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One hundred twenty proteins differing in kinetics of appearance were subjected to matrix-assisted laser desorption ionization time of flight mass spectrometry. These analyses provided peptide mass fingerprint data that identified 84 of them. Some of these proteins had previously been shown to accumulate during seed development in legumes (e.g. legumins, vicilins, convicilins, and lipoxygenases), confirming the validity of M. truncatula as a model for analysis of legume seed filling. The study also revealed proteins presumably involved in cell division during embryogenesis (β-tubulin and annexin). Their abundance decreased before the accumulation of the major storage protein families, which itself occurs in a specific temporal order: vicilins (14 d after pollination [DAP]), legumins (16 DAP), and convicilins (18 DAP). Furthermore, the study showed an accumulation of enzymes of carbon metabolism (e.g. sucrose synthase, starch synthase) and of proteins involved in embryonic photosynthesis (e.g. chlorophyll a\u002Fb binding), which may play a role in providing cofactors for protein\u002Flipid synthesis or for CO2 refixation during seed filling. Correlated with the reserve deposition phase was the accumulation of proteins associated with cell expansion (actin 7 and reversibly glycosylated polypeptide) and of components of the precursor accumulating vesicles, which give rise to a trypsin inhibitor on maturation. Finally, we revealed a differential accumulation of enzymes involved in methionine metabolism (S-adenosyl-methionine synthetase and S-adenosylhomo-cysteine hydrolase) and propose a role for these enzymes in the transition from a highly active to a quiescent state during seed development.\u003C\u002Fjats:p>",{"EN":468},"Proteomics of\u003Ci>Medicago truncatula\u003C\u002Fi>Seed Development Establishes the Time Frame of Diverse Metabolic Processes Related to Reserve Accumulation",{"VOID":470},"12972662",{"VOID":472},"10.1104\u002Fpp.103.025254",[174],"https:\u002F\u002Facademic.oup.com\u002Fplphys\u002Farticle\u002F133\u002F2\u002F664\u002F6111177",[476,495,512,529,546],{"id":477,"sortIndex":36,"researcher":18,"roles":478,"affiliations":479,"properties":490},"2338b064-fc70-40b9-a725-847896ffd9bd",[],[480],{"id":481,"sortIndex":19,"affiliation":482,"properties":18},"0b7d81db-d80c-4813-b2c7-e3f3fd12d2f7",{"id":483,"createTime":484,"updateTime":484,"relativeEntities":485,"slug":486,"properties":487,"entityType":191,"verifyStatus":17,"verifyTime":18,"verifyNote":18,"syncStatus":17,"languages":18,"translateLanguages":18,"viewCount":19},"e28e109a-e16d-481d-960a-a245db6c5b53","2024-10-08T23:50:59.452+00:00",[],"Unite-de-Ge-ne-tique-et-Ecophysiologie-des-Le-gumineuses-Institut-National-de-la-Recherche-Agronomique-INRA-Dijon-Domaine-d-Epoisses-21110-Bretenie-res-France-K-G-C-L-S-R-D-T-J-B-and-Flanders-Interuniversity-Institute-for-Biotechnology-and-Department-of-Biochemistry-Gent-University-Gent-Belgium-J-V-",{"title":488},{"EN":489},"Unité de Génétique et Ecophysiologie des Légumineuses, Institut National de la Recherche Agronomique (INRA)-Dijon, Domaine d'Epoisses, 21110 Bretenières, France (K.G., C.L.S., R.D.T., J.B); and Flanders Interuniversity Institute for Biotechnology and Department of Biochemistry, Gent University, Gent, Belgium (J.V.)",{"openalex":491,"title":493},{"VOID":492},"A5049291299",{"EN":494},"Joël