茶树CsBAP1基因的克隆与非生物胁迫和激素响应分析

Cloning and Expression Analysis of the Gene CsBAP1 under the Treatments of Hormones and Abiotic Stress in Camellia sinensis

该研究基于茶树转录组数据,从茶树‘龙井43'cDNA中克隆获得茶树CsBAP1基因,对其蛋白质序列特征和基因表达模式以及CsBAP1在茶树不同组织、不同激素处理及不同逆境胁迫下的表达水平进行荧光定量检测。结果显示:(1)克隆得到的茶树CsBAP1基因开放阅读框长度为543bp,编码180个氨基酸;CsBAP1蛋白为亲水性蛋白,理论相对分子质量为19 398Da,理论等电点为9.30,含有保守的Ca2+依赖性的C2结构域;茶树CsBAP1蛋白二级结构由8.89%的α-螺旋、7.78%的β-转角、35.56%的β-折叠和47.78%的随机卷曲组成;三级结构分析结果显示,CsBAP1包括螺旋和折叠结构,与二级结构吻合。(2)荧光定量分析结果显示,CsBAP1基因在茶树的花、花蕾、幼叶、老叶和成熟叶中均有表达,且在老叶中的表达量最高;外源施用ABA、IAA、MeJA、GA3以及SA均能够抑制茶树CsBAP1基因的表达;在高温(38℃)、低温(4℃)、干旱(200g·L^(-1) PEG)、高盐(200mmol·L^(-1) NaCl)胁迫下,CsBAP1的表达量均有所上调,且不同时间段的表达存在显著差异。该研究为进一步研究茶树CsBAP1基因的功能奠定了基础。

In this study,aCsBAP1 gene was cloned from the cDNA of‘Longjing 43＇based on the transcriptome data of tea plant.We analyzed its protein sequence characteristics and gene expression patterns.In addition,the expression profiles of the CsBAP1 gene in different tissues and under extreme temperatures and hormone treatments in tea plant was analyzed by quantitative real time PCR.（1）Sequence analysis showed that the length of the CsBAP1 open reading frame（ORF）is 543 bp,encoding 180 amino acids and contains a highly conserved region called C2 domain.Protein analysis suggested that the CsBAP1 is a hydrophilic protein with the theoretical relative molecular weight 19 398 Da and the theoretical isoelectric point 9.30.Secondary structure prediction showed that the CsBAP1 protein consists of 8.89% α-helix,7.78%β-turn,35.56%extended strand and 47.78%random coil.The three-dimensional structures indi-cated that CsBAP1 protein containsα-helix andβ-strands.（2）The expression profiles of the CsBAP1 was detected in flower,floral buds,young leaves,mature leaves and senescent leaves by the RT-qPCR.And the highest expression level was detected in the senescent leaves.The expression level of CsBAP1 was down-regulated by ABA,IAA,MeJA,GA3,and SA treatments,respectively.The expression level of CsBAP1 was up-regulated under high temperature（38℃）,low temperature（4℃）,drought（200 g·L（-1） PEG）,and high-salinity（200 mmol·L（-1） NaCl）treatments,respectively.These results provide a foundation for the function study of CsBAP1 gene in tea plants.