欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2009, Vol. 35 ›› Issue (2): 295-300.doi: 10.3724/SP.J.1006.2009.00295

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

青稞hblt4.2基因的克隆及功能分析

何涛1,2;贾敬芬2 *   

  1. 1青海大学生物科学系，青海西宁810016；2西北大学生命科学学院，陕西西安710069
  • 收稿日期:2008-05-26 修回日期:2008-10-24 出版日期:2009-02-12 网络出版日期:2008-12-11
  • 通讯作者: 贾敬芬
  • 基金资助:

    本研究由国家自然科学基金项目（30671082），陕西省教育厅重点项目（05JS48）资助。

Cloning and Function Analysis of hbltl4.2 Gene in Highland Barley(Hordeum vulgare L. var. nudum Hook. f.)

HE Tao1,2,JIA Jing-Fen2,*   

  1. 1Department of Biosciences, Qinghai University, Xining 810016,China; 2College of Life Sciences, Northwest University, Xi'an 710069,China
  • Received:2008-05-26 Revised:2008-10-24 Published:2009-02-12 Published online:2008-12-11
  • Contact: JIA Jing-Fen

PDF

1491

被引次数

3

摘要：

根­据大麦blt14.2基因序列设计引物, 用青稞的DNA为模板扩增低温反应基因的全长序列, 命名为hblt14.2, 在GenBank登录号为EF514912。该基因含470 bp, 包括249 bp的开放阅读框、69 bp的5'非翻译区和152 bp的3'非翻译区, 编码82个氨基酸残基, 富含Gly、Ala、Leu和Val氨基酸, 与其他冷诱导基因编码蛋白具相似性。与大麦blt14.2基因具98.9%同源性, 所编码的蛋白存在2个氨基酸的差别。将hblt14.2基因连接CaMV35S启动子和NOS终止子后定向插入质粒pCAMBIA1301的多克隆位点中, 构建植物表达载体pCAMBIA-BI-hblt, 通过农杆菌介导转化烟草, 并对转基因烟草进行抗寒性检测。结果表明, 青稞hblt14.2基因与植物的抗寒性有一定关系。

关键词: 青稞, 低温, hblt4.2基因, 功能分析

Abstract:

The response to low temperature stress in highland barley indicates that there is a potential gene bank for cold resistance, which is helpful to develop the genes associated with resistance to cold and to survey the regulation mechanism. The objective of this study was to exploit a new gene with cold resistance, to validate its function. A pair of primer was designed based on the blt14.2 gene sequence documented in GenBank, a low-temperature-responsive (LTR) gene hblt14.2 was cloned by PCR from highland barley with GenBank accession number EF514912. Its length was 470 bp, containing 249 bp ORF, 69 bp 5'untranslated region (UTR) and 152 bp 3'UTR. Nucleotide sequence of hblt14.2 gene shares 98.9% (only five base pairs difference)homology with that of blt14.2 in barley, which encoded a protein of 82 amino acids. Theprotein was a highly hydrophilic small protein with rich Gly, Ala, Leu and Val amino acids, which were similar to proteins encoded by other LTR genes. The hblt14.2 ligated with CaMV 35S promoter and NOS terminator was inserted into multiple clone site in plasmid CAMBIA1301 to construct plant expression vector pCAMBIA-BI-hblt, which was transformed into tobacco mediated by Agrobacterium tumefaciens. Transgenic plants response to low temperature were investigated. The results suggested that the hblt14.2 is related to cold resistance in highland barley.

