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在已构建的受条锈菌(Puccinia striiformis f.sp.tritici)诱导的小麦非亲和抑制差减杂交(suppression subtractive hybridization,SSH)文库中,筛选到1个与水稻丙氨酸氨基转移酶基因高度相似的表达序列标签(expressed sequence tag,EST),利用电子克隆与RT-PCR相结合的方法,从小麦中获得1个1563bp的cDNA序列,命名为TaAlaAT1。序列分析表明,TaAlaAT1包含1个完整的开放阅读框,长1440bp,推测编码479个氨基酸。该氨基酸序列具有氨基转移酶的保守特征,与水稻、葡萄、大豆、拟南芥、苜蓿等多种植物的丙氨酸氨基转移酶高度相似,其中与水稻的亲缘关系最近。半定量RT-PCR与实时定量PCR结果显示,TaAlaAT1在小麦与条锈菌互作的非亲和组合中总体呈上调表达,在亲和组合中呈下调表达,且在2个组合中的表达差异显著。推测TaAlaAT1参与了小麦对条锈菌的防御反应。
In the constructed SSH library of the suppression subtractive hybridization (SSH) induced by Puccinia striiformis f. Sp. Tritici, one mutant was isolated from the rice alanine aminotransferase gene A highly similar expressed sequence tag (EST) was obtained by a combination of electronic cloning and RT-PCR. A 1563bp cDNA sequence was obtained from wheat and named as TaAlaAT1. Sequence analysis showed that TaAlaAT1 contains a complete open reading frame, which is 1440 bp in length and presumably encodes 479 amino acids. This amino acid sequence has the conserved characteristics of aminotransferases and is highly similar to the alanine aminotransferases of many plants such as rice, grape, soybean, Arabidopsis thaliana, alfalfa and the like, of which the genetic relationship with rice is the closest. The results of semi-quantitative RT-PCR and real-time quantitative PCR showed that TaAlaAT1 was up-regulated in the non-affinity combination of wheat and stripe rust, and down-regulated in the affinity set, and the difference in expression between the two combinations Significant. It is speculated that TaAlaAT1 is involved in the defensive response of wheat to stripe rust.