欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2012, Vol. 38 ›› Issue (09): 1649-1656.doi: 10.3724/SP.J.1006.2012.01649

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

棉花S-腺苷甲硫氨酸脱羧酶基因的克隆及低温下的表达分析

耿卫东，李艳军，张新宇，朱华国*，孙杰*   

  1. 石河子大学农学院 / 新疆生产建设兵团绿洲生态农业重点实验室，新疆石河子832003
  • 收稿日期:2012-01-08 修回日期:2012-04-20 出版日期:2012-09-12 网络出版日期:2012-06-04
  • 通讯作者: 朱华国, Email: zhgroger@sohu.com; 孙杰, E-mail: sunjiezh@yahoo.com.cn
  • 基金资助:

    本研究由兵团博士资金项目(2011BB004)和国家转基因生物新品种培育重大专项子课题(2008ZX08005-005)资助。

Molecular Cloning and Expression Analysis of GhSAMDC at Low Temperature Stress in Cotton (Gossypium hirsutum L.)

GENG Wei-Dong,LI Yan-Jun,ZHANG Xin-Yu,ZHU Hua-Guo*,SUN Jie*   

  1. College of Agriculture / The key Laboratory of Oasis Eco-agriculture Xinjiang Production and Construction Group, Shihezi University, Shihezi 832003, China
  • Received:2012-01-08 Revised:2012-04-20 Published:2012-09-12 Published online:2012-06-04
  • Contact: 朱华国, Email: zhgroger@sohu.com; 孙杰, E-mail: sunjiezh@yahoo.com.cn

PDF

1551

被引次数

12

摘要： 根据棉花S-腺苷甲硫氨酸脱羧酶(S-adenosylmethionine decarboxylase, SAMDC)基因已知EST序列设计引物，采用RACE和RT-PCR技术克隆获得该基因的全长cDNA序列，命名为GhSAMDC (GenBank登录号为JN020148)。生物信息学分析表明，GhSAMDCcDNA序列全长1 874 bp，涵盖tiny ORF (tORF)、upstream ORF (uORF)和main ORF (mORF) 3个植物SAMDC基因特征ORF。其中mORF长1 064 bp，编码含355个氨基酸残基的SAMDC酶原，预测分子量为38.25 kD，该酶原含有高度保守的酶原剪切位点结构域(LSESSLF)和与SAMDC蛋白快速降解有关的PEST (TIHVTPEDGFSYAS)结构域。GhSAMDC基因组DNA序列全长2 743 bp，包含3个内含子，均位于5' UTR，其中一个位于uORF内。聚类分析表明，GhSAMDC与葡萄中该蛋白的同源关系最近，并且与其他双子叶植物聚为一类。荧光定量PCR分析结果表明，GhSAMDC表达受低温诱导，在冷敏感品种新陆早1号中基因表达水平明显高于在耐冷品种新陆早33号中。

关键词: 棉花, S-腺苷甲硫氨酸脱羧酶, 序列分析, 定量PCR, 低温表达

Abstract: S-adenosylmethionine decarboxylase is one of the key enzymes in polyamine biosynthesis. With the increase of polyamine accumulation in plants, SAMDC can improve crop tolerance to environment stresses. We cloned the S-adenosylmethionine decarboxylase gene from Gossypium hirsutum by RACE and RT-PCR, named GhSAMDC (GenBank accession number: JN020148). The cDNA full length was 1 874 bp, with three open reading frames, i.e. tiny ORF (tORF), upstream ORF (uORF) and main ORF (mORF). The mORF was 1 064 bp encoding 355 amino acids with a molecular mass of 38.25 kD, and its deduced protein had two highly conserved function domains (proenzyme cleavage site and PEST domain). The complete genomic DNA sequence of GhSAMDC was 2 743 bp in length with three introns, all of which were located in 5' UTR, and one lay in theuORF. Phylogenetic analysis showed GhSAMDC was the closest to that in Vitis vinifera, and Real-time PCR suggested that GhSAMDC was induced by cold stress. It showed lower gene expression level in cold resistant cultivar Xinluzao 33 than in cold sensitive cultivar Xinluzao 1.

