GERMPLASM STORAGE AT NCGRP

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GERMPLASM STORAGE AT NCGRP
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OUTLINE

• Seed Storage 900-940
• David Brenner
• Storage of Vegetatively Propagated Crops 940 -
  1020
• Phil Forsline
• Storage of Restricted Germplasm 1030 1045

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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

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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

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Seed storage

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

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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

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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

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Challenges to determining viability

• Low seed /seed fill
• Dormancy
• Germination information on wild relatives
• Limited resources
• Lack of facilities
• Lengthy germination methods
• Time

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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

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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 151-20
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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

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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?

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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?

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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?

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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?

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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 gt50 genera
• Answers were condensed for summarization

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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

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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, gt90 Sweet
  potato
• Short-term TC storage also limited

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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

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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

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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

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Cryopreservation

• Overview of dormant bud method

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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?

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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

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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?

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COFFEE BREAKYEAH!!!!!!

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Restricted Material

• Plant Variety Protection
• Crop Science Registration
• Safety back-up
• Quarantine
• Other material (GEO, patented, MTA)

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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

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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

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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

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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

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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

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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