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Isolation of pathogenesis-related proteins from TMV-infected tobacco and their influence on infectivity of TMV

机译:TMV感染烟草中病程相关蛋白的分离及其对TMV感染性的影响

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摘要

The composition of pathogenesis-related proteins (PR-proteins) in the intercellular fluid (ICF) and leaf tissue of the hypersensitive tobacco cultivar Xanthi-nc inoculated with Tobacco mosaic virus (TMV), and their inhibitory influence on TMV multiplication were studied. The ICF PR-proteins of infected plants were separated after solubilisation by decreasing gradient of ammonium sulphate, the cell PR-proteins were separated after acidic homogenisation of leaf tissues. The ICF and cell PR-proteins were further purified by ion exchange chromatography on DEAE cellulose. Using discontinuous non-denaturating polyacrylamide gel electrophoresis of DEAE cellulose fractions the PR-proteins were detected. Their molecular weights were estimated by SDS-PAGE. The ICF and cell proteins of infected leaves included PR-proteins of the molecular weights 15-16 kDa (Group 1), 27-28 kDa (Group 3: chitinases) and 36-40 kDa (Group 2a: beta-1,3-glucanases), Fractions with different PR-proteins were tested for their effect on infectivity of TMV. Particularly the PR3 and PR2a proteins seem to decrease the infectivity of TMV.
机译:研究了接种烟草花叶病毒(TMV)的高敏感性烟草品种Xanthi-nc的细胞间液(ICF)和叶片组织中的病程相关蛋白(PR-proteins)的组成及其对TMV增殖的抑制作用。溶解后,通过降低硫酸铵的梯度分离出被感染植物的ICF PR蛋白,并在叶片组织进行酸性均质化后分离出细胞PR蛋白。通过DEAE纤维素上的离子交换色谱法进一步纯化ICF和细胞PR蛋白。使用DEAE纤维素级分的不连续非变性聚丙烯酰胺凝胶电泳检测PR蛋白。通过SDS-PAGE估计它们的分子量。被感染叶片的ICF和细胞蛋白包括分子量为15-16 kDa(组1),27-28 kDa(组3:几丁质酶)和36-40 kDa(组2a:beta-1,3-)的PR蛋白。葡聚糖酶),测试了具有不同PR蛋白的级分对TMV感染力的影响。特别是PR3和PR2a蛋白似乎降低了TMV的传染性。

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