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OPTIMIZATION OF AGROBACTERIUM TUMEFACIENS MEDIATED TRANSFORMATION IN POPULUS DELTOIDES

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dc.contributor.author JOHN, EDWIN
dc.contributor.author MAQBOOL, ASMA
dc.contributor.author MALIK, KAUSER ABDULLA
dc.date.accessioned 2022-12-06T04:50:21Z
dc.date.available 2022-12-06T04:50:21Z
dc.date.issued 2014-07-11
dc.identifier.citation John, E., Maqbool, A., & Malik, K. A. (2014). Optimization of Agrobacterium tumefaciens mediated transformation in Populus deltoides. Pak. J. Bot, 46(3), 1079-1086. en_US
dc.identifier.issn 2070-3368
dc.identifier.uri http://142.54.178.187:9060/xmlui/handle/123456789/14707
dc.description.abstract The objective of the study was to develop an efficient protocol for Populus deltoides transformation through Agrobacterium tumefaciens LBA4404. Agrobacterium strain harboring binary plasmid pGA482 with Gus (uidA) gene under CamV35S promoter and Neomycin phosphotransferase (nptII) gene under Nos promoter was used for the transformation. Nodal, internodal and leaf explants from 4-5 months In vitro and fieldgrown plants were used for the transformation. Transformation was done under different conditions including, preculture time, optical density, acetosyringone concentration, infection time and co-cultivation time. Confirmation of transformation was done through GUS histochemical staining. Highest transformation efficiency was observed in one week precultured leaf explants from field grown source on preculture medium containing 200µM acetosyringone. Precultured explants from In vitro source also gave good results for transformation but the callus formation was found to be slow in leaf explant. Calli from the both sources did not show any transformation when infected with O.D A600nm range from 0.3-0.8. Node and internode though showed less transformation rate but the callogenesis was found to be highest in node and internode explants on CIM 1. Leaf explants from field source also gave high callus induction on CIM 5. A. tumefaciens O.D A600nm 0.3-0.5 was found to be effective. Infection time of 1-2 hour and co-cultivation time of 1day in dark were found to be optimum for the transformation. 200mg/l of timentin was found the best to control the overgrowth of Agrobacterium.100mg/l Kanamycin in growth medium was found to sufficient for selection for transformants. Selected transformants were confirmed through PCR for the presence of transgene. The present protocol for P. deltoides was found to be efficient for genetic ransformation and can be used to introduce novel traits in the P. deltoides. en_US
dc.language.iso en en_US
dc.publisher Karachi: Pakistan Journal of Botany , Botanical garden , University of Karachi en_US
dc.title OPTIMIZATION OF AGROBACTERIUM TUMEFACIENS MEDIATED TRANSFORMATION IN POPULUS DELTOIDES en_US
dc.type Article en_US


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