Cryopreservation of Cryopreservation of Clonally Maintained Clonally Maintained Germplasm Germplasm Preparation and Viability testing Preparation and Viability testing USDA-ARS, National Center for Genetic Resources USDA-ARS, National Center for Genetic Resources Preservation, Preservation, Fort Collins, CO Fort Collins, CO
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Cryopreservation of Clonally Maintained Germplasm Preparation and Viability testing
Cryopreservation of Clonally Maintained Germplasm Preparation and Viability testing. USDA-ARS, National Center for Genetic Resources Preservation, Fort Collins, CO. Maintenance of clonal germplasm. More vulnerable than seed-based accessions Requires more space - PowerPoint PPT Presentation
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Cryopreservation of Clonally Cryopreservation of Clonally Maintained GermplasmMaintained Germplasm
Preparation and Viability testingPreparation and Viability testing
USDA-ARS, National Center for Genetic Resources Preservation, USDA-ARS, National Center for Genetic Resources Preservation,
Fort Collins, COFort Collins, CO
Maintenance of clonal germplasmMaintenance of clonal germplasm
•More vulnerable than seed-based accessions
•Requires more space
•Requires special procedures and/or techniques
•Easily compromised and challenged (biotic + abiotic factors)
•Can require years to get established
•Distribution issues
•Yearly costs
Preserving and backing-up of Preserving and backing-up of clonally maintained germplasmclonally maintained germplasm
Primary site
•Field, greenhouse, laboratory/growth chamber
Back-up
•Field in another location, growth chamber in another location, cryopreservation
CryopreservationCryopreservation
•Requires species/genotype specific protocols
•Time and labor consuming
•Costly in initial stages
•HOWEVER, once in LN2 - cost of long-term storage is low
•Relatively safe back-up
What is needed to place clonal germplasm What is needed to place clonal germplasm in long-term LNin long-term LN22 storage at the NCGRP storage at the NCGRP
1. Procedures tested and modified (as needed) at the Center
2. Collaboration with Curators
Collaboration with Curators is Collaboration with Curators is CRITICALCRITICAL for for successsuccess
1. Decide which taxa are priority
2. Providing material/germplasm to be processed
Tissue culture, dormant budwood (DB), field derived material (i.e. garlic)
3. If DB are used – viability testing (grafting or rooting)
• During experimental protocol adjustments
• During routine cryopreservation
Dormant buds (DB)Dormant buds (DB)
Advantages:
•Large number of accessions able to process in one season
•Short (relatively) time from cryo to establishing a usable plant
Limitations:•Applicable only to woody and semi-woody accessions•Limited number of reliable cryo-procedures (apple, sour cherry)•Inconsistencies in viability testing/grafting
Species under protocol development in Species under protocol development in the Preservation Program (DB)the Preservation Program (DB)
•Juglans
black and English/Carpathian walnut; butternut
collaboration with UC Davis, Burchell Nursery, Forest Service, UM/MO
•Prunus
almond, apricot, peach, sweet cherry
collaboration with Davis repository, Burchell Nursery
•Pyrus, Ribes, Vaccinium
pear, black and red currant, highbush, lowbush and rabbiteye blueberry
collaboration with Corvallis repository
•Salix
collaboration with Ames (some willow accessions)
Example of material needed - DBExample of material needed - DB