title: A multiplex RT-PCR for simultaneous detection and identification of five viruses and two viroids infecting chrysanthemum.
abstract: Pathogens causing significant economic losses in chrysanthemum include tomato aspermy virus (TAV), chrysanthemum virus B (CVB), cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), potato virus Y (PVY), chrysanthemum stunt viroid (CSVd) and chrysanthemum chlorotic mottle viroid (CChMVd). A multiplex reverse transcription polymerase chain reaction (RT-PCR) method, using specific primer sets for each virus or viroid, was developed for simultaneous detection and differentiation of TAV, CVB, CMV, TMV, PVY, CChMVd, and CSVd. The RT-PCR method was validated by testing chrysanthemum samples collected from different regions of China. In this study, CVB, TAV, TMV, PVY, CSVd, CMV, and CChMVd were detected, respectively, in 24.7 %, 17.5 %, 4.4 %, 4.4 %, 2.9 %, 2.5 %, and 1.5 % of the samples tested. These results indicate that CVB and TAV (24.7 % and 17.5 %) are common, whereas CMV, TMV, CChMVd, CSVd, and PVY (all below 5 %) are less frequently encountered. This new multiplex RT-PCR method has potential to be used routinely in large-scale virus and viroid surveys.
authors.affiliation: Department of Biological Science, College of Life Sciences, Henan Normal University, Construction East Road No. 46, Muye District, Xinxiang, 453007, People's Republic of China.
authors.forename: Xiting
authors.initials: X
authors.lastname: Zhao
authors.forename: Xingliang
authors.initials: X
authors.lastname: Liu
authors.forename: Beibei
authors.initials: B
authors.lastname: Ge
authors.forename: Mingjun
authors.initials: M
authors.lastname: Li
authors.forename: Bo
authors.initials: B
authors.lastname: Hong
mesh_terms: D002681:China; D031202:Chrysanthemum / Q000821:virology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010935:Plant Diseases / Q000821:virology; D015995:Prevalence; D012328:RNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014772:Viroids / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*