Co-expression analysis identifies CRC and AP1 the regulator of Arabidopsis fatty acid biosynthesis.

Fatty acids (FAs) play crucial rules in signal transduction and plant development, however, the regulation of FA metabolism is still poorly understood. To study the relevant regulatory network, fifty-eight FA biosynthesis genes including de novo synthases, desaturases and elongases were selected as "guide genes" to construct the co-expression network. Calculation of the correlation between all Arabidopsis thaliana (L.) genes with each guide gene by Arabidopsis co-expression dating mining tools (ACT) identifies 797 candidate FA-correlated genes. Gene ontology (GO) analysis of these co-expressed genes showed they are tightly correlated to photosynthesis and carbohydrate metabolism, and function in many processes. Interestingly, 63 transcription factors (TFs) were identified as candidate FA biosynthesis regulators and 8 TF families are enriched. Two TF genes, CRC and AP1, both correlating with 8 FA guide genes, were further characterized. Analyses of the ap1 and crc mutant showed the altered total FA composition of mature seeds. The contents of palmitoleic acid, stearic acid, arachidic acid and eicosadienoic acid are decreased, whereas that of oleic acid is increased in ap1 and crc seeds, which is consistent with the qRT-PCR analysis revealing the suppressed expression of the corresponding guide genes. In addition, yeast one-hybrid analysis and electrophoretic mobility shift assay (EMSA) revealed that CRC can bind to the promoter regions of KCS7 and KCS15, indicating that CRC may directly regulate FA biosynthesis.

[1]  J. Browse,et al.  Fluxes through the prokaryotic and eukaryotic pathways of lipid synthesis in the '16:3' plant Arabidopsis thaliana. , 1986, The Biochemical journal.

[2]  Cindy Gustafson-Brown,et al.  Molecular characterization of the Arabidopsis floral homeotic gene APETALA1 , 1992, Nature.

[3]  Randall J Weselake,et al.  Gene coexpression clusters and putative regulatory elements underlying seed storage reserve accumulation in Arabidopsis , 2011, BMC Genomics.

[4]  Yoshiyuki Ogata,et al.  Approaches for extracting practical information from gene co-expression networks in plant biology. , 2007, Plant & cell physiology.

[5]  Stefan R. Henz,et al.  A gene expression map of Arabidopsis thaliana development , 2005, Nature Genetics.

[6]  H. Xue,et al.  Development of an efficient method for the isolation of factors involved in gene transcription during rice embryo development. , 2004, The Plant journal : for cell and molecular biology.

[7]  R. Mache Chloroplast ribosomal proteins and their genes. , 1990 .

[8]  E. Fehling,et al.  Acyl-CoA elongase from a higher plant (Lunaria annua): metabolic intermediates of very-long-chain acyl-CoA products and substrate specificity. , 1991, Biochimica et biophysica acta.

[9]  D. Roby,et al.  A MYB Transcription Factor Regulates Very-Long-Chain Fatty Acid Biosynthesis for Activation of the Hypersensitive Cell Death Response in Arabidopsis[W][OA] , 2008, The Plant Cell Online.

[10]  Staffan Persson,et al.  Co-expression tools for plant biology: opportunities for hypothesis generation and caveats. , 2009, Plant, cell & environment.

[11]  P. Moreau,et al.  The VLCFA elongase gene family in Arabidopsis thaliana: phylogenetic analysis, 3D modelling and expression profiling , 2008, Plant Molecular Biology.

[12]  Elliot M. Meyerowitz,et al.  Orchestration of Floral Initiation by APETALA1 , 2010, Science.

[13]  D. Shibata,et al.  Chloroplast development in Arabidopsis thaliana requires the nuclear‐encoded transcription factor Sigma B , 2000, FEBS letters.

[14]  T. Kuroiwa,et al.  Nuclear Encoding of a Chloroplast RNA Polymerase Sigma Subunit in a Red Alga , 1996, Science.

[15]  R. Upchurch Fatty acid unsaturation, mobilization, and regulation in the response of plants to stress , 2008, Biotechnology Letters.

[16]  Pil Joon Seo,et al.  The MYB96 Transcription Factor Regulates Cuticular Wax Biosynthesis under Drought Conditions in Arabidopsis[W] , 2011, Plant Cell.

[17]  M. Suh,et al.  The SebHLH transcription factor mediates trans-activation of the SeFAD2 gene promoter through binding to E- and G-box elements , 2007, Plant Molecular Biology.

[18]  John W. Pinney,et al.  Arabidopsis Co-expression Tool (ACT): web server tools for microarray-based gene expression analysis , 2006, Nucleic Acids Res..

[19]  P. McCourt,et al.  Fatty acid composition of leaf lipids determined after combined digestion and fatty acid methyl ester formation from fresh tissue. , 1986, Analytical biochemistry.

