A modified perlite protocol with a mixed dimethyl sulfoxide and trehalose cryoprotectant improves the viability of frozen cultures of ectomycorrhizal basidiomycetes.
Identifieur interne : 000267 ( Main/Exploration ); précédent : 000266; suivant : 000268A modified perlite protocol with a mixed dimethyl sulfoxide and trehalose cryoprotectant improves the viability of frozen cultures of ectomycorrhizal basidiomycetes.
Auteurs : Masanori Sato [Japon] ; Shigeki Inaba [Japon] ; Junji Sukenobe [Japon] ; Tomomi Sasaki [Japon] ; Ryutaro Inoue [Japon] ; Mariko Noguchi [Japon] ; Akira Nakagiri [Japon]Source :
- Mycologia [ 1557-2536 ]
Descripteurs français
- KwdFr :
- Basidiomycota (effets des médicaments et des substances chimiques), Basidiomycota (physiologie), Congélation (MeSH), Cryoconservation (méthodes), Cryoprotecteurs (pharmacologie), Diméthylsulfoxyde (pharmacologie), Mycorhizes (effets des médicaments et des substances chimiques), Mycorhizes (physiologie), Oxyde d'aluminium (pharmacologie), Phylogenèse (MeSH), Silice (pharmacologie), Tréhalose (pharmacologie), Viabilité microbienne (MeSH).
- MESH :
- effets des médicaments et des substances chimiques : Basidiomycota, Mycorhizes.
- méthodes : Cryoconservation.
- pharmacologie : Cryoprotecteurs, Diméthylsulfoxyde, Oxyde d'aluminium, Silice, Tréhalose.
- physiologie : Basidiomycota, Mycorhizes.
- Congélation, Phylogenèse, Viabilité microbienne.
English descriptors
- KwdEn :
- Aluminum Oxide (pharmacology), Basidiomycota (drug effects), Basidiomycota (physiology), Cryopreservation (methods), Cryoprotective Agents (pharmacology), Dimethyl Sulfoxide (pharmacology), Freezing (MeSH), Microbial Viability (MeSH), Mycorrhizae (drug effects), Mycorrhizae (physiology), Phylogeny (MeSH), Silicon Dioxide (pharmacology), Trehalose (pharmacology).
- MESH :
- chemical , pharmacology : Aluminum Oxide, Cryoprotective Agents, Dimethyl Sulfoxide, Silicon Dioxide, Trehalose.
- drug effects : Basidiomycota, Mycorrhizae.
- methods : Cryopreservation.
- physiology : Basidiomycota, Mycorrhizae.
- Freezing, Microbial Viability, Phylogeny.
Abstract
Homolka's perlite protocol (HPP) was evaluated for cryopreservation of a wide range of ectomycorrhizal basidiomycete cultures, then a modified perlite protocol (MPP), in which cryoprotectant was added just before freezing rather than during the culturing process, was applied to cryosensitive strains that failed to survive when HPP was used. Further modifications of MPP with various cryoprotectants were explored to improve the cryopreservation of hard-to-preserve strains. The efficacy of HPP was assessed in 111 strains of 38 species of basidiomycetes of various cryosensitivities. After freezing strains using HPP, the viability and colony diameter of the strains were examined after 2 wk, 6 mo, and 1 y of storage at -80 C. Of the 111 strains tested, 91 survived after 1 y of storage with high viability of 80% or more, whereas the remaining 20 strains exhibited low and unstable viability. For those selected cryosensitive strains that did not survive well when HPP was used, MPP was applied with a mixture of cryoprotectants, dimethyl sulfoxide (DMSO), glycerol, and trehalose, at different concentrations and combinations. Toxicity testing of the cryoprotectants in the nonfrozen state revealed that 12% (v/v) glycerol was highly toxic for six strains (four species), whereas DMSO (5% and 10% [v/v]) was less toxic than glycerol. The viability of the cryosensitive strains after freezing demonstrated that DMSO was more efficient than glycerol, and trehalose enhanced the cryoprotective effects of both glycerol and DMSO when MPP was used for cryopreservation. Our comparative analysis of MPP with various combinations and concentrations of cryoprotectants revealed that a mixture of 5% DMSO and 10% trehalose was the most effective cryoprotectant, and that using MPP with this cryoprotectant was applicable to many cryosensitive strains.
