茶树硝酸根转运蛋白基因NRT2.5的克隆及表达分析

doi:10.13305/j.cnki.jts.2014.04.007

茶叶科学 ›› 2014, Vol. 34 ›› Issue (4): 364-370.doi: 10.13305/j.cnki.jts.2014.04.007

茶树硝酸根转运蛋白基因NRT2.5的克隆及表达分析

冯素花¹, 王丽鸳^1,*, 陈常颂², 林郑和², 成浩^1,*, 韦康¹, 张成才¹

1. 中国农业科学院茶叶研究所,国家茶树改良中心,农业部茶树生物学与资源利用重点实验室,浙江杭州 310008;
2. 福建省农业科学院茶叶研究所,福建福安 355000

收稿日期:2014-03-05 修回日期:2014-04-04 出版日期:2014-08-15 发布日期:2019-09-03
通讯作者: *chenghao@mail.tricaas.com
作者简介:冯素花（1989— ）女,山东菏泽人,硕士研究生,主要从事茶树种质资源与育种研究。
基金资助:
浙江省茶树农业新品种选育重大科技专项（2012C12905）、国家茶叶产业技术体系（nycytx-23）

Cloning and Expressing Analysis of a Nitrogen Transporter 2.5 Gene from Tea Plant[Camellia sinensis (L.)]

FENG Suhua¹, WANG Liyuan^1,*, CHEN Changsong², LIN Zhenghe², CHENG Hao^1,*, WEI Kang¹, ZHANG Chengcai¹

1. Tea Research Institute, Chinese Academy of Agricultural Sciences, National Center for Tea Improvement, Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Hangzhou 310008, China;
2. Tea Research Institute, Fujian Academy of Agricultural Sciences, Fuan 355000, China

Received:2014-03-05 Revised:2014-04-04 Online:2014-08-15 Published:2019-09-03

摘要/Abstract

摘要： 通过RACE克隆从茶树[Camellia sinensis (L.)]叶片中获得NRT2.5基因cDNA全长序列。所得基因序列全长为2 457 bp,其中开放阅读框为1 362 bp,编码454个氨基酸,推测蛋白分子量为48.7 kD,理论等电点为9.63。生物信息学分析表明,该基因与可可树、拟南芥等的NRT2.5基因具有高度同源性,且其编码的蛋白质具有NRT家族共有的结构特征。实时定量PCR分析表明,NRT2.5基因在茶树不同组织中均有表达,但主要在成熟叶和根中表达;不同氮浓度处理后,茶树NRT2.5基因在低浓度下的表达量高于高浓度下。

关键词: 茶树, NRT2.5, 基因克隆, 定量表达分析

Abstract: The full cDNA length of NRT2.5 gene was obtained via rapid amplification of cDNA ends (RACE) from tea plant [Camellia sinessis (L.)] leaves. The cDNA sequence of this gene was 2 457 bp and opening reading frame (ORF) was 1 362 bp, encoding for 454 amino acids. The putative protein of this gene had an isoelectric point of 9.63 and a calculated molecular weight of 48.7 kD. Bioinformatics analysis showed that NRT2.5 of tea plant was highly homologous to the genes such as TcNRT2.5 and AtNRT2.5, and its encoded protein had the common structural characteristics of NRT family. Fluorescence quantitative real-time PCR analysis showed that NRT2.5 gene were detected in all the tested tissues of tea plant but was mainly detected in the mature leaves and roots. The expression level in different nitrogen concentration showed that the expression of NRT2.5 gene in low N concentration was significant higher than that in high concentration.

Key words: Camellia sinessis, NRT2.5 gene, clone, expression analysis

中图分类号:

S571.1
Q52

冯素花, 王丽鸳, 陈常颂, 林郑和, 成浩, 韦康, 张成才. 茶树硝酸根转运蛋白基因NRT2.5的克隆及表达分析[J]. 茶叶科学, 2014, 34(4): 364-370. doi: 10.13305/j.cnki.jts.2014.04.007.

FENG Suhua, WANG Liyuan, CHEN Changsong, LIN Zhenghe, CHENG Hao, WEI Kang, ZHANG Chengcai. Cloning and Expressing Analysis of a Nitrogen Transporter 2.5 Gene from Tea Plant[Camellia sinensis (L.)][J]. Journal of Tea Science, 2014, 34(4): 364-370. doi: 10.13305/j.cnki.jts.2014.04.007.

