白茶萎凋过程萜烯类合成相关基因的鉴定和表达分析

陈雪津; 王鹏杰; 林馨颖; 谷梦雅; 郑玉成; 郑知临; 叶乃兴

doi:10.13305/j.cnki.jts.2020.03.007

茶叶科学 >

2020 , Vol. 40 >Issue 3: 363 - 374

DOI: https://doi.org/10.13305/j.cnki.jts.2020.03.007

白茶萎凋过程萜烯类合成相关基因的鉴定和表达分析

陈雪津 ,
王鹏杰 ,
林馨颖 ,
谷梦雅 ,
郑玉成 ,
郑知临 ,
叶乃兴

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福建农林大学园艺学院/茶学福建省高校重点实验室,福建福州 350002

陈雪津,女,硕士研究生,主要从事茶树栽培育种和生物技术。

收稿日期: 2019-11-07

修回日期: 2019-12-13

网络出版日期: 2020-06-09

基金资助

国家自然科学基金项目（31270735）、福建农林大学科技创新专项基金项目（CXZX2017181、CXZX2016117）

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Identification and Expression Analysis of Terpene Synthesis Related Genes during the Withering of White Tea

CHEN Xuejin ,
WANG Pengjie ,
LIN Xinying ,
GU Mengya ,
ZHENG Yucheng ,
ZHENG Zhilin ,
YE Naixing

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College of Horticulture, Fujian Agriculture and Forestry University/Key Laboratory of Tea Science at Universities in Fujian, Fuzhou 350002, China

Received date: 2019-11-07

Revised date: 2019-12-13

Online published: 2020-06-09

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摘要

萜烯类化合物是植物中重要的次生代谢产物之一,对茶树挥发性香气的组成起着重要作用。从茶树基因组数据库中鉴定获得了141个茶树萜烯类合成相关基因,并对其不同组织表达特异性进行分析,筛选出16个在茶树顶芽和嫩叶中高表达的萜烯类合成代表基因。生物信息学分析结果表明,系统进化关系将茶树与拟南芥和葡萄的萜烯类合成相关基因分成了4个亚家族;茶树萜烯类合成相关基因含有5~14个外显子,在上游启动子区域分析发现了大量与光响应、植物生长发育、激素和胁迫响应密切相关的顺式作用元件。荧光定量检测发现,CsMVK、CsDXS和CsGGPS在白茶萎凋过程中的表达显著上调;CsDXR、CsMCT、CsCMK、CsMCS、CsHDS、CsGPPS和CsGGPPS在萎凋4 h和24 h的表达达到峰值。本研究结果为进一步挖掘茶叶萎凋过程中萜烯类合成相关基因对茶叶香气组分积累提供了理论依据。

关键词： 白茶; 香气; 萜烯类; 表达分析

本文引用格式

陈雪津 , 王鹏杰 , 林馨颖 , 谷梦雅 , 郑玉成 , 郑知临 , 叶乃兴 . 白茶萎凋过程萜烯类合成相关基因的鉴定和表达分析[J]. 茶叶科学, 2020 , 40(3) : 363 -374 . DOI: 10.13305/j.cnki.jts.2020.03.007

Abstract

Terpenes are the important secondary metabolites in plants and play an important role in the composition of the volatile aroma of tea plants. In this study, 141 tea plant terpenoid synthesis-related genes were identified from the tea plant genome database. Their expression specificities in different tissues were analyzed. Sixteen terpene synthetic genes which were highly expressed in the apical buds and young leaves of tea plants were screened. The results of bioinformatics methods show that the phylogenetic relationship divides the genes related to terpene synthesis of tea plant, Arabidopsis and grape into four subfamilies. The terpenoid synthesis related genes contain 5 to 14 exons and a large number of cis-related elements closely related to light response, plant growth and development, hormone and stress response according to the upstream promoter region analysis. Fluorescence quantitative detection showed that the expressions of CsMVK, CsDXS and CsGGPS were significantly up-regulated during the withering process of white tea. The expressions of CsDXR, CsMCT, CsCMK, CsMCS, CsHDS, CsGPPS and CsGGPPS showed the highest expressions at 4 h and 24 h after withering. The results of this study provided a theoretical basis for further exploring the functions of terpenoid synthesis related genes in tea.