DOI: 10.1080/00275514.2018.1520035
PubMed: 30714878
Affiliations:
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Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 </wicri:regionArea><wicri:noRegion>292-0818 </wicri:noRegion></affiliation></author><author><name sortKey="Sasaki, Tomomi" sort="Sasaki, Tomomi" uniqKey="Sasaki T" first="Tomomi" last="Sasaki">Tomomi Sasaki</name><affiliation wicri:level="1"><nlm:affiliation>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 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2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 </wicri:regionArea><wicri:noRegion>292-0818 </wicri:noRegion></affiliation></author><author><name sortKey="Nakagiri, Akira" sort="Nakagiri, Akira" uniqKey="Nakagiri A" first="Akira" last="Nakagiri">Akira Nakagiri</name><affiliation wicri:level="1"><nlm:affiliation>c Fungus/Mushroom Resource and Research Center, Faculty of Agriculture , Tottori University , 4-101 Koyama-Minami, Tottori 680-8553 , Japan.</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>c Fungus/Mushroom Resource and Research Center, Faculty of Agriculture , Tottori University , 4-101 Koyama-Minami, Tottori 680-8553 </wicri:regionArea><wicri:noRegion>Tottori 680-8553 </wicri:noRegion></affiliation></author></analytic><series><title level="j">Mycologia</title><idno type="eISSN">1557-2536</idno></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Aluminum Oxide (pharmacology)</term><term>Basidiomycota (drug effects)</term><term>Basidiomycota (physiology)</term><term>Cryopreservation (methods)</term><term>Cryoprotective Agents (pharmacology)</term><term>Dimethyl Sulfoxide (pharmacology)</term><term>Freezing (MeSH)</term><term>Microbial Viability (MeSH)</term><term>Mycorrhizae (drug effects)</term><term>Mycorrhizae (physiology)</term><term>Phylogeny (MeSH)</term><term>Silicon Dioxide (pharmacology)</term><term>Trehalose (pharmacology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Basidiomycota (effets des médicaments et des substances chimiques)</term><term>Basidiomycota (physiologie)</term><term>Congélation (MeSH)</term><term>Cryoconservation (méthodes)</term><term>Cryoprotecteurs (pharmacologie)</term><term>Diméthylsulfoxyde (pharmacologie)</term><term>Mycorhizes (effets des médicaments et des substances chimiques)</term><term>Mycorhizes (physiologie)</term><term>Oxyde d'aluminium (pharmacologie)</term><term>Phylogenèse (MeSH)</term><term>Silice (pharmacologie)</term><term>Tréhalose (pharmacologie)</term><term>Viabilité microbienne (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Aluminum Oxide</term><term>Cryoprotective Agents</term><term>Dimethyl Sulfoxide</term><term>Silicon Dioxide</term><term>Trehalose</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Basidiomycota</term><term>Mycorrhizae</term></keywords><keywords scheme="MESH" qualifier="effets des médicaments et des substances chimiques" xml:lang="fr"><term>Basidiomycota</term><term>Mycorhizes</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Cryopreservation</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Cryoconservation</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Cryoprotecteurs</term><term>Diméthylsulfoxyde</term><term>Oxyde d'aluminium</term><term>Silice</term><term>Tréhalose</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Basidiomycota</term><term>Mycorhizes</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Basidiomycota</term><term>Mycorrhizae</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Freezing</term><term>Microbial Viability</term><term>Phylogeny</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Congélation</term><term>Phylogenèse</term><term>Viabilité microbienne</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Homolka's perlite protocol (HPP) was evaluated for cryopreservation of a wide range of ectomycorrhizal basidiomycete cultures, then a modified perlite protocol (MPP), in which cryoprotectant was added just before freezing rather than during the culturing process, was applied to cryosensitive strains that failed to survive when HPP was used. Further modifications of MPP with various cryoprotectants were explored to improve the cryopreservation of hard-to-preserve strains. The efficacy of HPP was assessed in 111 strains of 38 species of basidiomycetes of various cryosensitivities. After freezing strains using HPP, the viability and colony diameter of the strains were examined after 2 wk, 6 mo, and 1 y of storage at -80 C. Of the 111 strains tested, 91 survived after 1 y of storage with high viability of 80% or more, whereas the