DNA去甲基化酶（dMTase）是主动去甲基化过程中具有糖基化酶和脱嘌呤裂合酶活性的双功能酶。基于茶树基因组数据,本研究利用生物信息学方法对茶树DNA去甲基化酶（CsdMTase）编码基因进行了全面鉴定和序列分析。全基因组鉴定结果表明,舒茶早基因组中包含4个CsdMTases,系统进化分析将CsdMTases分为DME和ROS两个分支,基因结构分析显示不同成员之间的基因序列长度和内含子数量存在差异。不同CsdMTases蛋白的理化性质相似,均包含ENDO3c和RRM_DME典型的保守结构域,且都定位在细胞核中。CsdMTases家族基因启动子区域包含大量光信号响应、植物激素响应、胁迫响应和生长发育等相关顺式作用元件。表达分析结果显示,CsdMTase基因在不同组织器官中均有表达,有一定的组织特异性;在炭疽菌（Colletotrichum camelliae）、拟盘多毛孢菌（Pseudopestalotiopsis camelliae-sinensis）和茶尺蠖（Ectropis oblique）等生物胁迫及干旱等非生物胁迫下,CsdMTase的表达均被显著诱导;冬季休眠过程中CsdMTases在不同品种成熟叶和越冬芽中的表达模式存在显著差异;CsDMEa、CsDMEb在花芽发育及种子萌发的不同阶段的表达存在显著上调过程。CsdMTases介导的主动去甲基化过程在调控茶树胁迫应答和生长发育中可能发挥了重要的作用,为深入揭示茶树表观调控机制提供理论依据。
DNA demethylase (dMTase) is a bifunctional enzyme with activities of glycosylase and apyrimidic lysase during active demethylation. Based on the available genome data of tea plant, bioinformatics analysis was used to comprehensively identify DNA demethylases (CsdMTase) in tea plant. The genome-wide identification results show that the tea cultivar Shuchazao contains 4 CsdMTases. These CsdMTases could be divided into two branches (DME and ROS) with differences in gene length and intron number by phylogenetic and gene structure analysis. Further analysis shows that the physical and chemical properties of different CsdMTase proteins are similar, which contain the typical conserved domains of ENCO3c and RRM_DME and are located in the nucleus. The promoter regions of CsdMTases contain a large number of cis-acting components related to light response, plant hormone response, stress response and growth and development. The CsdMTase genes are expressed in all detected tissues with certain tissue specificity. The expressions of CsdMTases were significantly up-regulated under biotic stresses, including Colletotrichum camelliae, Pseudopestalotiopsis camelliae-sinensis and Ectropis oblique treatments. During winter dormancy, different expression patterns of CsdMTases were detected in mature leaves and overwintering buds of different cultivars. The expressions of CsDMEa and CsDMEb at different stages of flower bud development and seed germination were significantly induced. The results show that active DNA demethylation process mediated by CsdMTases plays a crucial role in regulating the stress responses, growth and development of tea plant, providing a theoretical basis for discovering the epigenetic regulation mechanism in tea plant.
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