Simple methods to remove microbes from leaf surfaces.
Identifieur interne : 000118 ( Main/Exploration ); précédent : 000117; suivant : 000119Simple methods to remove microbes from leaf surfaces.
Auteurs : J Paola Saldierna Guzmán [États-Unis] ; Kennedy Nguyen [États-Unis] ; Stephen C. Hart [États-Unis]Source :
- Journal of basic microbiology [ 1521-4028 ] ; 2020.
Abstract
Endophytes have been defined as microorganisms living inside plant tissues without causing negative effects on their hosts. Endophytic microbes have been extensively studied for their plant growth-promoting traits. However, analyses of endophytes require complete removal of epiphytic microorganisms. We found that the established tests to evaluate surface sterility, polymerase chain reaction, and leaf imprints, are unreliable. Therefore, we used scanning electron microscopy (SEM) as an additional assessment of epiphyte removal. We used a diverse suite of sterilization protocols to remove epiphytic microorganisms from the leaves of a gymnosperm and an angiosperm tree to test the influence of leaf morphology on the efficacy of these methods. Additionally, leaf tissue damage was also evaluated by SEM, as damaging the leaves might have an impact on endophytes and could lead to inaccurate assessment of endophytic communities. Our study indicates, that complete removal of the leaf cuticle by the sterilization technique assures loss of epiphytic microbes, and that leaves of different tree species may require different sterilization protocols. Furthermore, our study demonstrates the importance of choosing the appropriate sterilization protocol to prevent erroneous interpretation of host-endophyte interactions. Moreover, it shows the utility of SEM for evaluating the effectiveness of surface sterilization methods and their impact on leaf tissue integrity.
DOI: 10.1002/jobm.202000035
PubMed: 32529642
Affiliations:
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Hart</name><affiliation wicri:level="2"><nlm:affiliation>Quantitative and Systems Biology, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Quantitative and Systems Biology, University of California, Merced</wicri:cityArea></affiliation><affiliation wicri:level="2"><nlm:affiliation>Sierra Nevada Research Institute, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Sierra Nevada Research Institute, University of California, Merced</wicri:cityArea></affiliation><affiliation wicri:level="2"><nlm:affiliation>Department of Life and Environmental Sciences, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Department of Life and Environmental Sciences, University of California, Merced</wicri:cityArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2020">2020</date><idno type="RBID">pubmed:32529642</idno><idno type="pmid">32529642</idno><idno type="doi">10.1002/jobm.202000035</idno><idno type="wicri:Area/Main/Corpus">000254</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000254</idno><idno type="wicri:Area/Main/Curation">000254</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000254</idno><idno type="wicri:Area/Main/Exploration">000254</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Simple methods to remove microbes from leaf surfaces.</title><author><name sortKey="Saldierna Guzman, J Paola" sort="Saldierna Guzman, J Paola" uniqKey="Saldierna Guzman J" first="J Paola" last="Saldierna Guzmán">J Paola Saldierna Guzmán</name><affiliation wicri:level="2"><nlm:affiliation>Quantitative and Systems Biology, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Quantitative and Systems Biology, University of California, Merced</wicri:cityArea></affiliation><affiliation wicri:level="2"><nlm:affiliation>Sierra Nevada Research Institute, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Sierra Nevada Research Institute, University of California, Merced</wicri:cityArea></affiliation></author><author><name sortKey="Nguyen, Kennedy" sort="Nguyen, Kennedy" uniqKey="Nguyen K" first="Kennedy" last="Nguyen">Kennedy Nguyen</name><affiliation wicri:level="2"><nlm:affiliation>Imaging and Microscopy Facility, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Imaging and Microscopy Facility, University of California, Merced</wicri:cityArea></affiliation></author><author><name sortKey="Hart, Stephen C" sort="Hart, Stephen C" uniqKey="Hart S" first="Stephen C" last="Hart">Stephen C. Hart</name><affiliation wicri:level="2"><nlm:affiliation>Quantitative and Systems Biology, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Quantitative and Systems Biology, University of California, Merced</wicri:cityArea></affiliation><affiliation wicri:level="2"><nlm:affiliation>Sierra Nevada Research Institute, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Sierra Nevada Research Institute, University of California, Merced</wicri:cityArea></affiliation><affiliation wicri:level="2"><nlm:affiliation>Department of Life and Environmental Sciences, University of California, Merced, California.</nlm:affiliation><country>États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Department of Life and Environmental Sciences, University of California, Merced</wicri:cityArea></affiliation></author></analytic><series><title level="j">Journal of basic microbiology</title><idno type="eISSN">1521-4028</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Endophytes have been defined as microorganisms living inside plant tissues without causing negative effects on their hosts. Endophytic microbes have been extensively studied for their plant growth-promoting traits. However, analyses of endophytes require complete removal of epiphytic microorganisms. We found that the established tests to evaluate surface sterility, polymerase chain reaction, and leaf imprints, are unreliable. Therefore, we used scanning electron microscopy (SEM) as an additional assessment of epiphyte removal. We used a diverse suite of sterilization protocols to remove epiphytic microorganisms from the leaves of a gymnosperm and an angiosperm tree to test the influence of leaf morphology on the efficacy of these methods. Additionally, leaf tissue damage was also evaluated by SEM, as damaging the leaves might have an impact on endophytes and could lead to inaccurate assessment of endophytic communities. Our study indicates, that complete removal of the leaf cuticle by the sterilization technique assures loss of epiphytic microbes, and that leaves of different tree species may require different sterilization protocols. Furthermore, our study demonstrates the importance of choosing the appropriate sterilization protocol to prevent erroneous interpretation of host-endophyte interactions. Moreover, it shows the utility of SEM for evaluating the effectiveness of surface sterilization methods and their impact on leaf tissue integrity.