Arabidopsis AtGPAT 1 , a Member of the Membrane-Bound Glycerol-3-Phosphate Acyltransferase Gene Family , Is Essential for Tapetum Differentiation and Male Fertility

Membrane-bound glycerol-3-phosphate acyltransferase (GPAT; EC 2.3.1.15) mediates the initial step of glycerolipid biosynthesis in the extraplastidic compartments of plant cells. Here, we report the molecular characterization of a novel GPAT gene family from Arabidopsis, designated AtGPAT . The corresponding polypeptides possess transmembrane domains and GPAT activity when expressed heterologously in a yeast lipid mutant. The functional significance of one isoform, AtGPAT1, is the focus of the present study. Disruption of the AtGPAT1 gene causes a massive pollen development arrest, and subsequent introduction of the gene into the mutant plant rescues the phenotype, illustrating a pivotal role for AtGPAT1 in pollen development. Microscopic examinations revealed that the gene lesion results in a perturbed degeneration of the tapetum, which is associated with altered endoplasmic reticulum profiles and reduced secretion. In addition to the sporophytic effect, AtGPAT1 also exerts a gametophytic effect on pollen performance, as the competitive ability of a pollen grain to pollinate is dependent on the presence of an AtGPAT1 gene. Deficiency in AtGPAT1 correlates with several fatty acid composition changes in flower tissues and seeds. Unexpectedly, however, a loss of AtGPAT1 causes no significant change in seed oil content.

[1]  Jonathan Pevsner,et al.  Basic Local Alignment Search Tool (BLAST) , 2005 .

[2]  A. Papini,et al.  Programmed-cell-death events during tapetum development of angiosperms , 1999, Protoplasma.

[3]  H. Owen,et al.  Ultrastructure of microsporogenesis and microgametogenesis inArabidopsis thaliana (L.) Heynh. ecotype Wassilewskija (Brassicaceae) , 1995, Protoplasma.

[4]  A. Cheung,et al.  Programmed cell death in plant reproduction , 2000, Plant Molecular Biology.

[5]  M. Frentzen,et al.  Substrate specificities of the membrane-bound and partially purified microsomal acyl-CoA:1-acylglycerol-3-phosphate acyltransferase from etiolated shoots of Pisum sativum (L.) , 1991, Planta.

[6]  Ettore Pacini,et al.  The tapetum: Its form, function, and possible phylogeny inEmbryophyta , 1985, Plant Systematics and Evolution.

[7]  M. Cresti,et al.  Secretory tapetum of Brassica oleracea L.: polarity and ultrastructural features , 2004, Sexual Plant Reproduction.

[8]  C. Jolly,et al.  Aging and acyl-CoA binding protein alter mitochondrial glycerol-3-phosphate acyltransferase activity. , 2003, Biochimica et biophysica acta.

[9]  J. Zou,et al.  Identification of a mitochondrial glycerol‐3‐phosphate dehydrogenase from Arabidopsis thaliana: evidence for a mitochondrial glycerol‐3‐phosphate shuttle in plants , 2003, FEBS letters.

[10]  J. Shaw,et al.  Mitochondrial GFA2 Is Required for Synergid Cell Death in Arabidopsis Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.002170. , 2002, The Plant Cell Online.

[11]  P. Mullineaux,et al.  Isolated Plant Mitochondria Import Chloroplast Precursor Proteinsin Vitro with the Same Efficiency as Chloroplasts* , 2002, The Journal of Biological Chemistry.

[12]  D. Bouchez,et al.  Pollen tube development and competitive ability are impaired by disruption of a Shaker K(+) channel in Arabidopsis. , 2002, Genes & development.

[13]  J. Zou,et al.  The Initial Step of the Glycerolipid Pathway , 2001, The Journal of Biological Chemistry.

[14]  Z. Zheng,et al.  Isolation and characterization of novel defence-related genes induced by copper, salicylic acid, methyl jasmonate, abscisic acid and pathogen infection in Brassica carinata. , 2001, Molecular plant pathology.

[15]  J. Balk,et al.  The PET 1-CMS Mitochondrial Mutation in Sunflower Is Associated with Premature Programmed Cell Death and Cytochrome c Release , 2001 .

[16]  R. Amasino,et al.  The Arabidopsis knockout facility at the University of Wisconsin-Madison. , 2000, Plant physiology.

