Cloning and Bioinformatics Analysis of LPAT4 in Rapeseed

  • XIAO Dan-wang ,
  • LIU Cong ,
  • HU Xue-fang ,
  • WU Ke-bin ,
  • GUAN Chun-yun ,
  • XIONG Xing-hua
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  • 1. Crop Gene Engineering Key Laboratory of Hunan Province, Hunan Agricultural University, Changsha 410128, China;
    2. Hunan Branch of National Oilseed Crops Improvement Centre, Hunan Agricultural University, Changsha 410128, China

Received date: 2014-03-12

  Online published: 2014-09-19

Abstract

To further study on LPAT4 of Brassica napus, a pair of primers were designed to obtain the full length CDs sequence of rapeseed LPAT4 gene, according to the homologous comparison of B.campestris, B.oleracea and assembled EST sequence, which was strong similarity to the LPAT4 gene of Arabidopsis thaliana by searching bioinformatics at Geboc database.In this paper, two copies of full length CDs sequences(1 143, 1 140 bp)have been cloned, these clones were designated as BnLPAT4-1 and BnLPAT4-2, respectively. BnLPAT4-1 encoded a protein of 380 amino acids with the molecular weights of 43.29 kDa and the isoelectric point of 8.74; BnLPAT4-2 encoded a polypeptide of 379 amino acids with the molecular weights of 43.16 kDa and the isoelectric point of 8.93.They belong to the LPLAT superfamily with LPLAT_LCLAT1 like domain.SNP analysis between different rapeseed cultivars revealed that LPAT4 exist 41 polymorphic sites, In which 7 were singleton polymorphic sites, and 34 were parsimony informative polymorphic sites.41 polymorphic sites leaded to 41 mutations, among them, 23 were synonymous changes and 18 were replacement changes which contribute to 16 amino acids changed.In terms of sequence composition, physico-chemical property, conserved domain and homologous-modeling analysis, the analyses showed that BnLPAT4-1 and BnLPAT4-2 were two copies of Xiangyou 15 LPAT4 gene, at the same time, they should be the members of LPLAT superfamily.

Cite this article

XIAO Dan-wang , LIU Cong , HU Xue-fang , WU Ke-bin , GUAN Chun-yun , XIONG Xing-hua . Cloning and Bioinformatics Analysis of LPAT4 in Rapeseed[J]. Acta Agriculturae Boreali-Sinica, 2014 , 29(3) : 52 -58 . DOI: 10.7668/hbnxb.2014.03.011

