Study On Anti-TMV Activity Of Secondary Substances From Walnut Seedcases

Tobacco mosaic virus, causing huge losses to the production of tobacco, has become one of the main diseases. The paper preliminary studied on the secondary material in Walnut Seedcases on inhibition of tobacco mosaic virus and anti-viral activity mechanism. The results were shown as follow:(1)To optimize the condition for extracting activity of antiviral substances from walnut peel, the antiviral substance of walnut (Juglans regia L.) peel were extracted by different solvents and methods, and its antiviral activities were determined, and the extraction process was studied by single factors and orthogonal experiment guided by the inhibition rate of antiviral substances from walnut peel. The results showed that antiviral substance activity is dissimilar based on extracts from walnut peel with different solvent and methods. When concentration inoculated is 10mg/mL, inhibition rate on Tobacco mosaic virus (TMV) of the extract with 95% (Volume part) ethanol is the best, 80.0%. The successive order of factors affecting the extracting was ethanol concentration, liquid-solid ratio, extract times and extracting time. When the method is cold soak, the optima conditions of extraction were determined to be 95% alcohol cold soaks in 26h for forth and liquid-solid ratio is 6:1。(2)Active tracking method is adopted to isolate antiviral activity substances from Walnut seedcases. According to optimum extraction, the Walnut seedcases extracts, suspended in distilled water, then extracted by petroleum ether, ethyl acetate, n-butanol, respectively received extraction of petroleum ether, ethyl acetate phase extracts, n-butanol phase and water phase. When concentration inoculated is 10mg/mL, inhibition rate on Tobacco Mosaic Virus (TMV) of the Ethyl acetate phase extraction is the best, 68.64%. The compound A was obtained from extraction of ethyl acetate through column chromatography, thin layer chromatography and recrystalization. When concentration inoculated is 5 mg/ml, the inhibition rate of the compound to TMV infecting Nicotiana glutinosa is 74.62%. The result indicated that the compound A is a main antiviral component from extraction of ethyl acetate.(3)Mix A with TMV and inoculated on the leaf of Nicotiana glutinosa after half hour, one hour and two hours by means of half-leaf, the concentration inoculated is 10mg/mL. The inhibition rate of necrotic local lesions was 64.97%,69.07% and 86.87%. Explain that the compound A had the ability of inactivating TMV in vitro. (4)The prevention experiment indicated: The inhibition rate is 50.645% when the compound A is inoculate before virus; the inhibition rate is 30.68% when virus is inoculated before the compound A. Explained that the compound A had the ability of preventing infectivity of TMV.(5)The compound A inoculated on the leaf of the Nicotiana tabacum. After the 3ds, 6ds, 9ds and 12ds, the content of Chlorophyll, MDA, Flavonoid, SOD, PPO and POD in Nicotiana tabacum was investigated. The results indicated that the compound could remarkedly increase the content of chlorophyll and flavonoid compared with check, enhances the activity of SOD, PPOand POD compared with check, and the content of MDA was remarkably reduced than check. Explained that the compound. A may through increase the content of chlorophyll and flavonoid, enhances the activity of SOD, PPOand POD, and reduces the content of MDA of lamina to strengthen the disease resistance of Nicotiana tabacum...