茶树CsCML24的克隆及表达分析  

Cloning and Expression Analysis of CsCML24 Gene in Camellia sinensis

作　　者：康芮 刘春晖 陈思文 赵仁亮[1] 周琼琼 KANG Rui;LIU Chun-hui;CHEN Si-wen;ZHAO Ren-liang;ZHOU Qiong-qiong(College of Horticulture,Henan Agricultural University,Zhengzhou 450000)

机构地区：[1]河南农业大学园艺学院,郑州450000

出　　处：《生物技术通报》2022年第3期22-30,共9页Biotechnology Bulletin

基　　金：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)。

摘　　要：类钙调素(calmodulin-like protein,CML)是植物体内一类钙受体蛋白,介导Ca^(2+)与下游靶蛋白的相互作用,在植物抗逆反应中发挥重要作用。探究茶树中CML蛋白在逆境胁迫中的功能,为进一步研究茶树CsCML24对逆境胁迫的响应机理提供理论依据。以龙井43一年生茶树扦插苗为材料,克隆得到类钙调蛋白基因CsCML24(GenBank登录号为MZ325391),并进行了生物信息学分析。同时,利用实时荧光定量PCR技术分析茶树CsCML24组织表达特异性以及不同非生物胁迫处理下的表达模式。结果显示,CsCML24的CDS长为480 bp,编码159个氨基酸。CsCML24为无信号肽及跨膜结构的稳定性亲水蛋白,含有EF-hand保守结构域,能与Ca^(2+)结合。qRT-PCR表明,CsCML24在茶树各组织中均表达,在成熟叶的表达量显著高于其他组织,茎中表达量较低;该基因在低温(10℃)、干旱(20%PEG 6000)、高盐胁迫(200 mmol/L NaCl)和ABA(100μmol/L)处理均能诱导表达,且在不同胁迫下表达差异显著,推测该基因可能通过ABA信号途径调节对低温和干旱的耐受性。Calmodulin-like protein(CML)is a type of calcium receptor protein,which mediates the interaction of Ca^(2+)with downstream target proteins,and plays an important role in plant abiotic stress response.Exploring the function of CML in tea plant(Camellia sinensis)under adversity stress could provide theoretical references for further studying the molecular regulation mechanism of CsCML24 in stress response of tea plants.The CsCML24 gene(GenBank accession number:MZ325391)of CML was cloned from the leaves of tea cultivar‘Longjing 43’and analyzed by bioinformatics.Meanwhile,the relative tissue-specific expressions and expression patterns under different abiotic stresses were analyzed by real-time quantitative PCR.Results of sequence analysis indicated that the full-length CDS sequence of the CsCML24 gene was 480 bp,encoding 159 amino acids.The CsCML24 had no signal peptide and transmembrane region,and belonged to hydrophilic protein,contained EF-hands,which can be combined with Ca^(2+).Tissue-specific expression results from qRT-PCR showed that the CsCML24 gene expressed in various tissues,and highly expressed in the mature leaves,and the least in the stem.The expression of CsCML24 was significantly induced under low temperature(10℃),drought stress(20%PEG 6000),salt stress(200 mmol/L NaCl)and ABA,and the expression varied in different stress.It is speculated that this gene regulates the low temperature and drought tolerance of plants via ABA signaling pathway.

关 键 词：茶树 类钙调蛋白 非生物胁迫 表达分析 

分 类 号：S571.1[农业科学—茶叶生产加工]