References

[1] 张 垚,李云昌,梅德圣,等.油菜油脂研究进展[J].植物学通报,2007,24(4):435-443.
[2] Vigeolas H,Waldeck P,Zank T,et al.Increasing seed oil content in oil-seed rape (Brassica napus L.) by over-expression of a yeast glycerol-3-phosphate dehydrogenase under the control of a seed-specific promoter[J].Plant Biotechnol J,2007,5(3):431-441.
[3] David C,Taylor Y Z A K,Dennis L,et al.Molecular modification of triacylglycerol accumulation by over-expression of DGAT1 to produce canola with increased seed oil content under field conditions[J].Botany,2009,87:533-543.
[4] Burns M J,Barnes S R,Bowman J G,et al.QTL analysis of an intervarietal set of substitution lines in Brassica napus:Seed oil content and fatty acid composition[J].Heredity (Edinb),2003,90(1):39-48.
[5] Chai G,Bai Z,Wei F,et al.Brassica GLABRA2 genes: analysis of function related to seed oil content and development of functional markers[J].TAG Theoretical and Applied Genetics,2010,120(8):1597.
[6] Zou J,Katavic V,Giblin E M,et al.Modification of seed oil content and acyl composition in the brassicaceae by expression of a yeast sn-2 acyltransferase gene[J].Plant Cell,1997,9(6):909-923.
[7] Rao S S,Hildebrand D.Changes in oil content of transgenic soybeans expressing the yeast SLC1 gene[J].Lipids,2009,44(10):945-951.
[8] Maisonneuve S,Bessoule J J,Lessire R,et al.Expression of rapeseed microsomal lysophosphatidic acid acyltransferase isozymes enhances seed oil content in Arabidopsis1 [J].Plant Physiol,2010,152(2):670-684.
[9] 戚维聪.油菜发育种子中油脂积累与Kennedy途径酶活性的关系研究[D].南京:南京农业大学,2008.
[10] 陈四龙,黄家权,雷 永,等.花生溶血磷脂酸酰基转移酶基因的克隆与表达分析[J].作物学报,2012,38(2):245-255.
[11] Kim H U,Li Y,Huang A H.Ubiquitous and endoplasmic reticulum-located lysophosphatidyl acyltransferase,LPAT2, is essential for female but not male gametophyte development in Arabidopsis[J].Plant Cell,2005,17(4):1073-1089.
[12] Arroyo-Caro J M,Chileh T,Kazachkov M,et al.The multigene family of lysophosphatidateacyltransferase (LPAT)- related enzymes in Ricinus communis.Cloning and molecular characterization of two LPAT genes that are expressed in castor seeds[J].Plant Sci,2013,199-200:29-40.
[13] Hanke C,Wolter F P,Coleman J,et al.A plant acyltransferase involved in triacylglycerol biosynthesis complements an Escherichia coli sn-1-acylglycerol-3-phosphate acyltransferase mutant[J].Eur J Biochem,1995,232(3):806-810.
[14] Knutzon D S,Lardizabal K D,Nelsen J S,et al.Cloning of a coconut endosperm cDNA encoding a 1-acyl-sn-glycerol-3-phosphate acyltransferase that accepts medium-chain-length substrates[J].Plant Physiol,1995,109(3):999-1006.
[15] Brown A P,Coleman J,Tommey A M,et al.Isolation and characterisation of a maize cDNA that complements a 1-acyl sn-glycer-ol-3-phosphate acyltransferase mutant of Escherichia coli and encodes a protein which has similarities to other acyltransferases[J].Plant Mol Biol,1994,26(1):211-223.
[16] Brown A P,Carnaby S,Brough C,et al.Limnanthes douglasii lysophosphatidic acid acyltransferases: immunological quantification,acyl selectivity and functional replacement of the Escherichia coli plsC gene[J].Biochem J,2002,364:795-805.
[17] Brown A P,Kroon J T,Swarbreck D,et al.Tissue-specific whole transcriptome sequencing in castor,directed at understanding triacylglycerol lipid biosynthetic pathways[J].PLoS One,2012,7(2):1-13.
[18] 姚行成.芸薹属栽培种与野生种Brassicamaurorum的种间杂交及基因组关系[D].武汉: 华中农业大学,2010.
[19] Chen X,Li X E,Zhang B,et al.Detection and genotyping of restriction fragment associated polymorphisms in polyploid crops with a pseudo-reference sequence: a case study in allotetraploid Brassica napus[J].BMC Genomics,2013,14(1):346.
[20] Kazachkova N,Fahleson J,Meijer J.Establishment of the amplified fragment length polymorphism (AFLP) technique for genotyping of pollen beetle (Meligethesaeneus)-a noxious insect pest on oilseed rape (Brassica napus)[J].MolBiol Rep,2004,31(1):37-42.
[21] Bus A,Hecht J,Huettel B,et al.High-throughput polymorphism detection and genotyping in Brassica napus using next-generation RAD sequencing[J].BMC Genomics,2012,13:281.
[22] Durstewitz G,Polley A,Plieske J,et al.SNP discovery by amplicon sequencing and multiplex SNP genotyping in the allopolyploid species Brassica napus[J].Genome,2010,53(11):948-956.
[23] Howell E C,Kearsey M J,Jones G H,et al.A and C genome distinction and chromosome identification in Brassica napus by sequential fluorescence in situ hybridization and genomic in situ hybridization[J].Genetics,2008,180(4):1849-1857.
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