Key words: Cotton (Gossypium hirsutum L.), S-adenosylmethionine decarboxylase, Sequence analysis, qPCR, Expression at low temperature

[1]Alcázar R, Marco F, Cuevas J C, Patron M, Ferrando A, Carrasco P, Tiburcio A F, Altabella T. Involvement of polyamines in plant response to abiotic stress. Biotechnol Lett, 2006, 28: 1867–1876

[2]Kusano T, Berberich T, Tateda C, Takahashi Y. Polyamines: essential factors for growth and survival. Planta, 2008, 228: 367–381

[3]Shen W Y, Nada K, Tachibana S. Involvement of polyamines in the chilling tolerance of cucumber cultivars. Plant Physiol, 2000, 124: 431–439

[4]Cuevas J C, López-Cobollo R, Alcázar R, Zarza X, Koncz C, Altabella T, Salinas J, Tiburcio A F, Ferrando A. Putrescine is involved in Arabidopsis freezing tolerance and cold acclimation by regulating abscisic acid levels in response to low temperature. Plant Physiol, 2008, 148: 1094–1105

[5]Franceschftti M, Hanfrey C, Scaramagli S, Torrigiani P, Bagni N, Burtin D, Anthony J M. Characterization of monocot and dicot plant S-adenosyl-L-methionine decarboxylase gene families including identification in the mRNA of a highly conserved pair of upstream overlapping open reading frames. Biochem J, 2001, 353: 403–409

[6]Bhavna W, Manchikatla V R. Effect of increased polyamine biosynthesis on stress responses in transgenic tobacco by introduction of human S-adenosylmethionine gene. Plant Sci, 2003, 164: 727–734

[7]Cheng L, Zou Y J, Ding S L, Zhang J J, Yu X L, Cao J S, Lu G. Polyamine accumulation in transgenic tomato enhances the tolerance to high temperature stress. J Integr Plant Biol, 2009, 51: 489–499

[8]Momtaz O A, Hussein E M, Fahmy E M, Ahmed S E. S-adenosylmethionine decarboxylase gene for polyamine accumulation in Egyptian cotton Giza 88 and Giza 90. GM Crops, 2010, 1: 257–266

[9]Zhao L L, Song L Q, You C X, Moriguchi T, Hao Y J. Functional characterization of the apple MdSAMDC2 gene by ectopic promoter analysis and over-expression in tobacco. Biol Plant, 2010, 54: 631–638

[10]Liu Z-Y(刘志勇), Wang X-X(王孝宣), Gao J-C(高建昌), Guo Y-M(国艳梅), Du Y-C(杜永臣), Ye X-L(叶雪凌). Cloning and sequence analysis of a S-adenosylmethionine decarboxylase gene (SlSAMDC1) in tomato. Acta Hort Sin (园艺学报), 2008, 35(8): 1137–1146 (in Chinese with English abstract)

[11]Liu J-X(刘金仙), Que Y-X(阙友雄), Guo J-L(郭晋隆), Xu L-P(许莉萍), Xu J-S(徐景升), Chen R-K(陈如凯). Molecular cloning of sugarcane S-adenosylmethionine decarboxylase gene (Sc-SAMDC) and its expression analysis. Sci Agric Sin (中国农业科学), 2010, 43(7): 1448–1457 (in Chinese with English abstract)

[12]Lu J-X(陆俊杏), Lu K(卢坤), Zhang K(张凯), Cao T(曹廷), Li J-N(李加纳), Liang Y(梁颖). Cloning and analysis of SAMDC3 genes and their promoters from Brassicanapus. Genom Appl Biol (基因组学与应用生物学), 2010, 29(2): 215–224 (in Chinese with English abstract)

[13]Zeng X-Q(曾兴权), Wang C-Y(王长有), Zhang H(张宏), Wei Z-X(韦泽秀), Liu X-L(刘新伦), Wang Y-J(王亚娟), Ji W-Q(吉万全). Cloning and characterization of an up-regulated S-adenosylmethionine decarboxylas (SAMDC) gene induced by stripe rust (Puccinia striiformisf. sp. tritici) in wheat. J Triticeae Crops (麦类作物学报), 2011, 31(5): 811–818 (in Chinese with English abstract)

[14]Jiang J-X(蒋建雄), Zhang T-Z(张天真). Extraction of total rna in cotton tissues with CTAB-acidic phenolic method. Cotton Sci (棉花学报), 2003, 15(3): 166–167 (in Chinese with English abstract)

[15]Zhao W-J(赵文静), Xu J(徐洁), Bao Q-H(包秋华), Chen Y-F(陈永福), Zhang H-P(张和平). Selection of reference genes for Real-time quantitative PCR. Microbiol China (微生物学通报), 2010, 37(12): 1825−1829 (in Chinese with English abstract)

[16]Livaka K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2–ΔΔCT method. Methods, 2001, 25: 402–408

[17]Stanley B A, PeggS A E, Holm I. Site of pyruvate formation and processing of mammalian S-adenosy-lmethionine decarboxylase preenzyme. J Biol Chem, 1989, 264: 21073–21079

[18]Rogers S, Wells R, Rechsteiner M. Amino acid sequence conunon to rapid degraded proteins: The PEST hypotheis. Science, 1986, 234: 364–368

