Genebank Conservation of Germplasm Collected from Wild Species

  • Christina WaltersEmail author
  • Christopher M. Richards
  • Gayle M. Volk


Crop genebanks are tasked with maintaining genetic resources that support agriculture. They must keep a diverse array of samples alive for decades to centuries. Controlled conditions within the genebank are necessary to maintain quality and ensure consistency of the sample through time. Challenges for providing quality and consistency increase with samples that are mostly unstudied and highly heterogeneous and respond in unpredicted ways, as is the case for samples collected from the wild. The task of genebanking will be facilitated by better definitions of the “conservation target,” meaning the level of diversity that the sample is intended to represent. With that definition, collectors will have better knowledge of what and where to collect – and when to stop – and “fit-for-purpose” samples will be preserved. Major uncertainties persist about the life expectancy of the sample and whether genebanking imposes genetic shifts. Standards have been recommended by the international community to ensure lasting quality of samples despite a large number of unknowns. We believe some of these standards will be counter-productive or unobtainable for wild-collected samples, and we have offered alternatives that stress documentation so future genebank users can predict whether a sample will suit their needs.


Conservation target Cryopreservation Germplasm Longevity Preservation Propagule Sampling Storage Ex situ conservation Genebank 


  1. Aubry C, Shoal R, Erickson VJ (2005) Grass cultivars: their origins, development and use on national forests and grasslands in the Pacific Northwest, U.S. Forest Service: 50Google Scholar
  2. Ballesteros D, Pence VC (2017) Survival and death of seeds during liquid nitrogen storage: a case study on seeds with short lifespans. CryoLetters 38:278–289PubMedGoogle Scholar
  3. Brockway LH (1979) Science and colonial expansion: the role of the British Royal Botanic Gardens. Am Ethnol 6:449–465CrossRefGoogle Scholar
  4. Burton PJ, Burton CM (2002) Promoting genetic diversity in the production of large quantities of native plant seed. Ecol Restor 20:117–123CrossRefGoogle Scholar
  5. Charlesworth D, Charlesworth B, McVean GA (2001) Genome sequences and evolutionary biology, a two-way interaction. Trends Ecol Evol 16:235–242CrossRefGoogle Scholar
  6. Clerkx EJ, El-Lithy ME, Vierling E, Ruys GJ, Blankestijn-De Vries H, Groot SP, Vreugdenhil D, Koornneef M (2004) Analysis of natural allelic variation of Arabidopsis seed germination and seed longevity traits between the accessions Landsberg erecta and Shakdara, using a new recombinant inbred line population. Plant Physiol 135:432–443CrossRefGoogle Scholar
  7. Colville L, Bradley EL, Lloyd AS, Pritchard HW, Castle L, Kranner I (2012) Volatile fingerprints of seeds of four species indicate the involvement of alcoholic fermentation, lipid peroxidation, and Maillard reactions in seed deterioration during ageing and desiccation stress. J Exp Bot 63:6519–6530CrossRefGoogle Scholar
  8. Dafni A, Firmage D (2000) Pollen viability and longevity: practical, ecological and evolutionary implications. In: Pollen and pollination. Springer Vienna, Vienna pp 113–132Google Scholar
  9. Daws MI, Lydall E, Chmielarz P, Leprince O, Matthews S, Thanos CA, Pritchard HW (2004) Developmental heat sum influences recalcitrant seed traits in Aesculus hippocastanum across Europe. New Phytol 162:157–166CrossRefGoogle Scholar
  10. Ellis RH, Roberts EH (1980) Improved equations for the prediction of seed longevity. Ann Bot 45:13–30CrossRefGoogle Scholar
  11. Endresen DTF, Knüpffer H (2012) The Darwin Core extension for genebanks opens up new opportunities for sharing genebank datasets. Biodivers Inform 8:12–29CrossRefGoogle Scholar
  12. Engelmann F (2011) Use of biotechnologies for the conservation of plant biodiversity. In Vitro Cell Dev Biol Plant 47:5–16CrossRefGoogle Scholar
