Establishment of a simple and efficient Agrobacterium-mediated transformation system for Phytophthora palmivora.
Identifieur interne : 000C45 ( Main/Exploration ); précédent : 000C44; suivant : 000C46Establishment of a simple and efficient Agrobacterium-mediated transformation system for Phytophthora palmivora.
Auteurs : Dongliang Wu [États-Unis] ; Natasha Navet [États-Unis] ; Yingchao Liu [États-Unis, République populaire de Chine] ; Janice Uchida [États-Unis] ; Miaoying Tian [États-Unis]Source :
- BMC microbiology [ 1471-2180 ] ; 2016.
Descripteurs français
- KwdFr :
- ADN bactérien (MeSH), ADN des protozoaires (MeSH), Agrobacterium (génétique), Biologie moléculaire (méthodes), Chlorure de calcium (MeSH), Kanamycin kinase (MeSH), Microscopie de fluorescence (MeSH), Oomycetes (génétique), Phytophthora (microbiologie), Plantes (microbiologie), Plantes (parasitologie), Plasmides (MeSH), Polyéthylène glycols (MeSH), Protoplastes (MeSH), Protéines à fluorescence verte (MeSH), Reproductibilité des résultats (MeSH), Régulation de l'expression des gènes (MeSH), Séquence nucléotidique (MeSH), Techniques de transfert de gènes (MeSH), Transformation génétique (génétique), Vecteurs génétiques (MeSH), Électroporation (méthodes), Éléments transposables d'ADN (MeSH).
- MESH :
- génétique : Agrobacterium, Oomycetes, Transformation génétique.
- microbiologie : Phytophthora, Plantes.
- méthodes : Biologie moléculaire, Électroporation.
- parasitologie : Plantes.
- ADN bactérien, ADN des protozoaires, Chlorure de calcium, Kanamycin kinase, Microscopie de fluorescence, Plasmides, Polyéthylène glycols, Protoplastes, Protéines à fluorescence verte, Reproductibilité des résultats, Régulation de l'expression des gènes, Séquence nucléotidique, Techniques de transfert de gènes, Vecteurs génétiques, Éléments transposables d'ADN.
English descriptors
- KwdEn :
- Agrobacterium (genetics), Base Sequence (MeSH), Calcium Chloride (MeSH), DNA Transposable Elements (MeSH), DNA, Bacterial (MeSH), DNA, Protozoan (MeSH), Electroporation (methods), Gene Expression Regulation (MeSH), Gene Transfer Techniques (MeSH), Genetic Vectors (MeSH), Green Fluorescent Proteins (MeSH), Kanamycin Kinase (MeSH), Microscopy, Fluorescence (MeSH), Molecular Biology (methods), Oomycetes (genetics), Phytophthora (microbiology), Plants (microbiology), Plants (parasitology), Plasmids (MeSH), Polyethylene Glycols (MeSH), Protoplasts (MeSH), Reproducibility of Results (MeSH), Transformation, Genetic (genetics).
- MESH :
- chemical : Calcium Chloride, DNA Transposable Elements, DNA, Bacterial, DNA, Protozoan, Green Fluorescent Proteins, Kanamycin Kinase, Polyethylene Glycols.
- genetics : Agrobacterium, Oomycetes, Transformation, Genetic.
- methods : Electroporation, Molecular Biology.
- microbiology : Phytophthora, Plants.
- parasitology : Plants.
- Base Sequence, Gene Expression Regulation, Gene Transfer Techniques, Genetic Vectors, Microscopy, Fluorescence, Plasmids, Protoplasts, Reproducibility of Results.
Abstract
BACKGROUND
As an agriculturally important oomycete genus, Phytophthora contains a large number of destructive plant pathogens that severely threaten agricultural production and natural ecosystems. Among them is the broad host range pathogen P. palmivora, which infects many economically important plant species. An essential way to dissect their pathogenesis mechanisms is genetic modification of candidate genes, which requires effective transformation systems. Four methods were developed for transformation of Phytophthora spp., including PEG(polyethylene glycol)/CaCl2 mediated protoplast transformation, electroporation of zoospores, microprojectile bombardment and Agrobacterium-mediated transformation (AMT). Among them, AMT has many advantages over the other methods such as easy handling and mainly generating single-copy integration in the genome. An AMT method previously reported for P. infestans and P. palmivora has barely been used in oomycete research due to low success and low reproducibility.
RESULTS
In this study, we report a simple and efficient AMT system for P. palmivora. Using this system, we were able to reproducibly generate over 40 transformants using zoospores collected from culture grown in a single 100 mm-diameter petri dish. The generated GFP transformants constitutively expressed GFP readily detectable using a fluorescence microscope. All of the transformants tested using Southern blot analysis contained a single-copy T-DNA insertion.