</div></front></TEI><pubmed><MedlineCitation Status="In-Process" Owner="NLM"><PMID Version="1">32529642</PMID><DateRevised><Year>2020</Year><Month>08</Month><Day>06</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1521-4028</ISSN><JournalIssue CitedMedium="Internet"><Volume>60</Volume><Issue>8</Issue><PubDate><Year>2020</Year><Month>Aug</Month></PubDate></JournalIssue><Title>Journal of basic microbiology</Title><ISOAbbreviation>J Basic Microbiol</ISOAbbreviation></Journal><ArticleTitle>Simple methods to remove microbes from leaf surfaces.</ArticleTitle><Pagination><MedlinePgn>730-734</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1002/jobm.202000035</ELocationID><Abstract><AbstractText>Endophytes have been defined as microorganisms living inside plant tissues without causing negative effects on their hosts. Endophytic microbes have been extensively studied for their plant growth-promoting traits. However, analyses of endophytes require complete removal of epiphytic microorganisms. We found that the established tests to evaluate surface sterility, polymerase chain reaction, and leaf imprints, are unreliable. Therefore, we used scanning electron microscopy (SEM) as an additional assessment of epiphyte removal. We used a diverse suite of sterilization protocols to remove epiphytic microorganisms from the leaves of a gymnosperm and an angiosperm tree to test the influence of leaf morphology on the efficacy of these methods. Additionally, leaf tissue damage was also evaluated by SEM, as damaging the leaves might have an impact on endophytes and could lead to inaccurate assessment of endophytic communities. Our study indicates, that complete removal of the leaf cuticle by the sterilization technique assures loss of epiphytic microbes, and that leaves of different tree species may require different sterilization protocols. Furthermore, our study demonstrates the importance of choosing the appropriate sterilization protocol to prevent erroneous interpretation of host-endophyte interactions. Moreover, it shows the utility of SEM for evaluating the effectiveness of surface sterilization methods and their impact on leaf tissue integrity.</AbstractText><CopyrightInformation>© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Saldierna Guzmán</LastName><ForeName>J Paola</ForeName><Initials>JP</Initials><Identifier Source="ORCID">http://orcid.org/0000-0003-1728-8154</Identifier><AffiliationInfo><Affiliation>Quantitative and Systems Biology, University of California, Merced, California.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Sierra Nevada Research Institute, University of California, Merced, California.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Nguyen</LastName><ForeName>Kennedy</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Imaging and Microscopy Facility, University of California, Merced, California.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hart</LastName><ForeName>Stephen C</ForeName><Initials>SC</Initials><AffiliationInfo><Affiliation>Quantitative and Systems Biology, University of California, Merced, California.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Sierra Nevada Research Institute, University of California, Merced, California.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Department of Life and Environmental Sciences, University of California, Merced, California.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>NNX15AQ01A</GrantID><Agency>NASA MIRO</Agency><Country></Country></Grant><Grant><Agency>UC MEXUS-CONACYT Doctoral Fellowship</Agency><Country></Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>06</Month><Day>11</Day></ArticleDate></Article><MedlineJournalInfo><Country>Germany</Country><MedlineTA>J Basic Microbiol</MedlineTA><NlmUniqueID>8503885</NlmUniqueID><ISSNLinking>0233-111X</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Pinus contorta</Keyword><Keyword MajorTopicYN="N">Populus fremontii</Keyword><Keyword MajorTopicYN="N">endophyte</Keyword><Keyword MajorTopicYN="N">leaf surface sterilization</Keyword><Keyword MajorTopicYN="N">scanning electron microscopy (SEM)</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2020</Year><Month>01</Month><Day>22</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2020</Year><Month>04</Month><Day>12</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2020</Year><Month>05</Month><Day>10</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">32529642</ArticleId><ArticleId IdType="doi">10.1002/jobm.202000035</ArticleId></ArticleIdList><ReferenceList><Title>REFERENCES</Title><Reference><Citation>Wilson D. 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Introduction to the scanning electron microscope: Theory, practice, & procedures. Facility for advanced instrumentation. Sacramento, CA: University of California, Davis; 1997. p. 1-51.</Citation></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country><region><li>Californie</li></region></list><tree><country name="États-Unis"><region name="Californie"><name sortKey="Saldierna Guzman, J Paola" sort="Saldierna Guzman, J Paola" uniqKey="Saldierna Guzman J" first="J Paola" last="Saldierna Guzmán">J Paola Saldierna Guzmán</name></region><name sortKey="Hart, Stephen C" sort="Hart, Stephen C" uniqKey="Hart S" first="Stephen C" last="Hart">Stephen C. Hart</name><name sortKey="Hart, Stephen C" sort="Hart, Stephen C" uniqKey="Hart S" first="Stephen C" last="Hart">Stephen C. Hart</name><name sortKey="Hart, Stephen C" sort="Hart, Stephen C" uniqKey="Hart S" first="Stephen C" last="Hart">Stephen C. Hart</name><name sortKey="Nguyen, Kennedy" sort="Nguyen, Kennedy" uniqKey="Nguyen K" first="Kennedy" last="Nguyen">Kennedy Nguyen</name><name sortKey="Saldierna Guzman, J Paola" sort="Saldierna Guzman, J Paola" uniqKey="Saldierna Guzman J" first="J Paola" last="Saldierna Guzmán">J Paola Saldierna Guzmán</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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