GERMPLASM GERMPLASM STORAGE AT NCGRPSTORAGE AT NCGRP

OUTLINE OUTLINE

Seed Storage – 9:00-9:40Seed Storage – 9:00-9:40 David BrennerDavid Brenner

Storage of Vegetatively Propagated Storage of Vegetatively Propagated Crops – 9:40 - 10:20Crops – 9:40 - 10:20 Phil ForslinePhil Forsline

Storage of Restricted Germplasm – Storage of Restricted Germplasm – 10:30 – 10:4510:30 – 10:45

Overview of Overview of Questionnaire Questionnaire

23 questionnaires returned23 questionnaires returned 15 for seed crops 15 for seed crops

8 for vegetatively propagated crops8 for vegetatively propagated crops

Most answers applied to multiple generaMost answers applied to multiple genera These grouped as single entry in summaryThese grouped as single entry in summary

NC-7 ornamental crops ~150 generaNC-7 ornamental crops ~150 genera

Pullman horticultural crops ~170 generaPullman horticultural crops ~170 genera

Answers were condensed for Answers were condensed for summarizationsummarization

Seed Drying/StorageSeed Drying/Storage

Majority use 23-26Majority use 23-26ooC for drying seedC for drying seed Field drying – grains and peanutsField drying – grains and peanuts

33- 3733- 37ooC for maize, mint, basil, chicory, C for maize, mint, basil, chicory, parsnips and parsnips and

55ooC for vegetable crops C for vegetable crops

Few control humidityFew control humidity

Dry time weeks to monthsDry time weeks to months

? The extent that MC is determined? The extent that MC is determined

Experience + facilities Experience + facilities

Seed storageSeed storage

Majority store between 4-10Majority store between 4-10ooCC RH 20-35%RH 20-35%

2 sites reported -202 sites reported -20ooC storageC storage

Basil, cucurbits, carrot, parsnip, chicory Basil, cucurbits, carrot, parsnip, chicory stored at 14stored at 14ooC 50%RHC 50%RH

Seed is generally sent to NCGRP for Seed is generally sent to NCGRP for back-up within one yearback-up within one year ““too long” too long”

Germination and Germination and viability monitoringviability monitoring

All sites are doing their own initial All sites are doing their own initial germination testsgermination tests Questionnaire unfortunately did not Questionnaire unfortunately did not

cover methods usedcover methods used i.e. AOSA rulesi.e. AOSA rules

Most sites are also doing monitor Most sites are also doing monitor testing testing Three crops have no monitor testing planThree crops have no monitor testing plan Every five years seemed to be the normEvery five years seemed to be the norm Range was 4-20 yearsRange was 4-20 years

Storability of seedStorability of seed

All indicated good storabilityAll indicated good storability 10+ years 10+ years

Parsnips and ornamentals were exceptionsParsnips and ornamentals were exceptions

This based mostly on experience with This based mostly on experience with only ½ the responses indicating dataonly ½ the responses indicating data

Storability variableStorability variable Storage conditionsStorage conditions

Species/cropsSpecies/crops

Lumping of crops Lumping of crops

Challenges to determining Challenges to determining viabilityviability

Low seed #/seed fillLow seed #/seed fill

DormancyDormancy

Germination information on wild Germination information on wild relativesrelatives

Limited resourcesLimited resources

Lack of facilitiesLack of facilities

Lengthy germination methodsLengthy germination methods

Time Time

What can NCGRP do to What can NCGRP do to help?help?

