ELISA法检测干旱下花生不同品种中AhNCED1蛋白表达变化

AhNCED1 protein expression in different peanut varieties in response to drought by ELISA

ABA合成关键酶Ah NCED1与花生抗旱性相关。优化ELISA法检测花生Ah NCED1蛋白体系,利用该体系检测不同产地7个花生品种幼苗干旱处理24h后叶片内Ah NCED1蛋白表达变化。结果表明,正常生长条件下,7个品种Ah NCED1蛋白表达量呈现较低水平,品种间无显著性差异;干旱24h后,所有品种的Ah NCED1蛋白表达量显著增加;Ah NCED1蛋白表达变化倍数从大到小依次是:中花16>濮花28>花育19>福花4>中花6>花育33>海花1,与QPCR检测Ah NCED1基因表达和Westernblot的检测结果比较一致。综合分析3种方法检测不同品种的Ah NCED1基因和蛋白结果表明,ELISA法及QPCR配合使用适用于大范围的快速筛选抗旱花生品种,为抗旱花生品种筛选提供技术支撑。

AhNCED1, the rate-limiting enzyme in the ABA biosynthetic pathway was found to be related with peanut drought resistance. The ELISA detection system was optimized to analyze the expression of AhNCED1 protein. The expression of AhNCEDI in leaves of 7 different peanut varieties under drought stress of 24 h was detected by ELISA system. No significant difference was found among different peanut varieties under normal condition. On the contrary, the expression of AhNCED 1 proteins of seven peanut varieties remarkably increased under drought stress. The expression change of AhNCED1 of peanut varieties was Zhonghua 16〉 Puhua 28〉Huayu 19〉Fuhua 4〉Zhonghua 6〉Huayu 33〉Haihua 1, respectively, which was consistent with the results of QPCR and WB. The ELISA and QPCR for the detecting AhNCED1 expression in different peanut varieties were recommended for selection of drought- resistant peanut varieties rapidly and largely, providing technology support for screening peanut varieties with drought resistant.