紫芽六堡茶花青素合成相关基因的转录组测序及表达分析  

Transcriptome Analysis on Anthocyanin Synthesis-related Genes in Liupao Tea Plants

作　　者：梁燕妮[1] 魏诗琴 乔如颖 梁剑锋 谭花桂 LIANG Yanni;WEI Shiqin;QIAO Ruying;LIANG Jianfeng;TAN Huagui(School of Food and Pharmaceutical Engineering,Wuzhou University(Guangxi Liupao Tea Modern Industry College),Wuzhou,Guangxi 543002,China)

机构地区：[1]梧州学院食品与制药工程学院(六堡茶现代产业学院),广西梧州543002

出　　处：《福建农业学报》2024年第6期711-719,共9页Fujian Journal of Agricultural Sciences

基　　金：广西自然科学基金面上项目(2023GXNSFAA026221);广西高校中青年教师基础能力提升项目(2021KY0680);广西大学生创新创业项目(2024年)。

摘　　要：【目的】紫芽六堡茶(Camellia sinensis var. sinensis cv. Liupao)是六堡群体种茶树中芽叶呈紫色且富含花青素的特异品系,研究紫芽六堡茶花青素合成相关基因可为深入了解紫芽六堡茶花青素积累的分子机制奠定基础,为高花青素六堡茶树的分子育种提供遗传资源。【方法】利用盐酸乙醇分别提取紫芽和绿芽六堡茶芽中花青素,测定其含量;通过Illumina Hiseq 2 500高通量测序平台对紫芽和绿芽六堡茶进行转录组测序,分析花青素合成相关基因的表达水平,找出差异表达基因,进一步进行GO功能分析和KEGG富集分析;并通过荧光定量PCR对转录组测序结果进行验证。【结果】紫芽六堡茶花青素含量是绿芽的7倍;转录组测序共获得165 570条Unigene,平均长度1 450bp;对测序结果进行分析,筛选出与紫芽六堡茶花青素生物合成通路相关的基因243条,进一步筛选出43个差异显著表达基因,这些基因共编码14个关键酶,包括苯丙氨酸氨裂解酶(phenylalanine ammonialyase, PAL)、查尔酮合成酶(chalconesynthase,CHS)、查尔酮异构酶(chalconeisomerase,CHI)、肉桂酸4-羟化酶(cinnamateacid4-hydroxylase, C4H)、花色素苷还原酶(anthocyanidin reductase, ANR)、4-香豆酸CoA连接酶(4-coumarate-Co A ligase,4CL)、乙酰辅酶A羧化酶(acetyl-CoA carboxylase, ACC)、黄酮醇合成酶(flavonol synthase, FLS)、黄酮3′,5′-羟化酶(flavonoid-3′,5′-hydroxylase, F3'5'H)、黄酮3′-羟化酶(flavonoid 3'-hydroxylase, F3'H)、黄酮3-羟化酶(flavanone-3-hydroxylase, F3H)、二氢黄酮醇还原酶(dihydroflavonol 4-reductase, DFR)、花青素成合酶(anthocyanidin synthase,ANS)、无色花青素还原酶(leucoanthocyanidin reductase, LAR)。【结论】编码花青素生物合成的14个关键酶基因中有34个基因在紫芽六堡茶中上调表达,推测这些基因在紫芽花青素积累中起重要作用。Objective Liupao Tea(Camellia sinensis var.sinensis cv.Liupao)was studied for the genes associated with anthocyanin synthesis for target breeding.Methods Anthocyanin were extracted with ethanol hydrochloride from Liupao Tea plants bearing purple or green buds.Transcriptome sequencing was performed using Illumina Hiseq 2500 high-throughput platform to identify differentially expressed genes,determine expression of the genes related to anthocyanin synthesis,and conduct GO and KEGG enrichment analyses on the extracts.The sequencing results were subsequently verified by fluorescence quantification PCR.Results The young leaves on the tea plants borne with purple buds contained sevenfold higher anthocyanin than those on the plants with green buds.The transcriptome of the genes of the purple buds had 165570 unigenes with an average length of 1450 bp.Of them,243 related to the anthocyanin biosynthesis pathway,and 43 significantly differentially expressed between the two types of plants.The 43 differentiated genes encoded 14 key enzymes,i.e.,phenylalanine ammonia-lyase(PAL),chalcone synthase(CHS),chalcone isomerase(CHI),cinnamate acid 4-hydroxylase(C4H),anthocyanidin reductase(ANR),4-coumarate-CoA ligase(4CL),acetyl-CoA carboxylase(ACC),flavonol synthase(FLS),flavonoid-3′,5′-hydroxylase(F3′5′H),flavonoid 3′-hydroxylase(F3′H),flavanone-3-hydroxylase(F3H),dihydroflavonol 4-reductase(DFR),anthocyanidin synthase(ANS),and leucoanthocyanidin reductase(LAR).Conclusion Thirty-four genes significantly upregulated in Liupao Tea plants with purple buds were found to be associated with 14 key enzymes encoding anthocyanin biosynthesis.They were speculated to play an important role in the bud color differentiation between the two varieties.

关 键 词：紫芽 六堡茶 花青素 转录组 差异表达基因 

分 类 号：S571.1[农业科学—茶叶生产加工]