Show simple item record V. en_US B. V. en_US S. en_US Sue J. en_US Phillip en_US M. G. en_US 2009-11-25T12:14:16Z 2009-11-25T12:14:16Z 2001 en_US
dc.identifier.citation Bhat , V , Bhat , B V , Kumar , S , Dalton , S J , Morris , P & Gupta , M G 2001 , ' Particle-inflow-gun-mediated genetic transformation of buffel grass (Cenchrus ciliaris L.): optimizing biological and physical parameters ' Journal of Applied Genetics , pp. 405-412 . en_US
dc.identifier.other PURE: 129822 en_US
dc.identifier.other dspace: 2160/3606 en_US
dc.description.abstract The present study was conducted to optimize various biological and physical parameters for developing an efficient and reproducible gene transfer method for genetic transformation of buffel grass. Transformation was carried out using a helium-driven particle inflow gun (PIG). Embryogenic calli produced from mature seeds of buffel grass cv. CC-119 were separately bombarded with four plasmids, containing Actin (pAct1DX), Ubiquitin (pAHC-25; pAHC-27) and CaMV-35S (pCaMVGUS) promoters, coated on tungsten and gold particles. The efficiency of transformation was monitored through transient GUS expression. Different parameters, viz., the type of promoter, type and size of microcarrier, helium gas pressure, distance and time of bombardment, were standardized for delivering DNA into embryogenic calli. Bombardment with plasmid DNA carrying the actin promoter coated on 1.6 m gold particles, at a helium pressure of 4 bars, a distance of 10 cm for 10 m sec and 28 mm Hg vacuum in the chamber, produced the best result in transient GUS expression. The Actin promoter was found to be more efficient in driving expression of the GUS gene in buffel grass, followed by Ubiquitin and CaMV-35S promoters. Lower helium pressure was found to be sub-optimal, while higher pressure produced a smaller number of blue spots, probably due to excessive damage to the cells. Maximum of 385 blue spots was observed with gold particles of 1.6 m size, whereas only 213 blue spots were recorded for tungsten particles of 1.0 m size. The optimized parameters can be employed for genetic transformation of buffel grass with genes of agronomic importance. en_US
dc.format.extent 8 en_US
dc.relation.ispartof Journal of Applied Genetics en_US
dc.title Particle-inflow-gun-mediated genetic transformation of buffel grass (Cenchrus ciliaris L.): optimizing biological and physical parameters en_US
dc.contributor.pbl Aberystwyth University en_US
dc.contributor.pbl Institute of Biological, Environmental and Rural Sciences en_US

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