刺葡萄愈伤组织褪黑素生物合成基因SNAT2、COMT3的克隆及其对光质的响应  

Cloning of SNAT2 and COMT3 genes involved in melatonin biosynthesis in Vitis davidii callus and their response to light quality

作　　者：贺丽媛 赖恭梯 李思雨 许恒 林俊璇 郭奥琳 赖钟雄[1] 赖呈纯[2,3] HE Liyuan;LAI Gongti;LI Siyu;XU Heng;LIN Junxuan;GUO Aolin;LAI Zhongxiong;LAI Chengchun(Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002,China 2;Institute of Food Science and Technology,Fujian Academy of Agricultural Sciences,Fuzhou 350003,China;Key Laboratory of Processing of Subtropical Characteristic Fruits,Vegetables and Edible Fungi,Ministry of Agriculture and Rural Affairs of China,Fuzhou 350003,China)

机构地区：[1]福建农林大学园艺植物生物工程研究所,福州350002 [2]福建省农业科学院农产品加工研究所,福州350003 [3]农业农村部亚热带特色果蔬菌加工重点实验室,福州350003

出　　处：《应用与环境生物学报》2024年第3期569-578,共10页Chinese Journal of Applied and Environmental Biology

基　　金：福建省自然科学基金项目(2021J05091,2022J01482);福建省属公益类科研院所基本科研专项(2021R1032009,2021R10320013)资助。

摘　　要：5-羟色胺-N-乙酰基转移酶(serotonin N-acetyltransferase,SNAT)和咖啡酸-O-甲基转移酶(caffeic acid O-methyltransferase,COMT)是植物褪黑素合成途径的关键酶,对褪黑素的生物合成有重要的调控作用.以刺葡萄愈伤组织为材料,分别克隆获得了VdSNAT2和VdCOMT3,对其生物信息学及启动子顺式作用元件进行预测,结合实时荧光定量PCR技术,分析不同光质处理下刺葡萄愈伤组织中褪黑素含量与VdSNAT2和VdCOMT3基因表达的相关性.结果表明,VdSNAT2和VdCOMT3基因ORF分别为549 bp和1158 bp,分别编码182和385个氨基酸,其编码蛋白前者为亲水蛋白,后者为疏水蛋白,亚细胞均定位于叶绿体;蛋白保守基序和进化树分析表明,刺葡萄VdSNAT2和VdCOMT3蛋白与葡萄的SNAT2和COMT3具有相同的保守基序,且两者进化相似、亲缘关系近.启动子顺式作用元件预测发现,VdSNAT2和VdCOMT3启动子序列中均有大量光响应等多种生物/非生物响应元件.qRT-PCR分析表明,光质显著影响VdSNAT2和VdCOMT3基因的表达,其中红光对刺葡萄愈伤组织中VdSNAT2和VdCOMT3基因表达的促进作用最为显著,与褪黑素含量具有正相关性,同时VdSNAT2和VdCOMT3基因对褪黑素的生物合成调控具有协同作用.本研究表明光质可以影响VdSNAT2和VdCOMT3基因的表达,从而定向调控刺葡萄愈伤组织褪黑素的生物合成.(图9表1参41)Serotonin N-acetyltransferase(SNAT)and caffeic acid O-methyltransferase(COMT)are pivotal enzymes that play important regulatory roles in melatonin biosynthesis in plants.VdSNAT2 and VdCOMT3 genes were isolated from spine grape(Vitis davidii)callus.Bioinformatics and promoter cis-acting elements were predicted,and correlations between melatonin content and VdSNAT2 and VdCOMT3 gene expression in the callus of spine grapes was determined using real-time fluorescent quantitative PCR.The open reading frames(ORFs)of VdSNAT2 and VdCOMT3 were 549 and 1158 bp,respectively,encoding 182 and 385 amino acids.The encoded proteins were hydrophilic and hydrophobic proteins,respectively,localized in chloroplasts.Protein domain and evolutionary tree analyses showed that VdSNAT2 and VdCOMT3 proteins in grapes have similar evolution and conserved motifs.Analysis of cis-acting promoter elements revealed the presence of biological/abiotic response elements in both VdSNAT2 and VdCOMT3 promoter sequences.qRT-PCR analysis showed that light quality exerts a significant impact on VdSNAT2 and VdCOMT3 expression,with red light significantly enhancing expression in spine grape calli and expression correlating positively with melatonin content.This study showed that light quality can affect VdSNAT2 and VdCOMT3 expression,thereby regulating melatonin biosynthesis in the calli of spine grapes.

关 键 词：刺葡萄 愈伤组织 褪黑素 光质 5-羟色胺-N-乙酰基转移酶 咖啡酸-O-甲基转移酶 

分 类 号：S663.1[农业科学—果树学]