参考文献 23

[1]	Lee R B, Clarkson D T.Nitrogen-13 studies of nitrate fluxes in barley roots. Compartmental analysis from measurements of 13N efflux[J]. The EMBO Journal, 1986, 5: 1753-1767.
[2]	Tsay Y F, Chu C C, Tsai C B, et al. Nitrate transporters and peptide transporters[J]. Febs Letters, 2001, 581: 2290-2300.
[3]	Pao SS, Paulsen IT, Saier MH.Major facilitator super-family[J]. Molecular Biology Reviews, 1998, 62(1): 1-34.
[4]	Galvan A, Fernandez E.Eukaryotic nitrate and nitrite transporters[J]. Cellular and Molecular Life Science, 2001, 58: 225-233.
[5]	Okamoto M, Jone Vidmar J, Glass A D M. Regulation of NRT1 and NRT2 gene families of Arabidosis thaliana: Responses to nitrate provision[J]. Plant Cell Physoil, 2003, 44(3): 304-317.
[6]	Brian G.Nitrate transporters in plants: Structure, function and regulation[J]. Forde Biochimica er Biophysica Acta, 2000, 1465(1/2): 219-235.
[7]	Orsel M, Krapp A, Daniel-Vedele F.Analysis of the NRT2 nitrate transporter family in Arabidopsis. Structure and gene expression.[J]. Plant Physiol, 2002, 129: 886-896.
[8]	Ryoichi A, Hiroshi H.Expression of rice gene involved in high-affinity nitrate transport during the period of nitrate induction[J]. Breeding Sci, 2006, 56: 295-402.
[9]	Trueman L J, Richardson A, Forde B G.Molecular cloning of higher plant homologues of the high-affinity nitrate transporters of Chlamydomonas reinhardtii and Aspergillus nidulans[J]. Gene, 1996, 175(1/2): 223-231.
[10]	Vidmar J J, Zhuo D, Siddiqi M Y, et al. Isolation and characterization of HvNRT2.3 and HvNRT2.4, cDNAs encoding high-affinity nitrate transporters from roots of barley[J]. Plant Physiology, 2000, 122(3): 783-792.
[11]	Amarasinghe B H, de Bruxelles G L, Braddon M, et al. Regulation of Gmnrt2 expression and nitrate transport activity in roots of soybean (Glycine max)[J]. Planta, 1998, 206(1): 44-52.
[12]	Quaggiotti S, Ruperti B, Borsa P, et al. Expression of a putative high-affinity NO3- transporter and of an H+-ATPase in relation to whole plant nitrate transport physiology in two maize genotypes differently responsive to low nitrogen availability[J]. Journal of Experiment Botany, 2003, 54: 1023-1031.
[13]	赵学强, 李玉京, 刘建中, 等. 小麦NO3-转运蛋白基因TaNRT2.3的克隆和表达分析[J]. 植物学报, 2004, 46(3): 347-354.
[14]	Forde B G.Nitrate transporters in plants: structure, function and regulation[J]. Biochim Biophys Acta, 2000, 1465: 219-235.
[15]	赖灯妮. 高亲和性硝酸盐转运基因型水稻品种初筛及该基因的诱导表达研究[D]. 长沙: 湖南农业大学, 2009.
[16]	蔡超. 禾谷类作物高亲和力NO3-吸收系统基因表达调控研究[D]. 北京: 中国科学院生态环境研究中心, 2007.
[17]	王新亮. 果树根系硝态氮信号响应关键基因的克隆与功能分析[D]. 泰安: 山东农业大学, 2012.
[18]	石乐松, 刘进平. 文心兰RNA不同提取方法比较[J]. 生物技术, 2012, 22(6): 42-45.
[19]	Kivak KJ, Schmittgen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J]. Methods, 2001, 25(4): 402-408.
[20]	许金兰, 谢小东, 冯爽, 等. 烟草硝酸盐转运蛋白基因的克隆及表达分析[J]. 烟草农学, 2013, 312(7): 68-71.
[21]	孔敏, 杨学东, 候喜林, 等. 白菜NRT2基因的克隆及表达模式分析[J]. 园艺学报, 2011, 38(12): 2309-2316.
[22]	徐海荣, 谷俊涛, 路文静, 等. 水稻新型硝酸盐转运蛋白基因OsTNrt1的编码蛋白特征和表达[J]. 作物学报, 2007, 33(5): 723-730.
[23]	钟丽华, 宋世威, 陈日远. 菜薹硝酸盐转运蛋白基因NRT2的克隆与表达分析[J]. 园艺学报, 2013, 40: 2650.

