000			03289nam a22002417a 4500
100			_aRekha, K.
245			_aGM rubber expressing tobacco osmotin gene: Towards the confined field trial.
260			_bPLACROSYM XXV: Building Smart & Resilient Farming and Systems Approaches for Prosperity in Plantation Crops Sector, 12-14 December 2023, ICAR-Indian Institute of Oil Palm Research, Pedavegi, Eluru, Andhra Pradesh, p. 51.
300			_bAbstract
520			_aImportance of genetic improvement in a perennial tree crop like rubber facing the vagaries of environment needs no emphasis in the context of climate change and global warming. Genetic transformation using Agrobacterium was attempted in Hevea for the integration of osmotin gene for imparting stress tolerance. Agrobacterium strain GV 2260 harbouring the plasmid osm/BinAR under the control of CaMV35S promoter containing kanamycin resistance as the selectable marker was employed for genetic transformation. Transgene detection through PCR analysis using gene specific primers could successfully amplify the 0.75 kb gene insert coding for osmotin protein and 0.7kb nptIIgene. The osmotin amplicon was cloned and sequenced and the sequence showed 100% similarity with the inserted gene, confirming the transgene integration. For further confirmation of gene integration, Southern analysis was performed using Osmotin gene amplified from the plasmid as probe and positive signals were obtained in the autoradiogram. For further confirmation of gene integration as well as its proper expression, Reverse Transcription PCR was performed using gene specific primers. Successful amplification of this gene confirmed that the Osmotin gene is integrated and expressed in Hevea. In order to assess the level of expression in the transgenics developed compared to control plants, relative gene expression studies were performed on the 5 transgenic plants and control through real time PCR. Tobacco Osmotin gene being a foreign gene with very low sequence homology to native osmotin gene of Hevea, its expression could not be detected in the control plants, whereas significant level of expression was observed in all the transgenics. These results confirmed that the Osmotin gene is stably integrated and expressed in the transgenic plants developed. Stress tolerance studies on transgenic leaf discs showed positive indications towards drought and cold as indicated by the less membrane damage under cold and PEG treatments. Application was submitted to RCGM for initiating the confined field trial with these plants in order to assess their field performance with respect to stress tolerance traits. The application was verified by RCGM and recommended to GEAC for further approval. After considering the environmental aspects, GEAC recommended the proposal. After proving the merit of the inserted traits, these plants will be cultivated under stressful environments in the non –traditional areas.
650			_aTobacco osmotin gene
650			_aAmplicon
650			_aGM rubber
650			_aField trials
700			_aThomas, K.U.
700			_aJayashree, R.
700			_aJayasree, P.K.
700			_aSumesh, K.V.
700			_aRatheena, K.T.
700			_aAthira, P.V.
700			_aKala, R.G.
942			_cJS
999			_c75176 _d75176