1个新棉花bHLH类基因GhbHLH-130的克隆及表达分析.pdf

棉 花 学 报 Cotton Science 20 14袁26 4 冤院363~370 1 新棉花bHLH 类基因 130 的克隆及表达分析 杨其 宋桂成 张金凤 王晓楠 唐灿明* 京农业大学农学院/ 作物遗传与种质创新国家重点实验室袁江苏南京2 10095 冤 摘要院近年来袁已发现碱性/ 螺旋- 环- 螺旋 basic/Helix-Loop-Helix, bHLH 冤 类转录因子家族在植物的生长发 育调控中发挥重要作用遥 本研究通过电子克隆方法得到1 个新的棉花bHLH 类转录因子基因的全长序列袁利 用反转录-PCR 技术从陆地棉中分离出1 个bHLH 类转录因子基因袁并命名为 130遥 该基因包含 1 个 552 bp 的开放阅读框袁 编 184 个氨基酸残基遥 序列分析表明GhbHLH 130 蛋白包含 1 个螺旋- 环- 螺旋 (Helix-Loop-Helix) 保守结构域袁属于bHLH 转录因子家族遥 实时荧光定量分析结果表明 130 基因的 表达受干旱尧高盐尧低温以及外源脱落酸的显著诱导遥亚细胞定位结果表明 130 基因在细胞核内表达遥 本研究结果表明 130 基因可能作为调控基因参与棉花的逆境应答响应过程遥 关键词院bHLH 转录因子曰亚细胞定位曰基因表达曰非生物胁迫曰棉花 中图分类号院S562.035 文献标志码院A 文章编号院1002-7807 20 14冤04-0363-08 Guang Yangqi, Song Guicheng, Zhang Jinfeng, Wang Xiaonan, Tang Canming* ( 2 10095 ) In recent years, it has been found that bHLH (basic/Helix-Loop-Helix) family transcription factors play an important role in regulation of plant growth and development. In the present study, a novel cDNA encoding a bHLH transcription factor was cloned in upland cotton ( L.) by silico cloning approach and RT-PCR. And the gene was designated as 130. The bioinformatics analyses demonstrated that this full-length cDNA contained a 552 bp ORF (Open reading frame) which encoded a novel bHLH protein with 184 amino acids residues, and contained an Helix-Loop-Helix conserved do- main, and belonged to the bHLH subfamily. Quantitative RT-PCR analysis showed that expression of 130 was highly induced by dehydration, NaCl and cold stresses. In addition, it was observed that 130 was up-regulated by exogenous ABA treatment. Moreover, subcellular localization analysis using onion epidermal cells showed that GhbHLH 130-GFP fusion protein was distribut