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根据几种丝状真菌Hog1 MAPK(丝裂原活化蛋白激酶)的保守氨基酸序列设计简并引物,从番茄早疫病菌Alternaria solani中扩增出MAPK同源基因的部分片段,命名为AsHog1。用Real-timeRT-PCR技术比较了AsHog1在异菌脲敏感菌株和抗性菌株中的表达特性。结果表明,敏感菌株QX21经0.8mol/L NaCl和2mg/L异菌脲处理后,AsHog1相对表达量持续升高,在12h达到最大值,分别为对照的4.12倍和25.23倍,16h略有降低;而室内诱导抗性突变体和田间抗性菌株经相同处理后的AsHog1表达量变化不大,且明显低于敏感菌株。由此推测,番茄早疫病菌AsHog1基因表达特征与其对异菌脲抗药性相关。研究结果为深入研究病原菌对异菌脲抗药性的分子机理、建立抗药性分子监测技术、延缓和防止抗药性的产生及发展奠定了一定的理论基础。
Based on the conserved amino acid sequence of several filamentous fungi Hog1 MAPK (mitogen-activated protein kinase), degenerate primers were designed and a partial fragment of MAPK homologous gene was amplified from Alternaria solani, named AsHog1. The expression characteristics of AsHog1 in isonicotinate-resistant and resistant strains were compared using Real-time RT-PCR. The results showed that the relative expression of AsHog1 in QX21 strain treated with 0.8 mol / L NaCl and 2 mg / L iprodione continuously increased and reached the maximum at 12 h, which was 4.12 and 25.23 fold higher than that of the control and slightly decreased at 16 h While the expression of AsHog1 in induced mutants and field resistant mutants did not change much, which was significantly lower than that of susceptible strains. Therefore, it was speculated that the expression of AsHog1 gene in Alternaria solani was related to its resistance to iprodione. The results provide a theoretical basis for further research on the molecular mechanism of drug resistance to iprodione and establishment of drug resistance molecular monitoring techniques to delay and prevent the emergence and development of drug resistance.