Abstract
Production of Agrobacterium tumefaciens-mediated transgenic plants, via direct shoot bud organogenesis from leaves of Catharanthus roseus, is reported. A. tumefaciens harbouring the plasmid pBI121 with GUS gene uidA and kanamycin resistance gene nptII was used. Highest transformation efficiency of 1.4 transgenic shoots/responded explant was obtained when pre-plasmolysed leaves, pre-incubated on shoot bud induction medium for 10 days, were subjected to sonication for 30 s prior to transformation. Using a selection medium containing 50 mg kanamycin l−1, transformants grew into micro-shoots and formed roots on a hormone-free half strength MS medium. The transgenic nature of the regenerated plants was confirmed by PCR amplification of uidA gene and GUS histochemical assay.
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Acknowledgments
This work was financially supported by The Council of Scientific and Industrial Research (CSIR), New Delhi (India) through its Network Project COR-02. Authors are also grateful to Dr D.V. Amla, National Botanical Research Institute, Lucknow (India) for providing the gene constructs and to Dr A.K. Gupta and Dr A.K. Shasany, CIMAP, Lucknow for providing the seeds and plant material of C. roseus genotype. The help rendered by Dr A. Mathur in PCR characterization work is also acknowledged. PV also thanks CSIR for awarding a senior research fellowship to her during the course of this investigation. We are also grateful to Director, CIMAP for providing facilities and support.
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Verma, P., Mathur, A.K. Agrobacterium tumefaciens-mediated transgenic plant production via direct shoot bud organogenesis from pre-plasmolyzed leaf explants of Catharanthus roseus. Biotechnol Lett 33, 1053–1060 (2011). https://doi.org/10.1007/s10529-010-0515-2
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DOI: https://doi.org/10.1007/s10529-010-0515-2