Key words： white tea; aroma; terpenes; expression analysis

参考文献

[1] 叶乃兴. 白茶:科学·技术与市场[M]. 北京: 中国农业出版社, 2010: 136-137.
Ye N X.The science technology and market of white tea [M]. Beijing: China agriculture press, 2010: 136-137.
[2] Chen Q C, Zhu Y, Dai W D, et al.Aroma formation and dynamic changes during white tea processing[J]. Food Chemistry, 2019, 274: 915-924.
[3] Wang Y, Zheng P C, Liu P P, et al.Novel insight into the role of withering process in characteristic flavor formation of teas using transcriptome analysis and metabolite profiling[J]. Food Chemistry, 2018, 272: 313-322.
[4] Deng W W, Wang R, Yang T, et al.Functional characterization of salicylic acid carboxyl methyltransferase fromCamellia sinensis, providing the aroma compound of methyl salicylate during withering process of white tea[J]. Journal of Agricultural and Food Chemistry, 2017, 65(50): 11036-11045.
[5] 徐燕. 茶树萜类合成途径关键基因克隆及表达研究[D]. 杭州: 浙江大学, 2013.
Xu Y.Studies in cloning and expression of the key genes involved in the terpenoids metabolic pathway of tea plant [D]. Hangzhou: Zhejiang University, 2013.
[6] Yahyaa M, Tholl D, Cormier G, et al.Identification and characterization of terpene synthases potentially involved in the formation of volatile terpenes in carrot (Daucus carotaL.) roots[J]. Journal of Agricultural and Food Chemistry, 2015, 63(19): 4870-4878.
[7] 贺志荣, 项威, 徐燕, 等. 茶树挥发性萜类物质及其糖苷化合物生物合成的研究进展[J]. 茶叶科学, 2012, 32(1): 1-8.
He Z R, Xiang W, Xu Y, et al.Progress in the research biosynthesis of volatile terpenoids and their glycosides in the tea plant[J]. Journal of Tea Science, 2012, 32(1): 1-8.
[8] 张冬桃, 孙君, 叶乃兴, 等. 茶树萜烯类香气物质合成相关酶研究进展[J]. 茶叶学报, 2016, 56(2): 68-79.
Zhang D T, Sun J, Ye N X, et al.Research progress of enzymes associated with terpene synthesis inCamellia sinensis[J]. Journal of Tea, 2016, 56(2): 68-79.
[9] 王鹏杰, 陈丹, 曹红利, 等. 茶树甲羟戊酸焦磷酸脱羧酶基因CsMVD的克隆与表达分析[J]. 西北植物学报, 2017, 37(12):2342-2349.
Wang P J, Chen D, Cao H L, et al.Cloning and expression of mevalonate diphosphate decarboxylase geneCsMVDin tea plant (Camellia sinensis)[J]. Journal of Northwest Botany, 2017, 37(12): 2342-2349.
[10] 姚雪倩, 岳川, 杨国一, 等. 茶树牻牛儿基牻牛儿基焦磷酸合成酶基因CsGGDPS的克隆及表达分析[J]. 茶叶科学, 2017, 37(1): 86-96.
Yao X Q, Yue C, Yang G Y, et al.Cloning and expression analysis of geranygeranyl diphosphate synthase gene CsGGDPS in tea plant (Camellia sinensis)[J]. Journal of Tea Science, 2017, 37(1): 86-96.
[11] 王鹏杰, 陈丹, 俞滢, 等. 茶树单萜合成酶CsTPS基因的克隆及表达分析[J]. 西北植物学报, 2017, 37(8): 1465-1473.
Wang P J, Chen D, Yu Y, et al.Cloning and expression analysis of Monoterpene Synthase GCsTPSin tea plant (Camellia sinensis)[J]. Northwest Journal of Botany, 2017, 37(8): 1465-1473.
[12] Hu C J, Li D, Ma Y X, et al.Formation mechanism of the oolong tea characteristic aroma during bruising and withering treatment[J]. Food Chemistry, 2018, 269: 202-211.
[13] Zheng Y C, Wang P J, Chen X J, et al.Transcriptome and metabolite profiling reveal novel insights into volatile heterosis in the tea plant (Camellia sinensis)[J]. Molecules, 2019, 24(18): 3380. doi: org/10.3390/molecules24183380
[14] 陈静, 俞滢, 张丹丹, 等. 白茶萎凋过程中儿茶素合成关键酶基因表达分析[J]. 南方农业学报, 2016, 47(8): 1364-1369.
Chen J, Yu Y, Zhang D D, et al.Expression of genes encoding of key enzymes in biosynthesis pathways of catechins in the withering process of white tea[J]. Southern Journal of Agriculture, 2016, 47(8): 1364-1369.
