Liu Yashi, Bian Zhiyuan, Jiang Shan, Wang Xiao, Jiao Lin, Shao Yun, Ma Chengmei, Chu Mingyu
College of Horticulture, Gansu Agricultural University, Lanzhou 730070, China.
Plants (Basel). 2025 Jul 7;14(13):2079. doi: 10.3390/plants14132079.
Caffeic acid-O-methyltransferase (COMT) is a key enzyme in lignin synthesis and secondary metabolism in plants, and it participates in the regulation of plant growth and development as well as plants' stress response. To further investigate the function of COMT in grapevine, a total of 124 family genes were identified from three species in this study, namely Pinot noir ( L.), , and . The amino acid sequence encoded by these genes ranged from 55 to 1422 aa, and their molecular mass ranged from 6640.82 to 77,034.43 Da. Subcellular localization prediction inferred that they were mainly located in the plasma membrane and cytoplasm. The prediction of secondary structures showed that α-helix and irregular coiled-coil were primary structural elements. These genes were unevenly distributed across 10 different chromosomes, respectively. Phylogenetic tree analysis of the amino acid sequences of VvCOMT, VaCOMT, VrCOMT, and AtCOMT proteins showed that they were closely related and were divided into four subgroups. The motif distribution was similar among the cluster genes, and the gene sequence was notably conserved. The 124 members of the COMT gene family possessed a variable number of exons, ranging from 2 to 13. The promoter region of all of these genes contained multiple cis-acting elements related to hormones (e.g., ABA, IAA, MeJA, GA, and SA), growth and development (e.g., endosperm, circadian, meristem, light response), and various stress responses (e.g., drought, low temperature, wounding, anaerobic, defense, and stress). The intraspecies collinearity analysis suggested that there were one pair, three pairs, and six pairs of collinear genes in , Pinot noir, and , respectively, and that tandem duplication contributed more to the expansion of these gene family members. In addition, interspecific collinearity revealed that the had the strongest homology with the followed by the , and the weakest homology with the . The expression patterns of different tissues and organs at different developmental stages indicated that the genes had obvious tissue expression specificity. The majority of genes were only expressed at higher levels in certain tissues. Furthermore, we screened 13 genes to conduct qRT-PCR verification according to the transcriptome data of under abiotic stresses (NaCl, PEG, and cold). The results confirmed that these genes were involved in the responses to NaCl, PEG, and cold stress. This study lays a foundation for the exploration of the function of the COMT genes, and is of great importance for the genetic improvement of abiotic stress resistance in grapes.
咖啡酸-O-甲基转移酶(COMT)是植物木质素合成和次生代谢中的关键酶,它参与植物生长发育的调控以及植物的胁迫响应。为了进一步研究COMT在葡萄中的功能,本研究从三个物种中总共鉴定出124个家族基因,即黑比诺(Pinot noir)、[此处原文缺失两个物种名称]。这些基因编码的氨基酸序列长度在55至1422个氨基酸之间,分子量在6640.82至77034.43道尔顿之间。亚细胞定位预测表明它们主要位于质膜和细胞质中。二级结构预测显示α-螺旋和不规则卷曲螺旋是主要结构元件。这些基因分别不均匀地分布在10条不同的染色体上。对VvCOMT、VaCOMT、VrCOMT和AtCOMT蛋白的氨基酸序列进行系统发育树分析表明,它们亲缘关系密切,分为四个亚组。聚类基因中的基序分布相似,基因序列明显保守。COMT基因家族的124个成员所含外显子数量不等,从2个到13个。所有这些基因的启动子区域都含有多个与激素(如脱落酸、生长素、茉莉酸甲酯、赤霉素和水杨酸)、生长发育(如胚乳、昼夜节律、分生组织、光响应)以及各种胁迫响应(如干旱、低温、创伤、厌氧、防御和胁迫)相关的顺式作用元件。种内共线性分析表明,[此处原文缺失两个物种名称]、黑比诺和[此处原文缺失一个物种名称]中分别有一对、三对和六对共线基因,串联重复对这些基因家族成员的扩增贡献更大。此外,种间共线性分析表明,[此处原文缺失一个物种名称]与[此处原文缺失一个物种名称]的同源性最强,其次是与[此处原文缺失一个物种名称],与[此处原文缺失一个物种名称]的同源性最弱。不同发育阶段不同组织和器官的表达模式表明,[此处原文缺失一个物种名称]基因具有明显的组织表达特异性。大多数[此处原文缺失一个物种名称]基因仅在某些组织中高水平表达。此外,我们根据[此处原文缺失一个物种名称]在非生物胁迫(NaCl、PEG和低温)下的转录组数据筛选了13个[此处原文缺失一个物种名称]基因进行qRT-PCR验证。结果证实这些基因参与了对NaCl、PEG和低温胁迫的响应。本研究为探索COMT基因的功能奠定了基础,对葡萄非生物胁迫抗性的遗传改良具有重要意义。