Serological detection of ‘Candidatus Liberibacter asiaticus’ in citrus, and identification by GeLC-MS/MS of a chaperone protein responding to cellular pathogens
Identifieur interne : 000056 ( Main/Merge ); précédent : 000055; suivant : 000057Serological detection of ‘Candidatus Liberibacter asiaticus’ in citrus, and identification by GeLC-MS/MS of a chaperone protein responding to cellular pathogens
Auteurs : Fang Ding [République populaire de Chine, États-Unis] ; Yongping Duan [États-Unis] ; Qing Yuan [République populaire de Chine] ; Jonathan Shao [États-Unis] ; John S. Hartung [États-Unis]Source :
- Scientific Reports [ 2045-2322 ] ; 2016.
Abstract
We describe experiments with antibodies against ‘
Url:
DOI: 10.1038/srep29272
PubMed: 27381064
PubMed Central: 4933950
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Serological detection of ‘<italic>Candidatus</italic> Liberibacter asiaticus’ in citrus, and identification by GeLC-MS/MS of a chaperone protein responding to cellular pathogens</title><author><name sortKey="Ding, Fang" sort="Ding, Fang" uniqKey="Ding F" first="Fang" last="Ding">Fang Ding</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Hubei Key Laboratory of Plant Pathology, Huazhong Agricultural University</institution>, Wuhan 430070<country>P. R. China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>USDA ARS MPPL 10300 Baltimore Ave.</institution>, Beltsville, MD 20705<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Duan, Yongping" sort="Duan, Yongping" uniqKey="Duan Y" first="Yongping" last="Duan">Yongping Duan</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>USDA ARS USHRL 2001 South Rock Road</institution>, Fort Pierce, FL 34945<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yuan, Qing" sort="Yuan, Qing" uniqKey="Yuan Q" first="Qing" last="Yuan">Qing Yuan</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Sichuan Medical University</institution>, Luzhou, Sichuan, 646000<country>P. R. China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Shao, Jonathan" sort="Shao, Jonathan" uniqKey="Shao J" first="Jonathan" last="Shao">Jonathan Shao</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>USDA ARS MPPL 10300 Baltimore Ave.</institution>, Beltsville, MD 20705<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hartung, John S" sort="Hartung, John S" uniqKey="Hartung J" first="John S." last="Hartung">John S. Hartung</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>USDA ARS MPPL 10300 Baltimore Ave.</institution>, Beltsville, MD 20705<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">27381064</idno><idno type="pmc">4933950</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4933950</idno><idno type="RBID">PMC:4933950</idno><idno type="doi">10.1038/srep29272</idno><date when="2016">2016</date><idno type="wicri:Area/Pmc/Corpus">000470</idno><idno type="wicri:Area/Pmc/Curation">000469</idno><idno type="wicri:Area/Pmc/Checkpoint">000042</idno><idno type="wicri:Area/Ncbi/Merge">002098</idno><idno type="wicri:Area/Ncbi/Curation">002098</idno><idno type="wicri:Area/Ncbi/Checkpoint">002098</idno><idno type="wicri:Area/Main/Merge">000056</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Serological detection of ‘<italic>Candidatus</italic> Liberibacter asiaticus’ in citrus, and identification by GeLC-MS/MS of a chaperone protein responding to cellular pathogens</title><author><name sortKey="Ding, Fang" sort="Ding, Fang" uniqKey="Ding F" first="Fang" last="Ding">Fang Ding</name><affiliation wicri:level="1"><nlm:aff id="a1"><institution>Hubei Key Laboratory of Plant Pathology, Huazhong Agricultural University</institution>, Wuhan 430070<country>P. R. China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="a2"><institution>USDA ARS MPPL 10300 Baltimore Ave.</institution>, Beltsville, MD 20705<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Duan, Yongping" sort="Duan, Yongping" uniqKey="Duan Y" first="Yongping" last="Duan">Yongping Duan</name><affiliation wicri:level="1"><nlm:aff id="a3"><institution>USDA ARS USHRL 2001 South Rock Road</institution>, Fort Pierce, FL 34945<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Yuan, Qing" sort="Yuan, Qing" uniqKey="Yuan Q" first="Qing" last="Yuan">Qing Yuan</name><affiliation wicri:level="1"><nlm:aff id="a4"><institution>Sichuan Medical University</institution>, Luzhou, Sichuan, 646000<country>P. R. China</country></nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Shao, Jonathan" sort="Shao, Jonathan" uniqKey="Shao J" first="Jonathan" last="Shao">Jonathan Shao</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>USDA ARS MPPL 10300 Baltimore Ave.</institution>, Beltsville, MD 20705<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Hartung, John S" sort="Hartung, John S" uniqKey="Hartung J" first="John S." last="Hartung">John S. Hartung</name><affiliation wicri:level="1"><nlm:aff id="a2"><institution>USDA ARS MPPL 10300 Baltimore Ave.</institution>, Beltsville, MD 20705<country>USA</country></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2016">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>We describe experiments with antibodies against ‘<italic>Candidatus</italic> Liberibacter asiaticus used to detect the pathogen in infected plants. We used scFv selected to bind epitopes exposed on the surface of the bacterium in tissue prints, with secondary monoclonal antibodies directed at a FLAG epitope included at the carboxyl end of the scFv. Unexpectedly, the anti-FLAG secondary antibody produced positive results with <italic>C</italic>aLas diseased samples when the primary scFv were not used. The anti-FLAG monoclonal antibody (Mab) also identified plants infected with other vascular pathogens. We then identified a paralogous group of secreted chaperone proteins in the HSP-90 family that contained the amino acid sequence DDDDK identical to the carboxy-terminal sequence of the FLAG epitope. A rabbit polyclonal antibody against one of the same epitopes combined with a goat anti-rabbit secondary antibody produced very strong purple color in individual phloem cells, as expected for this pathogen. These results were entirely specific for <italic>C</italic>aLas-infected citrus. The simplicity, cost and ability to scale the tissue print assay makes this an attractive assay to complement PCR-based assays currently in use. 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