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茶叶科学 ›› 2015, Vol. 35 ›› Issue (5): 481-490.doi: 10.13305/j.cnki.jts.2015.05.012

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茶树CsbHLH2基因克隆及表达分析

韩永涛, 肖斌*, 钱文俊, 梁少茹   

  1. 农业部西北地区园艺作物生物学与种质创制重点实验室 西北农林科技大学园艺学院,陕西 杨凌712100
  • 收稿日期:2014-12-08 修回日期:2015-01-18 出版日期:2015-10-15 发布日期:2019-08-26
  • 通讯作者: *1647785720@qq.com
  • 作者简介:韩永涛,男,硕士研究生,主要从事茶树分子生物学研究。
  • 基金资助:
    国家茶叶产业技术体系(CARS-23)、陕西省科技统筹创新(KTZB02-01)

Cloning and Expression Analysis of CsbHLH2 in Tea Plant (Camellia sinensis)

HAN Yongtao, XIAO Bin*, QIAN Wenjun, LIANG Shaoru   

  1. Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, College of Horticulture, Northwest A&F University, Yangling 712100, China
  • Received:2014-12-08 Revised:2015-01-18 Online:2015-10-15 Published:2019-08-26

摘要: 植物bHLH(碱性螺旋环螺旋)转录因子是植物体内一类重要的转录因子,在植物的生长发育以及胁迫应答过程中发挥着重要的调控作用。本研究以茶树品种陕茶一号为材料,采用同源克隆法从陕茶一号叶片cDNA中克隆了1个bHLH转录因子CsbHLH2。生物信息学分析表明,CsbHLH2开放阅读框(ORF)为714βbp,编码1个297个氨基酸的蛋白,预测分子量58.4βkD,等电点为5.14,氨基酸序列比对显示该蛋白与其他高等植物的bHLH蛋白具有较高同源性。拟南芥原生质体亚细胞定位表明,CsbHLH2编码蛋白定位在细胞核上。实时荧光定量PCR结果分析表明,CsbHLH2基因在茶树不同组织部位中均有表达,但是在幼叶中表达量最高;不同激素处理结果显示,CsbHLH2受SA、ETH、MEJA诱导。

关键词: 茶树, bHLH转录因子, 亚细胞定位, 进化分析, 定量表达分析

Abstract: The bHLH (basic Helix-Loop-Helix) transcription factor is one of the most important transcription factor in plants, which play important roles in plant growth and stress regulation. The bHLH transcription factor CsbHLH2 was cloned from tea plant (Camellia sinensis) cultivars ‘Shanchayihao’ by homologous cloning technology using cDNA template. Bioinformatics analysis showed that the CsbHLH2 contains 714 bp ORF and was predicted encoding 297 amino acid, the deduced protein molecular weight was 58.4 kD and its theoretical isoelectric point was 5.14. Amino acid sequence alignment revealed that CsbHLH2 was highly homologous to other higher plant bHLH proteins. Transient expression of recombinant plasmid CsbHLH2/PBI221-GFP in Arabidopsis protoplasts showed that CsbHLH2 was located in cell nuclei. Quantitative real-time PCR analysis of the expression profiles showed that the CsbHLH2 gene was expressed in bud, leaf, stem and root. The highest expression level of the CsbHLH2 was found in the young leaf. Different hormone treatments results showed that the CsbHLH2 was induced by ETH, MEJA and SA treatment, respectively.

Key words: Camellia sinensis (L.), bHLH transcription factor, subcelleular locatlization, phylogenetic analysis, expression analysis

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