Report more germination testsReport more germination tests

Studies on store temps for seed that Studies on store temps for seed that do not store well under present do not store well under present conditionsconditions

Studies on recalcitrant species (i.e. Studies on recalcitrant species (i.e. Hydrastis & goldenseal)Hydrastis & goldenseal)

Viability monitoring guidelinesViability monitoring guidelines

Comparisons on seed storage temps Comparisons on seed storage temps

Germination of wild relativesGermination of wild relatives

Comparison of Seed Comparison of Seed Storability TimesStorability Times

crop PI site Justice & Bass (index) Walters et. al. (P50)Small grains 15-25 yrs med >50 yrsRice 15-25 yrs med 34 yrsCucurbits 20+ yrs med >50 yrsAnnual clovers 10-25 yrs med - high 16 - >100 yrsChicory 10+ yrs med >39 - 50 yrsSafflower 15-20 yrs N/A 88 yrsPhaseolis >50 yrs med 31 yrsMaize >20 yrs poor - med 49 yrsSoybeans 10 yrs poor 32 yrsBrassica 10-15 yrs med 23 - 76 yrsAllium 10-15 yrs poor 21-70 yrs

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

DiscussionDiscussionGerm testing, shippingGerm testing, shipping

Difficulties?Difficulties?

How is seed stored prior to shipping How is seed stored prior to shipping to NCGRPto NCGRP

Why lag in shipping seed to NCGRPWhy lag in shipping seed to NCGRP

Germ testing – AOSA rulesGerm testing – AOSA rules

Imaging – is there a need for NCGRP Imaging – is there a need for NCGRP to be doing this?to be doing this? when, whywhen, why

DiscussionDiscussionStorage of seedStorage of seed

Have any comparisons of seed Have any comparisons of seed storage conditions been done by the storage conditions been done by the sites?sites?

What interesting things have your What interesting things have your observed?observed? What data exists at the sites?What data exists at the sites?

What do you want to do with viability What do you want to do with viability data?data?

What challenges do you have with What challenges do you have with seed storage?seed storage?

DiscussionDiscussionCommunication with NCGRPCommunication with NCGRP

Who should be contacted regarding Who should be contacted regarding accession questions?accession questions?

What should go to the RL vs the What should go to the RL vs the curator?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 No germ/seed number – how best to deal with this data?deal with this data?

DiscussionDiscussionSeed DistributionSeed Distribution

NCGRP will strive to notify curator NCGRP will strive to notify curator when NSSL-only accession is when NSSL-only accession is requestedrequested Is a cc on an email adequate?Is a cc on an email adequate?

NCGRP will strive to no longer ship NCGRP will strive to no longer ship accessions in the active collectionaccessions in the active collection

What other processes need to be put What other processes need to be put into place?into place?

What are some of the distribution What are some of the distribution challenges encountered?challenges encountered?

DiscussionDiscussionChallenges/Needs for Challenges/Needs for Germplasm StorageGermplasm Storage

What can NCGRP do from a storage What can NCGRP do from a storage standpoint that they are not?standpoint that they are not?

How many recalcitrant/intermediate How many recalcitrant/intermediate seeds do you encounter?seeds do you encounter? How do you handle them?How do you handle them?

What are you doing to increase the What are you doing to increase the longevity of poor storing seed?longevity of poor storing seed?

If you could have other storage If you could have other storage conditions, what would they be?conditions, what would they be?

Switch gears!Switch gears!Non-Seed CropsNon-Seed Crops

23 questionnaires returned23 questionnaires returned 15 for seed crops 15 for seed crops

8 for vegetatively propagated crops8 for vegetatively propagated crops

Most answers applied to multiple Most answers applied to multiple generagenera These grouped as single entry in These grouped as single entry in

summarysummary

NC-7 ornamental crops >50 generaNC-7 ornamental crops >50 genera

Answers were condensed for Answers were condensed for summarizationsummarization

Germplasm StorageGermplasm Storage

Field is primary storage locationField is primary storage location A few crops have tissue culture back-A few crops have tissue culture back-

upup Musa, sweet potato, mint, strawberry, Musa, sweet potato, mint, strawberry,

pelargonium, hopspelargonium, hops

Greenhouse/Screenhouse also back-upGreenhouse/Screenhouse also back-up % backed up varies% backed up varies

