茶树CsCML16基因的克隆及其低温胁迫下的表达分析

陈思文; 康芮; 郭志远; 周琼琼; 冯建灿

doi:10.13305/j.cnki.jts.20210514.001

茶叶科学 >

2021 , Vol. 41 >Issue 3: 315 - 326

DOI: https://doi.org/10.13305/j.cnki.jts.20210514.001

研究报告

茶树CsCML16基因的克隆及其低温胁迫下的表达分析

陈思文 ,
康芮 ,
郭志远 ,
周琼琼 ,
冯建灿

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河南农业大学园艺学院,河南郑州 450003

陈思文,女,硕士研究生,主要从事茶树遗传育种与抗逆机理研究。

收稿日期: 2020-09-02

修回日期: 2020-12-07

网络出版日期: 2021-06-15

基金资助

河南省重点研发与推广专项（科技攻关）（202102110204）、河南农业大学科技创新基金（KJCX2019A13）、河南省大学生创新创业训练计划项目（S202010466008）

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Cloning and Expression Analysis of CsCML16 in Tea Plants (Camellia sinensis) under Low Temperature Stress

CHEN Siwen ,
KANG Rui ,
GUO Zhiyuan ,
ZHOU Qiongqiong ,
FENG Jiancan

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College of Horticulture, Henan Agricultural University, Zhengzhou 450003, China

Received date: 2020-09-02

Revised date: 2020-12-07

Online published: 2021-06-15

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摘要

类钙调蛋白CMLs（CaM-like proteins）是植物体内一类重要的钙信号转导蛋白,在抗非生物胁迫中发挥重要作用。以龙井43、福鼎大白茶和黄金芽的一年生茶树扦插苗为材料,通过低温处理（10℃和4℃）分析茶树在低温胁迫下的生理变化;通过克隆茶树类钙调蛋白CsCML16,分析其在低温胁迫下不同抗寒性茶树品种中的表达模式。结果表明,低温胁迫下龙井43的抗寒性较强,福鼎大白茶次之,黄金芽品种较弱。以龙井43的cDNA为模板,克隆获得CsCML16基因,序列分析表明该基因CDS全长为480 bp,编码160个氨基酸,具有钙受体蛋白EF-Hand保守结构域,为小分子蛋白,相对分子量17.58 kDa;亚细胞定位结果显示CsCML16定位于细胞核和细胞膜。荧光定量结果表明,低温胁迫诱导CsCML16基因的上调表达,且在不同的茶树品种中表达量有差异。为进一步揭示CsCML16基因的生物学功能提供依据。

关键词： 茶树; 类钙调蛋白; 低温胁迫; 生理分析; 表达分析

本文引用格式

陈思文 , 康芮 , 郭志远 , 周琼琼 , 冯建灿 . 茶树CsCML16基因的克隆及其低温胁迫下的表达分析[J]. 茶叶科学, 2021 , 41(3) : 315 -326 . DOI: 10.13305/j.cnki.jts.20210514.001

Abstract

Calmodulin-like proteins (CMLs) are important calcium signaling proteins in plants, which play a pivotal role in abiotic stress. In this study, one-year-old tea cutting seedlings of ‘Longjing43’, ‘Fuding Dabai’ and ‘Huangjinya’ were used as research materials. The physiological changes of the three tea cultivars were explored to preliminarily screen the cold-resistance tea cultivar. The CsCML16 gene was cloned to analyze its expression patterns in different cold-tolerance tea cultivars under low temperature stress. The results show that ‘Longjing43’ had the highest cold tolerance under low temperature stress, followed by ‘Fuding Dabai’ and ‘Huangjinya’. CsCML16 gene was cloned from ‘Longjing43’. Sequence analysis shows that the CDS length of CsCML16 is 480 bp, encodes 160 amino acids, contains EF-hand conserved domains with molecular weight of 17.58 kDa. Subcellular localization assay indicates that CsCML16 localized in the nucleus and plasma membrane. Quantitative real-time PCR analysis reveals that CsCML16 gene was involved in the regulation of tea plants in response to low temperature stress, and its expression levels varied with tea cultivars with different cold tolerance. This study provided a basis for further study on the biological function of CsCML16 gene.

Key words： Camellia sinensis; calmodulin-like protein; low temperature stress; physiological analysis; expression analysis

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