Biophysical Characteristics of Successful Oilseed Embryo Cryoprotection and Cryopreservation Using Vacuum Infiltration Vitrification: An Innovation in Plant Cell Preservation
Identifieur interne : 001200 ( Pmc/Curation ); précédent : 001199; suivant : 001201Biophysical 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
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DOI: 10.1371/journal.pone.0096169
PubMed: 24788797
PubMed Central: 4006905
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">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><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Pritchard, Hugh W" sort="Pritchard, Hugh W" uniqKey="Pritchard H" first="Hugh W." last="Pritchard">Hugh W. Pritchard</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">24788797</idno><idno type="pmc">4006905</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4006905</idno><idno type="RBID">PMC:4006905</idno><idno type="doi">10.1371/journal.pone.0096169</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">001201</idno><idno type="wicri:Area/Pmc/Curation">001200</idno></publicationStmt><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><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Pritchard, Hugh W" sort="Pritchard, Hugh W" uniqKey="Pritchard H" first="Hugh W." last="Pritchard">Hugh W. Pritchard</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><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></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Sakai, A" uniqKey="Sakai A">A Sakai</name></author><author><name sortKey="Fuller, Bj" uniqKey="Fuller B">BJ Fuller</name></author><author><name sortKey="Lane, N" uniqKey="Lane N">N Lane</name></author><author><name sortKey="Benson, Ee" uniqKey="Benson E">EE Benson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Grout, Bww" uniqKey="Grout B">BWW Grout</name></author><author><name sortKey="Shelton, K" uniqKey="Shelton K">K Shelton</name></author><author><name sortKey="Pritchard, Hw" uniqKey="Pritchard H">HW 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<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS One</journal-id><journal-id journal-id-type="iso-abbrev">PLoS ONE</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosone</journal-id><journal-title-group><journal-title>PLoS ONE</journal-title></journal-title-group><issn pub-type="epub">1932-6203</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">24788797</article-id><article-id pub-id-type="pmc">4006905</article-id><article-id pub-id-type="publisher-id">PONE-D-13-52950</article-id><article-id pub-id-type="doi">10.1371/journal.pone.0096169</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Biology and Life Sciences</subject><subj-group><subject>Biotechnology</subject><subj-group><subject>Plant Biotechnology</subject></subj-group></subj-group><subj-group><subject>Cell Biology</subject><subj-group><subject>Molecular Cell Biology</subject><subject>Plant Cell Biology</subject></subj-group></subj-group><subj-group><subject>Cryobiology</subject></subj-group><subj-group><subject>Plant Science</subject></subj-group></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Ecology and Environmental Sciences</subject><subj-group><subject>Conservation Science</subject></subj-group></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Research and Analysis Methods</subject><subj-group><subject>Model Organisms</subject><subj-group><subject>Plant and Algal Models</subject></subj-group></subj-group></subj-group></article-categories><title-group><article-title>Biophysical Characteristics of Successful Oilseed Embryo Cryoprotection and Cryopreservation Using Vacuum Infiltration Vitrification: An Innovation in Plant Cell Preservation</article-title><alt-title alt-title-type="running-head">Vacuum Infiltration Vitrification Cryoprotection</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Nadarajan</surname><given-names>Jayanthi</given-names></name><xref ref-type="aff" rid="aff1"></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib><contrib contrib-type="author"><name><surname>Pritchard</surname><given-names>Hugh W.</given-names></name><xref ref-type="aff" rid="aff1"></xref></contrib></contrib-group><aff id="aff1"><addr-line>Seed Conservation Department, Royal Botanic Gardens Kew, Wellcome Trust Millennium Building, Ardingly, West Sussex, United Kingdom</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>Heazlewood</surname><given-names>Joshua L.</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>Lawrence Berkeley National Laboratory, United States of America</addr-line></aff><author-notes><corresp id="cor1">* E-mail: <email>j.nadarajan@kew.org</email></corresp><fn fn-type="conflict"><p><bold>Competing Interests: </bold>The authors received funding from a commercial source, Orange Plc. However, this does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.</p></fn><fn fn-type="con"><p>Conceived and designed the experiments: JN HWP. Performed the experiments: JN. Analyzed the data: JN HWP. Contributed reagents/materials/analysis tools: JN HWP. Wrote the paper: JN HWP.</p></fn></author-notes><pub-date pub-type="collection"><year>2014</year></pub-date><pub-date pub-type="epub"><day>1</day><month>5</month><year>2014</year></pub-date><volume>9</volume><issue>5</issue><elocation-id>e96169</elocation-id><history><date date-type="received"><day>16</day><month>12</month><year>2013</year></date><date date-type="accepted"><day>3</day><month>4</month><year>2014</year></date></history><permissions><copyright-year>2014</copyright-year><copyright-holder>Nadarajan, Pritchard</copyright-holder><license><license-p>This is an open-access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><abstract><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></abstract><funding-group><funding-statement>This work was supported by the Millennium Commission, The Wellcome Trust and Orange plc. The Royal Botanic Gardens Kew receives grant-in-aid from DEFRA, UK. All funders acknowledged in our manuscript had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.</funding-statement></funding-group><counts><page-count count="11"></page-count></counts></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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