Vandekerckhove",{"id":496,"sortIndex":39,"researcher":18,"roles":497,"affiliations":498,"properties":505},"a4ecaaa6-b554-4723-b451-a69c378ca805",[],[499],{"id":500,"sortIndex":19,"affiliation":501,"properties":18},"b13bc9f0-2b0c-413b-b689-d24f891e1083",{"id":483,"createTime":484,"updateTime":484,"relativeEntities":502,"slug":486,"properties":503,"entityType":191,"verifyStatus":17,"verifyTime":18,"verifyNote":18,"syncStatus":17,"languages":18,"translateLanguages":18,"viewCount":19},[],{"title":504},{"EN":489},{"openalex":506,"orcid":508,"title":510},{"VOID":507},"A5044041414",{"VOID":509},"https:\u002F\u002Forcid.org\u002F0000-0002-3204-3537",{"EN":511},"Christine Le Signor",{"id":513,"sortIndex":19,"researcher":18,"roles":514,"affiliations":515,"properties":522},"d9fe0b32-b474-497e-b9f8-a802e501e69b",[],[516],{"id":517,"sortIndex":19,"affiliation":518,"properties":18},"3a759855-a38c-423e-9753-26d7d4114f8b",{"id":483,"createTime":484,"updateTime":484,"relativeEntities":519,"slug":486,"properties":520,"entityType":191,"verifyStatus":17,"verifyTime":18,"verifyNote":18,"syncStatus":17,"languages":18,"translateLanguages":18,"viewCount":19},[],{"title":521},{"EN":489},{"openalex":523,"orcid":525,"title":527},{"VOID":524},"A5001074812",{"VOID":526},"https:\u002F\u002Forcid.org\u002F0000-0002-2343-1941",{"EN":528},"Karine Gallardo",{"id":530,"sortIndex":41,"researcher":18,"roles":531,"affiliations":532,"properties":539},"a1c4cfb4-6bc3-4bc7-86b8-164fbe2f2305",[],[533],{"id":534,"sortIndex":19,"affiliation":535,"properties":18},"7e90fd8d-81c1-4daa-9e9c-037c3ab01662",{"id":483,"createTime":484,"updateTime":484,"relativeEntities":536,"slug":486,"properties":537,"entityType":191,"verifyStatus":17,"verifyTime":18,"verifyNote":18,"syncStatus":17,"languages":18,"translateLanguages":18,"viewCount":19},[],{"title":538},{"EN":489},{"openalex":540,"orcid":542,"title":544},{"VOID":541},"A5038285270",{"VOID":543},"https:\u002F\u002Forcid.org\u002F0000-0001-6212-6774",{"EN":545},"Judith Burstin",{"id":547,"sortIndex":40,"researcher":18,"roles":548,"affiliations":549,"properties":556},"4e2ae936-5f39-4b6d-8c83-bd92493f53f0",[],[550],{"id":551,"sortIndex":19,"affiliation":552,"properties":18},"fed76c8b-b2c2-471c-bb00-7693713f67ae",{"id":483,"createTime":484,"updateTime":484,"relativeEntities":553,"slug":486,"properties":554,"entityType":191,"verifyStatus":17,"verifyTime":18,"verifyNote":18,"syncStatus":17,"languages":18,"translateLanguages":18,"viewCount":19},[],{"title":555},{"EN":489},{"openalex":557,"title":559},{"VOID":558},"A5105989947",{"EN":560},"Richard D. 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In a previous study, we have shown, using heterologous expression in the yeast Saccharomyces cerevisiae, that in the presence of toxic metals, AtHMA3 was able to phenotypically complement the cadmium\u002Flead (Cd\u002FPb)-hypersensitive strain ycf1 but not the zinc (Zn)-hypersensitive strain zrc1. In this study, we demonstrate that AtHMA3 in planta is located in the vacuolar membrane, with a high expression level in guard cells, hydathodes, vascular tissues, and the root apex. Confocal imaging in the presence of the Zn\u002FCd fluorescent probe BTC-5N revealed that AtHMA3 participates in the vacuolar storage of Cd. A T-DNA insertional mutant was found more sensitive to Zn and Cd. Conversely, ectopic overexpression of AtHMA3 improved plant tolerance to Cd, cobalt, Pb, and Zn; Cd accumulation increased by about 2- to 3-fold in plants overexpressing AtHMA3 compared with wild-type plants. 