砂藓耐脱水相关类钙调蛋白基因RcCML1的克隆与表达分析  

Cloning and expression analysis of dehydration tolerance related calmodulin-like protein gene RcCML1 in Racomitrium canescens

作　　者：孙天国[1] 谷新颖 衣兰 沙伟[1] 张梅娟[1] 马天意 SUN Tian-guo;GU Xin-ying;YI Lan;SHA Wei;ZHANG Mei-juan;MA Tian-yi(College of Life Sciences,Agriculture and Forestry,Qiqihar University,Heilongjiang Qiqihar 161006,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《齐齐哈尔大学学报(自然科学版)》2025年第3期68-74,94,共8页Journal of Qiqihar University(Natural Science Edition)

基　　金：黑龙江省省属高等学校基本科研业务费科研项目(135409427)。

摘　　要：类钙调蛋白(calmodulin-like protein,CML)是植物的一类钙结合蛋白,在植物生长和抗逆反应中起重要作用。砂藓是具有强耐脱水性的藓类植物,从砂藓脱水胁迫转录组数据中选取一个CML基因,命名为RcCML1,对其进行克隆获得编码宛列,并对RcCML1进行功能预测。对RcCML1蛋白质的性质进行预测,利用qPCR技术检测RcCML1在砂藓复水、脱水、高温胁迫下的表达响应。结果显示,RcCML1编码宛列长714 bp,预测RcCML1蛋白质含4个EF-hand结构域,相对分子质量约为25.97 kDa,pI为5.30,无跨膜结构和信号肽,为不稳定的亲水性蛋白质,亚细胞定位在细胞质膜上。RcCML1与小立碗藓和角齿藓中的CML有较高的同源性。qPCR分析显示,RcCML1的表达量在干燥砂藓复水处理1 min时迅速上调,后整体下降;在砂藓脱水过程中呈波动变化趋势,脱水1 h时最高;在45℃高温处理中,处理12 h时表达量最高,推测RcCML1可能参与砂藓耐脱水和高温胁迫机制。Calmodulin-like protein(CML)is a kind of calcium-binding protein in plants,which plays important roles in plant growth and stress resistance.Racomitrium canescens is a moss with strong desiccation resistance.In this study,a CML gene was selected from the transcriptomics database of dehydration stress in R.canescens,named RcCML1,the coding sequence was cloned and acquired,the functions of RcCML1 were predicted.The properties of RcCML1 were predicted,and qPCR was used for examine the expression responses of RcCML1 when R.canescens were treated with rehydration,dehydration and high temperature stresses,respectively.The coding sequence of RcCML1 was with a length of 714 bp,RcCML1 was predicted containing 4 EF-hand domains,with a relative molecular mass of about 25.97 kDa and a pI of 5.30.There is no transmembrane structures and signal peptides in RcCML1,and RcCML1 is an unstable protein with hydrophility.RcCML1 has high homology with CMLs in Physcomitrium patens and Ceratodon purpureus.Results of qPCR showed the expression of RcCML1 was up-regulated rapidly after 1 min of rehydration treatment of desiccated R.canescens,upregulated to the highest at 1 h of the dehydration progress of R.canescens.In the 45℃high-temperature treatment,the expression was the highest when treated for 12 h,respectively,suggesting that RcCML1 might be involved in the mechanism of resistance to dehydration and high temperature stress of R.canescens.

关 键 词：砂藓 RcCML1 类钙调蛋白 耐脱水性 高温胁迫 表达分析 

分 类 号：Q943[生物学—植物学]