, 2014).

8. The very objective of germplasm conservation (or storage) is to preserve the genetic diversity of a particular plant or genetic stock for its use at any time in future. In recent years, many new plant species with desired and improved characteristics have started replacing the primitive and conventionally used agricultural plants . It is important to conserve the endangered plants or else some of the valuable genetic traits present in the primitive plants may be lost.

9. Conservation – global concern

10. A global body namely International Board of Plant Genetic Resources (IBPGR ) now named Bioversity International has been established for germplasm conservation. Its main objective is to provide necessary support for collection, conservation and utilization of plant genetic resources throughout the world.

11. Two basic methods :  - In-situ conservation - Ex-situ conservation Methods of conservation

12. In-situ conservation   is on-site   conservation or the conservation of genetic resources in natural populations of plant , such as forest genetic resources in natural populations of tree species. It is the process of protecting an endangered plant  in its natural habitat, either by protecting or cleaning up the habitat itself, or by defending the species from predators. It is applied to conservation of agricultural biodiversity in agro ecosystems by farmers, especially those using unconventional farming practices.

13. Methods of in-situ conservation : Protected areas Conservation on-farm Home gardens The major limitations of in-situ conservation are listed below : The risk of losing germplasm due to environmental hazards The cost of maintenance of a large number of genotypes is very high .

14. Today, there are about 37,000 protected areas, parks, sanctuaries and biosphere reserves all around the world!!!

15. Protected areas A network of 668 protected areas (PAs) has been established comprising 102 national parks 515 wild life sanctuaries 47 conservation reserves 4 community reserves Source: www.moef.nic.in

16. On-farm conservation and home gardens Conservation On-farm: Maintenance of useful species in the production systems where they are grown. The oldest agro biodiversity method of conservation ever practiced by humankind since the discover of agriculture. Farmers maintain crops through their continued cultivation, and although they may not be aware of the genetic diversity they harbour on their farms, they contribute through their work to informally safeguard traditional crops that meet local needs both for their own families and for their communities ( Mekbib et al., 2009 ).

17. The conservation of crops on-farm has several advantages compared with ex-situ conservation Advantages Allows for evolution of crops through continued natural and human-driven selection, which contribute to greater adaptation and resilience in cultivation. Makes possible the maintenance of crops whose seeds cannot be stored at the low temperatures of genebanks . Supports the maintenance of traditional knowledge (TK, often termed indigenous knowledge – IK) associated with their use. Disadvantages Limited access to germplasm for breeders and other users Vulnerability of crops to natural disasters The fact that less diversity can be stored at any single location .

18. In-situ conservation of unique land races of Rice by Shri. Shankar Langati Ratnachuda, a fine grain good quality rice Bangarakaddi, a fine grain good quality rice Karigajaville, an aromatic with good nutrition Dambarsali, a purple rice variety for weed identification Conservation of unique land races of Rice, vegetables and legumes by Shri. Shankar Langati, Gundenatti, Belguam, Karnataka

19. In-situ conservation of unique land races of Rice by Shri. Shankar Langati Ambemohr, a fine grain aromatic rice Raj Bhog, a fine grain aromatic rice Gandhasale, fine graine aromatic rice Kumkum sali, a good quality aromatic rice

20. Conservation of Unique land races of vegetables Seven leaf Bhendi Backyard Brinjal Hairy Cluster bean Table purpose pumpkin Desi Tomato (table purpose)

21. Conservation of Unique land races of vegetables Madiwala Bitterguard Gandigawada Brinjal Perinnal Bitter guard Local Ridge guard

22. Practice of in situ green manuring, organic farming and water conservation

23. Storage of different land races of rice and other crops Awards secured for conservation of traditional varieties.

24. Organisation of exhibitions, field days and Farmers’ field school

25. Participation in State level Seed saviour conference

26. 1. Indrani 16.Sonum 2. Hegge 17.Nadandar Sali 3. Kunkum Sali 18.Manila 4. Krishna Kumu 19.Jadagi 5. Ratna Chudi 20. Dodda batta 6. Chandrikya Mugad 21.Ganda Sali 7. Barma Black 22.Karigijivili 8. Honne Kattu 23.Intan 9. Shiddagiri-2 24.Belgum Basmati 10. Padma Rekha 25.Dodda byairenellu 11. Mugad Dodgya 26.Mapale Samba 12. Navali Sali 27.Ramagalli 13. Bangar Kaddi 28.Karihakkala Sali 14. Mysore Mallige 29.Dambar Sali 15. Mugad Suganda 30. Kalajeeva 31. Rajabhoga Traditional rice varieties conserved at Gundenhatti Village

