Transcript and proteomic analysis of developing white lupin (Lupinus albus L.) roots

BackgroundWhite lupin (Lupinus albus L.) roots efficiently take up and accumulate (heavy) metals, adapt to phosphate deficiency by forming cluster roots, and secrete antimicrobial prenylated isoflavones during development. Genomic and proteomic approaches were applied to identify candidate genes and proteins involved in antimicrobial defense and (heavy) metal uptake and translocation.ResultsA cDNA library was constructed from roots of white lupin seedlings. Eight thousand clones were randomly sequenced and assembled into 2,455 unigenes, which were annotated based on homologous matches in the NCBInr protein database. A reference map of developing white lupin root proteins was established through 2-D gel electrophoresis and peptide mass fingerprinting. High quality peptide mass spectra were obtained for 170 proteins. Microsomal membrane proteins were separated by 1-D gel electrophoresis and identified by LC-MS/MS. A total of 74 proteins were putatively identified by the peptide mass fingerprinting and the LC-MS/MS methods. Genomic and proteomic analyses identified candidate genes and proteins encoding metal binding and/or transport proteins, transcription factors, ABC transporters and phenylpropanoid biosynthetic enzymes.ConclusionThe combined EST and protein datasets will facilitate the understanding of white lupin's response to biotic and abiotic stresses and its utility for phytoremediation. The root ESTs provided 82 perfect simple sequence repeat (SSR) markers with potential utility in breeding white lupin for enhanced agronomic traits.

[1]  Jungwon Yoon,et al.  The Arabidopsis Information Resource (TAIR): a model organism database providing a centralized, curated gateway to Arabidopsis biology, research materials and community , 2003, Nucleic Acids Res..

[2]  P. A. Rea Plant ATP-binding cassette transporters. , 2007, Annual review of plant biology.

[3]  M. Sorrells,et al.  Data mining for simple sequence repeats in expressed sequence tags from barley, maize, rice, sorghum and wheat , 2002, Plant Molecular Biology.

[4]  F. Dédaldéchamp,et al.  Direct evidence for ribonucleolytic activity of a PR-10-like protein from white lupin roots , 2000, Plant Molecular Biology.

[5]  David Haussler,et al.  The UCSC genome browser database: update 2007 , 2006, Nucleic Acids Res..

[6]  S. Rhee,et al.  Functional Annotation of the Arabidopsis Genome Using Controlled Vocabularies1 , 2004, Plant Physiology.

[7]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[8]  H. Ling,et al.  Transgenic expression of DwMYB2 impairs iron transport from root to shoot in Arabidopsis thaliana , 2006, Cell Research.

[9]  B Herbert,et al.  Advances in protein solubilisation for two‐dimensional electrophoresis , 1999, Electrophoresis.

[10]  J. Vivanco,et al.  Altered Profile of Secondary Metabolites in the Root Exudates of Arabidopsis ATP-Binding Cassette Transporter Mutants1[C][W][OA] , 2007, Plant Physiology.

[11]  Liangjiang Wang,et al.  Mapping the Proteome of Barrel Medic (Medicago truncatula)1,212 , 2003, Plant Physiology.

[12]  M. Ganal,et al.  The tomato fer gene encoding a bHLH protein controls iron-uptake responses in roots , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[13]  D. Barron,et al.  Isoprenylated flavonoids—a survey , 1996 .

[14]  C. Vance,et al.  Overexpression of malate dehydrogenase in transgenic alfalfa enhances organic acid synthesis and confers tolerance to aluminum. , 2001, Plant physiology.

[15]  Jane M. F. Johnson,et al.  Acclimation of white lupin to phosphorus deficiency involves enhanced expression of genes related to organic acid metabolism , 2004, Plant and Soil.

[16]  Hartmut K. Lichtenthaler,et al.  THE 1-DEOXY-D-XYLULOSE-5-PHOSPHATE PATHWAY OF ISOPRENOID BIOSYNTHESIS IN PLANTS. , 1999, Annual review of plant physiology and plant molecular biology.

[17]  J. Harborne,et al.  The isopentenyl isoflavone luteone as a pre-infectional antifungal agent in the genus Lupinus , 1976 .

[18]  Patrick Xuechun Zhao,et al.  PLAN: a web platform for automating high-throughput BLAST searches and for managing and mining results , 2007, BMC Bioinformatics.

[19]  P. Ximénez-Embún,et al.  Uptake of Heavy Metals by Lupin Plants in Artificially Contaminated Sand: Preliminary Results , 2002 .

