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United States Department of Agriculture

Agricultural Research Service

Research Project: Management of Temperate-Adapted Fruit, Nut, and Specialty Crop Genetic Resources and Associated Information

Location: National Clonal Germplasm Repository (Corvallis, Oregon)

Title: New technique for more rapid cryopreservation of dormant vegetative tree buds

Authors
item Kovalchuk, Irina -
item Turdiev, Timur -
item Mukhitdinova, Zinat -
item Frolov, Sergey -
item Kairova, Gulsharya -
item Reed, Barbara

Submitted to: Acta Horticulturae
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 10, 2014
Publication Date: June 23, 2014
Citation: Kovalchuk, I., Turdiev, T., Mukhitdinova, Z., Frolov, S., Kairova, G., Reed, B.M. 2014. New technique for more rapid cryopreservation of dormant vegetative tree buds. Acta Horticulturae. 1039:137-146.

Interpretive Summary: Storing dormant buds of temperate trees in liquid nitrogen can provide a safe backup of field germplasm collections. However the process requires several months of preparation before buds can be cryopreserved. Cryopreservation at the natural moisture content would greatly accelerate the storage process. This study examined the effect of cold acclimation, moisture content and cryoprotectants on the viability of dormant buds after cryopreservation. All natural moisture buds died after liquid nitrogen exposure despite any treatments. Cold treated buds dried to a low moiture had 66-100% viability. Viability was 100% for the dried buds with either treatment. Six treatment combinations were tested on cold treated buds at the natural moisture. Viability was 100% for buds with natural moisture with several treatments for tart and sweet cherry and apple buds. Using these treatment combinations with a programmable freezer reduced the time required to store dormant buds from nearly two months to one week. A liquid nitrogen stored dormant bud collection was established at the Kazakhstan genebank with 116 apples, 42 tart cherries and 58 sweet cherries.

Technical Abstract: Cryopreservation of dormant buds of temperate trees in liquid nitrogen can provide a safe backup of field germplasm collections. However the process requires several months of preparation before buds can be cryopreserved. Cryopreservation at the natural moisture content (MC) would greatly accelerate the storage process. This study examined the effect of cold acclimation, moisture content and cryoprotectants on the viability of dormant buds after cryopreservation. All natural MC buds (40 to 55% MC), both cold acclimatized (CA) at the rate of 3°C each 24 h from -5°C to -25°C for 1 week and non-CA buds, died after liquid nitrogen exposure. Post-cryopreservation viability of CA 30% MC buds was 66-100%. Viability was 100% for the 30% MC buds with either standard CA for 1 wk, or 2° C/min to -30° C (6 h in a programmable freezer). In both cases the viability after liquid nitrogen exposure was high. Six pretreatment and cryoprotectant combinations were tested on CA buds at the natural MC. Viability was 100% for buds with natural MC with 3-h pretreatment and cryopreservation with PVS2, PGD or honey for tart cherry; and with PVS2, PVS3, PGD, honey + 15% DMSO, or IPBB-1 (50% glycerol and 50% glucose) for sweet cherry. For apple buds IPBB-1 gave 100% viability for all and PVS2 for most genotypes. Using these pretreatments and cryoprotectants with a programmable freezer reduced the time required to store dormant buds from nearly two months to one week. A cryogenic germplasm collection was established at the Kazakhstan genebank with 116 apples, 42 tart cherries and 58 sweet cherries.

Last Modified: 11/24/2014
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