Key words: Highland barley, Low temperature, hbltl4.2 gene, Function analysis

[1]Guo B-Z(郭本兆). Flora of Qinghai. Xining: Qinghai People’s Press, 1987. p 701 (in Chinese)
[2]Hughes M A, Dunn M A. The molecular biology of plant accli-mation to low temperature. J Exp Bot, 1996, 47: 291-305
[3]Dunn M A, Hughes M A, Zhang L, Pearce R S, Quigley A S, Jack P L. Nucleotide sequence and molecular analysis of the low temperature induced barley gene, BLT4. Mol Gen Genet, 1991, 229: 389-394
[4]Dunn M A, Hughes M A, Pearce R S. Molecular characterization of a barley gene induced by cold treatment. J Exp Bot, 1990, 41: 1405-1413
[5]Dunn M A, Brown K, Lightowlers R, Hughes M A. A low-tem-perature-responsive gene from barley encodes a protein with sing-stranded nucleic acid-binding activity which is phosphory-lated in vitro. Plant Mol Biol, 1996, 30: 947-959
[6]Phillips J R, Dunn M A, Hughes M A. mRNA stability and local-ization of the low-temperature- responsive barley gene family blt14. Plant Mol Biol, 1997, 33: 1013-1023
[7]Thomashow M F. Arabidopsis thaliana as A model for studying mechanisms of plant cold tolerance. In: Meyerowitz E, Somer-ville C, eds. Arabidopsis. New York: Cold Spring Harbor Labo-ratory Press, 1994. pp 807-834
[8]Kurkela S, Borg-Franck M. Cloning and characterization of a cold- and ABA- inducible Arabidopsis gene. Plant Mol Biol, 1990, 15: 137-144
[9]Artus N N, Uemura M, Steponkus P L, Gilmour S J, Lin C T, Thomashow M F. Constitutive expression of the cold-regulated Arabidopsis thaliana COR15a gene affects both chloroplast and protoplast freezing tolerance. Proc Natl Acad Sci USA, 1996, 93: 1340-1349
[10]Raymond M J, Smirnoff N. Proline metabolism and transport in maize seedlings at low water potential. Ann Bot, 2002, 89: 813-823
[11]Alia-Prasad K V S K, Saradhi P P. Effect of zinc on free radicals and proline in Brassica and Cajanus. Phytochemistry, 1995, 39: 45-47
[12]Purvis A C, Yelenosky G. Translocation of carbohydrates and proline in young grapefruit trees at low temperatures. Plant Physiol, 1983, 73: 877-880
[13]Hsieh T H, Lee J T, Yang P T, Chiu J H, Chang Y Y, Wang Y C, Chan M T. Heterology expression of the Arabidopsis C-repeat/ dehydration-responsive element binding factor 1 gene confers elevated tolerance to chilling and oxidative stresses in transgenic tomato. Plant Physiol, 2002, 129: 1086-1094
[14]Kavi K. Overexpression of pyrroline-5-carboxylate synthetase increases proline production and confers osmotolerance in trans-genic plants. Plant Physiol, 1995, 108: 1387
[15]Liu Y-H(刘艳红), An L-Z(安黎哲), Zhang M-X(张满效), Chen T(陈拓), Xu S-J(徐世健), Liu G(刘光), Wang X-L(王勋陵). Analysis of free amino acids of alpine periglacial plants. J Gla-ciol Geocryol (冰川冻土), 2005, 27(4): 608-614(in Chinese with English abstract)
[16]Hare P D, Cress W A. Metabolic implications of stress-induced proline accumulation in plants. Plant Growth Regul, 1997, 21: 79-102
[17]Wanner L A, Junttila O. Cold-induced freezing tolerance in Arabidopsis. Plant Physiol, 1999, 120: 391-399
[1] 王兴荣, 李玥, 张彦军, 李永生, 汪军成, 徐银萍, 祁旭升. 青稞种质资源成株期抗旱性鉴定及抗旱指标筛选[J]. 作物学报, 2022, 48(5): 1279-1287.
[2] 姚晓华, 王越, 姚有华, 安立昆, 王燕, 吴昆仑. 青稞新基因HvMEL1 AGO的克隆和条纹病胁迫下的表达[J]. 作物学报, 2022, 48(5): 1181-1190.
[3] 冯亚, 朱熙, 罗红玉, 李世贵, 张宁, 司怀军. 马铃薯StMAPK4响应低温胁迫的功能解析[J]. 作物学报, 2022, 48(4): 896-907.
[4] 宋仕勤, 杨清龙, 王丹, 吕艳杰, 徐文华, 魏雯雯, 刘小丹, 姚凡云, 曹玉军, 王永军, 王立春. 东北主推玉米品种种子形态及贮藏物质与萌发期耐冷性的关系[J]. 作物学报, 2022, 48(3): 726-738.
[5] 薛晓梦, 吴洁, 王欣, 白冬梅, 胡美玲, 晏立英, 陈玉宁, 康彦平, 王志慧, 淮东欣, 雷永, 廖伯寿. 低温胁迫对普通和高油酸花生种子萌发的影响[J]. 作物学报, 2021, 47(9): 1768-1778.
[6] 李洁, 付惠, 姚晓华, 吴昆仑. 不同耐旱性青稞叶片差异蛋白分析[J]. 作物学报, 2021, 47(7): 1248-1258.
[7] 苏亚春, 李聪娜, 苏炜华, 尤垂淮, 岑光莉, 张畅, 任永娟, 阙友雄. 甘蔗割手密种类甜蛋白家族鉴定及栽培种同源基因功能分析[J]. 作物学报, 2021, 47(7): 1275-1296.
[8] 唐锐敏, 贾小云, 朱文娇, 印敬明, 杨清. 马铃薯热激转录因子HsfA3基因的克隆及其耐热性功能分析[J]. 作物学报, 2021, 47(4): 672-683.
[9] 杨阳, 李淮琳, 胡利民, 范楚川, 周永明. 白菜型油菜srb多室性状的遗传分析与分子鉴定[J]. 作物学报, 2021, 47(3): 385-393.
[10] 解盼, 刘蔚, 康郁, 华玮, 钱论文, 官春云, 何昕. 甘蓝型油菜CBF基因家族的鉴定和表达分析[J]. 作物学报, 2021, 47(12): 2394-2406.
[11] 高芸, 张玉雪, 马泉, 苏盛楠, 李春燕, 丁锦峰, 朱敏, 朱新开, 郭文善. 春季低温对小麦花粉育性及粒数形成的影响[J]. 作物学报, 2021, 47(1): 104-115.
[12] 李瑞杰,唐会会,王庆燕,许艳丽,王琦,卢霖,闫鹏,董志强,张凤路. 5-氨基乙酰丙酸和乙烯利对东北春玉米源库碳平衡的调控效应[J]. 作物学报, 2020, 46(7): 1063-1075.
[13] 王丹丹, 柳洪鹃, 王红霞, 张鹏, 史春余. 甘薯蔗糖转运蛋白基因IbSUT3的克隆及功能分析[J]. 作物学报, 2020, 46(7): 1120-1127.
[14] 张瑞栋,肖梦颖,徐晓雪,姜冰,邢艺凡,陈小飞,李邦,艾雪莹,周宇飞,黄瑞冬. 高粱种子对萌发温度的响应分析与耐低温萌发能力鉴定[J]. 作物学报, 2020, 46(6): 889-901.
[15] 赵小红,白羿雄,王凯,姚有华,姚晓华,吴昆仑. 种植密度对2个青稞品种抗倒伏及秸秆饲用特性的影响[J]. 作物学报, 2020, 46(4): 586-595.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2