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Acta Agron Sin ›› 2014, Vol. 40 ›› Issue (07): 1304-1310.doi: 10.3724/SP.J.1006.2014.01304

• RESEARCH NOTES • Previous Articles     Next Articles

Cloning and Expression Analysis of Two Homologous Genes Coding sn-Glycerol-3-Phosphate Acyltransferase 6 in Brassica napus

LIU Cong1,XIAO Dan-Wang1,HU Xue-Fang1,WU Ke-Bin1,GUAN Chun-Yun1,2,XIONG Xing-Hua1,2,*   

  1. 1 Crop Gene Engineering Key Laboratory of Hunan Province, Hunan Agricultural University, Changsha 410128, China; 2 Oilseed Crops Institute, Hunan Agricultural University, Changsha 410128, China
  • Received:2013-11-16 Revised:2014-04-16 Online:2014-07-12 Published:2014-05-16
  • Contact: 熊兴华, E-mail: ndxiongene@yahoo.com, Tel: 13508487613

Abstract:

A key enzyme sn-glycerol-3-phosphate acyltransferase (GPAT) catalyzes the initial step of TAG biosynthetic pathway, and is involved in many processes including plant growth, development and response to abiotic stresses. In this study, two B. napus GPAT6 homologs were cloned from the leaf of the cultivar xiangyou 15 using RT-PCR, and designated as BnGPAT6-1 and BnGPAT6-2, respectively. The coding DNA sequences (CDS) of two BnGPAT6 genes are 1506 bp in length, encoding two different polypeptides of 501 amino acid residues. The proteins were predicted to consist of a haloacid dehalogenase-like hydrolase domain and a lysophospholipid acyltransferase domain. Multiple sequence alignments and phylogenetic analysis of GPAT proteins showed that BnGPAT6-1 and BnGPAT6-2 share high similarity with GPAT6 genes from B. rapa, B. oleracea, Arabidopsis thaliana,and A. lyrata. Tissue expression amounts of BnGPAT6 genes showed that their mRNA were more abundant in flower than the other organs, and the patterns rise up at first and then down in the developing embryo. The expression of BnGPAT6 genes are inhibited by ABA, while go up simultaneously under drought and 6-BA conditions. In the treatment of salt, the expression of BnGPAT6 genes are uptrend in a short time and then down, and there isn’t obvious change under stress of water logging.

Key words: Brassica napus, sn-glycerol-3-phosphate acyltransferase, Gene cloning, Expression analysis, Abiotic stresses

[1]Gupta S M, Pandey P, Grover A, Patade V Y, Singh S, Ahmed Z. Cloning and characterization of GPAT gene from Lepidium latifolium L.: a step towards translational research in agri-genomics for food and fuel. Mol Biol Rep, 2013, 40: 4235–4240



[2]Chen X, Snyder C L, Truksa M, Shah S, Weselake R J. sn-Glycerol-3-phosphate acyltransferases in plants. Plant Signal Behav, 2011, 6: 1695–1699



[3]Nishida I, Tasaka Y, Shiraishi H, Murata N. The gene and the RNA for the precursor to the plastid-located glycerol-3-phosphate acyltransferase of Arabidopsis thaliana. Plant Mol Biol, 1993, 21: 267–277



[4]Yang W, Simpson J P, Li-Beisson Y, Beisson F, Pollard M, Ohlrogge J B. A land-plant-specific glycerol-3-phosphate acyltransferase family in Arabidopsis: substrate specificity, sn-2 preference, and evolution. Plant Physiol, 2012, 160: 638–652



[5]Gidda S K, Shockey J M, Rothstein S J, Dyer J M, Mullen R T. Arabidopsis thaliana GPAT8 and GPAT9 are localized to the ER and possess distinct ER retrieval signals: functional divergence of the dilysine ER retrieval motif in plant cells. Plant Physiol Biochem, 2009, 47: 867–879



