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植物研究 ›› 2013, Vol. 33 ›› Issue (3): 317-324.doi: 10.7525/j.issn.1673-5102.2013.03.011

• 论文 • 上一篇    下一篇

盐生植物盐节木甜菜碱醛脱氢酶基因的克隆及表达

高天鹏1,4;王春燕2;徐红伟3;孙海丽1;张鸣4;张庆4;石晓妮4   

  1. 1.兰州城市学院城市生态与环境生物技术中心,兰州 730070;2.兰州大学学报编辑部,兰州 730000;3.西北民族大学实验中心,兰州 730030;4.兰州城市学院化学与环境科学学院,兰州 730000
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2013-05-20 发布日期:2013-05-20
  • 基金资助:
     

Molecular Cloning and Expression Analysis of Betaine Aldehyde Dehydrogenase Gene from the Halophyte Halocnermum strobilaceum(Pall.)(Chenopodiaceae)

GAO Tian-Peng;WANG Chun-Yan;XU Hong-Wei;SUN Hai-Li;ZHANG Ming;ZHANG Qing;SHI Xiao-Ni   

  1. 1.Urban Ecology & Environmental Biotechnology Center,Lanzhou City University,Lanzhou 730000;2.Editorial Department of Journal,Lanzhou University,Lanzhou 730000;3.Science Experimental Center,Northwest University for Nationalities,Lanzhou 730030;4.School Of Chemistry and Environmental Science,Lanzhou City University,Lanzhou 730000
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-05-20 Published:2013-05-20
  • Supported by:
     

摘要: 采用RT-PCR、RACE技术从盐生植物盐节木(Halocnermum strobilaceum(Pall.) Bieb.)中克隆得到BADH基因的cDNA序列(命名为HsBADH),3′RACE和 5′RACE测序结果用DNA Man软件拼接获得了1 849 bp全长的HsBADH cDNA序列(GenBank登录号:JN969894)。其开放阅读框为1 503 bp,推测编码501个氨基酸残基,并含有高度保守的十肽基序(VTLELGGKSP)和与酶功能有关的醛脱氢酶Cys残基。其核苷酸序列与几种藜科植物如梭梭、白梭梭、盐穗木、碱蓬、滨藜、菠菜、山菠菜的同源性为86%,与甜土植物水稻的同源性为66%。氨基酸序列与以上两类植物(盐生植物和甜土植物) 的同源性比对分别为89%和70%,说明BADH基因在藜科盐生植物中是一种较高保守的基因。用不同浓度的NaCl胁迫处理盐节木植株,BADH mRNA的表达水平比对照植株高,说明盐节木BADH基因的表达受盐诱导, 进一步说明甜菜碱醛脱氢酶催化合成的甜菜碱作为渗透调节的小分子物质,它的积累与盐胁迫存在紧密关联,本研究为进一步从生理和分子水平阐明盐节木的适应盐渍化生境的机制提供一定的参考。

关键词: 盐节木, 甜菜碱醛脱氢酶, 基因克隆, 表达分析

Abstract: According to the published sequences of BADH cDNA of several other plants of Chenopodiaceae, two primers have been designed to amplify the fragments of BADH cDNA from Halocnermum strobilaceum(Pall.) Bieb. through RT-PCR (reverse transcription polymerase chain reaction). A 1 503 bp fragment containing entire betaine aldehyde dehydrogenase (BADH) coding region of 501 amino acids (aa) has been obtained. Nucleotide sequence of HsBADH was similar to the corresponding fragment of BADH cDNA of several other plants, such as Kalidium foliatum, Atriplex centralasiatica, Atriplex hortensis, Spinacia oleracea, Suaeda liaotungensis, Beta vulgaris subsp. Vulgaris, Oryza sativa, etc. Encoded protein by HsBADH and BADH protein from above mentioned plants also shared 89% and 70% identity at the amino acid level. The result showed that BADH gene was conserved, especially in Chenopodiaceae and encoded functional protein may play an important role in high plants during salt stress. Real-time fluorescence quantitative gene expression analysis showed that the level of BADH mRNA in plants treated with different NaCl concentrations was higher than that in the control plants, suggesting that the accumulation of betaine catalyzed by betaine alde-hyde dehydrogenase as an effective osmolyte is important for H.strobilaceum(Pall.) Bieb. during salt stress. The study provided material for further exploring salt tolerant mechanisms of H.strobilaceum(Pall.) Bieb. in physiological and molecular aspects.

Key words: Halocnermum strobilaceum(Pall.) Bieb., betaine aldehydede dehydrogenase, gene cloning, expression analysis

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