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作物学报 ›› 2010, Vol. 36 ›› Issue (12): 2084-2090.doi: 10.3724/SP.J.1006.2010.02084

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

棉花4个脂肪酸合成相关基因的克隆和表达特征分析

董佳,魏利斌,胡艳,张天真,郭旺珍*   

  1. 南京农业大学 / 作物遗传与种质创新国家重点实验室, 江苏南京 210095
  • 收稿日期:2010-05-24 修回日期:2010-08-02 出版日期:2010-12-12 网络出版日期:2010-10-09
  • 通讯作者: 郭旺珍, E-mail: moelab@njau.edu.cn
  • 基金资助:

    本研究由国家重大基础研究项目(973计划) (2007CB108805)和教育部高等学校学科创新引智计划项目(111项目)(B08025)资助。

Molecular Cloning and Expression Analysis of Four Novel Fatty Acid Synthesis Related Genes in Gossypium hirsutum L.

DONG Jia,WEI Li-Bin,HU Yan,ZHANG Tian-Zhen,GUO Wang-Zhen*   

  1. National Key Laboratory of Crop Genetics & Germplasm Enhancement / Nanjing Agricultural University, Nanjing 210095, China
  • Received:2010-05-24 Revised:2010-08-02 Published:2010-12-12 Published online:2010-10-09
  • Contact: GUO Wang-Zhen,E-mail:moelab@njau.edu.cn

PDF

2174

被引次数

12

摘要： 脂肪酸合成相关代谢在控制油的合成和抗非生物胁迫中均起着重要作用。其脂肪酸合成相关基因的时空表达水平直接影响油的含量和脂肪酸合成相关酶的活性。本研究克隆了4个脂肪酸合成相关基因，分别命名为GhKASII、GhKASIII、GhFAD和GhGPAT，其中GhKASIII、GhFAD和GhGPAT基因cDNA全长通过电子克隆和同源克隆得到。而GhKASII通过筛库和5'-RACE途径得到。组织表达分析表明, 上述4个基因在根、茎、叶及纤维发育不同时期均有表达，属于组成性表达基因。其中GhKASII、GhKASIII在25 DPA种子中表达量最高，GhGPAT在0 DPA胚珠和15 DPA纤维中表达量很高，GhFAD在0DPA胚珠, 15 DPA种子，20 DPA纤维中表达量均很高。不同非生物胁迫的诱导表达分析表明，上述4个基因均不同程度被茉莉酸甲酯，ABA，创伤和冷害等逆境诱导表达。

关键词: 脂肪酸合成, 克隆, 表达, 非生物胁迫, 陆地棉

Abstract: Metabolism related to fatty acid synthesis plays an important role both in regulating oil biosynthesis and in plant abiotic stress tolerance. The spatial and temporal expression level of genes related to fatty acid synthesis influences the oil content and enzymes activity for fatty acid synthesis. In this study, four novel genes related to fatty acid synthesis, designated GhKASII, GhKASIII, GhFAD, and GhGPAT, were first isolated from upland cotton, respectively. The cDNA full-length of GhKASIII, GhFAD and GhGPAT were obtained by combining homologic cloning and sillico cloning, while GhKASII was obtained by screening cDNA library and 5'-RACE technique. The expression pattern analysis of different tissues and organs revealed that the transcripts of these genes were widely distributed in all the tested tissues and organs. GhKASII and GhKASIII showed the highest expression level in seeds at 25 DPA (day post anthesis), however, GhGPAT in 0 DPA ovules and 15DPA fiber tissues and GhFAD in 0 DPA ovules, 15 DPA seeds and 20 DPA fibers, respectively. Further, the analysis of expression induced by abiotic treatments indicated that the genes were differentially regulated under wounding, methyl jasmonate (Meja), cold and ABA (abscisic acid) treatments. The study will pave a way to develop further research in oil improvement of cotton seed and resistance to abiotic stress in cotton.

