外源水杨酸对稻瘟病菌效应蛋白BAS4过表达菌株耐受性的影响

Effects of exogenous salicylic acid on tolerance of effectorprotein BAS4 overexpression strain of Magnaporthe oryzae

  • 摘要: 目的研究不同水杨酸(SA)浓度对稻瘟病菌菌株形态发育及受侵染水稻防御相关基因表达的影响,探究SA对稻瘟病活体寄生相关效应蛋白BAS4在稻瘟病菌与水稻互作中的作用机制,为生产上有效防控稻瘟病提供理论依据.方法利用不同浓度SA(50、100、200、500、1000和2000 μmol/L)处理稻瘟病菌过表达菌株(35S:BAS4/Mo-1)和野生型菌株(A1343R-7),统计分析SA对过表达菌株和野生型菌株菌落生长速率、产孢量及孢子萌发和附着胞形成等形态发育的影响.调查200 μmol/L SA处理稻瘟病菌菌株侵染水稻的发病率,实时荧光定量PCR(qRT-PCR)分析受侵染水稻防御相关基因表达情况.结果不同SA浓度对过表达菌株菌落生长速率、产孢量、孢子萌发率和附着胞形成的影响低于SA对野生型菌株的影响,经200 μmol/L SA处理过表达菌株侵染水稻的发病率降低,但降低幅度小于200 μmol/L SA处理野生型菌株侵染水稻的发病率.200 μmol/L SA处理能提高过表达菌株侵染水稻病程相关基因PR1a早期上调表达及PR10a基因72 h时的高效表达;使SA途径PAL基因表达量下调,JA途径AOS2基因早期上调表达,LOX2基因各时间点下调表达;使PR5基因早期上调表达,后期急剧下调;HSP90基因早期上调表达,但低于野生型菌株侵染水稻的表达量.结论BAS4过表达菌株对外源SA刺激有较强的耐受性.

     

    Abstract: ObjectiveEffects of different salicylic acid(SA)concentrations on the morphological development of rice blast fungus and the expression of defense-related genes in infected rice were studied,and mechanism of SA on inter-action of biotrophy-associated secreted protein 4(BAS4)in rice blast and rice was revealed to lay a theoretical foundation for effective control of rice blast in cultivation.MethodMagnaporthe oryzae overexpression strain(35S:BAS4/Mo-1)and wild strain(A1343R-7)were pretreated by SA of various concentrations(50,100,200,500,1000 and 2000 μmol/L). Colony growth,spore number,spore germination and appressorium formation of overexpression strain and wild strain were analyzed.The incidence of rice inoculated by M.oryzae which was pretreated by 200 μmol/L SA was investigated, and rice defense-related genes expression in infected rice was analyzed by real-time fluorescence quantitative PCR(qRT-PCR).ResultThe effects of SA concentrations on the colony growth,spore number,spore germination and appressorium formation of overexpression strain were less than those on the wild strain.Disease incidence of rice inoculated by over-expression strain pretreated with 200 μmol/L SA decreased,but the range was smaller than those inoculated by wild strain pretreated with 200 μmol/L SA.After treated by 200 μmol/L SA,the expression level of pathogenesis-related gene PR1a in overexpression strain upregulated at early stage and the expresion was effective at 72 h.At the same time,expression of SA-path gene PAL downregulated,and JA-path gene AOS2 upregulated its expression at early stage,expression of gene LOX2 downregulated at all time points;expression of gene PR5 upregulated at early stage and downregulated sharply at later stage;gene HSP900 upregulated its expression at early stage,but it was lower than the expression of rice infected by wild strain.ConclusionBAS4 overexpression strain is highly resistant to exogenous SA stimulation.

     

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