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              \u003Cjats:p>Lipid analysis of rosette leaves from Arabidopsis has revealed an accumulation of triacylglycerol (TAG) with advancing leaf senescence coincident with an increase in the abundance and size of plastoglobuli. The terminal step in the biosynthesis of TAG in Arabidopsis is catalyzed by diacylglycerol acyltransferase 1 (DGAT1; EC 2.3.1.20). When gel blots of RNA isolated from rosette leaves at various stages of development were probed with the Arabidopsis expressed sequence tag clone, E6B2T7, which has been annotated as DGAT1, a steep increase in DGAT1 transcript levels was evident in the senescing leaves coincident with the accumulation of TAG. The increase in DGAT1 transcript correlated temporally with enhanced levels of DGAT1 protein detected immunologically. Two lines of evidence indicated that the TAG of senescing leaves is synthesized in chloroplasts and sequesters fatty acids released from the catabolism of thylakoid galactolipids. First, TAG isolated from senescing leaves proved to be enriched in hexadecatrienoic acid (16:3) and linolenic acid (18:3), which are normally present in thylakoid galactolipids. Second, DGAT1 protein in senescing leaves was found to be associated with chloroplast membranes. These findings collectively indicate that diacylglycerol acyltransferase plays a role in senescence by sequestering fatty acids de-esterified from galactolipids into TAG. 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Globules of the spinach-beet chloroplast., Biochim Biophys Acta, 78, 163, 10.1016\u002F0006-3002(63)91621-4",{"doi":1701},"10.1016\u002F0006-3002(63)91621-4",{"id":18,"text":1703,"url":18,"identifiers":1704},"Bao, 1999, Supply of fatty acid is one limiting factor in the accumulation of triacylglycerol in developing embryos., Plant Physiol, 120, 1057, 10.1104\u002Fpp.120.4.1057",{"doi":1705},"10.1104\u002Fpp.120.4.1057",{"id":18,"text":1707,"url":18,"identifiers":1708},"Bertrams, 1976, Experiments on enzymatic acylation of sn-glycerol 3-phosphate with enzyme preparations from pea and spinach leaves., Planta, 132, 161, 10.1007\u002FBF00388898",{"doi":1709},"10.1007\u002FBF00388898",{"id":18,"text":1711,"url":18,"identifiers":1712},"Bligh, 1959, A rapid method of total lipid extraction and purification., Can J Biochem Physiol, 37, 911, 10.1139\u002Fo59-099",{"doi":1713},"10.1139\u002Fo59-099",{"id":18,"text":1715,"url":18,"identifiers":1716},"Browse, 1986, Fluxes through the prokaryotic and eukaryotic pathways of lipid synthesis in the ‘16:3′ plant Arabidopsis thaliana., Biochem J, 235, 25, 10.1042\u002Fbj2350025",{"doi":1717},"10.1042\u002Fbj2350025",{"id":18,"text":1719,"url":18,"identifiers":1720},"Colas des Frans-Small, 1993, Identification of a major soluble protein in mitochondria from nonphotosynthetic tissues as NAD-dependent formate dehydrogenase., Plant Physiol, 102, 1171, 10.1104\u002Fpp.102.4.1171",{"doi":1721},"10.1104\u002Fpp.102.4.1171",{"id":18,"text":1723,"url":18,"identifiers":1724},"Collawn, 1989, Production of antipeptide antibodies., Current Protocols in Molecular Biology on CD., 11.15.1",{},{"id":18,"text":1726,"url":18,"identifiers":1727},"Davis, 1986, Basic Methods in Molecular Biology., 130, 10.1016\u002FB978-0-444-01082-7.50040-0",{"doi":1728},"10.1016\u002FB978-0-444-01082-7.50040-0",{"id":18,"text":1730,"url":18,"identifiers":1731},"De Bellis, 1990, Localization of glyoxylate-cycle marker enzymes in peroxisomes of senescent leaves and green cotyledons., Planta, 180, 435, 10.1007\u002FBF01160401",{"doi":1732},"10.1007\u002FBF01160401",{"id":18,"text":1734,"url":18,"identifiers":1735},"Drenckhahn, 1993, Production of polyclonal antibodies against proteins and peptides., Methods in Cell Biology, 37, 7, 10.1016\u002FS0091-679X(08)60242-3",{"doi":1736},"10.1016\u002FS0091-679X(08)60242-3",{"id":18,"text":1738,"url":18,"identifiers":1739},"El-Hafid, 1989, Enzymatic breakdown of polar lipids in cotton leaves under water stress: I. 