棉花精氨酸脱羧酶基因GhADC1克隆与表达分析  

Cloning and Expression Analysis of an Arginine Decarboxylase Gene from Gossypium hirsutum

作　　者：李亚栋[1] 默辉娟[1] 王省芬[1] 孙艳香[1,2] 马峙英[1] 

机构地区：[1]河北农业大学/河北省作物种质资源重点实验室,河北保定071001 [2]廊坊师范学院遗传与育种研究所,河北廊坊065000

出　　处：《棉花学报》2013年第4期291-299,共9页Cotton Science

基　　金：973前期研究专项(2011CB111609)

摘　　要：以抑制性消减杂交(SSH)文库序列为基础,采用生物信息学和RT-PCR方法从陆地棉中克隆出精氨酸脱羧酶基因,命名为GhADC1,GenBank登录号为KC851856。该基因全长2954 bp,其中5'非翻译区477 bp,具有多个参与胁迫响应的顺式作用元件和一个编码8个氨基酸的微型编码序列(uORF);主编码区(mORF)2181 bp,共编码726个氨基酸,其中N端具有叶绿体定位信号肽。推测的GhADC1蛋白具有PLP结合位点及Orn/Dap/Arg脱羧酶的信号位点,属于典型的III型PLP依赖型精氨酸脱羧酶家族成员。荧光定量RT-PCR结果表明,正常条件下,GhADC1在叶片中表达量高于茎和根部组织,且在抗黄萎病品种冀棉20中的表达水平高于耐病品种中521;黄萎病菌胁迫能够快速诱导抗病品种根部GhADC1表达,而在耐病品种中未见明显差异,推测在抗病品种中GhADC1可能参与根部对黄萎病菌胁迫的早期应答。Based on a sequence in a suppression subtractive hybridization library, GhADC1, GenBank accession no. KC851856, an arginine decarboxylase gene was cloned from upland cotton using bioinformatics and RT-PCR technologies. The full-length cDNA of GhADC1 was 2954-bp long. The 5＇ UTR was 477-bp and contained several eis-acting elements involved in stress re- sponses and a uORF encoding an 8-amino acid sequence. The main ORF was 2181-bp long, and encoded 726 amino acids with a putative N-terminal chloroplast-targeting signal peptide. Conserved domain analysis showed that GhADC1 contained a PLP binding site and an Om/Dap/Arg decarboxylase signal site. These characteristics indicated that it belongs to the type III PLP-de- pendent arginine decarboxylase protein family. Quantitative reverse transcription-polymerase chain reaction results showed that, under untreated conditions, the expression level of GhADCI was higher in leaves than that in stems and roots, respectively, and was higher in the leaves of the resistant cotton variety Jiman 20 than that in those of the tolerant cotton variety Zhong 521. Fur- thermore, the expression of GhADC1 was rapidly up-regulated in the roots of the resistant variety by infection with Verticillium dahliae, while the tolerant variety did not show remarkable changes in its expression atter infection. This indicated that the gene probably involved in the early defense responses to V. dahliae in the roots of the resistant cotton variety.

关 键 词：棉花 精氨酸脱羧酶基因 多胺 黄萎病菌 基因表达 

分 类 号：S562[农业科学—作物学]