title: Combination of transcriptomic and proteomic approaches helps to unravel the protein composition of Chelidonium majus L. milky sap.
abstract: A novel annotated Chelidonium majus L. transcriptome database composed of 23,004 unique coding sequences allowed to significantly improve the sensitivity of proteomic C. majus assessments, which showed novel defense-related proteins characteristic to its latex. To date, the composition of Chelidonium majus L. milky sap and biosynthesis of its components are poorly characterized. We, therefore, performed de novo sequencing and assembly of C. majus transcriptome using Illumina technology. Approximately, 119 Mb of raw sequence data was obtained. Assembly resulted in 107,088 contigs, with N50 of 1913 bp and N90 of 450 bp. Among 34,965 unique coding sequences (CDS), 23,004 obtained CDS database served as a basis for further proteomic analyses. The database was then used for the identification of proteins from C. majus milky sap, and whole plant extracts analyzed using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) approach. Of about 334 different putative proteins were identified in C. majus milky sap and 1155 in C. majus whole plant extract. The quantitative comparative analysis confirmed that C. majus latex contains proteins connected with response to stress conditions and generation of precursor metabolites and energy. Notable proteins characteristic to latex include major latex protein (MLP, presumably belonging to Bet v1-like superfamily), polyphenol oxidase (PPO, which could be responsible for browning of the sap after exposure to air), and enzymes responsible for anthocyanidin, phenylpropanoid, and alkaloid biosynthesis.
authors.affiliation: Department of Molecular Virology, Institute of Experimental Biology, Faculty of Biology, Adam Mickiewicz University in Poznań, Umultowska 89, 61-614, Poznań, Poland. rnawrot@amu.edu.pl.
authors.forename: Robert
authors.initials: R
authors.lastname: Nawrot
authors.affiliation: Department of Molecular Virology, Institute of Experimental Biology, Faculty of Biology, Adam Mickiewicz University in Poznań, Umultowska 89, 61-614, Poznań, Poland.
authors.forename: Jakub
authors.initials: J
authors.lastname: Barylski
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, 06466, Gatersleben, Germany.|Sandoz GmbH, Biochemiestraße 10, 6250, Kundl, Austria.
authors.forename: Rico
authors.initials: R
authors.lastname: Lippmann
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, 06466, Gatersleben, Germany.
authors.forename: Lothar
authors.initials: L
authors.lastname: Altschmied
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, 06466, Gatersleben, Germany.
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Mock
mesh_terms: D000470:Alkaloids / Q000378:metabolism; D000975:Antioxidants / Q000378:metabolism; D053898:Biosynthetic Pathways / Q000235:genetics; D028281:Chelidonium / Q000235:genetics* / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D020869:Gene Expression Profiling / Q000379:methods*; D018506:Gene Expression Regulation, Plant; D063990:Gene Ontology; D007840:Latex / Q000378:metabolism*; D058977:Molecular Sequence Annotation; D010936:Plant Extracts / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D064210:Secondary Metabolism / Q000235:genetics; D017423:Sequence Analysis, RNA; D013312:Stress, Physiological / Q000235:genetics; D059467:Transcriptome / Q000235:genetics
chemical_list: D000470:Alkaloids; D000975:Antioxidants; D007840:Latex; D010936:Plant Extracts; D010940:Plant Proteins
keywords: Greater celandine; Latex; Laticifers; Major latex protein; Papaveraceae; Polyphenol oxidase; Sequencing