一个新的陆地棉抗逆锌指蛋白基因GhCHP的克隆与分析

Cloning and Analysis of a New Zinc Finger Gene Gh CHP in Gossypium hirsutum L.

锌指蛋白是一类重要的转录因子,在植物应对外界生物和非生物胁迫的过程中起着极为重要的调节作用。基于二倍体雷蒙德氏棉基因组数据库,本研究从抗枯萎病、耐黄萎病的转基因抗虫棉品种鲁棉研32号中克隆到棉花锌指蛋白类基因Gh CHP。序列分析表明,Gh CHP基因的开放阅读框长度为720 bp,编码239个氨基酸,预测的蛋白质等电点为9.12。通过与其它植物锌指蛋白的序列进行聚类和比对分析,发现Gh CHP与拟南芥的AT2g16050和小麦的Ta CHP极为相近,其中两个保守的C1结构域呈典型的锌指结构。实时荧光定量PCR检测表明,Gh CHP在经过不同胁迫处理的棉花根和叶中的表达模式存在差异。PEG、Na Cl、H2O2、低温和黄萎病菌侵染处理均对Gh CHP的表达产生较大影响,表明Gh CHP可能在棉花生物和非生物胁迫响应中起重要的调控作用,是棉花品种抗逆性分子改良的重要候选基因。

Zinc finger proteins are important regulatory factors involving in plant responses to biotic and abiotic stresses. Based on genomic sequences of diploid cotton Gossypium raimondii, we cloned a new cotton zinc finger gene GhCHP from Lumianyan32, a transgenic Bt upland cotton variety characterized by Fusarium wilt resistance and Verticillium wilt tolerance. Sequence analysis revealed that the open reading frame is 720 bp by length, encoding a protein of 239 amino acids with an estimated isoelectric point of 9.12. Compared to other zinc finger protein sequences, GhCHP shows highest similarity to AT2g16050 in A rabidopsis thaliana and TaCHP in Triticum aestivum, with two highly conserved C1 domain. Further analysis by real time quantitative PCR demonstrated that the expression patterns of GhCHP in cotton leaves and roots were different under stress conditions such as PEG, NaC1, H2O2, low temperature and the defoliating Verticillium strain VD8, respectively, suggesting that GhCHP may play an important regulatory role in cotton responses to both biotic and abiotic stresses, with the potential value in cotton germplasm improvement.