茶树硝酸根转运蛋白基因NRT2.5的克隆及表达分析

Cloning and Expressing Analysis of a Nitrogen Transporter 2.5 Gene from Tea Plant[Camellia sinensis (L.)]

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献 23

相关文章 15

Metrics

本文评价

推荐阅读 10

[1]	王留彬, 黄丽蕴, 滕翠琴, 吴立赟, 成浩, 于翠平, 王丽鸳. 梧州茶树种质资源的遗传多样性及亲缘关系分析[J]. 茶叶科学, 2022, 42(5): 601-609.
[2]	周汉琛, 杨霁虹, 徐玉婕, 吴琼, 雷攀登. 香叶醇生物合成相关基因NUDX1的进化分析[J]. 茶叶科学, 2022, 42(5): 638-648.
[3]	陈琪予, 马建强, 陈杰丹, 陈亮. 利用图像特征分析茶树成熟叶表型的遗传多样性[J]. 茶叶科学, 2022, 42(5): 649-660.
[4]	孙悦, 吴俊, 韦朝领, 刘梦月, 高晨曦, 张灵枝, 曹士先, 余顺甜, 金珊, 孙威江. 抗小贯松村叶蝉和茶棍蓟马的茶树种质筛选及其抗性相关因素分析[J]. 茶叶科学, 2022, 42(5): 689-704.
[5]	王玉源, 刘任坚, 刘少群, 舒灿伟, 孙彬妹, 郑鹏. 茶树R2R3-MYB转录因子CsTT2表达分析及功能初步鉴定[J]. 茶叶科学, 2022, 42(4): 463-476.
[6]	刘建军, 张金玉, 彭叶, 刘晓博, 杨云, 黄涛, 温贝贝, 李美凤. 不同光质摊青对夏秋茶树鲜叶挥发性物质及其绿茶品质影响研究[J]. 茶叶科学, 2022, 42(4): 500-514.
[7]	邢安琪, 武子辰, 徐晓寒, 孙怡, 王艮梅, 王玉花. 茶树富集氟的特点及其机制的研究进展[J]. 茶叶科学, 2022, 42(3): 301-315.
[8]	王涛, 王艺清, 漆思雨, 周喆, 陈志丹, 孙威江. 茶树CLH基因家族的鉴定与转录调控研究及其在白化茶树中的表达分析[J]. 茶叶科学, 2022, 42(3): 331-346.
[9]	刘任坚, 王玉源, 刘少群, 舒灿伟, 孙彬妹, 郑鹏. 茶树CsbHLH024和CsbHLH133转录因子功能鉴定[J]. 茶叶科学, 2022, 42(3): 347-357.
[10]	欧阳珂, 张成, 廖雪利, 坤吉瑞, 童华荣. 基于感官组学分析玉米香型南川大茶树工夫红茶特征香气[J]. 茶叶科学, 2022, 42(3): 397-408.
[11]	刘富浩, 范延艮, 王域, 孟凡月, 张丽霞. 茶树黄金芽CsHIPP26.1蛋白螯合离子的筛选与鉴定[J]. 茶叶科学, 2022, 42(2): 179-186.
[12]	杨妮, 李逸民, 李静文, 滕瑞敏, 陈益, 王雅慧, 庄静. 外源5-ALA对干旱胁迫下茶树叶绿素合成和荧光特性及关键酶基因表达的影响[J]. 茶叶科学, 2022, 42(2): 187-199.
[13]	疏再发, 郑生宏, 邵静娜, 周慧娟, 吉庆勇, 刘瑜, 何卫中, 王丽鸳. 不同茶树品种（系）对减半施肥的响应研究[J]. 茶叶科学, 2022, 42(2): 277-289.
[14]	陈潇敏, 赵峰, 王淑燕, 邵淑贤, 吴文晞, 林钦, 王鹏杰, 叶乃兴. 福建野生茶树资源嘌呤生物碱构成评价及特异资源筛选[J]. 茶叶科学, 2022, 42(1): 18-28.
[15]	刘庆帅, 璩馥榕, 魏梦园, 钟红, 王熠, 陈亮, 金基强. 基于UPLC技术解析金萱×紫娟F₁分离群体代谢物的遗传变异[J]. 茶叶科学, 2022, 42(1): 29-40.