Comparison of genetic transformation in Morus alba L. via different regeneration systems.
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 beta-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.
Agarwal
Sandhya
S
Department of Biotechnology, Dr Y S Parmar University of Horticulture and Forestry, Solan, India.
Kanwar
Kamlesh
K
D016960:Agrobacterium tumefaciens / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics; D007612:Kanamycin / Q000494:pharmacology; D019868:Kanamycin Kinase / Q000235:genetics; D031625:Morus / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism; D018517:Plant Roots / Q000254:growth & development; D018520:Plant Shoots / Q000254:growth & development / Q000378:metabolism; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic
D007612:Kanamycin; D019868:Kanamycin Kinase; D005966:Glucuronidase