  13. Falk D, Richards CM, Montalvo A, Knapp E (2006) Population and ecological genetics in restoration ecology. In: Falk D, Palmer M, Zedler J (eds) Foundations of restoration biology. Island Press, Washington, DC, pp 14–41Google Scholar
  14. FAO (2001) International treaty on plant genetic resources for food and agriculture. Food and Agriculture Organisation of the United Nations. Available at:
  15. FAO (2014) Genebank standards for plant genetic resources for food and agriculture. Rev. ed, RomeGoogle Scholar
  16. Fleming MB, Richards CM, Walters C (2017) Decline in RNA integrity of dry-stored soybean seeds correlates with loss of germination potential. J Exp Bot 68:2219–2230CrossRefGoogle Scholar
  17. Franchi GG, Piotto B, Nepi M, Baskin CC, Baskin JM, Pacini E (2011) Pollen and seed desiccation tolerance in relation to degree of developmental arrest, dispersal, and survival. J Exp Bot 62:5267–5281CrossRefGoogle Scholar
  18. Franks SJ, Avise JC, Bradshaw WE, Conner JK, Etterson JR, Mazer SJ, Weis AE (2008) The resurrection initiative: storing ancestral genotypes to capture evolution in action. Bioscience 58:870–873CrossRefGoogle Scholar
  19. Gilligan DM, Frankham R (2003) Dynamics of genetic adaptation to captivity. Conserv Genet 4:189–197CrossRefGoogle Scholar
  20. Greene SL, Kisha TJ, Yu L-X, Parra-Quijano M (2014) Conserving plants in gene banks and nature: investigating complementarity with Trifolium thompsonii Morton. PLoS One 9.: art. no. e105145Google Scholar
  21. Guerrant EO Jr, Fiedler PL (2004) Accounting for sample decline during ex situ storage and reintroduction. In: Guerrant EO Jr, Havens K, Maunder M (eds) Ex situ plant conservation: supporting species survival in the wild. Island Press, Covelo, pp 365–385Google Scholar
  22. Guerrant EO Jr, Havens K, Maunder M (eds) (2004) Ex situ plant conservation: supporting species survival in the wild, vol 3. Island Press, CoveloGoogle Scholar
  23. Guerrant EO Jr, Havens K, Vitt P (2014) Sampling for effective ex situ plant conservation. Int J Plant Sci 175:11–20CrossRefGoogle Scholar
  24. Haidet M, Olwell P (2015) Seeds of success: a national seed banking program working to achieve long-term conservation goals. Nat Areas J 35:165–173CrossRefGoogle Scholar
  25. Harding K (2004) Genetic integrity of cryopreserved plant cells: a review. CryoLetters 25:3–22PubMedGoogle Scholar
  26. Havens K, Guerrant EO Jr, Maunder M, Vitt P (2004) Guidelines for ex situ conservation collection management: minimizing risks. In: Guerrant EO Jr, Havens K, Maunder M (eds) Ex situ plant conservation: supporting species survival in the wild. Island Press, Covelo, pp 454–473Google Scholar
  27. Hay FR, Probert RJ (1995) Seed maturity and the effects of different drying conditions on desiccation tolerance and seed longevity in foxglove (Digitalis purpurea L.). Ann Bot 76:639–647CrossRefGoogle Scholar
  28. Hay FR, Probert RJ (2013) Advances in seed conservation of wild plant species: a review of recent research. Conserv Physiol 1.
  29. Hay FR, Mead A, Manger K, Wilson FJ (2003) One-step analysis of seed storage data and the longevity of Arabidopsis thaliana seeds. J Exp Bot 54:993–1011CrossRefGoogle Scholar
  30. Hoban S, Schlarbaum S (2014) Optimal sampling of seeds from plant populations for ex-situ conservation of genetic biodiversity, considering realistic population structure. Biol Conserv 177:90–99CrossRefGoogle Scholar
  31. Hoekstra FA (1995) Collecting pollen for genetic resources conservation. In: Guarino L, Rao VR, Reid R (eds). IPGRI/FAO/UNEP/IUCN Collecting plant genetic diversity: technical guidelines. CABI Publishing, Wallingford, pp 527–550Google Scholar
  32. Hokanson SC, Szewc-McFadden AK, Lamboy WF, McFerson JR (1998) Microsatellite (SSR) markers reveal genetic identities, genetic diversity and relationships in a Malus×domestica Borkh. core subset collection. Theor Appl Genet 97:671–683Google Scholar
