title: A new high-frequency Agrobacterium-mediated transformation technique for Sesamum indicum L. using de-embryonated cotyledon as explant.
abstract: In spite of the economic importance of sesame (Sesamum indicum L.) and the recent availability of its genome sequence, a high-frequency transformation protocol is still not available. The only two existing Agrobacterium-mediated transformation protocols that are available have poor transformation efficiencies of less than 2%. In the present study, we report a high-frequency, simple, and reproducible transformation protocol for sesame. Transformation was done using de-embryonated cotyledons via somatic embryogenic stages. All the critical parameters of transformation, like incubation period of explants in pre-regeneration medium prior to infection by Agrobacterium tumefaciens, cocultivation period, concentrations of acetosyringone in cocultivation medium, kanamycin concentration, and concentration of plant hormones, including 6-benzylaminopurine, have been optimized. This protocol is superior to the two existing protocols in its high regeneration and transformation efficiencies. The transformed sesame lines have been tested by PCR, RT-PCR for neomycin phosphotransferase II gene expression, and β-glucuronidase (GUS) assay. The regeneration frequency and transformation efficiency are 57.33 and 42.66%, respectively. T0 and T1 generation transgenic plants were analyzed, and several T1 plants homozygous for the transgenes were obtained.
authors.affiliation: Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, 700019, India, rohansupriyo@gmail.com.
authors.forename: Supriyo
authors.initials: S
authors.lastname: Chowdhury
authors.forename: Arpita
authors.initials: A
authors.lastname: Basu
authors.forename: Surekha
authors.initials: S
authors.lastname: Kundu
mesh_terms: D000098:Acetophenones / Q000494:pharmacology; D016960:Agrobacterium tumefaciens / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001593:Benzyl Compounds; D018548:Cotyledon / Q000166:cytology; D004351:Drug Resistance / Q000235:genetics; D018014:Gene Transfer Techniques; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics*; D007612:Kanamycin / Q000494:pharmacology; D019868:Kanamycin Kinase / Q000096:biosynthesis / Q000235:genetics*; D007701:Kinetin / Q000494:pharmacology; D030821:Plants, Genetically Modified / Q000235:genetics; D011687:Purines; D031685:Sesamum / Q000187:drug effects / Q000235:genetics*; D014170:Transformation, Genetic
chemical_list: D000098:Acetophenones; D000900:Anti-Bacterial Agents; D001593:Benzyl Compounds; D011687:Purines; D007612:Kanamycin; C051667:acetosyringone; D019868:Kanamycin Kinase; D005966:Glucuronidase; C480551:benzylaminopurine; D007701:Kinetin