鴉膽子內(nèi)生真菌Br11代謝產(chǎn)物抗TMV活性研究
[Abstract]:The earliest virus found in the world is tobacco mosaic virus (Tobacco msaic virus,TMV), which seriously affects the yield and quality of tobacco in China, and causes huge losses to the production of tobacco in China every year. But so far there is no cure for tobacco mosaic virus (TMV). Brucetin D (BruceineD), a chemical substance that has been isolated from Brucea javanica, a medicinal plant, which has a good inhibitory effect on tobacco mosaic virus (TMV) (TMV), is based on the mutually beneficial symbiotic relationship between endophytic fungi and the host. Plant endophytic fungi can produce the same or similar compounds as the host. The endophytic fungi from Brucea javanica plants were isolated and purified in the early stage, and the metabolites of these fungi were tested for their anti- TMV activity. In this paper, one of the better endophytic fungi Br11 resistant to TMV was selected for further study. In this paper, the metabolites of an endophytic fungus Br11 from Brucea javanica were isolated and its anti TMV activity was studied. The fermentation products of the strain were separated and purified by a variety of chromatographic separation techniques, such as RP-C18 column Sephadex LH-20 gel column chromatography. Six monomers and one extracellular polysaccharide (Extracellular polysaccharide,EPS) were obtained, numbered 1-7, respectively. By 1H-NMR-13C-NMR technique, the structures of the six compounds were identified and compared in literature. The results showed that compounds 1-6 were DothioreLone Dothiorelone (C) Dothiorelone (H) and Cytosporone (C) to Cyporone (U). The anti- TMV activity of these isolated compounds was studied. The inhibitory activities of these compounds on TMV infection and proliferation were determined by semi-leaf blight spot method and leaf disc method respectively. The results showed that the extracellular polysaccharides (component 7) had a strong anti-infection effect of TMV, its IC50 was 0.93 渭 g / mL, but the anti-proliferation effect of TMV was not obvious. Compound 6 Cytosporone U had a strong anti-proliferation effect on TMV, with a IC50 of 144.60 渭 g / mL. The bioactivity of compound 6 against TMV in tobacco plants was further determined by Western-blot and Agrobacterium tumefaciens infiltration method. The results showed that compound 6 had a good effect on the expression of TMV-CP, proliferation and movement of TMV at a concentration of 200 渭 g/mL. The effective period of compound 6 was 7 days. In this paper, the Br11 strain was identified by combining morphology and molecular biology. The fungus grew rapidly on the PDA medium. The mycelium had long tube and septum, and the color changed from white to light yellow and then to dark yellow. Yellow soluble pigment was produced. Endophytic fungi universal primer ITS 1 and ITS 4 were used to amplify the conservative region sequence of 28s rRNA of Br11 strain by PCR and sequencing the amplified products. The homology of BLAST analysis showed that this strain had the highest homology with P. pseudostema, which was 99xuan. By constructing phylogenetic tree, it was found that the Br11 strain was on the same branch as the phyllostachys. At the same time, the spore morphology of Br11 strain was not found in the spore induction experiment under different culture conditions. According to the above and related literatures, Br11 strains were identified as (Phomopsis sp) fungi without sporulation.
【學(xué)位授予單位】:福建農(nóng)林大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2017
【分類號(hào)】:S476
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