Evaluation of a modified encapsulation-dehydration procedure incorporating sucrose pretreatments for the cryopreservation of <Emphasis Type="Italic">Ribes</Emphasis> germplasm |
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Authors: | Email author" target="_blank">Barbara?M?ReedEmail author Laura?Schumacher Dominique?Dumet Erica?E?Benson |
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Institution: | (1) National Clonal Germplasm Repository, USDA-ARS, 33447 Peoria Road, 97333-2521 Corvallis, OR;(2) Plant Conservation Group, School of Contemporary Science, University of Abertay Dundee, Kydd Building, Bell Street, DD1 1HG Dundee, UK;(3) Present address: International Institute of Tropical Agriculture, c/o Lambourn U.K. Limited, Carolyn House, 26 Dingwall Road, CR9 3EE Croydon, UK |
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Abstract: | Summary 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 eight genotypes: (1) pregrowth of shoot tips in sucrose-supplemented
solid growth medium for 1 wk; (2) pretreatment of alginate-encapsulated shoot tips in sucrose-supplemented liquid culture
medium for 21 h; (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 (32–67%), 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 nine Ribes species. The procedure was successful for 18 of the 22 genotypes in the gene bank in Scotland. Screening genotype responses
at the time of storage demonstrated regrowth ≥60% for 15 genotypes, and only four genotypes had regrowth of 0–28%. Additional
genotypes were also added to the USDA cryopreserved Ribes collection. |
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Keywords: | black currant conservation gene bank gooseberry liquid nitrogen |
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