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GERMPLASM STORAGE AT NCGRP. OUTLINE. Seed Storage – 9:00-9:40 David Brenner Storage of Vegetatively Propagated Crops – 9:40 - 10:20 Phil Forsline Storage of Restricted Germplasm – 10:30 – 10:45. Overview of Questionnaire. 23 questionnaires returned 15 for seed crops
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OUTLINE • Seed Storage – 9:00-9:40 • David Brenner • Storage of Vegetatively Propagated Crops – 9:40 - 10:20 • Phil Forsline • Storage of Restricted Germplasm – 10:30 – 10:45
Overview of Questionnaire • 23 questionnaires returned • 15 for seed crops • 8 for vegetatively propagated crops • Most answers applied to multiple genera • These grouped as single entry in summary • NC-7 ornamental crops ~150 genera • Pullman horticultural crops ~170 genera • Answers were condensed for summarization
Seed Drying/Storage • Majority use 23-26oC for drying seed • Field drying – grains and peanuts • 33- 37oC for maize, mint, basil, chicory, parsnips and • 5oC for vegetable crops • Few control humidity • Dry time weeks to months • ? The extent that MC is determined • Experience + facilities
Seed storage • Majority store between 4-10oC • RH 20-35% • 2 sites reported -20oC storage • Basil, cucurbits, carrot, parsnip, chicory stored at 14oC 50%RH • Seed is generally sent to NCGRP for back-up within one year • “too long”
Germination and viability monitoring • All sites are doing their own initial germination tests • Questionnaire unfortunately did not cover methods used • i.e. AOSA rules • Most sites are also doing monitor testing • Three crops have no monitor testing plan • Every five years seemed to be the norm • Range was 4-20 years
Storability of seed • All indicated good storability • 10+ years • Parsnips and ornamentals were exceptions • This based mostly on experience with only ½ the responses indicating data • Storability variable • Storage conditions • Species/crops • Lumping of crops
Challenges to determining viability • Low seed #/seed fill • Dormancy • Germination information on wild relatives • Limited resources • Lack of facilities • Lengthy germination methods • Time
What can NCGRP do to help? • Report more germination tests • Studies on store temps for seed that do not store well under present conditions • Studies on recalcitrant species (i.e. Hydrastis & goldenseal) • Viability monitoring guidelines • Comparisons on seed storage temps • Germination of wild relatives
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
DiscussionGerm testing, shipping • Difficulties? • How is seed stored prior to shipping to NCGRP • Why lag in shipping seed to NCGRP • Germ testing – AOSA rules • Imaging – is there a need for NCGRP to be doing this? • when, why
DiscussionStorage of seed • Have any comparisons of seed storage conditions been done by the sites? • What interesting things have your observed? • What data exists at the sites? • What do you want to do with viability data? • What challenges do you have with seed storage?
DiscussionCommunication with NCGRP • Who should be contacted regarding accession questions? • What should go to the RL vs the curator? • What is not being communicated? • What needs to be communicated? • No germ/seed number – how best to deal with this data?
DiscussionSeed Distribution • NCGRP will strive to notify curator when NSSL-only accession is requested • Is a cc on an email adequate? • NCGRP will strive to no longer ship accessions in the active collection • What other processes need to be put into place? • What are some of the distribution challenges encountered?
DiscussionChallenges/Needs for Germplasm Storage • What can NCGRP do from a storage standpoint that they are not? • How many recalcitrant/intermediate seeds do you encounter? • How do you handle them? • What are you doing to increase the longevity of poor storing seed? • If you could have other storage conditions, what would they be?
Switch gears!Non-Seed Crops • 23 questionnaires returned • 15 for seed crops • 8 for vegetatively propagated crops • Most answers applied to multiple genera • These grouped as single entry in summary • NC-7 ornamental crops >50 genera • Answers were condensed for summarization
Germplasm Storage • Field is primary storage location • A few crops have tissue culture back-up • Musa, sweet potato, mint, strawberry, pelargonium, hops • Greenhouse/Screenhouse also back-up • % backed up varies • 90% of apples and wild peanut • 10% of ornamentals and “pears, hazelnuts, berries, mint, hops”
In vitro Capabilities • Few crops have established protocols • Few sites have adequate facilities/ resources • Many protocols were established for propagation and not storage • 100% musa, 30% pelargonium, 10% “pears, hazelnuts, berries, mint, hops”, >90% Sweet potato • Short-term TC storage also limited
Cryopreservation • 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 • Relatively limited in other crops • For those, focus is on core collections only
Limitations/needs • Resources • Facilities/space – proper back-up and growing locations • Clean up internal contaminants/diseases • Tissue culture • Cryopreservation • Prioritization/evaluation of collections • Lack of true-to-type seed
What can NCGRP do to help? • Provide back-up storage facilities • Provide better inventories of back-up storage materials • Develop tissue culture methods • Develop cryopreservation methods • Research on difficult to grow and preserve
Cryopreservation • Overview of dormant bud method
DiscussionIn vitro material • Need robust system that works with huge numbers of genotypes • Need sustained and predictable growth • Need dedicated TC facilities (?personnel) • For short-term storage – need to evaluate effect of slowing growth • Cold • Media manipulations • Who has capabilities to develop methods?
DiscussionCryopreservation • How should crops be prioritized • TC method established • Cryo method published • Application of methods to 5-10 genotypes • Most crops need TC going in and/or out • Who has the capabilities to develop the techniques • Who should have the resources to develop techniques? • Dormant bud method for other crops
DiscussionChallenges • Can seed be stored for some crops/ accession • Genebanking vs genotype banking • What is needed to do this? • “Bonsai” back-up • What is need to allow other sites to back-up each others materials? • Do we need a focused effort on TC or cryo? • What else?
Restricted Material • Plant Variety Protection • Crop Science Registration • Safety back-up • Quarantine • Other material (GEO, patented, MTA)
PVP • While under protection the seed is the property of the PVPO • No distribution to site until protection expires after 20 yrs • At expiry • NCGRP cannot release seed without written permission from PVPO • NCGRP splits sample – 2.5K seed to site • Patents, GEO status, etc. unknown
Crop Science Registration • Donor distributes seed for first 5 yrs • Restricted samples have protection for a max of 20 yrs – donor distributes during restriction • Overseas samples – request immediate inclusion into NPGS • NCGRP release seed to site after 5 yrs (or after restricted period) • Seed split between site and NCGRP • Requesting information on patents and GEO status
Safety Back-up • Non-NPGS material • Remains the property of the donor • Stored under a NFCA or MTA • Reviewed and renewed every 5 years • Requests for storage initiated by wide range of characters • PEO, sites, NPS • Organizations – NGOs, private, bot gardens • International – CG centers, National genebanks
Safety Back-up Storage • Plant Gene Resources of Canada • International Maize and Wheat Improvement Center • Center for Plant Conservation • Lady Bird Johnson Wildflower Center • Desert Legume Program • National Park Service • Seed Savers Exchange • New Zealand Inst. for Crop & Food Research Limited • Bureau of National Parks and Wildlife - Asuncion, Paraguay • N.I. Vavilov All-Russian Research Institute of Plant Industry • International Rice Research Institute • Arab Republic of Egypt
Quarantine Material • For inclusion into the NPGS or CSR • Arrange with curator for grow-out in quarantine facility • Facilitate permits when needed • Secure storage until it can be grown-out • Until grown out, quarantined under permit to NCGRP • Safety back-up • No action
Patents, GEOs, other • NCGRP attempts to determine if some restriction to distribution might apply • Elemental patent search • Request information from donor • NCGRP cannot police this • 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 • For informational uses only