90% of apples and wild peanut90% of apples and wild peanut 10% of ornamentals and “pears, hazelnuts, 10% of ornamentals and “pears, hazelnuts,

berries, mint, hops”berries, mint, hops”

In vitro CapabilitiesIn vitro Capabilities

Few crops have established protocolsFew crops have established protocols Few sites have adequate facilities/ Few sites have adequate facilities/

resourcesresources Many protocols were established for Many protocols were established for

propagation and not storagepropagation and not storage 100% musa, 30% pelargonium, 10% 100% musa, 30% pelargonium, 10%

“pears, hazelnuts, berries, mint, “pears, hazelnuts, berries, mint, hops”, >90% Sweet potatohops”, >90% Sweet potato

Short-term TC storage also limitedShort-term TC storage also limited

CryopreservationCryopreservation

Over half of the questionnaires said Over half of the questionnaires said cryopreservation had been reported cryopreservation had been reported for their cropsfor their crops

Apple (100%) and sour cherry (60%) Apple (100%) and sour cherry (60%) (discussion to follow) are poster (discussion to follow) are poster childrenchildren

Relatively limited in other cropsRelatively limited in other crops

For those, focus is on core collections For those, focus is on core collections onlyonly

Limitations/needsLimitations/needs

ResourcesResources Facilities/space – proper back-up and Facilities/space – proper back-up and

growing locationsgrowing locations Clean up internal Clean up internal

contaminants/diseasescontaminants/diseases Tissue cultureTissue culture CryopreservationCryopreservation Prioritization/evaluation of collectionsPrioritization/evaluation of collections Lack of true-to-type seedLack of true-to-type seed

What can NCGRP do to What can NCGRP do to help?help?

Provide back-up storage facilitiesProvide back-up storage facilities

Provide better inventories of back-up Provide better inventories of back-up storage materialsstorage materials

Develop tissue culture methodsDevelop tissue culture methods

Develop cryopreservation methodsDevelop cryopreservation methods

Research on difficult to grow and Research on difficult to grow and preservepreserve

CryopreservationCryopreservation

Overview of dormant bud methodOverview of dormant bud method

DiscussionDiscussionIn vitro materialIn vitro material

Need robust system that works with Need robust system that works with huge numbers of genotypeshuge numbers of genotypes

Need sustained and predictable growthNeed sustained and predictable growth

Need dedicated TC facilities (?personnel)Need dedicated TC facilities (?personnel)

For short-term storage – need to For short-term storage – need to evaluate effect of slowing growthevaluate effect of slowing growth ColdCold

Media manipulationsMedia manipulations

Who has capabilities to develop Who has capabilities to develop methods?methods?

DiscussionDiscussionCryopreservationCryopreservation

How should crops be prioritizedHow should crops be prioritized

1.1. TC method establishedTC method established

2.2. Cryo method publishedCryo method published

3.3. Application of methods to 5-10 genotypesApplication of methods to 5-10 genotypes

Most crops need TC going in and/or outMost crops need TC going in and/or out Who has the capabilities to develop Who has the capabilities to develop

the techniquesthe techniques Who should have the resources to Who should have the resources to

develop techniques?develop techniques? Dormant bud method for other cropsDormant bud method for other crops

DiscussionDiscussionChallengesChallenges

Can seed be stored for some crops/ Can seed be stored for some crops/ accessionaccession Genebanking vs genotype bankingGenebanking vs genotype banking

What is needed to do this?What is needed to do this?

““Bonsai” back-upBonsai” back-up

What is need to allow other sites to What is need to allow other sites to back-up each others materials?back-up each others materials?

Do we need a focused effort on TC or Do we need a focused effort on TC or cryo?cryo?

What else?What else?

COFFEE BREAKCOFFEE BREAK

YEAH!!!!!!YEAH!!!!!!