  33. ISBER (International Society for Biological an Environmental Repositories) (2012) Best practices for repositories: collection, storage, retrieval, and distribution of biological materials for research. Biopreserv Biobank 10:79–161CrossRefGoogle Scholar
  34. Kahler AL, Kern AJ, Porter RA, Phillips RL (2014) Maintaining food value of wild rice (Zizania palustris L.) using comparative genomics. In: Genomics of plant genetic resources. Springer, Dordrecht, pp 233–248CrossRefGoogle Scholar
  35. Khoury CK, Greene S, Wiersema J, Maxted N, Jarvis A, Struik PC (2013) An inventory of crop wild relatives of the United States. Crop Sci 53:1496–1508CrossRefGoogle Scholar
  36. Khoury CK, Castañeda-Alvarez NP, Achicanoy HA, Sosa CC, Bernau V, Kassa MT, Norton SL, van der Maeseng LJG, Upadhyaya HD, Ramírez-Villegas J, Jarvis A, Struik PC, Jarvis A (2015) Crop wild relatives of pigeonpea [Cajanus cajan (L.) Millsp.]: distributions, ex situ conservation status, and potential genetic resources for abiotic stress tolerance. Biol Conserv 184:259–270CrossRefGoogle Scholar
  37. Kilian B, Graner A (2012) NGS technologies for analyzing germplasm diversity in genebanks. Brief Funct Genomics 11:38 elr046CrossRefGoogle Scholar
  38. Kochanek J, Steadman KJ, Probert RJ, Adkins SW (2009) Variation in seed longevity among different populations, species and genera found in collections from wild Australian plants. Aust J Bot 57:123–131CrossRefGoogle Scholar
  39. Lawrence MJ, Marshall DF, Davies P (1995) Genetics of genetic conservation. I. Sample size when collecting germplasm. Euphytica 84:89–99CrossRefGoogle Scholar
  40. Li D-Z, Pritchard HW (2009) The science and economics of ex situ plant conservation. Trends Plant Sci 14:614–621CrossRefGoogle Scholar
  41. Lockwood DR, Richards CM, Volk GM (2007a) Probabilistic models for collecting genetic diversity: comparisons, caveats, and limitations. Crop Sci 47:861–866CrossRefGoogle Scholar
  42. Lockwood DR, Richards CM, Volk GM (2007b) Wild plant sampling strategies: the roles of ecology and evolution. Plant Breed Rev 29:285–313Google Scholar
  43. Marshall DR, Brown AHD (1975) Optimum sampling strategies in genetic resources conservation. In: Frankel OH, Hawkes JG (eds) Crop genetic resources for today and tomorrow. Cambridge University Press, Cambridge, UK, pp 53–80Google Scholar
  44. Maschinski J, Haskins KE (2012) Plant reintroduction in a changing climate: promises and perils. Island Press, Washington, DCCrossRefGoogle Scholar
  45. Mazur P, Leibo SP, Seidel GE Jr (2008) Cryopreservation of the germplasm of animals used in biological and medical research: importance, impact, status, and future directions. Biol Reprod 78:2–12CrossRefGoogle Scholar
  46. Mead A, Gray D (1999) Prediction of seed longevity: a modification of the shape of the Ellis and Roberts seed survival curves. Seed Sci Res 9:63–73CrossRefGoogle Scholar
  47. Menard KP (2008) Dynamic mechanical analysis: a practical introduction. CRC press, Boca RatonCrossRefGoogle Scholar
  48. Meyer RS, DuVal AE, Jensen HR (2012) Patterns and processes in crop domestication: an historical review and quantitative analysis of 203 global food crops. New Phytol 196:29–48CrossRefGoogle Scholar
  49. Mira S, Hill LM, González-Benito ME, Ibáñez MA, Walters C (2016) Volatile emission in dry seeds as a way to probe chemical reactions during initial asymptomatic deterioration. J Exp Bot 67:1783–1793CrossRefGoogle Scholar
  50. Mondoni A, Orsenigo S, Donà M, Balestrazzi A, Probert RJ, Hay FR, Petraglia A, Abeli T (2014) Environmentally induced transgenerational changes in seed longevity: maternal and genetic influence. Ann Bot 113:1257–1263CrossRefGoogle Scholar
  51. Nagel M, Börner A (2010) The longevity of crop seeds stored under ambient conditions. Seed Sci Res 20:1–12CrossRefGoogle Scholar
  52. Nagel M, Rosenhauer M, Willner E, Snowdon RJ, Friedt W, Börner A (2011) Seed longevity in oilseed rape (Brassica napus L.)–genetic variation and QTL mapping. Plant Genet Resour 9:260–263CrossRefGoogle Scholar
