甘薯盐胁迫诱导IbDEAD1基因的克隆、生物信息学及表达分析

Cloning,bioinformatics and expression analysis of salt-induced IbDEAD1 gene from sweetpotato(Ipomoea batatas)

DEAD-box解旋酶是植物中最大的解旋酶家族,不仅在几乎所有涉及DNA/RNA代谢的细胞过程中发挥关键作用,而且还广泛参与植物生长发育与胁迫响应过程.目前,甘薯中有关DEAD-box基因的功能研究尚未见报道.本研究以耐盐性品种徐薯22与盐敏感性品种徐薯32为研究材料,通过RNA-seq及生物信息学分析,首次从甘薯中筛选出一个受盐胁迫诱导的DEAD-box基因,命名为IbDEAD1.根据转录组分析的测序结果设计引物,从徐薯22中克隆出该基因的全长序列(GenBank登录号:MF043568).序列分析表明,IbDEAD1基因包含一个1 701bp的开放阅读框,编码566个氨基酸.序列分析显示IbDEAD1蛋白具有一个DEAD-box蛋白典型的解旋酶中心和多个磷酸化位点,且为DEAD-box RNA解旋酶.系统进化树及qRT-PCR分析均表明,IbDEAD1基因可能在甘薯盐胁迫响应过程中发挥重要作用.这些结果为进一步研究IbDEAD1基因在甘薯盐胁迫响应中的功能奠定了基础.

DEAD-box helicase is the largest helicase family in plants,not only plays important roles in almost all cellular processes involved in DNA/RNA metabolism,but also widely involves in the regulation of plant growth and stress response.However,presently there is no report on the function of DEAD-box gene in sweetpotato.Here the salttolerantcultivar Xushu 22 and the saltsensitivecultivar Xushu 32 were employed,and the IbDEAD1 gene,induced by salt stress was screened for the first time by RNA-seq and bioinformatics analysis.The full length of IbDEAD1 gene was cloned from Xushu 22 and contained a 1 701 bp open reading frame（ORF）encoding a 566 amino acids.Multiple sequence alignment showed that IbDEAD1 contains a typical helicase center of DEAD-box protein and multiple phosphorylation sites,and belongs to the DEAD-box RNA helicase.Phylogenetic and qRT-PCR analysis showed that IbDEAD1 gene may play an important role in salt stress response.These results lay a foundation for further study on the function of IbDEAD1 gene in salt stress response of sweetpotato.