27. 1. Madaki 2. White Alasndi 3. Red Alasandi 4. Gurellu (Niger ) 5. Malenad Alasandi 6. Kari Hurali 7. Kari Hesaru 8. Kempu Hurali 9. Gadde Avare 10. Vatani 11. Kare Kadale 12. Green Hesaru 13. Chennangi Traditional pulse varieties Traditional vegetable varieties 1. Badane ( brinjal ) 2. Mulangi ( Raddish ) 3. Sorekai 4. Entene Bende 5. Chappara Avare 6. Kidney Beans 7. Heere 8. Tuppire 9. Chakkar Kumbala

28. Yenigar Sorghum White Sorghum Javari Sajje Kari Ragi Kempu Ragi Urupula Navane Hala Navane 1.Amrut Godi 2 . Kempu Godi Traditional millets varieties Traditional Wheat varieties

29. Contact numbers: Shankar langati - 9972150378 Kombli - 9845890411 Kallappa neginal ( bagewadi )- 9980634062

30. Benefit sharing amount deposited Contributions from National & International Organizations National Gene Fund Section 45 of the PPV&FR Act, 2001 In Flow Reimbursement of compensation Supporting conservation Reimbursement of Benefit shares Out Flow Annual Fee National Gene Fund Constituted by the Government of India Compensation amount deposited National Gene Fund established on 26 March 2007 For supporting the conservation and sustainable use of genetic resources including in-situ and ex-situ collections

31. Reward and Recognition from the Gene Fund Rules, 2012 - Section 39 (1) (iii) (Conferred Annually) Reward : Maximum of ten rewards to farmers consisting of citation, memento and cash of Rupees one lakh each. Recognition : Maximum of twenty recognitions to farmers. Maximum five awards consisting of a citation, memento and cash of Rupees Ten lakh each Plant Genome Saviour Community Awards - Rule 70 (2) a ( Conferred Annually) Awards / Reward / Recognition Awarded Since Plant Genome Saviour Community Awards 10 2009-10 Plant Genome Saviour Farmer Reward 10 2012 Plant Genome Saviour Farmer Recognition 15 2012 Plant Genome Saviour Recognition Certificates (prior to initiation of PGSCA) 16 2007-08 to 2010-11

32. Home gardens Smaller scale generally more species Diverse Mainly medicinal, flavouring , or vegetable spp. Eg . Chelsea Physic Garden, London

33. Community Seed Banks (CSBs) Farmers can be also organized as so-called ‘ community gene banks ’, whereby leader custodian farmers maintain the diversity on behalf of all others members ( Ramprasad , 2002). Some of these community-based approaches are further structured and include multiple objectives in their work. ‘gene-seed-grain’ banks being practised in some regions of India (MSSRF, 2010). Community seed bank (CSB) at Udaipur, Rajasthan

34. Establishment, operation and management of community seed banks ( Malik et al ., 2013)

35. Scientific management of CSBs Maintenance of seed purity at farmers field and at CSBs Precautions at farmers field Maintaining isolation (b) Rouging Precautions at seed banks Physical purity (b) Freedom from weed seeds 2. Seed handling, cleaning and grading Cleaning (b) Grading 3. Seed storage procedures for CSBs Natural drying (b) Artificial drying (c) Seed processing (d) Seed storage Treatment of seeds and storage containers Monitoring seed quality and health 6. Monitoring seed germination, viability and vigour before seed distribution

36. Community Seed Banks at Udaipur ( Malik et al., 2013) Locations of 15 Community Seed Banks in 3 Blocks of District Udaipur

37. Community Seed Bank at Udaipur Visit of farmers to gene bank at MPUAT, Udaipur CSB awareness camp at village level in Udaipur district

38. Ex-situ conservation

39. Ex-situ conservation   means literally, "off-site conservation". It is the process of protecting an endangered species of plant or animal outside of its natural habitat . For example , by removing part of the population from a threatened habitat  and placing it in a new location, which may be a wild area or within the care of humans.

40. What is the difference between  In-situ  and  Ex-situ  conservation? In-situ Ex-situ done in the natural habitats of the biodiversity components done outside of their natural habitats. more dynamic more static involves designation, management, and monitoring of target taxa in their natural habitats involves sampling, transfer, and storage of target taxa from their natural habitats. populations remain within the ecosystem involving the process of evolution they are not involving the natural evolution process. time consuming but more sustainable aimed to use in conserving genetic components, in immediate occasions.