  53. PCA (Plant Conservation Alliance) (2015) National Seed Strategy for Rehabilitation and Restoration 2015–2020. (visited 18 Feb 2017)
  54. Pence VC (2011) Evaluating costs for the in vitro propagation and preservation of endangered plants. In Vitro Cell Dev Biol Plant 47:176–187CrossRefGoogle Scholar
  55. Pence VC (2013) Tissue cryopreservation for plant conservation: potential and challenges. Int J Plant Sci 175:40–45CrossRefGoogle Scholar
  56. Pence VC, Philpott M, Culley TM, Plair B, Yorke SR, Lindsey K, Vanhove A-C, Ballesteros D (2017) Survival and genetic stability of shoot tips of Hedeoma todsenii after long-term cryostorage. In Vitro Cell Dev Biol Plant 53:328–338CrossRefGoogle Scholar
  57. Probert RJ, Adams J, Coneybeer J, Crawford A, Hay F (2007) Seed quality for conservation is critically affected by pre-storage factors. Aust J Bot 55:326–335CrossRefGoogle Scholar
  58. Probert RJ, Daws MI, Hay FR (2009) Ecological correlates of ex situ seed longevity: a comparative study on 195 species. Ann Bot 104:57–69CrossRefGoogle Scholar
  59. RBG (Royal Botanic Gardens Kew) (2017) Seed Information Database (SID). Version 7.1. Available from: (February 2017)
  60. Richards CM, Reilley A, Touchell D, Antolin MF, Walters C (2004) Microsatellite primers for Texas wild rice (Zizania texana), and a preliminary test of the impact of cryogenic storage on allele frequency at these loci. Conserv Genet 5:853–859CrossRefGoogle Scholar
  61. Richards CM, Antolin MF, Reilley A, Poole J, Walters C (2007) Capturing genetic diversity of wild populations for ex situ conservation: Texas wild rice (Zizania texana) as a model. Genet Resour Crop Evol 54:837–848CrossRefGoogle Scholar
  62. Richards CM, Lockwood DR, Volk GM, Walters C (2010) Modeling demographics and genetic diversity in ex situ collections during seed storage and regeneration. Crop Sci 50:2440–2447CrossRefGoogle Scholar
  63. Righetti K, Vu JL, Pelletier S, Vu, BL, Glaab E, Lalanne D Pasha A, Patel RV, Provart NJ, Verdier J,. Leprince O Buitink J (2015) Inference of longevity-related genes from a robust coexpression network of seed maturation identifies regulators linking seed storability to biotic defense-related pathways. Plant Cell 27: 2692–2708PubMedPubMedCentralGoogle Scholar
  64. SCBD (2010) Strategic plan for biodiversity 2011–2020. Secretariat of the convention on biological diversity. Available at:
  65. SCBD (2014) Global strategy for plant conservation. Secretariat of the convention on biological diversity. Available at:
  66. Schoen DJ, Brown AHD (2001) The conservation of wild plant species in seed banks. Bioscience 51:960–966CrossRefGoogle Scholar
  67. Schwember AR, Bradford KJ (2010) Quantitative trait loci associated with longevity of lettuce seeds under conventional and controlled deterioration storage conditions. J Exp Bot 61:4423–4436CrossRefGoogle Scholar
  68. Soulé M (1991) Conservation: tactics for a constant crisis. Science 253:744–750CrossRefGoogle Scholar
  69. Tarquis AM, Bradford KJ (1992) Prehydration and priming treatments that advance germination also increase the rate of deterioration of lettuce seeds. J Exp Bot 43:307–317CrossRefGoogle Scholar
  70. Thormann I, Reeves P, Thumm S, Reilley A, Engels JMM, Biradar CM, Lohwasser U, Börner A, Pillen K, Richards CM (2016) Genotypic and phenotypic changes in wild barley (Hordeum vulgare subsp. spontaneum) during a period of climate change in Jordan. Genet Resour Crop Evol 64:1–18. CrossRefGoogle Scholar
  71. Towill LE, Forsline PL, Walters C, Waddell JW, Laufmann J (2004) Cryopreservation of Malus germplasm using a winter vegetative bud method: results from 1915 accessions. CryoLetters 25:323–334PubMedGoogle Scholar
  72. Tweddle JC, Dickie JB, Baskin CC, Baskin JM (2003) Ecological aspects of seed desiccation sensitivity. J Ecol 91:294–304CrossRefGoogle Scholar
  73. Urban MC (2015) Accelerating extinction risk from climate change. Science 348:571–573CrossRefGoogle Scholar
  74. Van de Wouw M, Kik C, van Hintum T, van Treuren R, Visser B (2010) Genetic erosion in crops: concept, research results and challenges. Plant Genet Resour Charact Util 8(01):1–15CrossRefGoogle Scholar
