Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production
Identifieur interne : 000D32 ( Istex/Corpus ); précédent : 000D31; suivant : 000D33Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production
Auteurs : L. Chen ; D. BeversdorfSource :
- Plant Cell, Tissue and Organ Culture [ 0167-6857 ] ; 1992-11-01.
Abstract
Abstract: Microspore cryopreservation is a potentially powerful method for long-term storage of germplasm for in vitro embryo production in plant species. In this study, several factors influencing embryo production following the ultra-low temperature (−196 °C in liquid nitrogen) storage of isolated microspores of rapeseed (Brassica napus L.) were investigated. Microspores were prepared in cryogenic vials and subjected to various cooling treatments before immersion in liquid nitrogen for varying periods. Efficiency of microspore cryopreservation was reflected by in vitro embryo production from frozen microspores. Of all the cooling treatments, microspores treated with a cooling rate of 0.25% °C/min and a cooling terminal temperature of −35 °C before immersion in liquid nitrogen produced the highest embryo yields (18% and 40% of unfrozen controls in two genotypes, respectively). Fast thawing in a 35 °C water bath was necessary to recover a high number of embryos from microspore samples being frozen at a higher cooling rate, while thawing speed did not affect samples after freezing at a slower cooling rate. The storage density of cryopreserved microspores affected embryo production. Storage at the normal culture density (8×104 microspores/ml) was less efficient for embryo production than at high densities (4×106 microspores/ml and 1.6×107 microspores/ml), although no significant difference was found between the high densities. Evaluation of plant lines derived from frozen microspores indicated no variation in isozyme pattern and no enhanced cold tolerance of these lines. Isolated microspores of B. napus could be stored for extended period for in vitro embryo production.
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DOI: 10.1007/BF00037698
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production</title><author><name sortKey="Chen, L" sort="Chen, L" uniqKey="Chen L" first="L." last="Chen">L. Chen</name><affiliation><mods:affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</mods:affiliation></affiliation></author><author><name sortKey="Beversdorf, D" sort="Beversdorf, D" uniqKey="Beversdorf D" first="D." last="Beversdorf">D. Beversdorf</name><affiliation><mods:affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:BAC9606C7A3476A12590D593CE15226D70E6E2E1</idno><date when="1992" year="1992">1992</date><idno type="doi">10.1007/BF00037698</idno><idno type="url">https://api.istex.fr/document/BAC9606C7A3476A12590D593CE15226D70E6E2E1/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">000D32</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production</title><author><name sortKey="Chen, L" sort="Chen, L" uniqKey="Chen L" first="L." last="Chen">L. Chen</name><affiliation><mods:affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</mods:affiliation></affiliation></author><author><name sortKey="Beversdorf, D" sort="Beversdorf, D" uniqKey="Beversdorf D" first="D." last="Beversdorf">D. Beversdorf</name><affiliation><mods:affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Plant Cell, Tissue and Organ Culture</title><title level="j" type="sub">An International Journal on Biotechnology of Higher Plants</title><title level="j" type="abbrev">Plant Cell Tiss Organ Cult</title><idno type="ISSN">0167-6857</idno><idno type="eISSN">1573-5044</idno><imprint><publisher>Kluwer Academic Publishers</publisher><pubPlace>Dordrecht</pubPlace><date type="published" when="1992-11-01">1992-11-01</date><biblScope unit="volume">31</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="141">141</biblScope><biblScope unit="page" to="149">149</biblScope></imprint><idno type="ISSN">0167-6857</idno></series><idno type="istex">BAC9606C7A3476A12590D593CE15226D70E6E2E1</idno><idno type="DOI">10.1007/BF00037698</idno><idno type="ArticleID">BF00037698</idno><idno type="ArticleID">Art8</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-6857</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Microspore