Restricted MaterialRestricted Material

Plant Variety ProtectionPlant Variety Protection

Crop Science RegistrationCrop Science Registration

Safety back-upSafety back-up

QuarantineQuarantine

Other material (GEO, patented, MTA)Other material (GEO, patented, MTA)

PVPPVP

While under protection the seed is While under protection the seed is the property of the PVPOthe property of the PVPO

No distribution to site until protection No distribution to site until protection expires after 20 yrsexpires after 20 yrs

At expiryAt expiry NCGRP cannot release seed without NCGRP cannot release seed without

written permission from PVPOwritten permission from PVPO

NCGRP splits sample – 2.5K seed to siteNCGRP splits sample – 2.5K seed to site

Patents, GEO status, etc. unknownPatents, GEO status, etc. unknown

Crop Science Crop Science RegistrationRegistration

Donor distributes seed for first 5 yrs Donor distributes seed for first 5 yrs Restricted samples have protection for a Restricted samples have protection for a

max of 20 yrs – donor distributes during max of 20 yrs – donor distributes during restrictionrestriction

Overseas samples – request immediate Overseas samples – request immediate inclusion into NPGSinclusion into NPGS

NCGRP release seed to site after 5 yrs NCGRP release seed to site after 5 yrs (or after restricted period)(or after restricted period) Seed split between site and NCGRPSeed split between site and NCGRP

Requesting information on patents Requesting information on patents and GEO statusand GEO status

Safety Back-upSafety Back-up

Non-NPGS materialNon-NPGS material

Remains the property of the donorRemains the property of the donor

Stored under a NFCA or MTAStored under a NFCA or MTA Reviewed and renewed every 5 yearsReviewed and renewed every 5 years

Requests for storage initiated by wide Requests for storage initiated by wide range of charactersrange of characters PEO, sites, NPSPEO, sites, NPS

Organizations – NGOs, private, bot gardensOrganizations – NGOs, private, bot gardens

International – CG centers, National International – CG centers, National genebanksgenebanks

Safety Back-up StorageSafety Back-up Storage Plant Gene Resources of CanadaPlant Gene Resources of Canada International Maize and Wheat Improvement Center International Maize and Wheat Improvement Center Center for Plant ConservationCenter for Plant Conservation Lady Bird Johnson Wildflower CenterLady Bird Johnson Wildflower Center Desert Legume ProgramDesert Legume Program National Park Service National Park Service Seed Savers ExchangeSeed Savers Exchange New Zealand Inst. for Crop & Food Research LimitedNew Zealand Inst. for Crop & Food Research Limited Bureau of National Parks and Wildlife - Asuncion, Bureau of National Parks and Wildlife - Asuncion,

ParaguayParaguay N.I. Vavilov All-Russian Research Institute of Plant N.I. Vavilov All-Russian Research Institute of Plant

IndustryIndustry International Rice Research InstituteInternational Rice Research Institute Arab Republic of EgyptArab Republic of Egypt

Quarantine MaterialQuarantine Material

For inclusion into the NPGS or CSRFor inclusion into the NPGS or CSR Arrange with curator for grow-out in Arrange with curator for grow-out in

quarantine facilityquarantine facility Facilitate permits when neededFacilitate permits when needed Secure storage until it can be grown-outSecure storage until it can be grown-out

Until grown out, quarantined under Until grown out, quarantined under permit to NCGRPpermit to NCGRP

Safety back-upSafety back-up No actionNo action

Patents, GEOs, otherPatents, GEOs, other NCGRP attempts to determine if some NCGRP attempts to determine if some

restriction to distribution might applyrestriction to distribution might apply Elemental patent searchElemental patent search

Request information from donorRequest information from donor

NCGRP cannot police thisNCGRP cannot police this

Assumption is that crop curator knows Assumption is that crop curator knows about the crop and other restrictionsabout the crop and other restrictions

? Have all PVP post 1992 maize, soybean, ? Have all PVP post 1992 maize, soybean, cotton and canola checked for GEO prior to cotton and canola checked for GEO prior to release into NPGSrelease into NPGS For informational uses onlyFor informational uses only