[20]  K. Shinozaki,et al.  Plastidic RNA polymerase σ factors in Arabidopsis , 1999 .

[21]  Cai-Zhong Jiang,et al.  WIN1, a transcriptional activator of epidermal wax accumulation in Arabidopsis. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Stefan de Folter,et al.  trans meets cis in MADS science. , 2006, Trends in plant science.

[23]  D. W. James,et al.  Mutants of Arabidopsis deficient in the synthesis of alpha-linolenate. Biochemical and genetic characterization of the endoplasmic reticulum linoleoyl desaturase. , 1993, The Journal of biological chemistry.

[24]  A. Subramanian Molecular genetics of chloroplast ribosomal proteins. , 1993, Trends in biochemical sciences.

[25]  M. Pollard,et al.  A specific acyl-ACP thioesterase implicated in medium-chain fatty acid production in immature cotyledons of Umbellularia californica. , 1991, Archives of biochemistry and biophysics.

[26]  J. Ohlrogge,et al.  Both antisense and sense expression of biotin carboxyl carrier protein isoform 2 inactivates the plastid acetyl-coenzyme A carboxylase in Arabidopsis thaliana. , 2002, The Plant journal : for cell and molecular biology.

[27]  C. Benning,et al.  wrinkled1: A novel, low-seed-oil mutant of Arabidopsis with a deficiency in the seed-specific regulation of carbohydrate metabolism. , 1998, Plant physiology.

[28]  Jan Ihmels,et al.  Principles of transcriptional control in the metabolic network of Saccharomyces cerevisiae , 2004, Nature Biotechnology.

[29]  Q. Meng,et al.  Antisense-mediated depletion of tomato endoplasmic reticulum omega-3 fatty acid desaturase enhances thermal tolerance. , 2010, Journal of integrative plant biology.

[30]  M. Hirai,et al.  Omics-based identification of Arabidopsis Myb transcription factors regulating aliphatic glucosinolate biosynthesis , 2007, Proceedings of the National Academy of Sciences.

[31]  G. Church,et al.  Expression dynamics of a cellular metabolic network , 2005, Molecular systems biology.

[32]  Yuqing He,et al.  Oil body biogenesis during Brassica napus embryogenesis. , 2009, Journal of integrative plant biology.

[33]  T. Shiina,et al.  Blue light-induced transcription of plastid-encoded psbD gene is mediated by a nuclear-encoded transcription initiation factor, AtSig5. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[34]  S. Fields,et al.  The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[35]  A. Aharoni,et al.  The SHINE Clade of AP2 Domain Transcription Factors Activates Wax Biosynthesis, Alters Cuticle Properties, and Confers Drought Tolerance when Overexpressed in Arabidopsis w⃞ , 2004, The Plant Cell Online.

[36]  Jian Zhang,et al.  LEAFY COTYLEDON1 Is a Key Regulator of Fatty Acid Biosynthesis in Arabidopsis1[C][W][OA] , 2008, Plant Physiology.

[37]  J. Ohlrogge,et al.  REGULATION OF FATTY ACID SYNTHESIS. , 1997, Annual review of plant physiology and plant molecular biology.

[38]  S. Rawsthorne Carbon flux and fatty acid synthesis in plants. , 2002, Progress in lipid research.

[39]  J. Browse,et al.  Arabidopsis mutants deficient in polyunsaturated fatty acid synthesis. Biochemical and genetic characterization of a plant oleoyl-phosphatidylcholine desaturase. , 1992, The Journal of biological chemistry.

[40]  The biochemistry and molecular biology of plant lipid biosynthesis , 1992 .

[41]  Fang-fang Fu,et al.  Coexpression Analysis Identifies Rice Starch Regulator1, a Rice AP2/EREBP Family Transcription Factor, as a Novel Rice Starch Biosynthesis Regulator1[W][OA] , 2010, Plant Physiology.

[42]  Fatty Acid Metabolism , 1988 .

[43]  S. Fields,et al.  A novel genetic system to detect protein–protein interactions , 1989, Nature.

[44]  Z. N. Oltvai,et al.  Topological units of environmental signal processing in the transcriptional regulatory network of Escherichia coli , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[45]  O. Fursova,et al.  Identification of ICE2, a gene involved in cold acclimation which determines freezing tolerance in Arabidopsis thaliana. , 2009, Gene.

[46]  R. Jetter,et al.  Sealing plant surfaces: cuticular wax formation by epidermal cells. , 2008, Annual review of plant biology.

[47]  D. Botstein,et al.  Cluster analysis and display of genome-wide expression patterns. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[48]  P. Stumpf,et al.  Isolation and function of spinach leaf beta-ketoacyl-[acyl-carrier-protein] synthases. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Jaworski,et al.  A Cerulenin Insensitive Short Chain 3-Ketoacyl-Acyl Carrier Protein Synthase in Spinacia oleracea Leaves. , 1989, Plant physiology.