Genomic variability in potato virus M and the development of RT-PCR and RFLP procedures for the detection of this virus in seed potatoes.
Potato virus M (PVM, Carlavirus) is considered to be one of the most common potato viruses distributed worldwide. Sequences of the coat protein (CP) gene of several Canadian PVM isolates were determined. Phylogenetic analysis indicated that all known PVM isolates fell into two distinct groups and the isolates from Canada and the US clustered in the same group. The Canadian PVM isolates could be further divided into two sub-groups. Two molecular procedures, reverse transcription - polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) were developed in this study for the detection and identification of PVM in potato tubers. RT-PCR was highly specific and only amplified PVM RNA from potato samples. PVM RNAs were easily detected in composite samples of 400 to 800 potato leaves or 200 to 400 dormant tubers. Restriction analysis of PCR amplicons with MscI was a simple method for the confirmation of PCR tests. Thus, RT-PCR followed by RFLP analysis may be a useful approach for screening potato samples on a large scale for the presence of PVM.
Xu
Huimin
H
Canadian Food Inspection Agency, Charlottetown Laboratory, 93 Mount Edward Road, Charlottetown, PEI, C1A 5T1, Canada. huimin.xu@inspection.gc.ca
D'Aubin
Jeanette
J
Nie
Jingbai
J
D002170:Canada; D036022:Capsid Proteins / Q000235:genetics; D017788:Carlavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016000:Cluster Analysis; D014644:Genetic Variation; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012639:Seeds / Q000821:virology*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D011198:Solanum tuberosum / Q000821:virology*; D014481:United States
D036022:Capsid Proteins; D012367:RNA, Viral