cryopreservation is a potentially powerful method for long-term storage of germplasm for in vitro embryo production in plant species. In this study, several factors influencing embryo production following the ultra-low temperature (−196 °C in liquid nitrogen) storage of isolated microspores of rapeseed (Brassica napus L.) were investigated. Microspores were prepared in cryogenic vials and subjected to various cooling treatments before immersion in liquid nitrogen for varying periods. Efficiency of microspore cryopreservation was reflected by in vitro embryo production from frozen microspores. Of all the cooling treatments, microspores treated with a cooling rate of 0.25% °C/min and a cooling terminal temperature of −35 °C before immersion in liquid nitrogen produced the highest embryo yields (18% and 40% of unfrozen controls in two genotypes, respectively). Fast thawing in a 35 °C water bath was necessary to recover a high number of embryos from microspore samples being frozen at a higher cooling rate, while thawing speed did not affect samples after freezing at a slower cooling rate. The storage density of cryopreserved microspores affected embryo production. Storage at the normal culture density (8×104 microspores/ml) was less efficient for embryo production than at high densities (4×106 microspores/ml and 1.6×107 microspores/ml), although no significant difference was found between the high densities. Evaluation of plant lines derived from frozen microspores indicated no variation in isozyme pattern and no enhanced cold tolerance of these lines. Isolated microspores of B. napus could be stored for extended period for in vitro embryo production.</div></front></TEI><istex><corpusName>springer</corpusName><author><json:item><name>J. L. Chen</name><affiliations><json:string>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</json:string></affiliations></json:item><json:item><name>W. D. Beversdorf</name><affiliations><json:string>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</json:string></affiliations></json:item></author><articleId><json:string>BF00037698</json:string><json:string>Art8</json:string></articleId><language><json:string>eng</json:string></language><originalGenre><json:string>OriginalPaper</json:string></originalGenre><abstract>Abstract: Microspore cryopreservation is a potentially powerful method for long-term storage of germplasm for in vitro embryo production in plant species. In this study, several factors influencing embryo production following the ultra-low temperature (−196 °C in liquid nitrogen) storage of isolated microspores of rapeseed (Brassica napus L.) were investigated. Microspores were prepared in cryogenic vials and subjected to various cooling treatments before immersion in liquid nitrogen for varying periods. Efficiency of microspore cryopreservation was reflected by in vitro embryo production from frozen microspores. Of all the cooling treatments, microspores treated with a cooling rate of 0.25% °C/min and a cooling terminal temperature of −35 °C before immersion in liquid nitrogen produced the highest embryo yields (18% and 40% of unfrozen controls in two genotypes, respectively). Fast thawing in a 35 °C water bath was necessary to recover a high number of embryos from microspore samples being frozen at a higher cooling rate, while thawing speed did not affect samples after freezing at a slower cooling rate. The storage density of cryopreserved microspores affected embryo production. Storage at the normal culture density (8×104 microspores/ml) was less efficient for embryo production than at high densities (4×106 microspores/ml and 1.6×107 microspores/ml), although no significant difference was found between the high densities. Evaluation of plant lines derived from frozen microspores indicated no variation in isozyme pattern and no enhanced cold tolerance of these lines. Isolated microspores of B. napus could be stored for extended period for in vitro embryo production.