茶树甜菜碱醛脱氢酶基因(CsBADH1)的全长cDNA克隆与表达分析

doi:10.13305/j.cnki.jts.2013.02.009

茶叶科学 ›› 2013, Vol. 33 ›› Issue (2): 99-108.doi: 10.13305/j.cnki.jts.2013.02.009

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茶树甜菜碱醛脱氢酶基因(CsBADH1)的全长cDNA克隆与表达分析

曹红利^1,2, 郝心愿^1,3, 岳川^1,2, 马春雷¹, 王新超^1,*, 杨亚军^1,*

1. 中国农业科学院茶叶研究所国家茶树改良中心, 浙江杭州 310008;
2. 中国农业科学院研究生院, 北京 100081;
3. 西北农林科技大学园艺学院, 陕西杨凌 712100

收稿日期:2012-08-02 修回日期:2012-11-19 出版日期:2013-04-30 发布日期:2019-09-04
通讯作者: *
作者简介:曹红利（1987— ）,女,四川泸州人,硕士研究生,主要从事茶树分子生物学研究。
基金资助:
国家自然科学基金（31170650）、浙江省自然科学基金项目（Z3100473, Y3100291）、国家茶叶产业技术体系（CARS-23）

Cloning and Expression Analysis of Betaine Aldehyde Dehydragenase Gene (CsBADH1) from Tea Plant [Camellia sinensis (L.) O. Kuntze]

CAO Hong-li^1,2, HAO Xin-yuan^1,3, YUE Chuan^1,2, MA Chun-lei¹, WANG Xin-chao^1,*, YANG Ya-jun^1,*

1. Tea Research Institute of Chinese Academy of Agricultural Sciences; National Center for Tea Improvement, Hangzhou 310008, China;
2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China;
3. College of Horticulture, Northwest A&F University, Yangling 712100, China;

Received:2012-08-02 Revised:2012-11-19 Online:2013-04-30 Published:2019-09-04

摘要/Abstract

摘要： 甜菜碱是一种能在逆境下大量积累的无毒性渗透相溶物质,而甜菜碱醛脱氢酶（BADH）是甜菜碱合成途径中的关键酶之一。本实验采用SMART-RACE技术获得了茶树甜菜碱醛脱氢酶基因的全长cDNA序列,命名为CsBADH1（GenBank 登陆号：JX050145）,并对其进行相关的生物信息学分析。结果表明：CsBADH1的cDNA序列全长1β972βbp,其开放阅读框（ORF）为1β518βbp,编码505个氨基酸,预测分子量为54.8βkD,理论等电点（PI）为5.652,是疏水性很强的蛋白,且具有BADH基因典型的高度保守十肽基序（VTLELGGKSP）和与醛脱氢酶（ALDHs）功能有关的半胱氨酸残基（C）。系统进化树分析表明,茶树BADH氨基酸序列与人参（Panax ginseng）的亲缘关系最近,相似度达87%,其余相似性也大部分在80%以上。实时荧光定量PCR分析结果显示：该基因在经历一定时间的冷驯化后表达量升高,说明CsBADH1基因可能参与茶树的冷驯化作用。

关键词: 茶树, 冷驯化, 甜菜碱醛脱氢酶（BADH）, 基因克隆, 表达分析

Abstract: Betaine is nontoxic osmotic solute which can accumulate as cryoprotectant molecule in response to environment stress in plants. Betaine aldehyde dehydragenase (BADH) is one of the key enzymes involving in the betaine biosynthesis pathway. A full length cDNA sequence of BADH gene was cloned by SMART RACE-PCR from tea plant, which was named as CsBADH1(GenBank Accession No. JX050145). The bioinformatic characterization indicated that the full length cDNA of CsBADH1 was 1972 bp,which contained 1β518βbp complete CDS and encoded 505 amino acid residues with a putative molecular mass of 54.8βkD and an isoionic point of 5.652. It belongs to a hydrophobic protein. The deduced amino acid sequence contained the conserved decapeptide (VTLELGGKSP) and cysteine residue(C) in aldehyde dehydrogenase (ALDHs). Phylogenetic analysis showed that CsBADH1 was most closely to Panax ginseng with 87% amino acids similarity, the most of other plants were also above 80%. Quantitative real-time PCR analysis further showed that the expression levels of CsBADH1 in acclimated samples were higher than non-acclimated samples (CK). It indicates that CsBADH1 may mediate the regulation of cold acclimation in tea plant.

Key words: tea plant (Camellia sinensis), cold acclimation, betaine aldehyde dehydrogenase (BADH), gene clonging, expression analysis

中图分类号:

曹红利, 郝心愿, 岳川, 马春雷, 王新超, 杨亚军. 茶树甜菜碱醛脱氢酶基因(CsBADH1)的全长cDNA克隆与表达分析[J]. 茶叶科学, 2013, 33(2): 99-108. doi: 10.13305/j.cnki.jts.2013.02.009.