41. Ex situ conservation can be carried out using several methods   - Seed gene bank - In vitro storage - DNA storage - Pollen storage - Field gene bank   - Botanical gardens

42. A  seed bank stores  seeds  as a source for planting in case seed reserves elsewhere are destroyed. It is a type of gene bank. Storing seeds also guards against catastrophic events like natural disasters, outbreaks of disease, or war. Unlike seed libraries or seed swaps that encourage frequent reuse and sharing of seeds, seed banks are not typically open to the public.

43. A seed bank is a facility used to store seeds of various crops and wild plants, in an effort to maintain biodiversity. These structures can be found scattered all over the world, established by governments and organizations concerned about crop diversity. The Global Crop Diversity Trust proposed in 2007 that an Arctic seed bank be established, to preserve seeds of vital crops in safe bunker conditions in the event of catastrophic events.

44. Seed conservation- Why d o we h ave Seed Banks? The storage of material in the form of seeds is one of the most widespread and valuable  ex situ  approaches to conservation. Seed banking has considerable advantages over other methods of  ex situ conservation such as ease of storage, economy of space, relatively low labour demands and consequently, the capacity to maintain large samples at an economically viable cost.

45. Why Store Seeds and Not Whole Plants? Seeds are a convenient means of long term storage of genetic diversity, as the samples are small in size, are easily handled, require low maintenance and frequently remain viable for long periods . In general, conditions of low temperature and desiccation allow seeds to maintain viability, in many cases indefinitely. Seed banks take up little space, but can be expensive to run, both because of the need to maintain low temperatures and the necessity for germination tests, growth trials and regeneration. They are not suitable for species with recalcitrant seeds.

46. There are however, certain limitations in the conservation of seeds i . Viability of seeds is reduced or lost with passage of time. ii. Seeds are susceptible to insect or pathogen attack, often leading to their destruction. iii. This approach is exclusively confined to seed propagating plants, and therefore it is of no use for vegetatively propagated plants e.g. potato, Ipomoea, Dioscorea . iv. It is difficult to maintain clones through seed conservation. Certain seeds are heterogeneous and therefore, are not suitable for true genotype maintenance.

47. In vitro methods for germplasm conservation In vitro methods employing shoots, meristems and embryos are ideally suited for the conservation of germplasm of vegetatively propagated plants. The plants with recalcitrant seeds and genetically engineered materials can also be preserved by this in vitro approach.

48. There are several advantages associated with in vitro germplasm conservation i . Large quantities of materials can be preserved in small space. ii. The germplasm preserved can be maintained in an environment, free from pathogens. iii. It can be protected against the nature’s hazards. iv. From the germplasm stock, large number of plants can be obtained whenever needed. v. Obstacles for their transport through national and international borders are minimal.

49. There are mainly three approaches for the in vitro conservation of germplasm 1. Cryopreservation (freeze-preservation) 2. Cold storage 3. Low-pressure and low-oxygen storage

50. Cryopreservation Cryopreservation (Greek, krayos -frost) literally means preservation in the frozen state. The principle involved in cryopreservation is to bring the plant cell and tissue cultures to a zero metabolism or non-dividing state by reducing the temperature in the presence of cryoprotectants .

51. Cryopreservation broadly means the storage of germplasm at very low temperatures i . Over solid carbon dioxide (at -79°C) ii. Low temperature deep freezers (at -80°C) iii. In vapour phase nitrogen (at -150°C) iv. In liquid nitrogen (at -196°C) In fact, cryopreservation has been successfully applied for germplasm conservation of a wide range of plant species e.g. rice, wheat, peanut, cassava, sugarcane, strawberry, coconut. Several plants can be regenerated from cells, meristems and embryos stored in cryopreservation.

52. Freezing The sensitivity of the cells to low temperature is variable and largely depends on the plant species . Four different types of freezing methods are used : 1. Slow-freezing method (suspension cultures) 2. Rapid freezing method (shoot tips and somatic embryos) 3. Stepwise freezing method (suspension cultures, shoot apices and buds) 4. Dry freezing method

53. Cold storage basically involves germplasm conservation at a low and non-freezing temperatures (1-9°C ) The growth of the plant material is slowed down in cold storage in contrast to complete stoppage in cryopreservation . Hence, cold storage is regarded as a slow growth germplasm conservation method. The major advantage of this approach is that the plant material (cells/tissues) is not subjected to cryogenic injuries.

54. Low-Pressure and Low-Oxygen Storage: As alternatives to cryopreservation and cold storage, low-pressure storage (LPS) and low-oxygen storage (LOS) have been developed for germplasm conservation.