</abstract><qualityIndicators><score>7.257</score><pdfVersion>1.3</pdfVersion><pdfPageSize>540 x 735 pts</pdfPageSize><refBibsNative>false</refBibsNative><keywordCount>0</keywordCount><abstractCharCount>1696</abstractCharCount><pdfWordCount>4305</pdfWordCount><pdfCharCount>26971</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>246</abstractWordCount></qualityIndicators><title>Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production</title><genre><json:string>research-article</json:string></genre><host><volume>31</volume><pages><last>149</last><first>141</first></pages><issn><json:string>0167-6857</json:string></issn><issue>2</issue><subject><json:item><value>Plant Sciences</value></json:item><json:item><value>Plant Physiology</value></json:item></subject><journalId><json:string>11240</json:string></journalId><genre><json:string>journal</json:string></genre><language><json:string>unknown</json:string></language><eissn><json:string>1573-5044</json:string></eissn><title>Plant Cell, Tissue and Organ Culture</title><publicationDate>1992</publicationDate><copyrightDate>1992</copyrightDate></host><publicationDate>1992</publicationDate><copyrightDate>1992</copyrightDate><doi><json:string>10.1007/BF00037698</json:string></doi><id>BAC9606C7A3476A12590D593CE15226D70E6E2E1</id><score>0.23489869</score><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/BAC9606C7A3476A12590D593CE15226D70E6E2E1/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/BAC9606C7A3476A12590D593CE15226D70E6E2E1/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/BAC9606C7A3476A12590D593CE15226D70E6E2E1/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production</title><respStmt><resp>Références bibliographiques récupérées via GROBID</resp><name resp="ISTEX-API">ISTEX-API (INIST-CNRS)</name></respStmt></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>Kluwer Academic Publishers</publisher><pubPlace>Dordrecht</pubPlace><availability><p>Kluwer Academic Publishers, 1992</p></availability><date>1991-11-25</date></publicationStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production</title><author xml:id="author-1"><persName><forename type="first">J.</forename><surname>Chen</surname></persName><affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</affiliation></author><author xml:id="author-2"><persName><forename type="first">W.</forename><surname>Beversdorf</surname></persName><affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</affiliation></author></analytic><monogr><title level="j">Plant Cell, Tissue and Organ Culture</title><title level="j" type="sub">An International Journal on Biotechnology of Higher Plants</title><title level="j" type="abbrev">Plant Cell Tiss Organ Cult</title><idno type="journal-ID">11240</idno><idno type="pISSN">0167-6857</idno><idno type="eISSN">1573-5044</idno><idno type="issue-article-count">12</idno><idno type="volume-issue-count">3</idno><imprint><publisher>Kluwer Academic Publishers</publisher><pubPlace>Dordrecht</pubPlace><date type="published" when="1992-11-01"></date><biblScope unit="volume">31</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="141">141</biblScope><biblScope unit="page" to="149">149</biblScope></imprint></monogr><idno type="istex">BAC9606C7A3476A12590D593CE15226D70E6E2E1</idno><idno type="DOI">10.1007/BF00037698</idno><idno type="ArticleID">BF00037698</idno><idno type="ArticleID">Art8</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1991-11-25</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: Microspore cryopreservation is a potentially powerful method for long-term storage of germplasm for in vitro embryo production in plant species. In this study, several factors influencing embryo production following the ultra-low temperature (−196 °C in liquid nitrogen) storage of isolated microspores of rapeseed (Brassica napus L.) were investigated. Microspores were prepared in cryogenic vials and subjected to various cooling treatments before immersion in liquid nitrogen for varying periods. Efficiency of microspore cryopreservation was reflected by in vitro embryo production from frozen microspores. Of all the cooling treatments, microspores treated with a cooling rate of 0.25% °C/min and a cooling terminal temperature of −35 °C before immersion in liquid nitrogen produced the highest embryo yields (18% and 40% of unfrozen controls in two genotypes, respectively). Fast