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Summer Research Fellowship Programme of India's Science Academies

Agrobacterium mediated transformation of Oryza sativa using dual-specificity phosphatase gene construct

Arpit Das

Department of Botany, Deshbandhu College, Block E, Kalkaji Main Road, Kalkaji, New Delhi.

Dr. Girdhar K. Pandey

Department of Plant Molecular Biology, Delhi University, South Campus, Block C, South Moti Bagh, New Delhi.

Abstract

The most evident adverse effects of the current climatic fluctuations and global warming can be observed in phenotypic alteration of crops and vegetation all around the globe. Overexploitation of resources has accelerated soil salinity, loss of edaphic microcosm and deprivation of nutrients, which has raised an alarm amongst the contemporary biologists for increased crop production and species sustenance. Since 1980s, one of the most commonly employed solutions is the creation of transgenic plants, which are stress tolerant high yielding varieties and have a crucial responsibility in tackling the stressful environmental conditions and evolve alongside the fulfillment of anthropogenic development. Biotechnological methodologies are exploited in order to comprehend and analyze various signaling pathways, which inform about the various biochemical processes that direct the growth and maintenance of plants. In this project, we are dealing with the role of dual-specificity phosphatases (DSPs) in Oryza sativa varieties. DSPs are a heterogeneous group of protein phosphatases that can dephosphorylate both phosphotyrosine and phosphoserine/ phosphothreonine residues within one substrate and are implicated as major modulators of critical signaling pathways. Phosphoglucan phosphatases are a distinct subset of DSPs, which have specific phosphatase domains for glucan dephosphorylation. In plants, these play a major role in the metabolism of stored carbohydrate, i.e., starch. Starch is synthesized through polymerization of glucose and is stored in chloroplasts in a granular form, which is hydrolyzed by the plant during unavailability of photosynthetic carbon dioxide. For efficient starch degradation, removal of phosphate groups is a necessary step. Our interest lies in increasing this starch hydrolysis by regulating the phosphate groups involved in the biochemical downstream processes. Agrobacterium sp. has been utilized for delivering a specific DNA molecule from its own genome into the host cell genome for making useful transgenics. Through Agrobacterium, we would be inserting our gene of interest (OsPP117-pGKP479) in rice varieties (IR-64 and PB1) using tissue culture techniques for creating transgenics for effective starch degradation.

Keywords: Transgenics, tissue culture, mature seed-derived calli, In planta transformation, regeneration, somatic embryogenesis.

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