1. GERMPLASM STORAGE AT NCGRP

2. OUTLINE Seed Storage – 9:00-9:40 Seed Storage – 9:00-9:40 David Brenner David Brenner Storage of Vegetatively Propagated Crops – 9:40 - 10:20 Storage of Vegetatively Propagated Crops – 9:40 - 10:20 Phil Forsline Phil Forsline Storage of Restricted Germplasm – 10:30 – 10:45 Storage of Restricted Germplasm – 10:30 – 10:45

3. Overview of Questionnaire 23 questionnaires returned 23 questionnaires returned 15 for seed crops 15 for seed crops 8 for vegetatively propagated crops 8 for vegetatively propagated crops Most answers applied to multiple genera Most answers applied to multiple genera These grouped as single entry in summary These grouped as single entry in summary NC-7 ornamental crops ~150 genera NC-7 ornamental crops ~150 genera Pullman horticultural crops ~170 genera Pullman horticultural crops ~170 genera Answers were condensed for summarization Answers were condensed for summarization

4. Seed Drying/Storage Majority use 23-26 o C for drying seed Majority use 23-26 o C for drying seed Field drying – grains and peanuts Field drying – grains and peanuts 33- 37 o C for maize, mint, basil, chicory, parsnips and 33- 37 o C for maize, mint, basil, chicory, parsnips and 5 o C for vegetable crops 5 o C for vegetable crops Few control humidity Few control humidity Dry time weeks to months Dry time weeks to months ? The extent that MC is determined ? The extent that MC is determined Experience + facilities Experience + facilities

5. Seed storage Majority store between 4-10 o C Majority store between 4-10 o C RH 20-35% RH 20-35% 2 sites reported -20 o C storage 2 sites reported -20 o C storage Basil, cucurbits, carrot, parsnip, chicory stored at 14 o C 50%RH Basil, cucurbits, carrot, parsnip, chicory stored at 14 o C 50%RH Seed is generally sent to NCGRP for back- up within one year Seed is generally sent to NCGRP for back- up within one year “too long” “too long”

6. Germination and viability monitoring All sites are doing their own initial germination tests All sites are doing their own initial germination tests Questionnaire unfortunately did not cover methods used Questionnaire unfortunately did not cover methods used i.e. AOSA rules i.e. AOSA rules Most sites are also doing monitor testing Most sites are also doing monitor testing Three crops have no monitor testing plan Three crops have no monitor testing plan Every five years seemed to be the norm Every five years seemed to be the norm Range was 4-20 years Range was 4-20 years

7. Storability of seed All indicated good storability All indicated good storability 10+ years 10+ years Parsnips and ornamentals were exceptions Parsnips and ornamentals were exceptions This based mostly on experience with only ½ the responses indicating data This based mostly on experience with only ½ the responses indicating data Storability variable Storability variable Storage conditions Storage conditions Species/crops Species/crops Lumping of crops Lumping of crops

8. Challenges to determining viability Low seed #/seed fill Low seed #/seed fill Dormancy Dormancy Germination information on wild relatives Germination information on wild relatives Limited resources Limited resources Lack of facilities Lack of facilities Lengthy germination methods Lengthy germination methods Time Time

9. What can NCGRP do to help? Report more germination tests Report more germination tests Studies on store temps for seed that do not store well under present conditions Studies on store temps for seed that do not store well under present conditions Studies on recalcitrant species (i.e. Hydrastis & goldenseal) Studies on recalcitrant species (i.e. Hydrastis & goldenseal) Viability monitoring guidelines Viability monitoring guidelines Comparisons on seed storage temps Comparisons on seed storage temps Germination of wild relatives Germination of wild relatives

10. Comparison of Seed Storability Times Justin & Bass 1978 Principals and Practices of Seed Storage. USDA Ag handbook No. 506 Walters et al. 2005 Longevity of seeds stored in a genebank. Seed Science Research 15:1-20

11. Discussion Germ testing, shipping Difficulties? Difficulties? How is seed stored prior to shipping to NCGRP How is seed stored prior to shipping to NCGRP Why lag in shipping seed to NCGRP Why lag in shipping seed to NCGRP Germ testing – AOSA rules Germ testing – AOSA rules Imaging – is there a need for NCGRP to be doing this? Imaging – is there a need for NCGRP to be doing this? when, why when, why

12. Discussion Storage of seed Have any comparisons of seed storage conditions been done by the sites? Have any comparisons of seed storage conditions been done by the sites? What interesting things have your observed? What interesting things have your observed? What data exists at the sites? What data exists at the sites? What do you want to do with viability data? What do you want to do with viability data? What challenges do you have with seed storage? What challenges do you have with seed storage?