thawing in a 35 °C water bath was necessary to recover a high number of embryos from microspore samples being frozen at a higher cooling rate, while thawing speed did not affect samples after freezing at a slower cooling rate. The storage density of cryopreserved microspores affected embryo production. Storage at the normal culture density (8×104 microspores/ml) was less efficient for embryo production than at high densities (4×106 microspores/ml and 1.6×107 microspores/ml), although no significant difference was found between the high densities. Evaluation of plant lines derived from frozen microspores indicated no variation in isozyme pattern and no enhanced cold tolerance of these lines. Isolated microspores of B. napus could be stored for extended period for in vitro embryo production.</p></abstract><textClass><keywords scheme="Journal Subject"><list><head>Life Sciences</head><item><term>Plant Sciences</term></item><item><term>Plant Physiology</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1991-11-25">Created</change><change when="1992-11-01">Published</change><change xml:id="refBibs-istex" who="#ISTEX-API" when="2016-09-22">References added</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/BAC9606C7A3476A12590D593CE15226D70E6E2E1/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Springer, Publisher found" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="UTF-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//Springer-Verlag//DTD A++ V2.4//EN" URI="http://devel.springer.de/A++/V2.4/DTD/A++V2.4.dtd" name="istex:docType"></istex:docType><istex:document><Publisher><PublisherInfo><PublisherName>Kluwer Academic Publishers</PublisherName><PublisherLocation>Dordrecht</PublisherLocation></PublisherInfo><Journal><JournalInfo JournalProductType="ArchiveJournal" NumberingStyle="Unnumbered"><JournalID>11240</JournalID><JournalPrintISSN>0167-6857</JournalPrintISSN><JournalElectronicISSN>1573-5044</JournalElectronicISSN><JournalTitle>Plant Cell, Tissue and Organ Culture</JournalTitle><JournalSubTitle>An International Journal on Biotechnology of Higher Plants</JournalSubTitle><JournalAbbreviatedTitle>Plant Cell Tiss Organ Cult</JournalAbbreviatedTitle><JournalSubjectGroup><JournalSubject Type="Primary">Life Sciences</JournalSubject><JournalSubject Type="Secondary">Plant Sciences</JournalSubject><JournalSubject Type="Secondary">Plant Physiology</JournalSubject></JournalSubjectGroup></JournalInfo><Volume><VolumeInfo VolumeType="Regular" TocLevels="0"><VolumeIDStart>31</VolumeIDStart><VolumeIDEnd>31</VolumeIDEnd><VolumeIssueCount>3</VolumeIssueCount></VolumeInfo><Issue IssueType="Regular"><IssueInfo TocLevels="0"><IssueIDStart>2</IssueIDStart><IssueIDEnd>2</IssueIDEnd><IssueArticleCount>12</IssueArticleCount><IssueHistory><CoverDate><Year>1992</Year><Month>11</Month></CoverDate></IssueHistory><IssueCopyright><CopyrightHolderName>Kluwer Academic Publishers</CopyrightHolderName><CopyrightYear>1992</CopyrightYear></IssueCopyright></IssueInfo><Article ID="Art8"><ArticleInfo Language="En" ArticleType="OriginalPaper" NumberingStyle="Unnumbered" TocLevels="0" ContainsESM="No"><ArticleID>BF00037698</ArticleID><ArticleDOI>10.1007/BF00037698</ArticleDOI><ArticleSequenceNumber>8</ArticleSequenceNumber><ArticleTitle Language="En">Cryopreservation of isolated micropores of spring rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.) for <Emphasis Type="Italic">in vitro</Emphasis> embryo production</ArticleTitle><ArticleFirstPage>141</ArticleFirstPage><ArticleLastPage>149</ArticleLastPage><ArticleHistory><RegistrationDate><Year>2004</Year><Month>4</Month><Day>24</Day></RegistrationDate><Received><Year>1991</Year><Month>11</Month><Day>25</Day></Received><Accepted><Year>1992</Year><Month>3</Month><Day>31</Day></Accepted></ArticleHistory><ArticleCopyright><CopyrightHolderName>Kluwer Academic Publishers</CopyrightHolderName><CopyrightYear>1992</CopyrightYear></ArticleCopyright><ArticleGrants Type="Regular"><MetadataGrant Grant="OpenAccess"></MetadataGrant><AbstractGrant Grant="OpenAccess"></AbstractGrant><BodyPDFGrant Grant="Restricted"></BodyPDFGrant><BodyHTMLGrant Grant="Restricted"></BodyHTMLGrant><BibliographyGrant Grant="Restricted"></BibliographyGrant><ESMGrant Grant="Restricted"></ESMGrant></ArticleGrants><ArticleContext><JournalID>11240</JournalID><VolumeIDStart>31</VolumeIDStart><VolumeIDEnd>31</VolumeIDEnd><IssueIDStart>2</IssueIDStart><IssueIDEnd>2</IssueIDEnd></ArticleContext></ArticleInfo><ArticleHeader><AuthorGroup><Author AffiliationIDS="Aff1"><AuthorName DisplayOrder="Western"><GivenName>J.