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Physiol, 126, 861, 10.1104\u002Fpp.126.2.861",{"doi":1790},"10.1104\u002Fpp.126.2.861",{"id":18,"text":1792,"url":18,"identifiers":1793},"Katavic, 1995, Alteration of seed fatty acid composition by an ethyl methanesulfonate-induced mutation in Arabidopsis thaliana affecting diacylglycerol acyltransferase activity., Plant Physiol, 108, 399, 10.1104\u002Fpp.108.1.399",{"doi":1794},"10.1104\u002Fpp.108.1.399",{"id":18,"text":1796,"url":18,"identifiers":1797},"Kessler, 1999, Identification of proteins associated with plastoglobules isolated from pea (Pisum sativum L.) chloroplasts., Planta, 208, 107, 10.1007\u002Fs004250050540",{"doi":1798},"10.1007\u002Fs004250050540",{"id":18,"text":1800,"url":18,"identifiers":1801},"Kim, 2001, Thermal inactivation kinetics and application of phospho-and galactolipid-degrading enzyme for evaluation of quality changes in frozen vegetables., J Agric Food Chem, 49, 2241, 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gene family., J Biol Chem, 276, 38862, 10.1074\u002Fjbc.M106168200",{"doi":1818},"10.1074\u002Fjbc.M106168200",{"id":18,"text":1820,"url":18,"identifiers":1821},"Lee, 2000, Membrane lipids: it’s only a phase., Curr Biol, 10, R377, 10.1016\u002FS0960-9822(00)00477-2",{"doi":1822},"10.1016\u002FS0960-9822(00)00477-2",{"id":18,"text":1824,"url":18,"identifiers":1825},"Lichtenthaler, 1969, The plastoglobuli of spinach: their size, isolation and lipoquinone composition., Protoplasma, 68, 65, 10.1007\u002FBF01247897",{"doi":1826},"10.1007\u002FBF01247897",{"id":18,"text":1828,"url":18,"identifiers":1829},"Lichtenthaler, 1970, the correlation between lipoquinone accumulation and plastoglobuli formation in the chloroplasts of Ficus elastica Roxb., Z Naturforsch, 25b, 619, 10.1515\u002Fznb-1970-0614",{"doi":1830},"10.1515\u002Fznb-1970-0614",{"id":18,"text":1832,"url":18,"identifiers":1833},"Martin, 1983, Properties of diacylglycerol acyltransferase from spinach leaves., Lipids, 18, 1, 10.1007\u002FBF02534682",{"doi":1834},"10.1007\u002FBF02534682",{"id":18,"text":1836,"url":18,"identifiers":1837},"Martin, 1984, Subcellular localization of TAG synthesis in spinach leaves., Lipids, 19, 117, 10.1007\u002FBF02534501",{"doi":1838},"10.1007\u002FBF02534501",{"id":18,"text":1840,"url":18,"identifiers":1841},"Matile, 1992, Chloroplast senescence., Crop Photosynthesis: Spatial and Temporal Determinants., 413, 10.1016\u002FB978-0-444-89608-7.50025-3",{"doi":1842},"10.1016\u002FB978-0-444-89608-7.50025-3",{"id":18,"text":1844,"url":18,"identifiers":1845},"Miquel, 1998, A new class of Arabidopsis mutants with reduced hexadecatrienoic acid fatty acid levels., Plant Physiol, 117, 923, 10.1104\u002Fpp.117.3.923",{"doi":1846},"10.1104\u002Fpp.117.3.923",{"id":18,"text":1848,"url":18,"identifiers":1849},"Miranda, 1993, Instantaneous blocking for immunoblots., Anal Biochem, 209, 376, 10.1006\u002Fabio.1993.1138",{"doi":1850},"10.1006\u002Fabio.1993.1138",{"id":18,"text":1852,"url":18,"identifiers":1853},"Morrison, 1964, Preparation of fatty acid methyl esters and dimethylacetals from lipids with boron fluoride-methanol., J Lipid Res, 5, 600, 10.1016\u002FS0022-2275(20)40190-7",{"doi":1854},"10.1016\u002FS0022-2275(20)40190-7",{"id":18,"text":1856,"url":18,"identifiers":1857},"Peoples, 1980, Nitrogen redistribution during grain growth in wheat (Triticum aestivum L.): II. Chloroplast senescence and the degradation of ribulose-1,5-bisphosphate carboxylase., Planta, 149, 