基于转录组测序的紫芽茶树花青素合成相关基因分析
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1云南省农业科学院茶叶研究所/云南省茶树种质资源创新与配套栽培技术工程研究中心/云南省茶学重点实验室,云南勐海 666201;,1云南省农业科学院茶叶研究所/云南省茶树种质资源创新与配套栽培技术工程研究中心/云南省茶学重点实验室,云南勐海 666201;,1云南省农业科学院茶叶研究所/云南省茶树种质资源创新与配套栽培技术工程研究中心/云南省茶学重点实验室,云南勐海 666201;,2中国农业科学院茶叶研究所,浙江杭州 310008,1云南省农业科学院茶叶研究所/云南省茶树种质资源创新与配套栽培技术工程研究中心/云南省茶学重点实验室,云南勐海 666201;,1云南省农业科学院茶叶研究所/云南省茶树种质资源创新与配套栽培技术工程研究中心/云南省茶学重点实验室,云南勐海 666201;

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国家自然科学基金项目(31560220);茶树生物学与资源利用国家重点实验室开放基金项目(SKLTOF20150105);云南省人才培养计划项目(2015HB105)


Transcriptome Analysis of Anthocyanin Synthesis Related Genes in Purple Bud Tea Plant
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1Tea Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Engineering Research Center of Tea Germplasm Innovation and Matching Cultivation / Yunnan Provincial Key Laboratory of Tea Science, Yunnan Menghai 666201;,1Tea Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Engineering Research Center of Tea Germplasm Innovation and Matching Cultivation / Yunnan Provincial Key Laboratory of Tea Science, Yunnan Menghai 666201;,1Tea Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Engineering Research Center of Tea Germplasm Innovation and Matching Cultivation / Yunnan Provincial Key Laboratory of Tea Science, Yunnan Menghai 666201;,2Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008,1Tea Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Engineering Research Center of Tea Germplasm Innovation and Matching Cultivation / Yunnan Provincial Key Laboratory of Tea Science, Yunnan Menghai 666201;,1Tea Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Engineering Research Center of Tea Germplasm Innovation and Matching Cultivation / Yunnan Provincial Key Laboratory of Tea Science, Yunnan Menghai 666201;

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    摘要:

    为筛选调控茶树芽叶紫色性状相关候选基因,以紫芽茶树品种‘紫娟’、绿芽茶树品种‘云抗10号’和‘福鼎大白茶’为材料,利用液质联用法(HPLC-MS)、转录组测序(RNA-Seq)和实时荧光定量PCR技术(qRT-PCR),分析了紫芽和绿芽茶树品种芽叶花青素、儿茶素含量及合成相关基因的表达水平。结果表明,紫娟、云抗10号和福鼎大白茶芽叶花青素含量分别为5.05 mg/g、0.08 mg/g和0.15 mg/g,紫芽与绿芽茶树间花青素含量差异显著。3个茶树品种儿茶素含量分别为15.12 mg/g、19.52 mg/g和15.09 mg/g,差异不显著。转录组测序共获得255 079条转录本序列(Transcripts),组装出166 118条单基因簇(Unigenes)。所得单基因簇(Unigenes)注释为GO分类中54 446条,KOG分类中30 666条,KEGG分类中20 336条。DEG-Seq分析得到紫芽与绿芽茶树品种间共有434条差异表达基因,涉及到38个代谢通路。进一步筛选到类黄酮生物合成途径相关基因112条,有15条基因在紫芽与绿芽茶树品种间差异表达,其中PAL (Cluster-1850.70337)、CHS (Cluster-21971.0)、ANS (Cluster-1850.80848)和UFGT (Cluster-1850.77163)基因在紫芽茶树品种中上调表达,在绿芽茶树品种间表达差异不显著,表达趋势与花青素积累情况相似。FLS (Cluster-1850.81068)基因在绿芽茶树品种中均为上调表达,可能与儿茶素合成相关。qRT-PCR随机检测9个差异表达基因,表明不同茶树品种间差异表达基因的相对表达水平与转录组测序结果基本一致。因此,从基因表达模式推测,PAL、CHS、ANS和UFGT基因可能在紫芽茶树芽叶花青素生物合成途径中起关键作用。本研究探索了茶树芽叶紫色性状的转录组信息,筛选到一些差异表达基因,为进一步研究茶树芽叶呈色机制提供参考。

    Abstract:

    In order to screen the differential expression of young shoot pigment related genes of the tea plant. In this study, tea cultivars for example purple bud ‘Zijuan’, green bud ‘Yunkang No.10’ and ‘Fuding Dabaicha’ were used as materials. The contents and compositions of anthocyanin and catechin were analyzed by high performance liquid chromatography-mass (HPLC-MS), and the expression of synthesis-related genes were tested by RNA-sequencing (RNA-Seq) and quantitative real-time (qRT-PCR) technology. HPLC-MS analysis results showed that the contents of anthocyanin in ‘Zijuan’, ‘Yunkang No.10’ and ‘Fuding Dabaicha’ were 5.05 mg/g, 0.08 mg/g and 0.15 mg/g, respectively, and the variance was significantly difference among purple bud vs. green bud tea cultivars (P<0.05). Catechin contents of three tea cultivars were 15.12 mg/g, 19.52 mg/g and 15.09 mg/g, respectively, their difference were not significant (P?0.05). RNA-Seq results showed that a total of 255 079 reads were produced and assembled into a total of 166 118 unigenes. All unigenes were identified as putative homologs of annotated sequences in public protein databases, of which 54 446 were assigned to GO terms, 30 666 were assigned to KOG categories, and 20 336 were assigned to KEGG pathways. DEG-Seq analysis results indicated that 434 differentially expressed genes between purple bud vs. green bud tea cultivars, mainly related 38 metabolic pathways. Further analysis was found, 112 genes were expressed in anthocyanin biosynthesis pathway of tea plant, among them15 genes were differentially expressed between purple bud vs. green bud tea cultivars. We found that the relative expression of PAL (cluster-1850.70337), CHS (cluster-21971.0), ANS (cluster-1850.80848) and UFGT (cluster-1850.77163) were up-regulated expression in purple bud tea cultivar, but the relative expression of these genes showed no significantly difference among green bud tea cultivars. Which was consistent with the anthocyanin synthesis changes. FLS (Cluster-1850.81068) genes were up-regulated expression in green bud tea cultivars, may be related to the catechin accumulation in green bud tea cultivars. The expressed contents of the selected 9 differentially expressed genes by the qRT-PCR, were basically the same with the sequencing results by RNA-Seq technology. Therefore, based on the differential gene expression patterns, this study suggested that PAL、CHS、ANS and UFGT genes may plays an important role in anthocyanin synthesis of purple bud tea plant. Transcriptome information about leaf color was explored for tea plant, and the differentially expressed genes were identified in this study. It will provide a theoretical basis for further understanding of the leaf color in the tea plant.

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蒋会兵,夏丽飞,田易萍,等.基于转录组测序的紫芽茶树花青素合成相关基因分析[J].植物遗传资源学报,2018,19(5):967-978.

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  • 收稿日期:2018-01-26
  • 最后修改日期:2018-03-25
  • 录用日期:2018-04-18
  • 在线发布日期: 2018-07-18
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