remaining 20 strains exhibited low and unstable viability. For those selected cryosensitive strains that did not survive well when HPP was used, MPP was applied with a mixture of cryoprotectants, dimethyl sulfoxide (DMSO), glycerol, and trehalose, at different concentrations and combinations. Toxicity testing of the cryoprotectants in the nonfrozen state revealed that 12% (v/v) glycerol was highly toxic for six strains (four species), whereas DMSO (5% and 10% [v/v]) was less toxic than glycerol. The viability of the cryosensitive strains after freezing demonstrated that DMSO was more efficient than glycerol, and trehalose enhanced the cryoprotective effects of both glycerol and DMSO when MPP was used for cryopreservation. Our comparative analysis of MPP with various combinations and concentrations of cryoprotectants revealed that a mixture of 5% DMSO and 10% trehalose was the most effective cryoprotectant, and that using MPP with this cryoprotectant was applicable to many cryosensitive strains.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">30714878</PMID><DateCompleted><Year>2019</Year><Month>05</Month><Day>02</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>02</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1557-2536</ISSN><JournalIssue CitedMedium="Internet"><Volume>111</Volume><Issue>1</Issue><PubDate><MedlineDate>2019 Jan-Feb</MedlineDate></PubDate></JournalIssue><Title>Mycologia</Title><ISOAbbreviation>Mycologia</ISOAbbreviation></Journal><ArticleTitle>A modified perlite protocol with a mixed dimethyl sulfoxide and trehalose cryoprotectant improves the viability of frozen cultures of ectomycorrhizal basidiomycetes.</ArticleTitle><Pagination><MedlinePgn>161-176</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1080/00275514.2018.1520035</ELocationID><Abstract><AbstractText>Homolka's perlite protocol (HPP) was evaluated for cryopreservation of a wide range of ectomycorrhizal basidiomycete cultures, then a modified perlite protocol (MPP), in which cryoprotectant was added just before freezing rather than during the culturing process, was applied to cryosensitive strains that failed to survive when HPP was used. Further modifications of MPP with various cryoprotectants were explored to improve the cryopreservation of hard-to-preserve strains. The efficacy of HPP was assessed in 111 strains of 38 species of basidiomycetes of various cryosensitivities. After freezing strains using HPP, the viability and colony diameter of the strains were examined after 2 wk, 6 mo, and 1 y of storage at -80 C. Of the 111 strains tested, 91 survived after 1 y of storage with high viability of 80% or more, whereas the remaining 20 strains exhibited low and unstable viability. For those selected cryosensitive strains that did not survive well when HPP was used, MPP was applied with a mixture of cryoprotectants, dimethyl sulfoxide (DMSO), glycerol, and trehalose, at different concentrations and combinations. Toxicity testing of the cryoprotectants in the nonfrozen state revealed that 12% (v/v) glycerol was highly toxic for six strains (four species), whereas DMSO (5% and 10% [v/v]) was less toxic than glycerol. The viability of the cryosensitive strains after freezing demonstrated that DMSO was more efficient than glycerol, and trehalose enhanced the cryoprotective effects of both glycerol and DMSO when MPP was used for cryopreservation. Our comparative analysis of MPP with various combinations and concentrations of cryoprotectants revealed that a mixture of 5% DMSO and 10% trehalose was the most effective cryoprotectant, and that using MPP with this cryoprotectant was applicable to many cryosensitive strains.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Sato</LastName><ForeName>Masanori</ForeName><Initials>M</Initials><Identifier Source="ORCID">0000-0002-0568-6756</Identifier><AffiliationInfo><Affiliation>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Inaba</LastName><ForeName>Shigeki</ForeName><Initials>S</Initials><Identifier Source="ORCID">0000-0003-2741-9752</Identifier><AffiliationInfo><Affiliation>b Biological Resource Center, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sukenobe</LastName><ForeName>Junji</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sasaki</LastName><ForeName>Tomomi</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Inoue</LastName><ForeName>Ryutaro</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Noguchi</LastName><ForeName>Mariko</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>a International Patent Organism Depositary, National Institute of Technology and Evaluation , 2-5-8, Kazusakamatari, Kisarazu-shi , Chiba , 292-0818 , Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Nakagiri</LastName><ForeName>Akira</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>c Fungus/Mushroom Resource and Research Center, Faculty of Agriculture , Tottori University , 4-101 Koyama-Minami, Tottori 680-8553 , Japan.