title: Development and optimisation of a label-free quantitative proteomic procedure and its application in the assessment of genetically modified tomato fruit.
abstract: A key global challenge for plant biotechnology is addressing food security, whereby provision must be made to feed 9 billion people with nutritional feedstuffs by 2050. To achieve this step change in agricultural production new crop varieties are required that are tolerant to environmental stresses imposed by climate change, have better yields, are more nutritious and require less resource input. Genetic modification (GM) and marker-assisted screening will need to be fully utilised to deliver these new crop varieties. To evaluate these varieties both in terms of environmental and food safety and the rational design of traits a systems level characterisation is necessary. To link the transcriptome to the metabolome, quantitative proteomics is required. Routine quantitative proteomics is an important challenge. Gel-based densitometry and MS analysis after stable isotope labeling have been employed. In the present article, we describe the application of a label-free approach that can be used in combination with SDS-PAGE and reverse-phase chromatography to evaluate the changes in the proteome of new crop varieties. The workflow has been optimised for protein coverage, accuracy and robustness, then its application demonstrated using a GM tomato variety engineered to deliver nutrient dense fruit.
authors.affiliation: Centre for Systems and Synthetic Biology, School of Biological Sciences, Royal Holloway, University of London, Egham, UK.
authors.forename: Leticia
authors.initials: L
authors.lastname: Mora
authors.forename: Peter M
authors.initials: PM
authors.lastname: Bramley
authors.forename: Paul D
authors.initials: PD
authors.lastname: Fraser
mesh_terms: D000595:Amino Acid Sequence; D056148:Chromatography, Reverse-Phase; D004591:Electrophoresis, Polyacrylamide Gel; D005638:Fruit / Q000737:chemistry*; D016014:Linear Models; D018551:Solanum lycopersicum / Q000737:chemistry*; D008969:Molecular Sequence Data; D015999:Multivariate Analysis; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis / Q000737:chemistry; D030821:Plants, Genetically Modified / Q000737:chemistry*; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D010446:Peptide Fragments; D010940:Plant Proteins; D020543:Proteome
keywords: Food safety; Genetic modification; Label-free quantitation; Plant proteomics; Tomato