Biophysical Characteristics of Successful Oilseed Embryo Cryoprotection and Cryopreservation Using Vacuum Infiltration Vitrification: An Innovation in Plant Cell Preservation
Identifieur interne : 000B77 ( Main/Exploration ); précédent : 000B76; suivant : 000B78Biophysical Characteristics of Successful Oilseed Embryo Cryoprotection and Cryopreservation Using Vacuum Infiltration Vitrification: An Innovation in Plant Cell Preservation
Auteurs : Jayanthi Nadarajan ; Hugh W. PritchardSource :
- PLoS ONE [ 1932-6203 ] ; 2014.
Abstract
Heterogeneity in morphology, physiology and cellular chemistry of plant tissues can compromise successful cryoprotection and cryopreservation. Cryoprotection is a function of exposure time × temperature × permeability for the chosen protectant and diffusion pathway length, as determined by specimen geometry, to provide sufficient dehydration whilst avoiding excessive chemical toxicity. We have developed an innovative method of vacuum infiltration vitrification (VIV) at 381 mm (15 in) Hg (50 kPa) that ensures the rapid (5 min), uniform permeation of Plant Vitrification Solution 2 (PVS2) cryoprotectant into plant embryos and their successful cryopreservation, as judged by regrowth
Url:
DOI: 10.1371/journal.pone.0096169
PubMed: 24788797
PubMed Central: 4006905
Affiliations:
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Biophysical Characteristics of Successful Oilseed Embryo Cryoprotection and Cryopreservation Using Vacuum Infiltration Vitrification: An Innovation in Plant Cell Preservation</title>
<author><name sortKey="Nadarajan, Jayanthi" sort="Nadarajan, Jayanthi" uniqKey="Nadarajan J" first="Jayanthi" last="Nadarajan">Jayanthi Nadarajan</name>
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<author><name sortKey="Pritchard, Hugh W" sort="Pritchard, Hugh W" uniqKey="Pritchard H" first="Hugh W." last="Pritchard">Hugh W. Pritchard</name>
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<front><div type="abstract" xml:lang="en"><p>Heterogeneity in morphology, physiology and cellular chemistry of plant tissues can compromise successful cryoprotection and cryopreservation. Cryoprotection is a function of exposure time × temperature × permeability for the chosen protectant and diffusion pathway length, as determined by specimen geometry, to provide sufficient dehydration whilst avoiding excessive chemical toxicity. We have developed an innovative method of vacuum infiltration vitrification (VIV) at 381 mm (15 in) Hg (50 kPa) that ensures the rapid (5 min), uniform permeation of Plant Vitrification Solution 2 (PVS2) cryoprotectant into plant embryos and their successful cryopreservation, as judged by regrowth <italic>in vitro</italic>
. This method was validated on zygotic embryos/embryonic axes of three species (<italic>Carica papaya</italic>
, <italic>Passiflora edulis</italic>
and <italic>Laurus nobilis</italic>
) up to 1.6 mg dry mass and 5.6 mm in length, with varying physiology (desiccation tolerances) and 80°C variation in lipid thermal profiles, i.e., visco-elasticity properties, as determined by differential scanning calorimetry. Comparisons between the melting features of cryoprotected embryos and embryo regrowth indicated an optimal internal PVS2 concentration of about 60% of full strength. The physiological vigour of surviving embryos was directly related to the proportion of survivors. Compared with conventional vitrification, VIV-cryopreservation offered a ∼ 10-fold reduction in PVS2 exposure times, higher embryo viability and regrowth and greater effectiveness at two pre-treatment temperatures (0°C and 25°C). VIV-cryopreservation may form the basis of a generic, high throughput technology for the <italic>ex situ</italic>
conservation of plant genetic resources, aiding food security and protection of species from diverse habitats and at risk of extinction.</p>
</div>
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<tree><noCountry><name sortKey="Nadarajan, Jayanthi" sort="Nadarajan, Jayanthi" uniqKey="Nadarajan J" first="Jayanthi" last="Nadarajan">Jayanthi Nadarajan</name>
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