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本工作采用酿酒酵母细胞表达载体pESC和植物细胞表达载体pPZP211分析了拟南芥AZI1基因对真菌的抗性功能。半乳糖诱导产生的AZI1蛋白可以使酵母细胞的生长能力明显降低。DAB和台酚蓝染色结果显示用蒜薹灰霉菌孢子处理Col-0野生型植株叶片后被侵染部位只能产生少量H2O2,病原体可以扩散,而AZI1基因过表达植株叶片在侵染部位有大量H2O2产生,着色较深,表明转化体能够以局部细胞的死亡来阻止病原体侵染周围的细胞。在Col-0野生型植株中,AZI1基因的表达受外源水杨酸诱导,24h后达到峰值。以上结果说明AZI1基因在拟南芥对生物胁迫因素的应答过程中具有重要作用。
In this work, we used the Saccharomyces cerevisiae cell expression vector pESC and the plant cell expression vector pPZP211 to analyze the resistance of Arabidopsis AZI1 to fungi. Galactose-induced production of AZI1 protein can significantly reduce the growth capacity of yeast cells. DAB and testol blue staining showed that only a small amount of H2O2 could be produced in the infected parts of Col-0 wild type plants after spores of garlic sprouts treated with spores of Botrytis cinerea, and the pathogen could diffuse. However, leaves of AZI1 overexpressing plants had a large amount of H2O2 Produced, darker, suggesting that transformants are able to stop the pathogen from infecting the surrounding cells with local cell death. In Col-0 wild-type plants, the expression of AZI1 gene was induced by exogenous salicylic acid and peaked at 24 h. The above results indicate that AZI1 gene plays an important role in the response of Arabidopsis thaliana to biotic stress factors.