</GivenName><GivenName>L.</GivenName><FamilyName>Chen</FamilyName></AuthorName></Author><Author AffiliationIDS="Aff1"><AuthorName DisplayOrder="Western"><GivenName>W.</GivenName><GivenName>D.</GivenName><FamilyName>Beversdorf</FamilyName></AuthorName></Author><Affiliation ID="Aff1"><OrgDivision>Department of Crop Science</OrgDivision><OrgName>University of Guelph</OrgName><OrgAddress><Postcode>N1G 2W1</Postcode><State>Ontario</State><Country>Canada</Country></OrgAddress></Affiliation></AuthorGroup><Abstract ID="Abs1" Language="En"><Heading>Abstract</Heading><Para>Microspore cryopreservation is a potentially powerful method for long-term storage of germplasm for <Emphasis Type="Italic">in vitro</Emphasis> embryo production in plant species. In this study, several factors influencing embryo production following the ultra-low temperature (−196 °C in liquid nitrogen) storage of isolated microspores of rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.) were investigated. Microspores were prepared in cryogenic vials and subjected to various cooling treatments before immersion in liquid nitrogen for varying periods. Efficiency of microspore cryopreservation was reflected by <Emphasis Type="Italic">in vitro</Emphasis> embryo production from frozen microspores. Of all the cooling treatments, microspores treated with a cooling rate of 0.25% °C/min and a cooling terminal temperature of −35 °C before immersion in liquid nitrogen produced the highest embryo yields (18% and 40% of unfrozen controls in two genotypes, respectively). Fast thawing in a 35 °C water bath was necessary to recover a high number of embryos from microspore samples being frozen at a higher cooling rate, while thawing speed did not affect samples after freezing at a slower cooling rate. The storage density of cryopreserved microspores affected embryo production. Storage at the normal culture density (8×10<Superscript>4</Superscript> microspores/ml) was less efficient for embryo production than at high densities (4×10<Superscript>6</Superscript> microspores/ml and 1.6×10<Superscript>7</Superscript> microspores/ml), although no significant difference was found between the high densities. Evaluation of plant lines derived from frozen microspores indicated no variation in isozyme pattern and no enhanced cold tolerance of these lines. Isolated microspores of <Emphasis Type="Italic">B. napus</Emphasis> could be stored for extended period for <Emphasis Type="Italic">in vitro</Emphasis> embryo production.</Para></Abstract><KeywordGroup Language="En"><Heading>Key words</Heading><Keyword><Emphasis Type="Italic">Brassica napus</Emphasis></Keyword><Keyword>cryopreservation</Keyword><Keyword><Emphasis Type="Italic">in vitro</Emphasis> embryo</Keyword><Keyword>microspore</Keyword><Keyword>rapeseed</Keyword></KeywordGroup></ArticleHeader><NoBody></NoBody></Article></Issue></Volume></Journal></Publisher></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Cryopreservation of isolated micropores of spring rapeseed ( Brassica napus L.) for in vitro embryo production</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Cryopreservation of isolated micropores of spring rapeseed (Brassica napus L.) for in vitro embryo production</title></titleInfo><name type="personal"><namePart type="given">J.</namePart><namePart type="given">L.</namePart><namePart type="family">Chen</namePart><affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">W.</namePart><namePart type="given">D.