CONCLUSIONS
This system is highly effective and reproducible for transformation of P. palmivora and expected to be adaptable for transformation of additional Phytophthora spp. and other oomycetes. Its establishment will greatly accelerate their functional genomic studies.
DOI: 10.1186/s12866-016-0825-1
PubMed: 27599726
PubMed Central: PMC5012004
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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(microbiology)</term><term>Plants (parasitology)</term><term>Plasmids (MeSH)</term><term>Polyethylene Glycols (MeSH)</term><term>Protoplasts (MeSH)</term><term>Reproducibility of Results (MeSH)</term><term>Transformation, Genetic (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN bactérien (MeSH)</term><term>ADN des protozoaires (MeSH)</term><term>Agrobacterium (génétique)</term><term>Biologie moléculaire (méthodes)</term><term>Chlorure de calcium (MeSH)</term><term>Kanamycin kinase (MeSH)</term><term>Microscopie de fluorescence (MeSH)</term><term>Oomycetes (génétique)</term><term>Phytophthora (microbiologie)</term><term>Plantes (microbiologie)</term><term>Plantes (parasitologie)</term><term>Plasmides (MeSH)</term><term>Polyéthylène glycols (MeSH)</term><term>Protoplastes (MeSH)</term><term>Protéines à fluorescence verte (MeSH)</term><term>Reproductibilité des résultats (MeSH)</term><term>Régulation de l'expression des gènes (MeSH)</term><term>Séquence nucléotidique (MeSH)</term><term>Techniques de transfert de gènes (MeSH)</term><term>Transformation génétique (génétique)</term><term>Vecteurs génétiques (MeSH)</term><term>Électroporation (méthodes)</term><term>Éléments transposables d'ADN (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Calcium Chloride</term><term>DNA Transposable Elements</term><term>DNA, Bacterial</term><term>DNA, Protozoan</term><term>Green Fluorescent Proteins</term><term>Kanamycin Kinase</term><term>Polyethylene Glycols</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Agrobacterium</term><term>Oomycetes</term><term>Transformation, Genetic</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Agrobacterium</term><term>Oomycetes</term><term>Transformation génétique</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Electroporation</term><term>Molecular Biology</term></keywords><keywords scheme="MESH" qualifier="microbiologie" xml:lang="fr"><term>Phytophthora</term><term>Plantes</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Phytophthora</term><term>Plants</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Biologie moléculaire</term><term>Électroporation</term></keywords><keywords scheme="MESH" qualifier="parasitologie" xml:lang="fr"><term>Plantes</term></keywords><keywords scheme="MESH" qualifier="parasitology" xml:lang="en"><term>Plants</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Gene Expression Regulation</term><term>Gene Transfer Techniques</term><term>Genetic Vectors</term><term>Microscopy, Fluorescence</term><term>Plasmids</term><term>Protoplasts</term><term>Reproducibility of Results</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ADN bactérien</term><term>ADN des protozoaires</term><term>Chlorure de calcium</term><term>Kanamycin kinase</term><term>Microscopie de fluorescence</term><term>Plasmides</term><term>Polyéthylène glycols</term><term>Protoplastes</term><term>Protéines à fluorescence verte</term><term>Reproductibilité des résultats</term><term>Régulation de l'expression des gènes</term><term>Séquence nucléotidique</term><term>Techniques de transfert de gènes</term><term>Vecteurs génétiques</term><term>Éléments transposables d'ADN</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b></p><p>As an agriculturally important oomycete genus, Phytophthora contains a large number of destructive plant pathogens that severely threaten agricultural production and natural ecosystems. Among them is the broad host range pathogen P. palmivora, which infects many economically important plant species. An essential way to dissect their pathogenesis mechanisms is genetic modification of candidate genes, which requires effective transformation systems. Four methods were developed for transformation of Phytophthora spp., including PEG(polyethylene glycol)/CaCl2 mediated protoplast transformation, electroporation of zoospores, microprojectile bombardment and Agrobacterium-mediated transformation (AMT). Among them, AMT has many advantages over the other methods such as easy handling and mainly generating single-copy integration in the genome. An AMT method previously reported for P. infestans and P. palmivora has barely been used in oomycete research due to low success and low reproducibility.</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>In this study, we report a simple and efficient AMT system for P. palmivora. Using this system, we were able to reproducibly generate over 40 transformants using zoospores collected from culture grown in a single 100 mm-diameter petri dish. The generated GFP transformants constitutively expressed GFP readily detectable using a fluorescence microscope. All of the transformants tested using Southern blot analysis contained a single-copy T-DNA insertion.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSIONS</b></p><p>This system is highly effective and reproducible for transformation of P. palmivora and expected to be adaptable for transformation of additional Phytophthora spp. and other oomycetes. Its establishment will greatly accelerate their functional genomic studies.