[15] Wei C L, Yang H, Wang S B, et al.Draft genome sequence ofCamellia sinensisvar.sinensisprovides insights into the evolution of the tea genome and tea quality[J]. Proceedings of the National Academy of Sciences, 2018, 115(18): E4151-E4158.
[16] 王鹏杰, 郑玉成, 林浥, 等. 茶树GRF基因家族的全基因组鉴定及表达分析[J]. 西北植物学报, 2019, 39(3): 38-46.
Wang P J, Zheng Y C, Lin Y, et al.Genome-wide identification and expression analysis ofGRFgene Family inCamellia sinensis[J]. Journal of Northwest Botany, 2019, 39(3): 38-46.
[17] Trapnell C, Roberts A, Goff L, et al.Differential gene and transcript expression analysis of RNA-seq experiments with tophat and cufflinks[J]. Nature Protocols, 2012, 7(3): 562-578.
[18] Anders S, Pyl P T, Huber W, et al.Htseq-a python framework to work with high-throughput sequencing data[J]. Bioinformatics, 2015, 31(2): 166-169.
[19] Livak K, Schmittgen T.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta c(t)) method[J]. Methods, 2000, 25(4): 402-408.
[20] 岳跃冲, 范燕萍. 植物萜类合成酶及其代谢调控的研究进展[J]. 园艺学报, 2011, 38(2): 379-388.
Yue Y C, Fan Y P.The terpenoid synthase and Regulation of terpene metabolism in plants[J]. Horticultural Journal, 2011, 38(2): 379-388.
[21] Xu Y Y, Zhu C Q, Xu W P, et al.Integration of metabolite profiling and transcriptome analysis reveals genes related to volatile terpenoid metabolism in finger citron (C. medicavar.sarcodactylis). Molecules, 2019, 24(14), 2564. doi: 10.3390/molecules24142564.
[22] Shi J W, Fei X T, Hu Y, et al.Identification of key genes in the synthesis pathway of volatile terpenoids in fruit of zanthoxylum bungeanum maxim[J]. Forests, 2019, 10(4): 328. doi: org/10.3390/f10040328.
[23] Zeng X L, Cai L, Riru Z, et al.Emission and accumulation of monoterpene and the key terpene synthase (TPS) associated with monoterpene biosynthesis in osmanthus fragrans lour[J]. Frontiers in Plant Science, 2016, 6: 1232. doi: org/10.3389/fpls.2015.01232.
[24] Tholl D, Lee S.Terpene specialized metabolism inArabidopsis thaliana[J]. The Arabidopsis Book, 2011, 9: e0143. Doi: 10.1199/tab.0143.
[25] Wang Y, Zheng P C, Liu P P, et al.Novel insight into the role of withering process in characteristic flavor formation of teas using transcriptome analysis and metabolite profiling[J]. Food Chemistry, 2019, 272: 313-322.
[26] Han Z X, Rana M M, Liu G F, et al.Green tea flavour determinants and their changes over manufacturing processes[J]. Food Chemistry, 2016, 212: 739-748.
[27] Ying Y, Shi H L, Dan C, et al.Volatiles emitted at different flowering stages of jasminum sambac and expression of genes related toα-farnesene biosynthesis[J]. Molecules, 2017, 22(4): 546. doi: org/10.3390/molecules22040546.
[28] Xu Q S, He Y X, Yan X M, et al.Unraveling a crosstalk regulatory network of temporal aroma accumulation in tea plant (Camellia sinensis) leaves by integration of metabolomics and transcriptomics[J]. Environmental and Experimental Botany, 2018, 149: 81-94.
[29] Lorenzo C, Albert C, Patricia B, et al.Enhanced flux through the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing deoxyxylulose 5-phosphate reductoisomerase[J]. Plant Molecular Biology, 2006, 62(4): 683-695.
[30] Phillips M A, Walter M H, Ralph S G, et al.Functional identification and differential expression of 1-deoxy-d-xylulose 5-phosphate synthase in induced terpenoid resin formation of Norway spruce (Picea abies)[J]. Plant Molecular Biology, 2007, 65(3): 243-257.

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