  75. Volk G (2011) Collecting pollen for genetic resources conservation. In: Guarino L, Ramanatha Rao V, Goldberg E (eds) Collecting plant genetic diversity: technical guidelines—2011 updateGoogle Scholar
  76. Volk GM, Walters C (2004) Preservation of genetic resources in the national plant germplasm clonal collections. Plant Breed Rev 23:291–344Google Scholar
  77. Volk GM, Richards CM, Reilley AA, Henk AD, Forsline PL, Aldwinckle HS (2005) Ex situ conservation of vegetatively propagated species: development of a seed-based core collection for Malus sieversii. J Am Soc Hortic Sci 130:203–210Google Scholar
  78. Volk GM, Waddell J, Bonnart R, Towill L, Ellis D, Luffman M (2008) High viability of dormant Malus buds after 10 years of storage in liquid nitrogen vapour. CryoLetters 29:89–94PubMedGoogle Scholar
  79. Volk GM, Richards CM, Forsline PL (2009) A comprehensive approach toward conserving Malus germplasm. In international symposium on molecular markers in horticulture 859: 177–182Google Scholar
  80. Volk GM, Chao CT, Norelli J, Brown SK, Fazio G, Peace C, Mcferson J, Zhong G, Bretting P (2015) The vulnerability of US apple (Malus) genetic resources. Genet Resour Crop Evol 62:765–794CrossRefGoogle Scholar
  81. Volk GM, Henk AD, Forsline PL, Szewc-Mcfadden AK, Fazio G, Aldwinckle H, Richards CM (2016) Seeds capture the diversity of genetic resource collections of Malus sieversii maintained in an orchard. Genet Resour Crop Evol 64:1513. CrossRefGoogle Scholar
  82. Walters C (1998) Ultra-dry technology: perspective from the National Seed Storage Laboratory, USA. Seed Sci Res 8(suppl 1):11–14Google Scholar
  83. Walters C (2015a) Genebanking seeds from natural populations. Nat Areas J 35:98–105CrossRefGoogle Scholar
  84. Walters C (2015b) Orthodoxy, recalcitrance and in-between: describing variation in seed storage characteristics using threshold responses to water loss. Planta 242:397–406CrossRefGoogle Scholar
  85. Walters C, Hanner R (2006) Platforms for DNA banking. In: de Vicente MC, Andersson MS (eds) DNA banks: providing novel options for Genebanks. Bioversity InternationalGoogle Scholar
  86. Walters C, Wheeler L, Stanwood PC (2004) Longevity of cryogenically stored seeds. Cryobiology 48:229–244CrossRefGoogle Scholar
  87. Walters C, Wheeler LM, Grotenhuis JM (2005) Longevity of seeds stored in a genebank: species characteristics. Seed Sci Res 15:1–20CrossRefGoogle Scholar
  88. Walters C, Volk GM, Richards CM (2008) Genebanks in the post-genomic age: emerging roles and anticipated uses. Biodiversity 9:68–71CrossRefGoogle Scholar
  89. Walters C, Ballesteros D, Vertucci VA (2010) Structural mechanics of seed deterioration: standing the test of time. Plant Sci 179:65–573CrossRefGoogle Scholar
  90. Walters C, Berjak P, Pammenter N, Kennedy K, Raven P (2013) Preservation of recalcitrant seeds. Science 339:915–916CrossRefGoogle Scholar
  91. Wesley-Smith J, Berjak P, Pammenter NW, Walters C (2014) Intracellular ice and cell survival in cryo-exposed embryonic axes of recalcitrant seeds of Acer saccharinum: an ultrastructural study of factors affecting cell and ice structures. Ann Bot 113:695–709CrossRefGoogle Scholar
  92. Wieczorek J, Bloom D, Guralnick R, Blums, Döring M, Giovanni R, Robertson T, Vieglais D (2012) Darwin Core: an evolving community-developed biodiversity data standard. PLoS One 7(1):e29715CrossRefGoogle Scholar
  93. Zheng GH, Jing XM, Tao K-L (1998) Ultradry seed storage cuts cost of gene bank. Nature 393:2CrossRefGoogle Scholar

Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2018

Authors and Affiliations

  • Christina Walters
    • 1
    Email author
  • Christopher M. Richards
    • 1
  • Gayle M. Volk
    • 1
  1. 1.USDA, Agricultural Research Service, Center for Agricultural Resources Research, National Laboratory for Genetic Resource PreservationFort CollinsUSA

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