[19]Zhang J-J(张佳景), Ding S-L(丁淑丽), Zhou Y-J(邹宜静), Cheng L(程琳), Lu G(卢钢). Research advance on S-adenosylmethionine decarboxylase in Plants. Chin J Cell Biol (细胞生物学杂志), 2008, 30(5): 622–628 (in Chinese with English abstract)

[20]Wang X-L(王小利), Liu X-X(刘晓霞), Wang S-Y(王舒颖), Yang Y-C(杨义成), Wu J-H(吴佳海). Cloning and differential expression analysis of S–adenosylmethionine decarboxylase gene FaSAMDC in tall fescue. Acta Pratac Sin (草业学报), 2011, 20(4): 169–179 (in Chinese with English abstract)
[1] 周静远, 孔祥强, 张艳军, 李雪源, 张冬梅, 董合忠. 基于种子萌发出苗过程中弯钩建成和下胚轴生长的棉花出苗壮苗机制与技术[J]. 作物学报, 2022, 48(5): 1051-1058.
[2] 孙思敏, 韩贝, 陈林, 孙伟男, 张献龙, 杨细燕. 棉花苗期根系分型及根系性状的关联分析[J]. 作物学报, 2022, 48(5): 1081-1090.
[3] 闫晓宇, 郭文君, 秦都林, 王双磊, 聂军军, 赵娜, 祁杰, 宋宪亮, 毛丽丽, 孙学振. 滨海盐碱地棉花秸秆还田和深松对棉花干物质积累、养分吸收及产量的影响[J]. 作物学报, 2022, 48(5): 1235-1247.
[4] 郑曙峰, 刘小玲, 王维, 徐道青, 阚画春, 陈敏, 李淑英. 论两熟制棉花绿色化轻简化机械化栽培[J]. 作物学报, 2022, 48(3): 541-552.
[5] 张艳波, 王袁, 冯甘雨, 段慧蓉, 刘海英. 棉籽油分和3种主要脂肪酸含量QTL分析[J]. 作物学报, 2022, 48(2): 380-395.
[6] 张特, 王蜜蜂, 赵强. 滴施缩节胺与氮肥对棉花生长发育及产量的影响[J]. 作物学报, 2022, 48(2): 396-409.
[7] 赵文青, 徐文正, 杨锍琰, 刘玉, 周治国, 王友华. 棉花叶片响应高温的差异与夜间淀粉降解密切相关[J]. 作物学报, 2021, 47(9): 1680-1689.
[8] 岳丹丹, 韩贝, Abid Ullah, 张献龙, 杨细燕. 干旱条件下棉花根际真菌多样性分析[J]. 作物学报, 2021, 47(9): 1806-1815.
[9] 曾紫君, 曾钰, 闫磊, 程锦, 姜存仓. 低硼及高硼胁迫对棉花幼苗生长与脯氨酸代谢的影响[J]. 作物学报, 2021, 47(8): 1616-1623.
[10] 李增强, 丁鑫超, 卢海, 胡亚丽, 岳娇, 黄震, 莫良玉, 陈立, 陈涛, 陈鹏. 铅胁迫下红麻生理特性及DNA甲基化分析[J]. 作物学报, 2021, 47(6): 1031-1042.
[11] 马欢欢, 方启迪, 丁元昊, 池华斌, 张献龙, 闵玲. 棉花GhMADS7基因正调控棉花花瓣发育[J]. 作物学报, 2021, 47(5): 814-826.
[12] 许乃银, 赵素琴, 张芳, 付小琼, 杨晓妮, 乔银桃, 孙世贤. 基于GYT双标图对西北内陆棉区国审棉花品种的分类评价[J]. 作物学报, 2021, 47(4): 660-671.
[13] 周冠彤, 雷建峰, 代培红, 刘超, 李月, 刘晓东. 棉花CRISPR/Cas9基因编辑有效sgRNA高效筛选体系的研究[J]. 作物学报, 2021, 47(3): 427-437.
[14] 卢海, 李增强, 唐美琼, 罗登杰, 曹珊, 岳娇, 胡亚丽, 黄震, 陈涛, 陈鹏. 红麻DNA甲基化响应镉胁迫及甲基化差异基因的表达分析[J]. 作物学报, 2021, 47(12): 2324-2334.
[15] 卢合全, 唐薇, 罗振, 孔祥强, 李振怀, 徐士振, 辛承松. 商品有机肥替代部分化肥对连作棉田土壤养分、棉花生长发育及产量的影响[J]. 作物学报, 2021, 47(12): 2511-2521.
Viewed
Full text


Abstract

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