Comparison of genetic transformation in Morus alba L. via different regeneration systems |
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Authors: | Sandhya Agarwal Kamlesh Kanwar |
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Affiliation: | (1) Department of Biotechnology, Dr. Y. S. Parmar University of Horticulture and Forestry, Solan, 173230, India;(2) Present address: Division of Crop Improvement, Central Potato Research Institute, Shimla, 171001, India |
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Abstract: | Three different regeneration systems, viz. direct regeneration of adventitious shoot buds from explant, regeneration through callus cultures and somatic embryos were compared to see their effect on transfer of neomycin phosphotransferase (nptII) and β-glucuronidase (GUS) reporter gene (gus) to Morus alba clone M5, through Agrobacterium tumefaciens mediated transformation. Pre-conditioning and co-cultivation durations had a marked effect on transformation frequency. The highest transformation frequency of 18.6% was obtained using direct induction of adventitious shoot buds. Expression and presence of transgene were assayed histochemically and through polymerase chain reaction. Southern analysis of GUS and PCR positive transformants confirmed stable integration of transgenes with two to four copy numbers. The selected transformants showed normal phenotype under in vitro and field conditions. |
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Keywords: | Genetic transformation Morus alba L. Shoot regeneration Somatic embryogenesis Tissue culture |
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