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采用形态学观察和分子鉴定方法对2011年在陕西省发生的一种烟草未知病害的病原菌进行鉴定。从病叶组织分离纯化得到病原菌,通过致病性测定以及人工接种后再分离病菌,证明编号LJL007的菌株为该病的致病菌。依据病原菌的形态学和培养特征,将菌株LJL007鉴定为灰葡萄孢Botrytis cinerea Pers.,其有性型为富氏葡萄孢盘菌Botryotinia fuckeliana Whetzel。通过核糖体DNAITS序列分析,分离菌株LJL007序列(登录号:HM17900)与富氏葡萄孢盘菌序列(登录号:HM849615)同源性达100%,进一步证明该病原菌是灰葡萄孢Botrytis cinerea。云芝多糖在离体条件下,对灰葡萄孢的菌丝生长和孢子萌发均无直接抑制作用。云芝多糖对烟草灰霉病有较好预防保护作用,其预防效果可达56.29%。云芝多糖可显著提高烟草体内几丁质酶和β-1,3-葡聚糖酶活性,其活性峰值分别比对照提高56.89%和429.83%,说明云芝多糖可诱导植物产生抗病性。
Morphological observation and molecular identification methods were used to identify the pathogen of unknown tobacco disease in Shaanxi Province in 2011. Pathogenic bacteria were isolated and purified from the diseased leaf tissue, and the pathogenicity was confirmed by pathogenicity test and artificial inoculation. It was proved that the strain No. LJL007 was the pathogen of the disease. The strain LJL007 was identified as Botrytis cinerea Pers. According to the morphological and cultural characteristics of the pathogen, and its genus was Botryotinia fuckeliana Whetzel. The homology of the isolate LJL007 (accession number: HM17900) and Botrytis cinerea (accession number: HM849615) was 100% by ribosomal DNA ITS sequence analysis, which further proved that the pathogen was Botrytis cinerea. In vitro, polysaccharides from Coriolus versicolor did not directly inhibit mycelial growth and spore germination of Botrytis cinerea. Coriolus versicolor polysaccharide against tobacco gray mold have a good preventive and protective effect of its prevention effect up to 56.29%. PSK could significantly increase the activities of chitinase and β-1, 3-glucanase in tobacco, and their peak activity were increased by 56.89% and 429.83% respectively compared with the control, indicating that PSK could induce the plant to produce disease resistance.