丙酰紫草素防治植物病毒病作用机理研究

The antiviral mechanism of natural product propionylshikonin was studied. Tobacco mosaic virus(TMV ) was isolated from tocacco plants, its purity was analyzed by electroscope, UV/VIS, MALDI-TOFMS, SDS-PAGE. TMV -CP (coat protein) and RNA were isolated from TMV. Treated RNA with propionylshikonin and analyzed with UV/VIS spectrum, we found the specific peak of propionylshikonin in the UV/VIS spectrum of RNA. Inoculated tabocca leaves with TMV-RNA treated by propionylshikonin, it was found the inhibition rate of local lesions were about 85%. It shown that propionylshikonin bound with TMV-RNA. Treated TMV-CP with propionylshikonin solution, then removed un-combined propionylshikonin by dialysis and centrifugation. Analyzed the treated TMV-CP with UV/VIS, we found the specific peak of propionylshikonin in the spectrum. Analyzed this treated TMV-CP by MALDI-TOF MS. It was found the mass peak changed both in it's shape and m/z, which peptides mapping of trypsin was also changed , it was shown propionylshikon is binding with TMV-CP, too, and simulated the three-dimensional structure of binding site with SGI workstation. Propionylshikonin inhibited the combination of viral RNA with plant ribosome both in vitro and invivo. It also inhibited the synthesis of TMV-RNA and TMV protein in vivo. The results assumed that the main target of propionylshikonin was TMV-RNA. The FCM techniques were used to investigate the changes of infected tobacco cells in cycles and apoptosis. The results of cell cycles and apoptosis by FCM show that TMV makes cells mainly stop in G0/G1 phase, S phases decreases and apoptosis number increases with TMV infection time and TMV concentration. Antiviral have a firm reversal effect on the cell proliferating phases, make cells S and G2/M phases increases, and apoptosis number decreases. .It is an important research aspect in agriculture remote sensing to use hyper-spectral data for monitoring growth of infected plants, identifying disease type and evaluating effect of control diseases. The examination results show that the healthy and infected tobacco have different reflectance spectrum during the same growing period. Red edge location, red edge peak value and first derivatives integral can be used as the indicators to identify viral disease types and evaluate effect of control diseases.

在农药化学、植物病毒学、植物学理论的指导下，从寄主、病原菌（ＴＭＶ）三者之间关系入手，探讨左旋及右旋丙酰紫草素防治植物花叶病毒病的作用机理。观察化合物与病毒粒子结合的方式及结合位点，研究它们抑制病毒侵入及增殖的原因。了解它们对寄主症状的缓解作用方式及对寄主病生理变化的调节。为植物病毒病天然药物研制、应用提供理论依据。