55. In low-pressure storage, the atmospheric pressure surrounding the plant material is reduced. This results in a partial decrease of the pressure exerted by the gases around the germplasm . The lowered partial pressure reduces the in vitro growth of plants (of organized or unorganized tissues). Low-pressure storage systems are useful for short-term and long-term storage of plant materials. The short-term storage is particularly useful to increase the shelf life of many plant materials e.g. fruits, vegetables, cut flowers, plant cuttings. The germplasm grown in cultures can be stored for long term under low pressure. Besides germplasm preservation, LPS reduces the activity of pathogenic organisms and inhibits spore germination in the plant culture systems.

56. Low-Oxygen Storage (LOS ) In the low-oxygen storage, the oxygen concentration is reduced, but the atmospheric pressure is maintained by the addition of inert gases (particularly nitrogen). The partial pressure of oxygen below 50 mm Hg reduces plant tissue growth (organized or unorganized tissue). This is due to the fact that with reduced availability of O 2 , the production of CO 2  is low. As a consequence, the photosynthetic activity is reduced, thereby inhibiting the plant tissue growth and dimension.

57. Other biotechnological approaches in plant genetic resources conservation A number of biotechnological approaches are being used for the conservation and improvement of plant species for desired traits and each of them has some advantages and disadvantages. Some important techniques which have great potential in the determination of genetic diversity have been used in many plant species including rare and endangered species. ( Salim et al., 2012)

58. Molecular marker technology Biochemical marker Phytochemical markers DNA based markers Random amplified polymorphic DNA (RAPD) Sequence characterized amplified region (SCAR) Simple sequence repeat (SSR) Inter simple sequence repeat (ISSR) Restriction fragment length polymorphism (RFLP).

59. Amplified fragment length polymorphism (AFLP) Selective amplification of microsatellite polymorphic loci (SAMPL) Single nucleotide polymorphism (SNP) Expressed sequence tagged (EST) Single strand conformation polymorphism (SSCP) Cleaved amplified polymorphic sequence (CAPS) 2. Additional markers Internal transcribed spacer (ITS) sequences Chloroplast spacer sequences DNA microarray 3. Plant DNA bank

60. Table. List of several micropropagated medicinally important plants. ( Malabika and Mohammad, 2014)

61. Table. List of endangered and threatened plants regenerated via somatic embryogenesis and organogenesis by the process of plant tissue culture. ( Malabika and Mohammad, 2014)

62. How are Botanic Gardens Involved? Several botanic gardens have developed the capacity to store isolated embryos, minute seeds and tissues under conditions of cryopreservation, maintaining samples in liquid nitrogen at temperatures of – 196°C Such techniques offer great potential for the maintenance and conservation of biodiversity, particularly plant species that cannot be stored by conventional means

63. Gene bank procedures Collecting Registration Sample processing Germplasm testing Viability testing Health diagnosis Transgene detection Monitoring Conservation Seed bank Field bank In vitro bank Cryo bank Vegetative bank DNA bank Characterization Regeneration Distribution Safety duplication Equipment/supplies

64. Status of availability and safety duplication of CGIAR genebanks across years.

65. % Seed accessions available for international distribution from 2013 to 2015

66. NBPGR –A nodal agency for conservation of seeds and other propagules in India. NBPGR and its ten regional stations in different agro climatic zones of the country effectively support the germplasm collection, conservation and evaluation activities of their respective regions. Integrated conservation approach is carried out by a network of more than 50 National Active Germplasm Sites(NAGS) of the National Agricultural Research System spread throughout the country

67. Mandate To act as nodal institute at national level for acquisition and management of indigenous and exotic plant genetic resources for food and agriculture, and to carry out related research and human resource development, for sustainable growth of agriculture. Objectives of NBPGR To plan, organize, conduct and coordinate exploration and collection of indigenous and exotic plant genetic resources. To undertake introduction, exchange and quarantine of plant genetic resources. To characterize, evaluate, document and conserve crop genetic resources and promote their use, in collaboration with other national organizations. To develop information network on plant genetic resources. To conduct research, undertake teaching and training, develop guidelines and create public awareness on plant genetic resources.

68. The NAGS centres are responsible for crop specific collection , multiplication, evaluation, maintainance and conservation of active collections and are distributing germplasm based on indenters requests routed through NBPGR. Seed gene bank at NBPGR has capacity to store 0.75 million accessions, making India third largest gene bank after US and China.

