Proteome changes in tomato lines transformed with phytoene synthase-1 in the sense and antisense orientations

Abstract

The commercial cultivation of genetically engineered (GE) crops in Europe has met with considerable consumer resistance, which has led to vigorous safety assessments including the measurement of substantial equivalence between the GE and parent lines. This necessitates the identification and quantification of significant changes to the metabolome and proteome in the GE crop. In this study, the quantitative proteomic analysis of tomato fruit from lines that have been transformed with the carotenogenic gene phytoene synthase-1 (Psy-1), in the sense and antisense orientations, in comparison with a non-transformed, parental line is described. Multidimensional protein identification technology (MudPIT), with tandem mass spectrometry, has been used to identify proteins, while quantification has been carried out with isobaric tags for relative and absolute quantification (iTRAQ). Fruit from the GE plants showed significant alterations to their proteomes compared with the parental line, especially those from the Psy-1 sense transformants. These results demonstrate that MudPIT and iTRAQ are suitable techniques for the verification of substantial equivalence of the proteome in GE crops.

A comprehensive review on the colorless carotenoids phytoene and phytofluene

“…Being a key regulatory enzyme in carotenoid biosynthesis, PSY has been important for the modification of the carotenoid levels in crops by genetic engineering [70][71][72][73][74]. Psy from daffodil (along with the bacterial phytoene desaturase CrtI) was introduced into rice for the development of Golden Rice [75].…”

Metabolite analysis of Mycobacterium species under aerobic and hypoxic conditions reveals common metabolic traits

“…For the identification of proteins of the secreted layer, a previously published protocol (Robertson et al, 2012;Mora et al, 2013), which included precipitation of proteins and separation by SDS-PAGE followed by in-gel digestion of the four major protein bands and analysis with nano-LC/MS/MS, was followed. The proteins were identified through the detected peptides as previously published (Nogueira et al, 2013) with SwissProt protein database for taxonomy oM.…”

Subchromoplast Sequestration of Carotenoids Affects Regulatory Mechanisms in Tomato Lines Expressing Different Carotenoid Gene Combinations

“…Extraction, separation, immunodetection, and liquid chromatography-mass spectrometry (LC-MS) analysis of proteins were performed following the protocols of Robertson et al (2012) and Mora et al (2013).…”