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ARS Home » Pacific West Area » Corvallis, Oregon » National Clonal Germplasm Repository » Research » Publications at this Location » Publication #171307
Title: EVALUATION OF A MODIFIED ENCAPSULATION-DEHYDRATION PROCEDURE INCORPORATING SUCROSE PRETREATMENTS FOR THE CRYOPRESERVATION OF RIBES GERMPLASM

Author
item Reed, Barbara
item Schumacher, Laura
item DUMET, DOMINIQUE - U OF ABERTAY DUNDEE
item BENSON, ERICA - U OF ABERTAY DUNDEE

Submitted to: In Vitro Cellular and Developmental Biology - Plants
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/16/2005
Publication Date: 8/20/2005
Citation: Reed, B.M., Schumacher, L.D., Dumet, D., Benson, E. 2005. Evaluation of a modified encapsulation-dehydration procedure incorporating sucrose pretreatments for the cryopreservation of ribes germplasm. In Vitro Cellular and Developmental Biology - Plants. 41:431-436.

Interpretive Summary: A new protocol for storing plant shoot tips in liquid nitrogen using a sugar pretreatment was tested for the long-term storage of currants and gooseberries. Four steps in the procedure were examined for 8 different plant types. Recovery of growing shoots was high (72-100%) at all four steps for the five black currants tested. Air drying decreased viability slightly for some black currants and in some cases liquid nitrogen exposure reduced regrowth. In contrast, three gooseberry species had poor recovery from the initial sucrose culture step (50-67%), indicating sensitivity to stress, which predisposed these genotypes to poor survival after liquid nitrogen exposure (12-26%). We also screened 22 plant types from 9 species for storage in a liquid nitrogen genebank in Scotland. The procedure was successful for 18 of the 22 plant types in the genebank. Screening plant responses at the time of storage demonstrated high regrowth for 14 plants and only 4 had low regrowth. Additional plants were also added to the USDA cryopreserved collection.

Technical Abstract: A modified encapsulation-dehydration cryopreservation protocol based on the replacement of cold acclimation with high-sucrose pretreatment was assessed for the long-term storage of Ribes germplasm. Four steps in the procedure were examined for 8 genotypes: (1) Pregrowth of shoot tips in sucrose-supplemented solid growth medium for 1 week; (2) Pretreatment of alginate-encapsulated shoot tips in sucrose-supplemented liquid culture medium for 21 hours; (3) Evaporative desiccation of encapsulated-dehydrated shoot-tips; and (4) Exposure to liquid nitrogen (LN). Differential responses were observed for black currant and gooseberry genotypes. Recovery of growing shoots was high (72-100%) at all four steps for the five black currants tested. Evaporative desiccation slightly decreased viability for some black currants and in some cases LN exposure reduced regrowth. In contrast, three gooseberry species had poor recovery from the initial sucrose culture step (50-67%), indicating sensitivity to osmotic stress, which predisposed these genotypes to poor survival after LN exposure (12-26%). The effectiveness of the modified protocol for conserving a wider range of Ribes genotypes was further ascertained by screening 22 genotypes derived from 9 Ribes species. The procedure was successful for 18 of the 22 genotypes in the genebank. Screening genotype responses at the time of storage demonstrated regrowth >60% for 15 genotypes and only 4 genotypes had regrowth of 0 to 28%. Additional genotypes were also added to the USDA cryopreserved Ribes collection.