甘蔗CIPK23基因克隆及对低钾、干旱、盐胁迫的响应

Molecular Cloning and Expression Analysis of CIPK23 Gene from Sugarcane under Low Potassium, Drought and Salt Stress

本研究采用RT-PCR技术从低钾胁迫的甘蔗根系中克隆得到一个长度为1 749 bp的与CBL互作的丝氨酸/苏氨酸蛋白激酶基因,命名为Ss CIPK23(Gen Bank登录号为KM981737),包含1 350 bp的开放阅读框(ORF),可编码一个449氨基酸残基的多肽。Ss CIPK23具有CIPK基因家族典型的蛋白激酶结构域和NAF调控结构域。进化树分析显示Ss CIPK23与其他作物的CIPK23关系较近,具有较高的保守性。q PCR分析结果表明,在低钾、干旱和盐胁迫下,Ss CIPK23的表达都发生改变。在低钾胁迫24 h后Ss CIPK23相对表达量开始上升,而且随着胁迫时间的延长,相对表达量持续增加;而在干旱和盐胁迫下,相对表达量最高出现在48 h,随后表达量开始下降,暗示了该基因在低钾、干旱和盐胁迫下发挥重要的调控作用,可为深入研究Ss CIPK23基因功能奠定坚实的基础。

In this study, A CBL-interacting protein kinase gene named Ss CIPK23(Gen Bank accession number:KM981737) was cloned by reverse transcription-polymerase chain reaction(RT-PCR) from root of sugarcane under low potassium stress. The full length of Ss CIPK23 gene was 1 749 bp, containing a 1 350 bp complete open reading frame, which encoded 449 amino acids. The Ss CIPK23 contained a protein kinase and a NAF domain which are the typical domains of CIPK family. The phylogenetic analysis of Ss CIPK23 with CIPK family in various plants revealed that Ss CIPK23 was closely related with CIPK23 of various plants and the CIPK23 genes in various plants were highly conservative. The result of q PCR indicated that the expression of Ss CIPK23 was changed with low potassium, drought and salt stresses. Under low potassium stress, the expression of Ss CIPK23 was up-regulated at 24 h under, and the relative expression increased with the stress time; under drought and salt stress, the highest relative express of Ss CIPK23 appeared at 48 h and then relative express decreased at 92 h. The results revealed that the Ss CIPK23 might take part in responses to various stress. It builds a great base for researching functional of Ss CIPK23.