Detection Of Two Plant Virus CMLV And PRMV Using TaqMan Real-time PCR

Plant virus is a kind of the serious plant pathogens. In this research, we designed two TaqMan probes for real-time PCR to detect the pathogens of the diseases, Cherry mottle leaf and Peach rosette mosaic respectively. And here is the effect:1) Cherry mottle leaf virus (CMLV) is one of the most important diseases on cherry in South America. In this research, we successfully synthesized the sequences of CMLV 6741～7322 nt genes (Genbank AF170028.1) and constructed positive standard plasmids of this nucleic fragment on the vector. According to the result of the real-time PCR,23 copies/μL of the plasmid was detected as the detection limit of the real-time PCR. And the inhibiting substance in cherry is little effect the detection.2) Peach rosette mosaic virus (PRMV), the cause agent of Vitis labrusca, Prunus persica, Vitis vinifera and many other cultivars of grapevine, is a quarantine pest in EPPO Al list. In this research, we successfully synthesized the sequences of PRMV 988～1755 nt genes (Genbank AF016626) and constructed positive standard plasmids of this fragment on the vector. The sensitivity detection of the real-time RT-PCR shows that,37 copies/μL of the RNA was detected as the detection limit, which is 100×more sensitive than RT-PCR.Finally, The FQ-PCR in this research could detection the CMLV and PRMV sensitively and rapidly. The methods are significant for the detection and prevention of these two viruses.