241, 10.1007\u002FBF00384560",{"doi":1858},"10.1007\u002FBF00384560",{"id":18,"text":1860,"url":18,"identifiers":1861},"Post-Beittenmiller, 1996, Biochemistry and molecular biology of wax production in plants., Annu Rev Plant Physiol Plant Mol Biol, 47, 405, 10.1146\u002Fannurev.arplant.47.1.405",{"doi":1862},"10.1146\u002Fannurev.arplant.47.1.405",{"id":18,"text":1864,"url":18,"identifiers":1865},"Pozueta-Romero, 1997, A ubiquitous plant housekeeping gene, PAP, encodes a major protein component of bell pepper chromoplasts., Plant Physiol, 115, 1185, 10.1104\u002Fpp.115.3.1185",{"doi":1866},"10.1104\u002Fpp.115.3.1185",{"id":18,"text":1868,"url":18,"identifiers":1869},"Rey, 2000, Over-expression of a pepper plastid lipid-associated protein in tobacco leads to changes in plastid ultrastructure and plant development upon stress., Plant J, 21, 483, 10.1046\u002Fj.1365-313x.2000.00699.x",{"doi":1870},"10.1046\u002Fj.1365-313x.2000.00699.x",{"id":18,"text":1872,"url":18,"identifiers":1873},"Routaboul, 1999, The TAG1 locus of Arabidopsis encodes for a diacylglycerol acyltransferase., Plant Physiol Biochem, 37, 831, 10.1016\u002FS0981-9428(99)00115-1",{"doi":1874},"10.1016\u002FS0981-9428(99)00115-1",{"id":18,"text":1876,"url":18,"identifiers":1877},"Sakaki, 1990, Free fatty acids regulate two galactosyltransferases in chloroplast envelope membranes isolated from spinach leaves., Plant Physiol, 94, 781, 10.1104\u002Fpp.94.2.781",{"doi":1878},"10.1104\u002Fpp.94.2.781",{"id":18,"text":1880,"url":18,"identifiers":1881},"Sakaki, 1990, Pathway for the synthesis of triacylglycerols from monogalactosyldiacylglycerols in ozone-fumigated spinach leaves., Plant Physiol, 94, 773, 10.1104\u002Fpp.94.2.773",{"doi":1882},"10.1104\u002Fpp.94.2.773",{"id":18,"text":1884,"url":18,"identifiers":1885},"Sakaki, 1990, Conversion of monogalactosyldiacylglycerols to triacylglycerols in ozone-fumigated spinach leaves., Plant Physiol, 94, 766, 10.1104\u002Fpp.94.2.766",{"doi":1886},"10.1104\u002Fpp.94.2.766",{"id":18,"text":1888,"url":18,"identifiers":1889},"Sanchez, 1981, Synthesis of acyl CoAs by isolated spinach chloroplasts in relation to added CoA and ATP., Planta, 153, 519, 10.1007\u002FBF00385535",{"doi":1890},"10.1007\u002FBF00385535",{"id":18,"text":1892,"url":18,"identifiers":1893},"Shimakata, 1982, Fatty ace synthetase of Spinacia oleracea leaves., Plant Physiol, 69, 1257, 10.1104\u002Fpp.69.6.1257",{"doi":1894},"10.1104\u002Fpp.69.6.1257",{"id":18,"text":1896,"url":18,"identifiers":1897},"Siebertz, 1979, Characterization of lipids from chloroplast envelopes., Eur J Biochem, 101, 429, 10.1111\u002Fj.1432-1033.1979.tb19736.x",{"doi":1898},"10.1111\u002Fj.1432-1033.1979.tb19736.x",{"id":18,"text":1900,"url":18,"identifiers":1901},"Smart, 1994, Gene expression during leaf senescence., New Phytol, 126, 419, 10.1111\u002Fj.1469-8137.1994.tb04243.x",{"doi":1902},"10.1111\u002Fj.1469-8137.1994.tb04243.x",{"id":18,"text":1904,"url":18,"identifiers":1905},"Smith, 2000, Co-association of cytochrome f catabolites and plastid-lipid-associated protein with chloroplast lipid particles., Plant Physiol, 124, 211, 10.1104\u002Fpp.124.1.211",{"doi":1906},"10.1104\u002Fpp.124.1.211",{"id":18,"text":1908,"url":18,"identifiers":1909},"Sprey, 1966, Zur frage der beziehungen zwischen plastoglobuli und thylakoidgense in gertenkeimlingen., Z Naturforsch, 21b, 697, 10.1515\u002Fznb-1966-0717",{"doi":1910},"10.1515\u002Fznb-1966-0717",{"id":18,"text":1912,"url":18,"identifiers":1913},"Steinmüller, 1985, Composition and function of plastoglobuli: I. 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