69. Ongoing Research Activities Supported by IPGRI Ultra-dry Seed Storage Sun-drying: an Alternative Way to Lower Seed Moisture Content Cryopreservation: a Potential Alternative for Conserving Seed of "Problem" Species Germplasm Health: a Vital Consideration Over the years, the Institute has developed strong collaborative partnerships with several research centres including the Seed Laboratory of the University of Reading (U.K.), Royal Botanic Gardens, Kew (U.K.), the Boyce Thompson Institute (U.S.A.), Cornell University (U.S.A.), the National Seed Science Laboratory at Fort Collins (U.S.A.), the University of Wageningen (The Netherlands) and the Beijing Botanical Garden, Institute of Botany, Chinese Academy of Sciences (China).

70. 15 Seed Saving Initiatives Protecting Biodiversity for Future Generations 1. AVRDC – The World Vegetable Center 2. Camino Verde 3. Great Lakes Bioneers Chicago (GLBC) Seed Saving Initiative 4. Hawai’s Public Seed Initiative  5. International Center for Tropical Agriculture 6. Louisiana Native Plant Initiative 7. Man and the Biosphere Programme

71. 8 . Millennium Seed Bank Partnership 9. Native Seed / SEARCH  10. Navdanya   11. New York City Native Plant Conservation Initiative  12. The NSW Seed bank 13. Seed Savers Exchange  14. Slow Food International 15. Svalbard Global Seed Vault 

72. Did you know!!!... There are botanic gardens and arboreta in 148 countries worldwide and they maintain more than 4 million living plant collections . Amongst their collections are representatives of more than 100,000 species, almost one third of the known vascular plant species of the world . There are a total of 142 million herbarium specimens in botanic garden herbaria and 6.13 million accessions in their living collections . Over 500 botanic gardens occur in Western Europe, more than 350 in North America and over 200 in East and Southeast Asia, of which the majority are in China. Most of the southern Asian botanic gardens are to be found in India.

73. The Svalbard global seed vault: 'Doomsday   Seed Vault ' The Global Crop Diversity Trust proposed in 2007 that an Arctic seed bank be established, to preserve seeds of vital crops in safe bunker conditions in the case of catastrophic events.

74. Svalbard global seed vault is a secure seed bank on the Norwegian island of Spitsbergen  near  Longyearbyen  in the remote Arctic Svalbard archipelago, about 1,300 kilometres (810 mi) from the North Pole . Conservationist  Cary Fowler, in association with the Consultative Group on International Agricultural Research (CGIAR),started the vault to preserve a wide variety of plant seeds.

75.  I t is built right into a sandstone mountain and covered in a thick layer of permafrost, are kept at an icy minus 0.4 degrees Fahrenheit (minus 18 degrees Celsius).

76. Contains global accessions

77. Soil Seed Bank Soil seed bank is a natural storage of viable seeds (often dormant) within the soil of most ecosystems. Soil seed bank is generally been defined as the amount of viable seeds present in the soil of defined area (Zhang zhiquan,1996 ). The study of soil seed bank started in 1859 when Charles Darwin observed the emergence of seedlings using soil samples from the bottom of a lake. Important fields in soil seed banks are Weed seed bank Forest regeneration and Restoration ecology

78. Environmental significance : Play an important role in the natural environment of many ecosystems. Rapid re-vegetation of sites disturbed by wildfire, catastrophic weather, agricultural operations, and timber harvesting. Forest ecosystems and wetlands contain a number of specialized plant species forming persistent soil seed bank. A knowledge of the soil seed bank is of great value in the agriculture, forestry, conservation management and vegetation of mining wastelands ( Zhang zhiquan ,)

79. Soil diaspore bank and Soil bud bank: The term soil diaspore bank can be used to include non-flowering plants such as ferns and bryophytes Many plants have vegetative propagules to facilitate forming new plants, migration into new ground, or reestablishment after being top-killed. These propagules are collectively called as the soil bud bank , which includes dormant and adventitious buds on stolons , rhizomes and, roots .

80. Similarity between the seed bank and the aboveground vegetation species composition (Guillaume et al ., 2004) Forest of Le Nouvion , Northern France In SC (Selective cutting system), pre-commercial thinning treatments are conducted every 4 years (removal of almost all shrubs and unsuitable trees at 23m3 per ha) and commercial fellings every 8 years (selective cutting of mature trees at 10m3 per ha). In CWS(Coppice-With-oak Standards), commercial felling usually occurs every 30 years, removing the whole coppice timber and about three quarters of the standards (i.e., from 150 to 250m3 per ha).

81. Seed security for Food security