13. Discussion Communication with NCGRP Who should be contacted regarding accession questions? Who should be contacted regarding accession questions? What should go to the RL vs the curator? What should go to the RL vs the curator? What is not being communicated? What is not being communicated? What needs to be communicated? What needs to be communicated? No germ/seed number – how best to deal with this data? No germ/seed number – how best to deal with this data?

14. Discussion Seed Distribution NCGRP will strive to notify curator when NSSL-only accession is requested NCGRP will strive to notify curator when NSSL-only accession is requested Is a cc on an email adequate? Is a cc on an email adequate? NCGRP will strive to no longer ship accessions in the active collection NCGRP will strive to no longer ship accessions in the active collection What other processes need to be put into place? What other processes need to be put into place? What are some of the distribution challenges encountered? What are some of the distribution challenges encountered?

15. Discussion Challenges/Needs for Germplasm Storage What can NCGRP do from a storage standpoint that they are not? What can NCGRP do from a storage standpoint that they are not? How many recalcitrant/intermediate seeds do you encounter? How many recalcitrant/intermediate seeds do you encounter? How do you handle them? How do you handle them? What are you doing to increase the longevity of poor storing seed? What are you doing to increase the longevity of poor storing seed? If you could have other storage conditions, what would they be? If you could have other storage conditions, what would they be?

16. Switch gears! Non-Seed Crops 23 questionnaires returned 23 questionnaires returned 15 for seed crops 15 for seed crops 8 for vegetatively propagated crops 8 for vegetatively propagated crops Most answers applied to multiple genera Most answers applied to multiple genera These grouped as single entry in summary These grouped as single entry in summary NC-7 ornamental crops >50 genera NC-7 ornamental crops >50 genera Answers were condensed for summarization Answers were condensed for summarization

17. Germplasm Storage Field is primary storage location Field is primary storage location A few crops have tissue culture back-up A few crops have tissue culture back-up Musa, sweet potato, mint, strawberry, pelargonium, hops Musa, sweet potato, mint, strawberry, pelargonium, hops Greenhouse/Screenhouse also back-up Greenhouse/Screenhouse also back-up % backed up varies % backed up varies 90% of apples and wild peanut 90% of apples and wild peanut 10% of ornamentals and “pears, hazelnuts, berries, mint, hops” 10% of ornamentals and “pears, hazelnuts, berries, mint, hops”

18. In vitro Capabilities Few crops have established protocols Few crops have established protocols Few sites have adequate facilities/ resources Few sites have adequate facilities/ resources Many protocols were established for propagation and not storage Many protocols were established for propagation and not storage 100% musa, 30% pelargonium, 10% “pears, hazelnuts, berries, mint, hops”, >90% Sweet potato 100% musa, 30% pelargonium, 10% “pears, hazelnuts, berries, mint, hops”, >90% Sweet potato Short-term TC storage also limited Short-term TC storage also limited

19. Cryopreservation Over half of the questionnaires said cryopreservation had been reported for their crops Over half of the questionnaires said cryopreservation had been reported for their crops Apple (100%) and sour cherry (60%) (discussion to follow) are poster children Apple (100%) and sour cherry (60%) (discussion to follow) are poster children Relatively limited in other crops Relatively limited in other crops For those, focus is on core collections only For those, focus is on core collections only

20. Limitations/needs Resources Resources Facilities/space – proper back-up and growing locations Facilities/space – proper back-up and growing locations Clean up internal contaminants/diseases Clean up internal contaminants/diseases Tissue culture Tissue culture Cryopreservation Cryopreservation Prioritization/evaluation of collections Prioritization/evaluation of collections Lack of true-to-type seed Lack of true-to-type seed

21. What can NCGRP do to help? Provide back-up storage facilities Provide back-up storage facilities Provide better inventories of back-up storage materials Provide better inventories of back-up storage materials Develop tissue culture methods Develop tissue culture methods Develop cryopreservation methods Develop cryopreservation methods Research on difficult to grow and preserve Research on difficult to grow and preserve

22. Cryopreservation Overview of dormant bud method Overview of dormant bud method

23. Discussion In vitro material Need robust system that works with huge numbers of genotypes Need robust system that works with huge numbers of genotypes Need sustained and predictable growth Need sustained and predictable growth Need dedicated TC facilities (?personnel) Need dedicated TC facilities (?personnel) For short-term storage – need to evaluate effect of slowing growth For short-term storage – need to evaluate effect of slowing growth Cold Cold Media manipulations Media manipulations Who has capabilities to develop methods? Who has capabilities to develop methods?

