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Generation And Proteomic Profiling Of Transgenic Mentha Overexpressing Glutathione Biosynthesis Pathway Gene/s And Comparative Proteomic Profiling of Arabidopsis Under Altered Glutathione Conditions

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Title Generation And Proteomic Profiling Of Transgenic
Mentha Overexpressing Glutathione Biosynthesis
Pathway Gene/s And Comparative Proteomic
Profiling of Arabidopsis Under
Altered Glutathione Conditions
 
Creator Sinha, Ragini
 
Subject Drug Development/Diagnostics & Biotechnology
 
Description Generation And Proteomic Profiling Of Transgenic Mentha Overexpressing
Glutathione Biosynthesis Pathway Gene/s And Comparative Proteomic Profiling of
Arabidopsis Under Altered Glutathione Conditions
Glutathione (GSH) is a nearly ubiquitous, most abundant non-protein tripeptide thiol
compound found in both prokaryotes and eukaryotes. Recent years have witnessed
considerable progress in understanding GSH synthesis, degradation and transport,
particularly in relation to cellular redox homeostasis and related signalling under optimal and
stress conditions. The role of GSH in plant defence has long been known in addition to its
substantial role in stress tolerance and antioxidant signalling. Function of GSH has been
reported vividly in various types of biotic and abiotic stress tolerance in plants. In the present
study the role of GSH in plant defence has been further explored using two systems. First
deals with the enhancement of GSH content in target plant viz. Mentha arvensis using
transgenic approach. A constitutive overexpression of g-ECS (rate limiting enzyme of GSH
biosynthetic pathway) is performed through Agrobacterium tumefaciens mediated
transformation. The transgenic plants showed an enhancement of total GSH content and
displayed stress tolerant ability against metal toxicity and pathogen stress. A comparative
protein profile between wild type and transgenic plants under non-infected and Alternaria
alternata infected conditions showed differential accumulation of protein species related to
stress and defense, redox regulation, transcription factors and energy and metabolism. Protein
species related to signalling and gene regulation of defence related proteins showed
differential accumulation specifically in infected transgenic plants. Thus it can be postulated
that transgenic plants with enhanced GSH content could be a promising approach for raising
stress tolerant plants. Owing to the limited database information with Mentha, the study was
further continued with Arabidopsis thaliana. Hence in the second part of the study, the role of
GSH was explored using in-vivo feeding of GSH to Col 0 and pad2-1, an A. thaliana mutant
for g-ECS allele with Col 0 as control. Proteins related to stress, defence signalling, redox
related proteins, protein synthesis and degradation, DNA repair proteins were identified as
differentially regulated through comparative proteomics. Likewise genes related to biotic
stress, abiotic stress, redox, signalling, RNA regulation, protein modification, development,
cell organisation, protein synthesis and degradation were identified as differentially expressed
through microarray analysis. The study demonstrated that GSH has a role in defence in target
plant which was further validated in model plant. The bio-informatics analyses of integrated
proteome and transcriptome was further performed to gain an in depth insight into
involvement of GSH in defense signalling network. The interplay between GSH and salicylic
acid (SA) is noted by differential expression of SA-mediated signalling related genes. In
addition, a new insight into the interaction between GSH and ethylene in defence signalling
pathway is observed. The effect of GSH on glucosinolate (GSL) biosynthetic pathways is
also observed which might be an important information linking the GSH to the GSL mediated
defence response. Furthermore, present dataset serves as a resource of altered genes and
protein under changed GSH content in A. thaliana and offers clues for future investigation on
GSH as signalling molecule.
 
Date 2014
 
Type Thesis
NonPeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/2005/1/Ragini_Sinha.pdf
Sinha, Ragini (2014) Generation And Proteomic Profiling Of Transgenic Mentha Overexpressing Glutathione Biosynthesis Pathway Gene/s And Comparative Proteomic Profiling of Arabidopsis Under Altered Glutathione Conditions. PhD thesis, University of Calcutta.
 
Relation http://www.eprints.iicb.res.in/2005/