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Urumqi, Xinjiang seed potato, an important production base of commodity potato. The production of potato virus disease of widespread and serious, to become the main obstacles restricting production. The high-quality virus-free seed potato industry an important guarantee for the development of healthy, efficient, and to establish a rapid, sensitive, accurate and potato virus detection technology is the key to the production of virus-free seed. This study investigated the type of potato virus identification Urumqi, study the establishment of important pathogenic viruses and viroids molecular biology technology, viral kinds of materials screening and cultivation of potatoes, seed transporting quarantine inspection, the implementation of effective prevention and treatment of viral diseases and to provide scientific basis and technical support. Symptoms survey, electron microscopy, RT-PCR Amplification and cloning, sequence analysis, clearly the leading cause of infection Urumqi potato virus PVX, PVY, PVA, PLRV, like virus PSTVd. PVY and PLRV infection rate is the highest, followed by PVX, PVA, PSTVd ??infection rate is low. Homology analysis showed that the, PVY separation was 5 for PVYO lines, one for PVYN lines; the PSTVd ??Urumqi isolates with the Wisconsin Department of weak homology was 98.4%, with the X76844 middle of the French lines M (mild ) homology of 98.8%. Urumqi major pathogen of potato virus isolates most highly homologous nucleotide sequence between isolates and some other regions at home and abroad, there is a big difference. For Urumqi major pathogen of potato viruses PVX, PVY, PVA, PLRV, design virus coat protein gene-specific primers, the establishment of a synchronous detection of the virus one-step multiplex RT-PCR detection method. Target band size were amplified by the method of PVX, PVY, PVA, PLRV 732bp, 422bp, 132bp and 336bp. The lowest limit of detection of the viral RNA 7.8pg/μL. PVM, PVS, AMV, TMV and PSTVd ??amplification was negative. The results show that the method than the two-step multiplex RT-PCR, fast, easy and efficient. This study were designed according to the conserved sequence of PVA and PVY isolates CP gene, two pairs of specific primers and two TaqMan probes labeled with different fluorophores. Optimization of reaction conditions and reagent concentrations, synchronous detection of PVA and PVY double fluorescence quantitative PCR method. The detection sensitivity reach 0.51fg/μL, than conventional RT-PCR is 100 times higher. Assembled into kits based on the optimized one-step multiplex RT-PCR detection system developed and used in various types of sample testing. The results show that a specific, sensitive, simple, efficient, low cost and so on, the detection of a large number of samples for virus-free seedlings and virus-free seed potato production units do.
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