24. Discussion Cryopreservation How should crops be prioritized How should crops be prioritized 1. TC method established 2. Cryo method published 3. Application of methods to 5-10 genotypes Most crops need TC going in and/or out Most crops need TC going in and/or out Who has the capabilities to develop the techniques Who has the capabilities to develop the techniques Who should have the resources to develop techniques? Who should have the resources to develop techniques? Dormant bud method for other crops Dormant bud method for other crops

25. Discussion Challenges Can seed be stored for some crops/ accession Can seed be stored for some crops/ accession Genebanking vs genotype banking Genebanking vs genotype banking What is needed to do this? What is needed to do this? “Bonsai” back-up “Bonsai” back-up What is need to allow other sites to back- up each others materials? What is need to allow other sites to back- up each others materials? Do we need a focused effort on TC or cryo? Do we need a focused effort on TC or cryo? What else? What else?

26. COFFEE BREAK YEAH!!!!!!

27. Restricted Material Plant Variety Protection Plant Variety Protection Crop Science Registration Crop Science Registration Safety back-up Safety back-up Quarantine Quarantine Other material (GEO, patented, MTA) Other material (GEO, patented, MTA)

28. PVP While under protection the seed is the property of the PVPO While under protection the seed is the property of the PVPO No distribution to site until protection expires after 20 yrs No distribution to site until protection expires after 20 yrs At expiry At expiry NCGRP cannot release seed without written permission from PVPO NCGRP cannot release seed without written permission from PVPO NCGRP splits sample – 2.5K seed to site NCGRP splits sample – 2.5K seed to site Patents, GEO status, etc. unknown Patents, GEO status, etc. unknown

29. Crop Science Registration Donor distributes seed for first 5 yrs Donor distributes seed for first 5 yrs Restricted samples have protection for a max of 20 yrs – donor distributes during restriction Restricted samples have protection for a max of 20 yrs – donor distributes during restriction Overseas samples – request immediate inclusion into NPGS Overseas samples – request immediate inclusion into NPGS NCGRP release seed to site after 5 yrs (or after restricted period) NCGRP release seed to site after 5 yrs (or after restricted period) Seed split between site and NCGRP Seed split between site and NCGRP Requesting information on patents and GEO status Requesting information on patents and GEO status

30. Safety Back-up Non-NPGS material Non-NPGS material Remains the property of the donor Remains the property of the donor Stored under a NFCA or MTA Stored under a NFCA or MTA Reviewed and renewed every 5 years Reviewed and renewed every 5 years Requests for storage initiated by wide range of characters Requests for storage initiated by wide range of characters PEO, sites, NPS PEO, sites, NPS Organizations – NGOs, private, bot gardens Organizations – NGOs, private, bot gardens International – CG centers, National genebanks International – CG centers, National genebanks

31. Safety Back-up Storage Plant Gene Resources of Canada Plant Gene Resources of Canada International Maize and Wheat Improvement Center International Maize and Wheat Improvement Center Center for Plant Conservation Center for Plant Conservation Lady Bird Johnson Wildflower Center Lady Bird Johnson Wildflower Center Desert Legume Program Desert Legume Program National Park Service National Park Service Seed Savers Exchange Seed Savers Exchange New Zealand Inst. for Crop & Food Research Limited New Zealand Inst. for Crop & Food Research Limited Bureau of National Parks and Wildlife - Asuncion, Paraguay Bureau of National Parks and Wildlife - Asuncion, Paraguay N.I. Vavilov All-Russian Research Institute of Plant Industry N.I. Vavilov All-Russian Research Institute of Plant Industry International Rice Research Institute International Rice Research Institute Arab Republic of Egypt Arab Republic of Egypt

32. Quarantine Material For inclusion into the NPGS or CSR For inclusion into the NPGS or CSR Arrange with curator for grow-out in quarantine facility Arrange with curator for grow-out in quarantine facility Facilitate permits when needed Facilitate permits when needed Secure storage until it can be grown-out Secure storage until it can be grown-out Until grown out, quarantined under permit to NCGRP Until grown out, quarantined under permit to NCGRP Safety back-up Safety back-up No action No action

33. Patents, GEOs, other NCGRP attempts to determine if some restriction to distribution might apply NCGRP attempts to determine if some restriction to distribution might apply Elemental patent search Elemental patent search Request information from donor Request information from donor NCGRP cannot police this NCGRP cannot police this Assumption is that crop curator knows about the crop and other restrictions Assumption is that crop curator knows about the crop and other restrictions ? Have all PVP post 1992 maize, soybean, cotton and canola checked for GEO prior to release into NPGS ? Have all PVP post 1992 maize, soybean, cotton and canola checked for GEO prior to release into NPGS For informational uses only For informational uses only