title: New nodes and edges in the glucosinolate molecular network revealed by proteomics and metabolomics of Arabidopsis myb28/29 and cyp79B2/B3 glucosinolate mutants.
abstract: Glucosinolates present in Brassicales are important for human health and plant defense against insects and pathogens. Here we investigate the proteomes and metabolomes of Arabidopsis myb28/29 and cyp79B2/B3 mutants deficient in aliphatic glucosinolates and indolic glucosinolates, respectively. Quantitative proteomics of the myb28/29 and cyp79B2/B3 mutants led to the identification of 2785 proteins, of which 142 proteins showed significant changes in the two mutants compared to wild type (WT). By mapping the differential proteins using STRING, we detected 59 new edges in the glucosinolate metabolic network. These connections can be classified as primary with direct roles in glucosinolate metabolism, secondary related to plant stress responses, and tertiary involved in other biological processes. Gene Ontology analysis of the differential proteins showed high level of enrichment in the nodes belonging to metabolic process including glucosinolate biosynthesis and response to stimulus. Using metabolomics, we quantified 292 metabolites covering a broad spectrum of metabolic pathways, and 89 exhibited differential accumulation patterns between the mutants and WT. The changing metabolites (e.g., γ-glutamyl amino acids, auxins and glucosinolate hydrolysis products) complement our proteomics findings. This study contributes toward engineering and breeding of glucosinolate profiles in plants in efforts to improve human health, crop quality and productivity.
  
  
  CONCLUSIONS
  Glucosinolates in Brassicales constitute an important group of natural metabolites important for plant defense and human health. Its biosynthetic pathways and transcriptional regulation have been well-studied. Using Arabidopsis mutants of important genes in glucosinolate biosynthesis, quantitative proteomics and metabolomics led to identification of many proteins and metabolites that are potentially related to glucosinolate metabolism. This study provides a comprehensive insight into the molecular networks of glucosinolate metabolism, and will facilitate efforts toward engineering and breeding of glucosinolate profiles for enhanced crop defense, and nutritional value.
authors.affiliation: Department of Biology, University of Florida, Gainesville, FL 32610, USA; Genetics Institute, University of Florida, Gainesville, FL 32610, USA; Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt.
authors.forename: Islam
authors.initials: I
authors.lastname: Mostafa
authors.affiliation: Department of Biology, University of Florida, Gainesville, FL 32610, USA; Genetics Institute, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Ning
authors.initials: N
authors.lastname: Zhu
authors.affiliation: Department of Biology, University of Florida, Gainesville, FL 32610, USA; Genetics Institute, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Mi-Jeong
authors.initials: MJ
authors.lastname: Yoo
authors.affiliation: Department of Biology, University of Florida, Gainesville, FL 32610, USA; Genetics Institute, University of Florida, Gainesville, FL 32610, USA; Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Balmant
authors.affiliation: Department of Biology, University of Florida, Gainesville, FL 32610, USA; Genetics Institute, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Biswapriya B
authors.initials: BB
authors.lastname: Misra
authors.affiliation: Thermo Fisher Scientific, West Palm Beach, FL 33407, USA.
authors.forename: Craig
authors.initials: C
authors.lastname: Dufresne
authors.affiliation: Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt.
authors.forename: Maged
authors.initials: M
authors.lastname: Abou-Hashem
authors.affiliation: Department of Biology, University of Florida, Gainesville, FL 32610, USA; Genetics Institute, University of Florida, Gainesville, FL 32610, USA; Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, FL 32610, USA; Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, FL 32610, USA. Electronic address: schen@ufl.edu.
authors.forename: Sixue
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt.
authors.forename: Maher
authors.initials: M
authors.lastname: El-Domiaty
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D005961:Glucosinolates / Q000096:biosynthesis / Q000235:genetics; D051548:Histone Acetyltransferases / Q000235:genetics*; D006801:Humans; D055432:Metabolomics; D009154:Mutation; D040901:Proteomics; D014157:Transcription Factors / Q000235:genetics*; D000099224:p300-CBP-Associated Factor
chemical_list: D029681:Arabidopsis Proteins; D003577:Cytochrome P-450 Enzyme System; D005961:Glucosinolates; D051548:Histone Acetyltransferases; D014157:Transcription Factors; C523552:Myb29 protein, Arabidopsis; C530432:cytochrome P-450 79B2 protein, Arabidopsis; C530433:cytochrome P-450 79B3, Arabidopsis; C474636:GCN5 protein, Arabidopsis; D000099224:p300-CBP-Associated Factor
keywords: Aliphatic and indolic glucosinolates; Arabidopsis; Genetic perturbation; Stress and defense; iTRAQ