Key words: Fatty acid synthesis, Cloning, Expression, Abiotic stress, Upland cotton

[1]Seki M, Kamei A, Shinozaki K Y. Molecular responses to drought, salinity and frost: common and different paths for plant protection. Curr Opin Biotechnol, 2003, 14: 194–199
[2]Pirtle I L, Kongcharoensuntorn W, Nampaisansuk M, Knesek J E, Chapman K D, Pirtle R M. Molecular cloning and functional expression of the gene for a cotton Δ12 fatty acid desaturase (FAD2). Biochimica et Biophysica Acta, 2001, 2: 122–129
[3]Wolter F P, Schmidt R, Heinz E. Chilling sensitivity of Arabidopsis thaliana with genetically engineered membrane lipids. EMBO Journal, 1992, 11: 4685–4692
[4]Ohlrogge J B, Kuhn D N, Stumpf P K. Subcellular localization of acyl carrier protein in leaf protoplasts of Spinacia oleracea. Proc Natl Acad Sci USA, 1979, 76: 1194–1198
[5]Anai T, Koga M, Tanaka H, Kinoshita T, Rahman S M, Takagi Y. Improvement of rice (Oryza sativa L.) seed oil quality through introduction of a soybean microsomal omega-3 fatty acid desaturase gene. Plant Cell Rep, 2003, 21: 988–992
[6]Wu Y-T(武耀廷), Liu J-Y(刘进元). A modified hot borate method for efficient isolation of total RNA from different cotton tissues. Cotton Sci (棉花学报), 2004, 16(2): 67–71 (in Chinese with English abstract)
[7]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)
[8]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real­time quantitative PCR and the 2(­delta delta C(T)) method. Methods, 2001, 25: 402–408
[9]Hwang S K, Hwang Y S. Molecular cloning and functional expression of perilla frutescens 3-ketoacyl-
[acyl carrier protein] synthase III. Mol Cells, 2000, 10: 375–381
[10]Hwang S K, Kim K H, Hwang Y S. Molecular cloning and expression analysis of 3-ketoacyl-acp synthases in the immature seeds of Perilla frutescens. Mol cells, 2000, 10: 533–539
[11]Pidkowich M S, Nguyen H T, Heilmann I, Ischebeck T, Shanklin J. Modulating seed β-ketoacyl-acyl carrier protein synthase II level converts the composition of a temperate seed oil to that of a palm-like tropical oil. Proc Natl Acad Sci USA, 2007, 104: 4742–4747
[12]Wanjie S W, Welti R, Moreau R A, Chapman K D. Identi?cation and quanti?cation of glycerolipids in cotton fibers: reconciliation with metabolic pathway predictions from DNA databases. Lipids, 2005, 40: 773–785
[13]Qin Y M, Hu C Y, Pang Y, Kastaniotis A J, Hiltunen J K, Zhu Y X. Saturated very-long-chain fatty acids promote cotton fiber and Arabidopsis cell elongation by activating ethylene biosynthesis. Plant Cell, 2007, 19: 3692–3704
[14]Ji S J, Lu Y C, Feng J X, Wei G, Li J, Shi Y H, Fu Q, Liu D, Luo J C, Zhu Y X. Isolation and analyses of genes preferentially expressed during early cotton fiber development by subtractive PCR and cDNA array. Nucleic Acids Res, 2003, 31: 2534–2543
[15]Nishiuchi T, Hamada T, Kodama H. Wounding changes the spatial expression pattern of the Arabidopsis plastid ω-3 fatty acid desaturase gene (FAD7) through different signal transduction pathways. Plant Cell, 1997, 9: 1701–1712
[16]Hamada T, Nishiuchi T, Kodama H, Nishimura M, Iba K. cDNA cloning of a wounding-inducible gene encoding a plastid ω-3 fatty acid desaturase from tobacco. Plant Cell Physiol, 1996, 37: 606–611
[17]Zou J, Abrams G D, Barton D L, Taylor D C, Pomeroy M K, Abrams S R. lnduction of lipid and oleosin biosynthesis by (+)-abscisic acid and its metabolites in microspore-derived embryos of Brassica napus L. cv. Reston (biological responses in the presence of 8
[prime]-hydroxyabscisic acid). Plant Physiol, 1995, 108: 563–571
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