[6]Murata N, Tasaka Y. Glycerol-3-phosphate acyltransferase in plants. Biochim Biophys Acta, 1997, 1348: 10–16



[7]Wendel A A, Lewin T M, Coleman R A. Glycerol-3-phosphate acyltransferases: Rate limiting enzymes of triacylglycerol biosynthesis. Biochimica et Biophysica Acta (BBA). Mol Cell Biol Lipids, 2009, 1791: 501–506



[8]Zhang Y M, Rock C O. Thematic review series: glycerolipids. Acyltransferases in bacterial glycerophospholipid synthesis. J Lipid Res, 2008, 49: 1867–1874



[9]Yang W, Pollard M, Li-Beisson Y, Beisson F, Feig M, Ohlrogge J. A distinct type of glycerol-3-phosphate acyltransferase with sn-2 preference and phosphatase activity producing 2-monoacylglycerol. Proc Natl Acad Sci USA, 2010, 107: 12040–12045



[10]Bourgis F, Kader J C, Barret P, Renard M, Robinson D, Robinson C, Delseny M, Roscoe T J. A plastidial lysophosphatidic acid acyltransferase from oilseed rape. Plant Physiol, 1999, 120: 913–922



[11]Lung S C, Weselake R J. Diacylglycerol acyltransferase: a key mediator of plant triacylglycerol synthesis. Lipids, 2006, 41: 1073–1088



[12]Jain R K, Coffey M, Lai K, Kumar A, Mackenzie S L. Enhancement of seed oil content by expression of glycerol-3-phosphate acyltransferase genes. Biochem Soc Trans, 2000, 28: 958–961



[13]i Y, Beisson F, Koo A J, Molina I, Pollard M, Ohlrogge J. Identification of acyltransferases required for cutin biosynthesis and production of cutin with suberin-like monomers. Proc Natl Acad Sci USA, 2007, 104: 18339–18344



[14]Li X C, Zhu J, Yang J, Zhang G R, Xing W F, Zhang S, Yang Z N. Glycerol-3-phosphate acyltransferase 6 (GPAT6) is important for tapetum development in Arabidopsis and plays multiple roles in plant fertility. Mol Plant, 2012, 5: 131–142



[15]Chen Y Q, Kuo M S, Li S, Bui H H, Peake D A, Sanders P E, Thibodeaux S J, Chu S, Qian Y W, Zhao Y, Bredt D S, Moller D E, Konrad R J, Beigneux A P, Young S G, Cao G. AGPAT6 is a novel microsomal glycerol-3-phosphate acyltransferase. J Biol Chem, 2008, 283: 10048–10057



[16]Chen X, Truksa M, Snyder C L, El-Mezawy A, Shah S, Weselake R J. Three homologous genes encoding sn-glycerol-3-phosphate acyltransferase 4 exhibit different expression patterns and functional divergence in Brassica napus. Plant Physiol, 2011, 155: 851–865



[17]Manas-Fernandez A, Li-Beisson Y, Alonso D L, Garcia-Maroto F. Cloning and molecular characterization of a glycerol-3-phosphate O-acyltransferase (GPAT) gene from Echium (Boraginaceae) involved in the biosynthesis of cutin polyesters. Planta, 2010, 232: 987–997



[18]Li-Beisson Y, Pollard M, Sauveplane V, Pinot F, Ohlrogge J, Beisson F. Nanoridges that characterize the surface morphology of flowers require the synthesis of cutin polyester. Proc Natl Acad Sci USA, 2009, 106: 22008–22013



[19]Tamada T, Feese M D, Ferri S R, Kato Y, Yajima R, Toguri T, Kuroki R. Substrate recognition and selectivity of plant glycerol-3-phosphate acyltransferases (GPATs) from Cucurbita moscata and Spinacea oleracea. Acta Crystallogr D Biol Crystallogr, 2004, 60: 13–21



