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茶叶科学 ›› 2017, Vol. 37 ›› Issue (6): 575-585.

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茶树MADS-box转录因子基因的克隆与非生物胁迫响应分析

沈威, 滕瑞敏, 李辉, 刘志薇, 王永鑫, 王文丽, 庄静*   

  1. 南京农业大学园艺学院,茶叶科学研究所,江苏 南京 210095
  • 收稿日期:2017-04-28 修回日期:2017-07-03 出版日期:2017-12-15 发布日期:2019-08-23
  • 通讯作者: *zhuangjing@njau.edu.cn
  • 作者简介:沈威,男,硕士研究生,主要从事茶树分子生物学研究。
  • 基金资助:
    国家自然科学基金面上项目(31570691)

Cloning of a MADS-box Transcription Factor Gene from Camellia sinensis and its Response to Abiotic Stresses

SHEN Wei, TENG Ruimin, LI Hui, LIU Zhiwei, WANG Yongxin, WANG Wenli, ZHUANG Jing*   

  1. Tea Science Research Institute, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2017-04-28 Revised:2017-07-03 Online:2017-12-15 Published:2019-08-23

摘要: 本研究基于茶树转录组数据库,以茶树龙井43为试验材料,通过RT-PCR方法从该茶树的cDNA中克隆得到1个CsMADS1基因。序列分析表明:茶树CsMADS1基因开放阅读框长度为657βbp,编码218个氨基酸,是典型的植物MADS-box家族转录因子。序列多重比对显示,该序列与多个相关物种的MADS-box序列一致性为65.65%,含有高度保守的MADS结构域和半保守的K结构域。氨基酸理化性质、亲疏水性、亚细胞定位预测、无序化分析,以及二级和三级结构分析显示,CsMADS1转录因子是亲水性蛋白,可能定位于细胞核中,无序化程度明显,以α-螺旋结构为主,并与人MEF2蛋白具有相似的三级结构。利用实时荧光定量PCR方法分析了茶树龙井43中CsMADS1基因在非生物胁迫下的表达。结果表明,茶树中CsMADS1基因对高温、低温、干旱和高盐等不同非生物胁迫有响应,且表达存在差异。

关键词: 茶树, MADS-box, 转录因子, 进化分析, 非生物胁迫, 表达分析

Abstract: In this study, based on the transcriptome database of tea plant, the CsMADS1 gene was cloned from cDNA of ‘Longjing43’ by RT-PCR method. The length of open reading frame of CsMADS1 gene was 657βbp, encoding 218 amino acids, which was a typical transcription factor of MADS-box family. Multiple alignments of CsMADS1 with related species showed that the identity of them was 65.65%, with a highly conserved MADS domain and a semi-conserved K domain. The physicochemical properties, hydrophilicity and hydrophobicity, subcellular localization, disordered feature, secondary and tertiary structure were also analyzed. CsMADS1 transcription factor is a hydrophilic protein, may be located in nucleus. The disordered feature of CsMADS1 protein was obvious, which was mainly composed of alpha helix structure, and had similar tertiary structure with MEF2 of human. Quantitative real-time PCR was used to analyze the expression profiles of the CsMADS1 gene under abiotic stress treatments of tea cultivar ‘Longjing43’. The results showed that the CsMADS1 gene responded to high and low temperatures, drought and high salinity. Different expression patterns of CsMADS1 gene were observed under those abiotic stress treatments.

Key words: Camellia sinensis, MADS-box, transcription factor, phylogenetic analysis, abiotic stress, expression analysis

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