[17]  J. Ohlrogge,et al.  Accumulation of palmitate in Arabidopsis mediated by the acyl-acyl carrier protein thioesterase FATB1. , 2000, Plant physiology.

[18]  A. Jones,et al.  Does the plant mitochondrion integrate cellular stress and regulate programmed cell death? , 2000, Trends in plant science.

[19]  K. Athenstaedt,et al.  Phosphatidic acid, a key intermediate in lipid metabolism. , 1999, European journal of biochemistry.

[20]  Benning,et al.  The TAG1 locus of Arabidopsis encodes for a diacylglycerol acyltransferase. , 1999, Plant physiology and biochemistry : PPB.

[21]  H. Sul,et al.  Acyltransferases of de novo glycerophospholipid biosynthesis. , 1999, Progress in lipid research.

[22]  A. Kumar,et al.  The Arabidopsis thaliana TAG1 mutant has a mutation in a diacylglycerol acyltransferase gene. , 1999, The Plant journal : for cell and molecular biology.

[23]  Voelker,et al.  Lysophosphatidic acid acyltransferase from coconut endosperm mediates the insertion of laurate at the sn-2 position of triacylglycerols in lauric rapeseed oil and can increase total laurate levels , 1999, Plant physiology.

[24]  J. Harwood,et al.  ACYL-CoA: GLYCEROL-3-PHOSPHATE ACYLTRANSFERASE FROM OIL PALM ( Elaeis guineensis ) TISSUES , 1999 .

[25]  D. Murphy,et al.  Composition and role of tapetal lipid bodies in the biogenesis of the pollen coat of Brassica napus , 1999, Planta.

[26]  T. M. Lewin,et al.  Analysis of amino acid motifs diagnostic for the sn-glycerol-3-phosphate acyltransferase reaction. , 1999, Biochemistry.

[27]  J. Vance,et al.  Mechanisms of lipid-body formation. , 1999, Trends in biochemical sciences.

[28]  K. Platt,et al.  Ultrastructural Study of Lipid Accumulation in Tapetal Cells of Brassica napus L. Cv. Westar during Microsporogenesis , 1998, International Journal of Plant Sciences.

[29]  D. Murphy,et al.  Biogenesis and function of the lipidic structures of pollen grains , 1998, Sexual Plant Reproduction.

[30]  P. Schnable,et al.  The molecular basis of cytoplasmic male sterility and fertility restoration , 1998 .

[31]  C. Benning,et al.  wrinkled 1 : A Novel , Low-Seed-Oil Mutant of Arabidopsis with a Deficiency in the Seed-Specific Regulation of Carbohydrate Metabolism 1 , 1998 .

[32]  W. Wilkison,et al.  sn-Glycerol-3-phosphate acyltransferase from Escherichia coli. , 1997, Biochimica et biophysica acta.

[33]  D. Taylor,et al.  Modification of seed oil content and acyl composition in the brassicaceae by expression of a yeast sn-2 acyltransferase gene. , 1997, The Plant cell.

[34]  D. Murphy,et al.  Intra- and extracellular lipid composition and associated gene expression patterns during pollen development in Brassica napus. , 1997, The Plant journal : for cell and molecular biology.

[35]  M. Montagu,et al.  Oleosin gene expression in Arabidopsis thaliana tapetum coincides with accumulation of lipids in plastids and cytoplasmic bodies , 1997 .

[36]  G. Wullems,et al.  Tapetum-specific genes : What role do they play in male gametophyte development? , 1996 .

[37]  H. Davies,et al.  Lysophosphatidic Acid Acyltransferase from Meadowfoam Mediates Insertion of Erucic Acid at the sn-2 Position of Triacylglycerol in Transgenic Rapeseed Oil , 1995, Plant physiology.

[38]  H. Davies,et al.  Cloning of a Coconut Endosperm cDNA Encoding a 1-Acyl-sn-Glycerol-3-Phosphate Acyltransferase That Accepts Medium-Chain-Length Substrates , 1995, Plant physiology.

[39]  M. Uemura,et al.  Cold Acclimation of Arabidopsis thaliana (Effect on Plasma Membrane Lipid Composition and Freeze-Induced Lesions) , 1995, Plant physiology.

