Cloning and Bioinformatics Analysis of GPDH in Rapeseed

doi:10.7668/hbnxb.2017.02.017

Abstract

Abstract: To further study on GPDH of Brassica napus,through the collinearity analysis of the GPDH from Arabidopsis thaliana and Brassica napus:there are 4 copies of GPDH of A genome and C genome in the Brassica napus called as GSBRNA2T00132785001,GSBRNA2T00024258001,GSBRNA2T00058395001,GSBRN-A2T00019118001.Four cDNA were downloaded from GenBank,are named as:GPDH1,GPDH2,GPDH3 and GPDH4. Four pairs of primers were designed according to the sequence informations,and four nucleotide fragments were amplified from Xiangyou 15 full-length cDNA,designated as BnGPDH1,BnGPDH2,BnGPDH3,BnGPDH4. The result of nucleotide sequences and protein structure analysis by bioinformatics software and online system showed:Four CDS sequences length were 1 395,1 395,1 389,1 536 bp respectively.When BnGPDH4 is translated into the amino acid sequence,we found a stop codon in the middle of BnGPDH4. Which isn't an active protein.The number of amino acid of three other BnGPDH are 464,464 and 462 respectively.Secondary structure are consistant of the alpha helix (42%),random coil (42%)and extended chain (15%).The transmembrane structure of the three proteins was uniform.Their conservative domains consist of GpsA,NADB_Rossmann superfamily and NAD_Gly3P_dh_C superfamily,which belong to the GPDH superfamily.Homology modeling analysis showed that they were of model of Glycerol-3-phosphate dehydrogenase and located in cytoplasm by subcellular localization analysis.In terms of sequence composition,phylogenic tree,physico-chemical property,secondary structure and tertiary structure and subcellular localization,they have the characteristics of GPDH superfamily.They should be the members of GPDH superfamily.

Key words: GPDH, Brassica napus L., Bioinformatics analysis

CLC Number:

Q78
S635.03

LIU Shaofeng, WANG Huihui, WU Kebin, XIONG Xinghua. Cloning and Bioinformatics Analysis of GPDH in Rapeseed[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(2): 109-116. doi: 10.7668/hbnxb.2017.02.017.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2017.02.017

http://www.hbnxb.net/EN/Y2017/V32/I2/109

References

[1] 王汉中,殷艳.我国油料产业形势分析与发展对策建议[J].中国油料作物学报,2014,36(3):414-421.
[2] Shen W,Wei Y,Dauk M,et al.Involvement of a glycerol-3-phosphate dehydrogenase in modulating the NADH[J].Plant Cell,2006,18(2):422-441.
[3] Yao Y,Yang L,Peng K T,et al.Glycerol and neutral lipid production in the oleaginous marine diatom Phaeodactylum tricornutum promoted by over expression of glycerol-3-phosphate dehydrogenase[J].Biotechnology for Biofuels,2014,7(1):1-9.
[4] 秦萌,徐敏,刘倩楠,等.酿酒酵母细胞GUT2基因的研究进展[J].中国医学工程,2015,23(1):199-200.
[5] Hubmann G,Guillouet S,Nevoigt E.Gpd1 and Gpd2 fine-tuning for sustainable reduction of glycerol formation in Saccharomyces cerevisiae[J].Applied and Environmental Microbiology,2011,77(17):5857-5867.
[6] Kachroo A,Venugopal S,Lapchyk L,et al.Oleic acid levels regulated by glycerolipid metabolism modulate defense gene expression in Arabidopsis[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(14):5152-5157.
[7] Chandra-Shekara A,Venugopal S,Barman S,et al.Plastidial fatty acid levels regulate resistance gene-dependent defense signaling in Arabidopsis[J].Proceedings of the National Academy of Sciences of the United States of America,2007,104(17):7277-7282.
[8] Vigeolas H,Geigenberger P.Increased levels of glycerol-3-phosphate Lead to a stimulation of flux into triacylglycerol synthesis after supplying glycerol to developing seeds of Brassica napus L.in planta[J].Planta,2004,219(5):827-835.
[9] Roesler K,Shintani D,Savage L,et al.Targeting of the Arabidopsis homomeric acetyl-coenzyme a carboxylase to plastids of rapeseeds[J].Plant Physiology,1997,113(1):75-81.
[10] Taylor D C,Katavic V,Zou J T,et al.Field testing of transgenic rapeseed cv.Hero transformed with a yeast sn-2 acyltransferase results in increased oil content,erucic acid contentand seed yield[J].Molecular Breeding:New Strategies in Plant Improvement,2002,8(4):317-322.
[11] Jako C,Kumar A,Wei Y D,et al.Seed-specific over-expression of an Arabidopsis cDNA encoding a diacylglycerol acyltransferase enhances seed oil content and seed weight[J].Plant Physiology,2001,126(2):861-874.
[12] Mu J Y,Tan H L,Zheng Q,et al.LEAFY COTYLEDON1 is a key regulator of fatty acid biosynthesis in Arabidopsis[J].Plant Physiology,2008,148(2):1042-1054.
[13] Tan H,Yang X,Zhang F,et al.Enhanced seed oil production in canola by conditional expression of Brassica napus LEAFY COTYLEDON1 and LEC1-LIKE in developing seeds[J].Plant Physiology,2011,156(3):1577-1588.
[14] 刘野.海滨锦葵茎尖再生体系建立及GPD1和DGAT植物表达载体构建[D].大连:辽宁师范大学,2010.
[15] 弭宪杰,徐荣华,刘爱忠,等.蓖麻三磷酸甘油脱氢酶基因(RcGPDH)的克隆及功能分析[J].中国油料作物学报,2011,33(5):451-458.
[16] 胡军.三磷酸甘油脱氢酶基因和甘油代谢对拟南芥种子含油量与根系发育的影响[D].武汉:华中农业大学,2013.
[17] 张超.油菜甘油-3-磷酸脱氢酶基因的克隆及功能研究[D].南京:南京农业大学,2012.
[18] Chalhoub B,Denoeud F,Liu S,et al.Plant genetics.early allopolyploid evolution in the post-Neolithic Brassica napus oilseed genome[J].Science,2014,345(6199):950-953.
[19] Shen W,Wei Y,Dauk M,et al.Identification of a mitochondrial glycerol-3-phosphate dehydrogenase from Arabidopsis thaliana:evidence for a mitochondrial glycerol-3-phosphate shuttle in plants[J].FEBS Letters,2003,536(1/3):92-96.
[20] Kurkdjian A,Guern J.Intracellular PH:measurement and importance in cell activity[J].Plant Biology,1989,40(40):271-303.
[21] Shen W,Li J,Dauk M,et al.Metabolic and transcriptional responses of glycerolipid pathways to a perturbation of glycerol 3-phosphate metabolism in Arabidopsis[J].The Journal of Biological Chemistry,2010,285(30):22957-22965.

Please choose a citation manager

Content to export