烟草盐诱导Abc1基因NtSIA1的克隆及表达分析

Cloning and Expression Analysis of NtSIA1 from Salt-Induced Abc1 Gene in Nicotiana tabacum

为了研究烟草Abc1基因家族成员在植物非生物胁迫应答过程中的作用,根据烟草转录组数据,利用巢式PCR技术克隆得到1个烟草盐诱导Abc1基因NtSIA1。序列分析表明,该基因与拟南芥AtSIA1基因具有67.57%的一致性,开放阅读框长度为2109bp,编码702个氨基酸,含有一个典型的ABC1结构域、两个激酶结构域和两个跨膜结构域。采用实时荧光定量PCR技术,对NtSIA1基因在烟草不同组织以及盐胁迫、Cd胁迫等处理下的表达分析表明:该基因主要在花和叶中表达;200mmol/LNaCl处理6h以及60μmol/L和100μmol/LCdCl2处理48h后,NtSIA1基因的表达量分别为对照组的2.27、2.9和3.1倍。结果表明烟草NtSIA1基因的表达具有组织特异性,且受到盐胁迫和Cd2+的诱导。

In order to investigate the function of Abcl from Nicotiana tabacum in non biological adversity response, a salt-induced Abc 1 gene NtSIA 1 was cloned by PCR. Sequence analysis results showed that the NtSIA l gene shares 67.57% identity with AtSIA1 gene, and the open reading frame （ORF） of the NtS1A 1 gene was 2 109 bp which encoded a deduced protein including 702 amino acid residues. The protein sequence of NtSIA 1 possessed a conserved ABC1 domain, two kinase domains and two transmembrane domains. The relative expression of NtSIA 1 under the treatments of salt stress and cd stress were determined by quantitative real-time PCR （qRT-PCR）. The results showed that NtSIA 1 gene was mainly expressed in flowers and leaves. After treatment of 200 mmol/L NaCl for 6 h and 60 μmol/L and 100 μmol/L CdClz for 48 h, the expression of NtSIA 1 gene was 2.27, 2.9 and 3.1 times as great as than those of the control group. These results indicated that NtSIA 1 was a gene of tissue specific expression and could be induced by salt stress and Cd2＋.