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Molecular identification of <i>Neofabraea </i>species associated with bull's eye rot on apple using rolling-circle amplification of partial <i>EF-1α</i> sequence.

Authors
  • Lin, Huijiao1, 2
  • Jiang, Xiang3
  • Yi, Jianping4
  • Wang, Xinguo5
  • Zuo, Ranling6
  • Jiang, Zide7
  • Wang, Weifang8
  • Zhou, Erxun9
  • 1 South China Agricultural University, 12526, Guangdong Province Key Laboratory of Microbial Signals and Disease Control, Guangzhou 510642, Guangdong, China. , (China)
  • 2 Huangpu Entry-Exit Inspection and Quarantine Bureau, Inspection and Test Center, Guangzhou 510730, Guangdong, China ; [email protected] , (China)
  • 3 Huangpu Entry-Exit Inspection and Quarantine Bureau, Inspection and Test Center, Guangzhou 510730, Guangdong, China ; [email protected] , (China)
  • 4 Shanghai Entry-Exit Inspection and Quarantine Bureau, Technical Center for Amimal, Plant and Food Inspection and Quarantine, Shanghai 200135, China ; [email protected] , (China)
  • 5 Huangpu Entry-Exit Inspection and Quarantine Bureau, Inspection and Test Center, Guangzhou 510730, Guangdong, China ; [email protected] , (China)
  • 6 Huangpu Entry-Exit Inspection and Quarantine Bureau, Inspection and Test Center, Guangzhou 510730, Guangdong, China ; [email protected] , (China)
  • 7 South China Agricultural University, Guangdong Province Key Laboratory of Microbial Signals and Disease Control, Guangzhou 510642, Guangdong, China ; [email protected] , (China)
  • 8 Guangdong Inspection and Quarantine Technology Center, Guangdong Key Laboratory of Import and Export Technical Measures of Animal, Plant and Food, Guangzhou 510623, Guangdong, China ; [email protected] , (China)
  • 9 South China Agricultural University, Guangdong Province Key Laboratory of Microbial Signals and Disease Control, Guangzhou 510642, Guangdong, China ; [email protected] , (China)
Type
Published Article
Journal
Canadian Journal of Microbiology
Publisher
Canadian Science Publishing
Publication Date
Oct 30, 2017
Identifiers
DOI: 10.1139/cjm-2017-0448
PMID: 29084390
Source
Medline
License
Unknown

Abstract

A rolling-circle amplification (RCA) method with padlock probes targeted on <i>EF-1α</i> regions was developed for rapid detection of apple bull's eye rot (BER) pathogens, including<i> Neofabraea malicorticis</i>,<i> N. perennans</i>, <i>N. kienholzii</i> and <i>N. vagabunda</i> (synonym:<i> N. alba</i>). Four padlock probes (PLP-Nm, PLP-Np, PLP-Nk and PLP-Nv) were designed and tested against 28 samples, including 22 BER pathogen cultures, 4 closely related species and 2 unrelated species which may cause serious apple decays. The assay successfully identified all the BER pathogenic fungi at the level of species, while no cross reaction was observed in all target species and no false positive reaction was observed with all strains used for reference. This study showed that the use of padlock probes and the combination of probe signal amplification by RCA provided an effective and sensitive method for the rapid identification of <i>Neofabraea</i> spp.. The method could therefore be a useful tool for monitoring bull's eye rot pathogens in port quarantine and orchard epidemiological studies.

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