BMC Plant Biology · 2009 · 322 citations · 46 references
Life SciencesBiochemistryMolecular BiologyGenomicsProteomicsTranscriptomicsMass Spectrometry Data AnalysisMass Spectrometry-based ProteomicsPlant ProteomicsProtein Database SearchProtein IdentificationProtein Sequence AnalysisAbc TransportersCdna CloningCdna Library ConstructionHeavy Metal UptakeMetal-binding ProteinsPeptide Mass SpectrometryPeptide-mass FingerprintingPlant Protein Identification
BACKGROUND: White 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. RESULTS: A 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. CONCLUSION: The 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.
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