A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, Phytophthora hibernalis.
Identifieur interne : 000593 ( Main/Exploration ); précédent : 000592; suivant : 000594A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, Phytophthora hibernalis.
Auteurs : Tingting Dai [République populaire de Chine] ; Tao Hu [République populaire de Chine] ; Xiao Yang [États-Unis] ; Danyu Shen [République populaire de Chine] ; Binbin Jiao [République populaire de Chine] ; Wen Tian [République populaire de Chine] ; Yue Xu [République populaire de Chine]Source :
- PeerJ [ 2167-8359 ] ; 2019.
Abstract
Phytophthora hibernalis, the causal agent of brown rot of citrus fruit, is an important worldwide pathogen and a quarantine pest in China. Current diagnosis of the disease relies on disease symptoms, pathogen isolation and identification by DNA sequencing. However, symptoms caused by P. hibernalis can be confused with those by other Phytophthora and fungal species. Moreover, pathogen isolation, PCR amplification and sequencing are time-consuming. In this study, a rapid assay including 20-min recombinase polymerase amplification targeting the Ypt1 gene and 5-min visualization using lateral flow dipsticks was developed for detecting P. hibernalis. This assay was able to detect 0.2 ng of P. hibernalis genomic DNA in a 50-µL reaction system. It was specific to P. hibernalis without detection of other tested species including P. citrophthora, P. nicotianae, P. palmivora and P. syringae, four other important citrus pathogens. Using this assay, P. hibernalis was also detected from artificially inoculated orange fruits. Results in this study indicated that this assay has the potential application to detect P. hibernalis at diagnostic laboratories and plant quarantine departments of customs, especially under time- and resource-limited conditions.
DOI: 10.7717/peerj.8083
PubMed: 31763074
PubMed Central: PMC6870529
Affiliations:
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wicri:level="1"><nlm:affiliation>Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu</wicri:regionArea><wicri:noRegion>Jiangsu</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Xiao" sort="Yang, Xiao" uniqKey="Yang X" first="Xiao" last="Yang">Xiao Yang</name><affiliation wicri:level="2"><nlm:affiliation>United States Department of Agriculture, Agricultural Research Service, Foreign Disease-Weed Science Research Unit, Fort Detrick, MD, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>United States Department of Agriculture, Agricultural Research Service, Foreign Disease-Weed Science Research Unit, Fort Detrick, MD</wicri:regionArea><placeName><region type="state">Maryland</region></placeName></affiliation><affiliation wicri:level="2"><nlm:affiliation>Oak Ridge Institute for Science and Education, ARS Research Participation Program, Oak Ridge, TN, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Oak Ridge Institute for Science and Education, ARS Research Participation Program, Oak Ridge, TN</wicri:regionArea><placeName><region type="state">Tennessee</region></placeName></affiliation></author><author><name sortKey="Shen, Danyu" sort="Shen, Danyu" uniqKey="Shen D" first="Danyu" last="Shen">Danyu Shen</name><affiliation wicri:level="1"><nlm:affiliation>Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Plant Pathology, Nanjing Agricultural University, Nanjing</wicri:regionArea><wicri:noRegion>Nanjing</wicri:noRegion></affiliation></author><author><name sortKey="Jiao, Binbin" sort="Jiao, Binbin" uniqKey="Jiao B" first="Binbin" last="Jiao">Binbin Jiao</name><affiliation wicri:level="1"><nlm:affiliation>Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs, Shanghai, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs, Shanghai</wicri:regionArea><wicri:noRegion>Shanghai</wicri:noRegion></affiliation></author><author><name sortKey="Tian, Wen" sort="Tian, Wen" uniqKey="Tian W" first="Wen" last="Tian">Wen Tian</name><affiliation wicri:level="1"><nlm:affiliation>Jiangyin Customs House, Jiangyin, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Jiangyin Customs House, Jiangyin</wicri:regionArea><wicri:noRegion>Jiangyin</wicri:noRegion></affiliation></author><author><name sortKey="Xu, Yue" sort="Xu, Yue" uniqKey="Xu Y" first="Yue" last="Xu">Yue Xu</name><affiliation wicri:level="1"><nlm:affiliation>Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu</wicri:regionArea><wicri:noRegion>Jiangsu</wicri:noRegion></affiliation></author></analytic><series><title level="j">PeerJ</title><idno type="ISSN">2167-8359</idno><imprint><date when="2019" type="published">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><i>Phytophthora hibernalis</i>, the causal agent of brown rot of citrus fruit, is an important worldwide pathogen and a quarantine pest in China. Current diagnosis of the disease relies on disease symptoms, pathogen isolation and identification by DNA sequencing. However, symptoms caused by <i>P. hibernalis</i> can be confused with those by other <i>Phytophthora</i> and fungal species. Moreover, pathogen isolation, PCR amplification and sequencing are time-consuming. In this study, a rapid assay including 20-min recombinase polymerase amplification targeting the <i>Ypt1</i> gene and 5-min visualization using lateral flow dipsticks was developed for detecting <i>P. hibernalis</i>. This assay was able to detect 0.2 ng of <i>P. hibernalis</i> genomic DNA in a 50-µL reaction system. It was specific to <i>P. hibernalis</i> without detection of other tested species including <i>P. citrophthora</i>, <i>P. nicotianae</i>, <i>P. palmivora</i> and <i>P. syringae</i>, four other important citrus pathogens. Using this assay, <i>P. hibernalis</i> was also detected from artificially inoculated orange fruits. Results in this study indicated that this assay has the potential application to detect <i>P. hibernalis</i> at diagnostic laboratories and plant quarantine departments of customs, especially under time- and resource-limited conditions.