Journal Title
Title of Journal: Plant Cell Tiss Organ Cult
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Abbravation: Plant Cell, Tissue and Organ Culture (PCTOC)
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Publisher
Springer Netherlands
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Authors: Manoharan Rajesh Murugaraj Jeyaraj Ganeshan Sivanandhan Kondeti Subramanyam Thankaraj Salammal Mariashibu Subramanian Mayavan Gnanajothi Kapil Dev Vasudevan Ramesh Anbazhagan Markandan Manickavasagam Andy Ganapathi
Publish Date: 2013/02/27
Volume: 114, Issue: 1, Pages: 71-82
Abstract
An efficient Agrobacteriummediated genetic transformation method has been developed for the medicinal plant Podophyllum hexandrum Royle an important source of the anticancer agent podophyllotoxin Highly proliferating embryogenic cells were infected with Agrobacterium tumefaciens harbouring pCAMBIA 2301 which contains npt II and gusA as selection marker and reporter genes respectively The transformed somatic embryos and plantlets were selected on Murashige and Skoog MS basal medium containing kanamycin and germination medium respectively GUS histochemical analysis polymerase chain reaction and Southern blot hybridisation confirmed that gusA was successfully integrated and expressed in the P hexandrum genome Compared with cefotaxime 200 mg l−1 timentin completely arrested Agrobacterium growth and favoured somatic embryo development from embryogenic cells Among the different Agrobacterium strains acetosyringone concentrations and cocultivation durations tested embryogenic callus infected with A tumefaciens EHA 105 and cocultivated for 3 days on MS basal medium containing 100 μM acetosyringone proved to be optimal and produced a transformation efficiency of 2964 with respect to germinated GUSpositive plantlets The Agrobacteriummediated genetic transformation method developed in the present study facilitates the transference of desirable genes into P hexandrum to improve the podophyllotoxin content and to enhance other useful traitsThe authors are thankful to the Life Science Research Board Defence Research and Development Organisation DRDO Govt of India for financial support DLS/81/48222/LSRB–171 BTB/2008 used to carry out the present work The corresponding author is thankful to the University Grants Commission UGC Govt of India for providing an Emeritus fellowship under the BSR scheme The authors are also thankful to Prof SK Nandi GB Pant Institute of Himalayan Environment and Development Almora Uttarakhand India and Prof MC Nautiyal HNB Garhwal University Garhwal Srinagar Uttarakhand India for help with the collection of Podophyllum hexandrum seeds The authors are thankful to Dr AS Rao for his valuable suggestions to improve the manuscript
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