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 |
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Creator |
Sinha, Ragini
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Subject |
Drug Development/Diagnostics & Biotechnology
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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. |
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Date |
2014
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Type |
Thesis
NonPeerReviewed |
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Format |
application/pdf
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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. |
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Relation |
http://www.eprints.iicb.res.in/2005/
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