基于高通量测序筛选&#x02018;紫娟&#x02019;花青素合成相关的miRNA

陈林波; 夏丽飞; 刘悦; 孙云南; 蒋会兵; 田易萍; 陈亮

doi:10.13305/j.cnki.jts.2019.06.007

茶叶科学 >

2019 , Vol. 39 >Issue 6: 681 - 691

DOI: https://doi.org/10.13305/j.cnki.jts.2019.06.007

基于高通量测序筛选‘紫娟’花青素合成相关的miRNA

陈林波 ,
夏丽飞 ,
刘悦 ,
孙云南 ,
蒋会兵 ,
田易萍 ,
陈亮

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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;
2. 中国农业科学院茶叶研究所,浙江杭州 310008

陈林波,男,研究员,主要从事茶树资源育种研究。

收稿日期: 2019-03-04

修回日期: 2019-05-25

网络出版日期: 2019-12-24

基金资助

国家自然科学基金项目（No.31560220）、国家重点研发项目（2016YFD0200903）、国家茶叶额产业体系（CARS-19）、云南省人才培养计划项目（2015HB105）

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Screening of miRNA Related to Anthocyanin Synthesis in Tea Cultivar ‘Zijuan’ Based on High Throughput Sequencing

CHEN Linbo ,
XIA Lifei ,
LIU Yue ,
SUN Yunnan ,
JIANG Huibing ,
TIAN Yiping ,
CHEN Liang

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1. Tea Research Institute, Yunnan Academy of Agricultural Sciences/Yunnan Engineering Research Centerof Tea Germplasm Innovation and Matching Cultivation /Yunnan Provincial Key Laboratory of Tea Science, Menghai 666201, China;
2. Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China

Received date: 2019-03-04

Revised date: 2019-05-25

Online published: 2019-12-24

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摘要

为筛选调控紫娟茶树花青素合成相关miRNA,以茶树品种紫娟（ZJ）、云抗10号（YK）和福鼎大白茶（FD）为材料,构建了miRNA文库。鉴定出46种已知的miRNAs和67种新的miRNAs,预测到具有注释的靶基因765个。通过差异表达分析,筛选出在ZJ与YK、ZJ与FD共有差异表达的miRNA 24个。通过对24个差异表达miRNA的靶基因分析,筛选出可能参与调控花青素合成的miRNA 4个,包括miR828a、miR845c、novel_14和novel_87,其预测的靶基因包括转录因子基因MYB4、MYB23、MYB26、MYB82、bHLH74以及4-香豆酰辅酶A链接酶（4CL）、二氢黄酮醇4-还原酶（DFR）和UDP-葡萄糖黄酮3-O-葡糖基转移酶（UFGT）等基因。利用RT-PCR分析8个差异表达的miRNA,其结果与转录组分析一致。本研究结果为进一步开展茶树花青素生物合成的调控机制研究奠定基础。

关键词： 茶树; 花青素合成; 高通量测序; miRNA; 靶基因

本文引用格式

陈林波 , 夏丽飞 , 刘悦 , 孙云南 , 蒋会兵 , 田易萍 , 陈亮 . 基于高通量测序筛选‘紫娟’花青素合成相关的miRNA[J]. 茶叶科学, 2019 , 39(6) : 681 -691 . DOI: 10.13305/j.cnki.jts.2019.06.007

Abstract

In order to screen the miRNAs related to anthocyanin synthesis in 'Zijuan', the miRNA library was constructed by using the tea cultivars 'Zijuan' (ZJ), 'Yunkang 10' (YK) and 'Fuding Dabaicha' (FD). In this study, 46 known miRNAs and 67 unknown miRNAs were identified, and 765 annotated target genes were predicted. A total of 24 miRNAs were screened out, which were differentially expressed between ZJ and YK, and between ZJ and FD. Four target miRNAs involved in the regulation of anthocyanin synthesis, namely miR828a, miR845c, novel_14 and novel_87, were identified by target gene analysis of 24 differentially expressed miRNAs. The predicted target genes include transcription factor genes MYB4, MYB23, MYB26, MYB82, bHLH74 and 4CL (4-coincyl-CoA-linked enzyme), DFR (dihydroflavonol 4-reductase) and UFGT (UDP-glucoside flavonoid 3-O-glucosyltransferase). Eight differentially expressed miRNAs were analyzed by RT-PCR and the results were consistent with transcriptome analysis. Finally, a theoretical basis for further study of regulation mechanisms related to anthocyanin biosynthesis in tea plant was provided.

Key words： Camellia sinensis; anthocyanin synthesis; high throughput sequencing; microRNAs; target genes

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