[20]Ferri S R, Toguri T. Substrate specificity modification of the stromal glycerol-3-phosphate acyltransferase. Arch Biochem Biophys, 1997, 337: 202–208



[21]Sui N, Li M, Zhao S J, Li F, Liang H, Meng Q W. Overexpression of glycerol-3-phosphate acyltransferase gene improves chilling tolerance in tomato. Planta, 2007, 226: 1097–1108



[22]Zhu S Q, Zhao H, Liang J S, Ji B H, Jiao D M. Relationships between phosphatidylglycerol molecular species of thylakoid membrane lipids and sensitivities to chilling-induced photoinhibition in rice. J Integr Plant Biol, 2008, 50: 194–202



[23]Szalontai B, Kota Z, Nonaka H, Murata N. Structural consequences of genetically engineered saturation of the fatty acids of phosphatidylglycerol in tobacco thylakoid membranes. An FTIR study. Biochemistry, 2003, 42: 4292–4299



[24]Bertrams M, Heinz E. Positional specificity and fatty acid selectivity of purified sn-glycerol 3-phosphate acyltransferases from chloroplasts. Plant Physiol, 1981, 68: 653-657.



[25]Weber S, Wolter F P, Buck F, Frentzen M, Heinz E. Purification and cDNA sequencing of an oleate-selective acyl-ACP:sn-glycerol-3-phosphate acyltransferase from pea chloroplasts. Plant Mol Biol, 1991, 17: 1067–1076



[26]Cronan J E, Roughan P G. Fatty acid specificity and selectivity of the chloroplast sn-glycerol 3-phosphate acyltransferase of the chilling sensitive plant. Amaranthus lividus Plant Physiol, 1987, 83: 676–680



[27]Yan K, Chen N, Qu Y Y, Dong X C, Meng Q W, Zhao S J. Overexpression of sweet pepper glycerol-3-phosphate acyltransferase gene enhanced thermotolerance of photosynthetic apparatus in transgenic tobacco. J Integr Plant Biol, 2008, 50: 613–621



[28]Zheng Z, Xia Q, Dauk M, Shen W, Selvaraj G, Zou J. Arabidopsis AtGPAT1, a member of the membrane-bound glycerol-3-phosphate acyltransferase gene family, is essential for tapetum differentiation and male fertility. Plant Cell, 2003, 15: 1872–1887



[29]华玮, 李荣俊, 梁述平, 吕应堂. 烟草幼苗两种钙调素结合蛋白激酶的活性调节及基因表达. 植物生理与分子生物学学报, 2005, 31: 305–310



Hua W, Li R J, Liang S P, Lu Y T. Gene expression and activity regulation of two calmodulin binding protein kinases in tobacco seedling. J Plant Physiol Mol Biol, 2005, 31: 305–310 (in Chinese with English abstract)



[30]Hua W, Zhang L, Liang S, Jones R L, Lu Y T. A tobacco calcium/calmodulin-binding protein kinase functions as a negative regulator of flowering. J Biol Chem, 2004, 279: 31483–31494



[31]郭彦, 杨洪双, 李清旭, 孙学彬. 激素对野生大豆幼苗抗旱能力的影响. 河南农业科学, 2007, (4): 37–39



Guo Y, Yang H S, Li Q X, Sun X B. Effects of hormones on the tolerance of wild soybean seedlings against water stress. J Henan Agric Sci, 2007, (4): 37–39 (in Chinese with English abstract)



[32]童晋, 詹高淼, 王新发, 刘贵华, 华玮, 王汉中. 油菜柠檬酸合酶基因的克隆及在逆境下的表达. 作物学报, 2009, 35: 33–40



Tong J, Zhan G M, Wang, X F, Liu G H, Hua W, Wang H Z. Cloning of citrate synthase gene in rapeseed (Brassica napus L.) and its expression under stresses. Acta Agron Sin, 2009: 33–40 (in Chinese with English abstract)

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