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">27599726</PMID><DateCompleted><Year>2017</Year><Month>08</Month><Day>31</Day></DateCompleted><DateRevised><Year>2018</Year><Month>12</Month><Day>02</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1471-2180</ISSN><JournalIssue CitedMedium="Internet"><Volume>16</Volume><Issue>1</Issue><PubDate><Year>2016</Year><Month>09</Month><Day>06</Day></PubDate></JournalIssue><Title>BMC microbiology</Title><ISOAbbreviation>BMC Microbiol</ISOAbbreviation></Journal><ArticleTitle>Establishment of a simple and efficient Agrobacterium-mediated transformation system for Phytophthora palmivora.</ArticleTitle><Pagination><MedlinePgn>204</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1186/s12866-016-0825-1</ELocationID><Abstract><AbstractText Label="BACKGROUND">As an agriculturally important oomycete genus, Phytophthora contains a large number of destructive plant pathogens that severely threaten agricultural production and natural ecosystems. Among them is the broad host range pathogen P. palmivora, which infects many economically important plant species. An essential way to dissect their pathogenesis mechanisms is genetic modification of candidate genes, which requires effective transformation systems. Four methods were developed for transformation of Phytophthora spp., including PEG(polyethylene glycol)/CaCl2 mediated protoplast transformation, electroporation of zoospores, microprojectile bombardment and Agrobacterium-mediated transformation (AMT). Among them, AMT has many advantages over the other methods such as easy handling and mainly generating single-copy integration in the genome. An AMT method previously reported for P. infestans and P. palmivora has barely been used in oomycete research due to low success and low reproducibility.</AbstractText><AbstractText Label="RESULTS">In this study, we report a simple and efficient AMT system for P. palmivora. Using this system, we were able to reproducibly generate over 40 transformants using zoospores collected from culture grown in a single 100 mm-diameter petri dish. The generated GFP transformants constitutively expressed GFP readily detectable using a fluorescence microscope. All of the transformants tested using Southern blot analysis contained a single-copy T-DNA insertion.</AbstractText><AbstractText Label="CONCLUSIONS">This system is highly effective and reproducible for transformation of P. palmivora and expected to be adaptable for transformation of additional Phytophthora spp. and other oomycetes. Its establishment will greatly accelerate their functional genomic studies.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Wu</LastName><ForeName>Dongliang</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, 3190 Maile Way, St. John 317, Honolulu, HI, 96822, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Navet</LastName><ForeName>Natasha</ForeName><Initials>N</Initials><AffiliationInfo><Affiliation>Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, 3190 Maile Way, St. John 317, Honolulu, HI, 96822, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Yingchao</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, 3190 Maile Way, St. John 317, Honolulu, HI, 96822, USA.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Present Address: College of Plant Protection, Agricultural University of Hebei, Baoding, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Uchida</LastName><ForeName>Janice</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, 3190 Maile Way, St. John 317, Honolulu, HI, 96822, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tian</LastName><ForeName>Miaoying</ForeName><Initials>M</Initials><Identifier Source="ORCID">0000-0001-8900-6414</Identifier><AffiliationInfo><Affiliation>Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, 3190 Maile Way, St. John 317, Honolulu, HI, 96822, USA. mtian@hawaii.edu.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2016</Year><Month>09</Month><Day>06</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>BMC Microbiol</MedlineTA><NlmUniqueID>100966981</NlmUniqueID><ISSNLinking>1471-2180</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D004251">DNA Transposable Elements</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D004269">DNA, Bacterial</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D016054">DNA, Protozoan</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C036483">T-DNA</NameOfSubstance></Chemical><Chemical><RegistryNumber>147336-22-9</RegistryNumber><NameOfSubstance UI="D049452">Green Fluorescent 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de Chine</li><li>États-Unis</li></country></list><tree><country name="États-Unis"><noRegion><name sortKey="Wu, Dongliang" sort="Wu, Dongliang" uniqKey="Wu D" first="Dongliang" last="Wu">Dongliang Wu</name></noRegion><name sortKey="Liu, Yingchao" sort="Liu, Yingchao" uniqKey="Liu Y" first="Yingchao" last="Liu">Yingchao Liu</name><name sortKey="Navet, Natasha" sort="Navet, Natasha" uniqKey="Navet N" first="Natasha" last="Navet">Natasha Navet</name><name sortKey="Tian, Miaoying" sort="Tian, Miaoying" uniqKey="Tian M" first="Miaoying" last="Tian">Miaoying Tian</name><name sortKey="Uchida, Janice" sort="Uchida, Janice" uniqKey="Uchida J" first="Janice" last="Uchida">Janice Uchida</name></country><country name="République populaire de Chine"><noRegion><name sortKey="Liu, Yingchao" sort="Liu, Yingchao" uniqKey="Liu Y" first="Yingchao" last="Liu">Yingchao Liu</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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