TaCDPK2基因在小麦与叶锈菌互作过程中的表达分析

在本实验室前期构建的非亲和互作的SSH文库基础上,从叶锈菌侵染的小麦叶片中发现在接种后8 h表现高表达量的CDPK2-EST。CDPKs即钙离子依赖蛋白激酶,是植物细胞应对各种生物和非生物胁迫过程中承接Ca2+流变化的重要因素。为进一步研究CDPK2在小麦与叶锈菌互作过程中的表达特性,采用RT-PCR和Western-Blotting技术分别在mRNA水平和蛋白质水平对小麦受叶锈菌侵染后不同亲和性组合中CDPK2的表达谱进行了检测。结果表明,小麦CDPK2基因受叶锈菌侵染诱导,不亲和组合在接种后4 h无论在mRNA水平还是蛋白质水平该基因均表现上调表达,而转录水平的表达量在接种后16 h降至对照水平;而在蛋白水平其表达量在接种16 h达到最大,之后又恢复至对照水平。亲和组合中该基因在接种后8 h在mRNA水平略有表达,在蛋白水平其表达量在接种后8 h和16 h有上调表达但其表达量低于不亲和组合,之后又趋于对照水平。这一结果表明,小麦CDPK2基因参与了小麦与叶锈菌的互作过程,并在该过程中对提高小麦抗叶锈能力有一定作用。这一结果为深入探讨小麦CDPK2基因在小麦抵抗叶锈菌侵染中的作用及作用机制奠定了基础。

Expression Analysis of TaCDPK2 Gene in the Process of Wheat Infected by Puccinia triticina

The CDPK2-EST was isolated from the wheat SSH cDNA library induced by leaf rust,which has a high level expression in 8h after inoculated leaf rust in incompatible combination.CDPKs(calcium-dependent protein kinases)are key elements in decoding the changes in cytosolic calcium concentrations during the plant defense responses.The Western Blotting analysis and semi-QRT-PCR analysis were used to further investigate relative expression of the CDPK2 in different time points after wheat inoculated leaf rust.These results showed that the expression of CDPK2 was obviously increased in the incompatible combination compared with the control whether at the levels of mRNA or protein at 4 h after inoculation,and the level of mRNA was declined to that in control level at 16 h which the protein level reached the maximum at the meantime.In compatible combination,TaCDPK2 was detectable at 8 h after inoculation,then declined to the normal level.These results implied that TaCDPK2 was involved in leaf rust resistance in wheat,,and laid the foundation for further function research of TaCDPK2 in the further.