</namePart><namePart type="family">Beversdorf</namePart><affiliation>Department of Crop Science, University of Guelph, N1G 2W1, Ontario, Canada</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="OriginalPaper"></genre><originInfo><publisher>Kluwer Academic Publishers</publisher><place><placeTerm type="text">Dordrecht</placeTerm></place><dateCreated encoding="w3cdtf">1991-11-25</dateCreated><dateIssued encoding="w3cdtf">1992-11-01</dateIssued><copyrightDate encoding="w3cdtf">1992</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">Abstract: Microspore cryopreservation is a potentially powerful method for long-term storage of germplasm for in vitro embryo production in plant species. In this study, several factors influencing embryo production following the ultra-low temperature (−196 °C in liquid nitrogen) storage of isolated microspores of rapeseed (Brassica napus L.) were investigated. Microspores were prepared in cryogenic vials and subjected to various cooling treatments before immersion in liquid nitrogen for varying periods. Efficiency of microspore cryopreservation was reflected by in vitro embryo production from frozen microspores. Of all the cooling treatments, microspores treated with a cooling rate of 0.25% °C/min and a cooling terminal temperature of −35 °C before immersion in liquid nitrogen produced the highest embryo yields (18% and 40% of unfrozen controls in two genotypes, respectively). Fast thawing in a 35 °C water bath was necessary to recover a high number of embryos from microspore samples being frozen at a higher cooling rate, while thawing speed did not affect samples after freezing at a slower cooling rate. The storage density of cryopreserved microspores affected embryo production. Storage at the normal culture density (8×104 microspores/ml) was less efficient for embryo production than at high densities (4×106 microspores/ml and 1.6×107 microspores/ml), although no significant difference was found between the high densities. Evaluation of plant lines derived from frozen microspores indicated no variation in isozyme pattern and no enhanced cold tolerance of these lines. Isolated microspores of B. napus could be stored for extended period for in vitro embryo production.</abstract><relatedItem type="host"><titleInfo><title>Plant Cell, Tissue and Organ Culture</title><subTitle>An International Journal on Biotechnology of Higher Plants</subTitle></titleInfo><titleInfo type="abbreviated"><title>Plant Cell Tiss Organ Cult</title></titleInfo><genre type="journal" displayLabel="Archive Journal"></genre><originInfo><dateIssued encoding="w3cdtf">1992-11-01</dateIssued><copyrightDate encoding="w3cdtf">1992</copyrightDate></originInfo><subject><genre>Life Sciences</genre><topic>Plant Sciences</topic><topic>Plant Physiology</topic></subject><identifier type="ISSN">0167-6857</identifier><identifier type="eISSN">1573-5044</identifier><identifier type="JournalID">11240</identifier><identifier type="IssueArticleCount">12</identifier><identifier type="VolumeIssueCount">3</identifier><part><date>1992</date><detail type="volume"><number>31</number><caption>vol.</caption></detail><detail type="issue"><number>2</number><caption>no.</caption></detail><extent unit="pages"><start>141</start><end>149</end></extent></part><recordInfo><recordOrigin>Kluwer Academic Publishers, 1992</recordOrigin></recordInfo></relatedItem><identifier type="istex">BAC9606C7A3476A12590D593CE15226D70E6E2E1</identifier><identifier type="DOI">10.1007/BF00037698</identifier><identifier type="ArticleID">BF00037698</identifier><identifier type="ArticleID">Art8</identifier><accessCondition type="use and reproduction" contentType="copyright">Kluwer Academic Publishers, 1992</accessCondition><recordInfo><recordContentSource>SPRINGER</recordContentSource><recordOrigin>Kluwer Academic Publishers, 1992</recordOrigin></recordInfo></mods></metadata><enrichments><json:item><type>multicat</type><uri>https://api.istex.fr/document/BAC9606C7A3476A12590D593CE15226D70E6E2E1/enrichments/multicat</uri></json:item><istex:refBibTEI uri="https://api.istex.fr/document/BAC9606C7A3476A12590D593CE15226D70E6E2E1/enrichments/refBib"><teiHeader></teiHeader><text><front></front><body></body><back><listBibl><biblStruct xml:id="b0"><analytic><title level="a" type="main">Regeneration of plants from pollenembryos of Arachis, Brassica and Triticum spp. cryopreserved for one year</title><author><persName><forename type="first">Yps</forename><surname>Bajaj</surname></persName></author></analytic><monogr><title level="j">Curr. 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