CAO Hong-li, HAO Xin-yuan, YUE Chuan, MA Chun-lei, WANG Xin-chao, YANG Ya-jun. Cloning and Expression Analysis of Betaine Aldehyde Dehydragenase Gene (CsBADH1) from Tea Plant [Camellia sinensis (L.) O. Kuntze][J]. Journal of Tea Science, 2013, 33(2): 99-108. doi: 10.13305/j.cnki.jts.2013.02.009.

参考文献 20

[1]	Hanson AD, May AM, Grument R.Betaine synthesis in chenopods: localization in chloroplasts[J]. Proceedings of National Academy of Sciences of the United States of America, 1985, 82: 3678-3682.
[2]	Jagendorf AT, Takabe T.Inducers of glycine betaine synthesis in barley[J]. Plant Physiology, 2001, 127(4): 1827-1835.
[3]	Arakawa K, Katayama M, Takabe T.Level of betaine and betaine aldehydede hydrogenase activity in the green leaves and etiolated leaces and roots of barley[J]. Plant Cell Physiology, 1990, 31: 797-803.
[4]	Nakamura T, Yokota S, Muramoto Y, et al. Expression of a betaine aldehyde dehydrogenase gene in rice, a glycinebetaine nonaccumulator, and possible localization of its protein in peroxisomes[J]. The Plant Journal, 1997, 11(5): 1115-1120.
[5]	Sophos NA, Vasiliou V. Aldehyde dehydrogenase gene superfamily: the2002 update[J]. Chemico-Biological Interactions, 2003, 143-144(1): 5-22.
[6]	Weretilnyk EA, Hanson AD.Molecular cloning of a plant betaine aldehyde dehydrogenase, an enzyme implicated in adaptation to salinity and drought[J]. Proceedings of National Academy of Sciences of the United States of America, 1990, 87: 2745-2749.
[7]	王新超, 杨亚军. 茶树抗性育种研究现状[J]. 茶叶科学, 2003, 23(2): 94-98.
[8]	杨亚军, 郑雷英, 王新超. 冷驯化和ABA对茶树抗寒力及其体内脯氨酸含量的影响[J]. 茶叶科学, 2004, 24(3): 177-182.
[9]	Yin XJ, Zhao YX, Luo D, et al. Isolating the promoter of a stress-induced gene encoding betaine aldehyde dehydrogenase from the halophyte Atriplex centralasiatica Iljin[J]. Biochimica et Biophysica Acta (BBA)-Gene Structure and Expression, 2002, 1577: 452-456.
[10]	Bendtsen J D, Nielsen H, Heijne G, et al. Improved prediction of signal peptides: SignalP 3.0[J]. Journal of Molecular Biology, 2004, 340: 783-795.
[11]	Tamura K, Dudley J, Nei M, et al. MEGA4: Molecular evolutionary genetics analysis(MEGA)software version 4.0[J]. Molecular Biology and Evolution, 2007, 2415: 1596-1599.
[12]	Combet C, Jambon M, Deléage G, et al. Geno3D: Automatic comparative molecular modelling of protein[J]. Bioinformatics, 2002, 18: 213-214.
[13]	赵丽萍, 陈亮, 王新超, 等. 茶树新梢不同叶片中β-葡萄糖苷酶和β-樱草糖苷酶基因表达的实时定量PCR分析[J]. 茶叶科学, 2006, 26(1): 11-16.
[14]	Nakamura T, Nomura M, Mori H, et al. An isozyme of betaine aldehyde dehydrogenase in barley[J]. Plant Cell Physiology, 2001, 42(10): 1088-1092.
[15]	Hibino T, Meng YL, Kawamitsu Y, et al. Molecular cloning and function characterization of two kinds of betaine-accumulating mangrove Avicennia marina(Forsk) Vierh[J]. Plant Molecular Biology, 2001, 45: 353-363.
[16]	Li QL, Gao XR, Yu XH, et al. Molecular cloning and characterization of betaine aldehyde dehydrogenase gene from Suaeda liaotungensis and its use in improved tolerance to salinity in transgenic tobacco[J]. Biotechnology Letters, 2003, 25: 1431-1436.
[17]	Ton HH, Chen, Norio Murata.Glycinebetaine protects plants against abiotic stress: mechanisms and biotechnological applications[J]. Plant, Cell and Environment, 2011, 34: 1-20.
[18]	Jia G X, Zhu Z Q, Chang F Q, et al. Transformation of tomato with the BADH gene from Atriplex improves salt tolerance[J]. Plant Cell Reports, 2002, 21: 141-146.
[19]	Supaporn H, Valentine N, Kanyaratt S, et al. Expression of Indica rice OsBADH1 gene under salinity stress in transgenic tobacco[J]. Plant Biotechnology Reports, 2010, 4: 75-83.
[20]	Zhang K, Wang J, Lian L J, et al. Increased chilling tolerance following transfer of a betA gene enhancing glycine betaine synthesis in cotton(Gossypium hirsutum L.)[J]. Plant Molecular Biology Reporter, 2012, 30(5): 1158-1171.

茶树甜菜碱醛脱氢酶基因(CsBADH1)的全长cDNA克隆与表达分析

Cloning and Expression Analysis of Betaine Aldehyde Dehydragenase Gene (CsBADH1) from Tea Plant [Camellia sinensis (L.) O. Kuntze]

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