| Lanzhou lily(Lilium davidii var.unicolor) is a variant of Sichuan lily(Lilium davidii Duchautre).It is a kind of perennial herb and bulb edible,widely cultivated and as a economic crop in Lanzhou area.The cultivating region of Lanzhou lily is mostly distributed in Xiguoyuan town of South Suburb in Lanzhou.In recent years,the quality fluctuated frequently in the production of this crop.Viral infections other than human factor,such as field management of the perennial crop were presumed as one of the main reasons to induce such fluctuations.The epidemiological situation of viruses which infected Lanzhou lily has not been well reported.In this thesis,the viral infection of Lanzhou lily has been primarily investigated,several molecular biologic and immunologic methods had been employed to determine the viral infections in Lanzhou lily,and some elementary plant pathologic analysis have been done.It has been found in this thesis that Lily symptomless virus (LSV),Cucumber mosaic virus(CMV) and Lily mottle virus(LMoV) can infect Lanzhou lily,and infection rate of LSV and CMV reach to 100%.We applied two gel filtration chromatography methods to purify the viral particles of LSV and CMV by using infected Lanzhou lily as a material.After analysed by SDS-PAGE,MALDI-Tof mass spectrum,the isolated product has been identified to be LSV and CMV respectively. By adapting a RT-PCR protocol,the coat protein gene of LSV and CMV has been cloned into E.coli and sequenced.Recombinant coat protein(CP) was expressed and purified.A phylogenetic tree has been built after a blast searching in NCBI using the coat protein animo acid sequence of isolated LSV and CMV.We studied the viral disease of Lanzhou lily preliminarily,isolating and purifying the infected virus.Meanwhile,as a basis for further study on a double antibody sandwich ELISA method,we also provided some new direction and thinking on detection of LSV and CMV. |