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">31763074</PMID><DateRevised><Year>2020</Year><Month>10</Month><Day>01</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">2167-8359</ISSN><JournalIssue CitedMedium="Print"><Volume>7</Volume><PubDate><Year>2019</Year></PubDate></JournalIssue><Title>PeerJ</Title><ISOAbbreviation>PeerJ</ISOAbbreviation></Journal><ArticleTitle>A recombinase polymerase amplification-lateral flow dipstick assay for rapid detection of the quarantine citrus pathogen in China, <i>Phytophthora hibernalis</i>.</ArticleTitle><Pagination><MedlinePgn>e8083</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.7717/peerj.8083</ELocationID><Abstract><AbstractText><i>Phytophthora hibernalis</i>, the causal agent of brown rot of citrus fruit, is an important worldwide pathogen and a quarantine pest in China. Current diagnosis of the disease relies on disease symptoms, pathogen isolation and identification by DNA sequencing. However, symptoms caused by <i>P. hibernalis</i> can be confused with those by other <i>Phytophthora</i> and fungal species. Moreover, pathogen isolation, PCR amplification and sequencing are time-consuming. In this study, a rapid assay including 20-min recombinase polymerase amplification targeting the <i>Ypt1</i> gene and 5-min visualization using lateral flow dipsticks was developed for detecting <i>P. hibernalis</i>. This assay was able to detect 0.2 ng of <i>P. hibernalis</i> genomic DNA in a 50-µL reaction system. It was specific to <i>P. hibernalis</i> without detection of other tested species including <i>P. citrophthora</i>, <i>P. nicotianae</i>, <i>P. palmivora</i> and <i>P. syringae</i>, four other important citrus pathogens. Using this assay, <i>P. hibernalis</i> was also detected from artificially inoculated orange fruits. Results in this study indicated that this assay has the potential application to detect <i>P. hibernalis</i> at diagnostic laboratories and plant quarantine departments of customs, especially under time- and resource-limited conditions.</AbstractText><CopyrightInformation>© 2019 Dai et al.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Dai</LastName><ForeName>Tingting</ForeName><Initials>T</Initials><Identifier Source="ORCID">0000-0003-0534-6724</Identifier><AffiliationInfo><Affiliation>Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hu</LastName><ForeName>Tao</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Xiao</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>United States Department of Agriculture, Agricultural Research Service, Foreign Disease-Weed Science Research Unit, Fort Detrick, MD, USA.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Oak Ridge Institute for Science and Education, ARS Research Participation Program, Oak Ridge, TN, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Shen</LastName><ForeName>Danyu</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jiao</LastName><ForeName>Binbin</ForeName><Initials>B</Initials><AffiliationInfo><Affiliation>Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs, Shanghai, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tian</LastName><ForeName>Wen</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>Jiangyin Customs House, Jiangyin, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Xu</LastName><ForeName>Yue</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, Jiangsu, China.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>11</Month><Day>18</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>PeerJ</MedlineTA><NlmUniqueID>101603425</NlmUniqueID><ISSNLinking>2167-8359</ISSNLinking></MedlineJournalInfo><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Exclusivity</Keyword><Keyword MajorTopicYN="N">Inclusivity</Keyword><Keyword MajorTopicYN="N">Isothermal diagnostic assay</Keyword><Keyword MajorTopicYN="N">Oomycetes</Keyword><Keyword MajorTopicYN="N">Plant destroyers</Keyword><Keyword MajorTopicYN="N">RPA–LFD</Keyword></KeywordList><CoiStatement>Binbin Jiao is employed by the Technical Center for Animal, Plant and Food Inspection and Quarantine of Shanghai Customs. Wen Tian is employed by Jiangyin Customs House. The authors declare that they have no competing interests.</CoiStatement></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>07</Month><Day>24</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>10</Month><Day>22</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2019</Year><Month>11</Month><Day>26</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>11</Month><Day>26</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2019</Year><Month>11</Month><Day>26</Day><Hour>6</Hour><Minute>1</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">31763074</ArticleId><ArticleId IdType="doi">10.7717/peerj.8083</ArticleId><ArticleId IdType="pii">8083</ArticleId><ArticleId 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last="Hu">Tao Hu</name><name sortKey="Jiao, Binbin" sort="Jiao, Binbin" uniqKey="Jiao B" first="Binbin" last="Jiao">Binbin Jiao</name><name sortKey="Shen, Danyu" sort="Shen, Danyu" uniqKey="Shen D" first="Danyu" last="Shen">Danyu Shen</name><name sortKey="Tian, Wen" sort="Tian, Wen" uniqKey="Tian W" first="Wen" last="Tian">Wen Tian</name><name sortKey="Xu, Yue" sort="Xu, Yue" uniqKey="Xu Y" first="Yue" last="Xu">Yue Xu</name></country><country name="États-Unis"><region name="Maryland"><name sortKey="Yang, Xiao" sort="Yang, Xiao" uniqKey="Yang X" first="Xiao" last="Yang">Xiao Yang</name></region><name sortKey="Yang, Xiao" sort="Yang, Xiao" uniqKey="Yang X" first="Xiao" last="Yang">Xiao Yang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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