[40]  J. Harwood,et al.  Solubilisation, partial purification and properties of acyl-CoA: glycerol-3-phosphate acyltransferase from avocado (Persea americana) fruit mesocarp. , 1995, Biochimica et biophysica acta.

[41]  P. Covello,et al.  Alteration of Seed Fatty Acid Composition by an Ethyl Methanesulfonate-Induced Mutation in Arabidopsis thaliana Affecting Diacylglycerol Acyltransferase Activity , 1995, Plant physiology.

[42]  P. Chourey,et al.  Epistatic interaction and functional compensation between the two tissue- and cell-specific sucrose synthase genes in maize. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[43]  C. Caelles,et al.  Arabidopsis thaliana contains two differentially expressed 3-hydroxy-3-methylglutaryl-CoA reductase genes, which encode microsomal forms of the enzyme. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[44]  H. Sul,et al.  Expression and identification of p90 as the murine mitochondrial glycerol-3-phosphate acyltransferase. , 1993, Biochemistry.

[45]  Kay Hofmann,et al.  Tmbase-A database of membrane spanning protein segments , 1993 .

[46]  J. Harwood,et al.  Glycerol 3-phosphate acylation by microsomal fractions from avocado mesocarp. , 1992, Biochemical Society transactions.

[47]  G von Heijne,et al.  Membrane protein structure prediction. Hydrophobicity analysis and the positive-inside rule. , 1992, Journal of molecular biology.

[48]  M. Bafor,et al.  Properties of the glycerol acylating enzymes in microsomal preparations from the developing seeds of safflower (Carthamus tinctorius) and turnip rape (Brassica campestris) and their ability to assemble cocoa-butter type fats , 1990 .

[49]  M. Frentzen Comparison of certain properties of membrane bound and solubilized acyltransferase activities of plant microsomes , 1990 .

[50]  R. Bell,et al.  Phospholipid dependence of homogeneous, reconstituted sn-glycerol-3-phosphate acyltransferase of Escherichia coli. , 1989, The Journal of biological chemistry.

[51]  A. Huang,et al.  Acyl coenzyme a preference of the glycerol phosphate pathway in the microsomes from the maturing seeds of palm, maize, and rapeseed. , 1988, Plant physiology.

[52]  R. Peterson,et al.  A comparative light and electron microscopic study of microspore and tapetal development in male fertile and cytoplasmic male sterile oilseed rape (Brassica napus) , 1986 .

[53]  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.

[54]  C. A. Robertson,et al.  Cocoa butter biosynthesis. Purification and characterization of a soluble sn-glycerol-3-phosphate acyltransferase from cocoa seeds. , 1986, The Journal of biological chemistry.

[55]  S. Stymne,et al.  The acylation of sn-glycerol 3-phosphate and the metabolism of phosphatidate in microsomal preparations from the developing cotyledons of safflower (Carthamus tinctorius L.) seed. , 1985, The Biochemical journal.

[56]  K. Ichihara,et al.  sn-Glycerol-3-phosphate acyltransferase in a particulate fraction from maturing safflower seeds. , 1984, Archives of biochemistry and biophysics.

[57]  E. Heinz,et al.  Similarities and differences in lipid metabolism of chloroplasts isolated from 18:3 and 16:3 plants. , 1983, Plant physiology.

[58]  P. Roughan,et al.  CELLULAR ORGANIZATION OF GLYCEROLIPID METABOLISM , 1982 .

[59]  Y. Fujiki,et al.  Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum , 1982, The Journal of cell biology.

[60]  H. Horner,et al.  A COMPARATIVE LIGHT‐ AND ELECTRON‐MICROSCOPIC STUDY OF MICROSPOROGENESIS IN MALE‐FERTILE AND CYTOPLASMIC MALE‐STERILE SUNFLOWER (HELIANTHUS ANNUUS) , 1977 .

[61]  H. E. Warmke,et al.  Cytoplasmic Male Sterility in Sorghum , 1972 .

[62]  M. Zenkteler MICROSPOROGENESIS AND TAPETAL DEVELOPMENT IN NORMAL AND MALE‐STERILE CARROTS (DAUCUS CAROTA) , 1962 .

[63]  W. J. Dyer,et al.  A rapid method of total lipid extraction and purification. , 1959, Canadian journal of biochemistry and physiology.