[20]  V. Page,et al.  Heavy metals in white lupin: uptake, root-to-shoot transfer and redistribution within the plant. , 2006, The New phytologist.

[21]  S Rozen,et al.  Primer3 on the WWW for general users and for biologist programmers. , 2000, Methods in molecular biology.

[22]  M. Grusak,et al.  Identification and Characterization of Several New Members of the ZIP Family of Metal Ion Transporters in Medicago Truncatula , 2004, Plant Molecular Biology.

[23]  Karen Schlauch,et al.  Cytosolic Ascorbate Peroxidase 1 Is a Central Component of the Reactive Oxygen Gene Network of Arabidopsisw⃞ , 2005, The Plant Cell Online.

[24]  V. Walbot,et al.  A Multidrug Resistance–Associated Protein Involved in Anthocyanin Transport in Zea mays , 2004, The Plant Cell Online.

[25]  D. Ro,et al.  Functional characterization and subcellular localization of poplar (Populus trichocarpa x Populus deltoides) cinnamate 4-hydroxylase. , 2001, Plant physiology.

[26]  Kanako Sasaki,et al.  Cloning and Characterization of Naringenin 8-Prenyltransferase, a Flavonoid-Specific Prenyltransferase of Sophora flavescens1[W] , 2008, Plant Physiology.

[27]  C. Vance,et al.  Transgenic proteoid roots of white lupin: a vehicle for characterizing and silencing root genes involved in adaptation to P stress. , 2005, The Plant journal : for cell and molecular biology.

[28]  Ting Wang,et al.  The UCSC Genome Browser Database: update 2009 , 2008, Nucleic Acids Res..

[29]  Zhentian Lei,et al.  A Two-dimensional Electrophoresis Proteomic Reference Map and Systematic Identification of 1367 Proteins from a Cell Suspension Culture of the Model Legume Medicago truncatula*S , 2005, Molecular & Cellular Proteomics.

[30]  C. Vance,et al.  Nylon Filter Arrays Reveal Differential Gene Expression in Proteoid Roots of White Lupin in Response to Phosphorus Deficiency , 2003, Plant Physiology.

[31]  M. M. Lucas,et al.  An unusual infection mechanism and nodule morphogenesis in white lupin (Lupinus albus). , 2004, The New phytologist.

[32]  C. Vance,et al.  A phosphate transporter with enhanced expression in proteoid roots of white lupin (Lupinus albus L.) , 2001, Plant and Soil.

[33]  H. Phan,et al.  The First Genetic and Comparative Map of White Lupin (Lupinus albus L.): Identification of QTLs for Anthracnose Resistance and Flowering Time, and a Locus for Alkaloid Content , 2007, DNA research : an international journal for rapid publication of reports on genes and genomes.

[34]  P. A. Rea,et al.  Alternate Energy-Dependent Pathways for the Vacuolar Uptake of Glucose and Glutathione Conjugates1 , 2002, Plant Physiology.

[35]  Andreas Graner,et al.  Genic microsatellite markers in plants: features and applications. , 2005, Trends in biotechnology.

[36]  M. Erbaş,et al.  Some chemical properties of white lupin seeds (Lupinus albus L.) , 2005 .

[37]  Thomas V. O'Halloran,et al.  Metallochaperones, an Intracellular Shuttle Service for Metal Ions* , 2000, The Journal of Biological Chemistry.

[38]  H. Ling,et al.  AtbHLH29 of Arabidopsis thaliana is a functional ortholog of tomato FER involved in controlling iron acquisition in strategy I plants , 2005, Cell Research.

[39]  P. Reay,et al.  Mineral-element composition ofLupinus albus andLupinus angustifolius in relation to manganese accumulation , 1981, Plant and Soil.

[40]  A. Sugiyama,et al.  Involvement of a Soybean ATP-Binding Cassette-Type Transporter in the Secretion of Genistein, a Signal Flavonoid in Legume-Rhizobium Symbiosis1 , 2007, Plant Physiology.

[41]  R. Dixon Natural products and plant disease resistance , 2001, Nature.

[42]  Miguel Lara,et al.  Sequencing and Analysis of Common Bean ESTs. Building a Foundation for Functional Genomics1[w] , 2005, Plant Physiology.

[43]  K. Niyogi,et al.  Two P-Type ATPases Are Required for Copper Delivery in Arabidopsis thaliana Chloroplasts , 2005, The Plant Cell Online.

[44]  A. Kohler,et al.  The poplar root transcriptome: analysis of 7000 expressed sequence tags , 2003, FEBS letters.

[45]  M. Ashburner,et al.  Gene Ontology: tool for the unification of biology , 2000, Nature Genetics.