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>02</Month><Day>04</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Mycologia</MedlineTA><NlmUniqueID>0400764</NlmUniqueID><ISSNLinking>0027-5514</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D003451">Cryoprotective Agents</NameOfSubstance></Chemical><Chemical><RegistryNumber>0SG101ZGK9</RegistryNumber><NameOfSubstance UI="C003076">Perlite</NameOfSubstance></Chemical><Chemical><RegistryNumber>7631-86-9</RegistryNumber><NameOfSubstance UI="D012822">Silicon Dioxide</NameOfSubstance></Chemical><Chemical><RegistryNumber>B8WCK70T7I</RegistryNumber><NameOfSubstance UI="D014199">Trehalose</NameOfSubstance></Chemical><Chemical><RegistryNumber>LMI26O6933</RegistryNumber><NameOfSubstance UI="D000537">Aluminum Oxide</NameOfSubstance></Chemical><Chemical><RegistryNumber>YOW8V9698H</RegistryNumber><NameOfSubstance UI="D004121">Dimethyl Sulfoxide</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000537" MajorTopicYN="N">Aluminum Oxide</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="N">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D001487" MajorTopicYN="N">Basidiomycota</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="Y">drug effects</QualifierName><QualifierName UI="Q000502" MajorTopicYN="N">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015925" MajorTopicYN="N">Cryopreservation</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D003451" MajorTopicYN="N">Cryoprotective Agents</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="Y">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004121" MajorTopicYN="N">Dimethyl Sulfoxide</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="Y">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005615" MajorTopicYN="N">Freezing</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D050296" MajorTopicYN="N">Microbial Viability</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D038821" MajorTopicYN="N">Mycorrhizae</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="Y">drug effects</QualifierName><QualifierName UI="Q000502" MajorTopicYN="N">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010802" MajorTopicYN="N">Phylogeny</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012822" MajorTopicYN="N">Silicon Dioxide</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="N">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014199" MajorTopicYN="N">Trehalose</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="N">pharmacology</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">Basidiomycete</Keyword><Keyword MajorTopicYN="Y">Homolka’s perlite protocol</Keyword><Keyword MajorTopicYN="Y">cryopreservation</Keyword><Keyword MajorTopicYN="Y">hard-to-preserve fungal culture</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>2</Month><Day>5</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2019</Year><Month>5</Month><Day>3</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2019</Year><Month>2</Month><Day>5</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">30714878</ArticleId><ArticleId IdType="doi">10.1080/00275514.2018.1520035</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Japon</li></country></list><tree><country name="Japon"><noRegion><name sortKey="Sato, Masanori" sort="Sato, Masanori" uniqKey="Sato M" first="Masanori" last="Sato">Masanori Sato</name></noRegion><name sortKey="Inaba, Shigeki" sort="Inaba, Shigeki" uniqKey="Inaba S" first="Shigeki" last="Inaba">Shigeki Inaba</name><name sortKey="Inoue, Ryutaro" sort="Inoue, Ryutaro" uniqKey="Inoue R" first="Ryutaro" last="Inoue">Ryutaro Inoue</name><name sortKey="Nakagiri, Akira" sort="Nakagiri, Akira" uniqKey="Nakagiri A" first="Akira" last="Nakagiri">Akira Nakagiri</name><name sortKey="Noguchi, Mariko" sort="Noguchi, Mariko" uniqKey="Noguchi M" first="Mariko" last="Noguchi">Mariko Noguchi</name><name sortKey="Sasaki, Tomomi" sort="Sasaki, Tomomi" uniqKey="Sasaki T" first="Tomomi" last="Sasaki">Tomomi Sasaki</name><name sortKey="Sukenobe, Junji" sort="Sukenobe, Junji" uniqKey="Sukenobe J" first="Junji" last="Sukenobe">Junji Sukenobe</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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