title: Assessment of factors influencing the Agrobacterium-mediated in planta seed transformation of brinjal (Solanum melongena L.).
abstract: An efficient and reproducible in planta transformation method was developed for brinjal using seed as an explant. The brinjal seeds were infected with Agrobacterium tumefaciens EHA 105 harbouring pCAMBIA 1301-bar plasmid, and the transformants were selected against BASTA®. Several parameters influencing the in planta seed transformation such as pre-culture duration, acetosyringone concentration, surfactants, duration of sonication, vacuum pressure and vacuum duration have been evaluated. The putatively transformed (T 0) brinjal plants were screened by GUS histochemical analysis. Among the different combinations and concentrations tested, when the 18-h pre-cultured brinjal seeds were sonicated for 20 min and vacuum infiltered for 3 min at 500 mm of Hg in Agrobacterium suspension containing 100 μM acetosyringone, 0.2 % Silwett L-77 favoured the Agrobacterium infection and showed maximum transformation efficiency. Among the five brinjal varieties evaluated, Arka Samhitha showed maximum transformation efficiency at 45.66 %. The transgene was successfully transmitted to progeny plants (T 1) which was evidenced by GUS histochemical analysis, polymerase chain reaction and Southern hybridisation. The in planta protocol developed in the present study would be beneficial to transfer the economically and nutritionally important genes into different varieties of brinjal, and the transgenic brinjal plants can be produced in less time (approximately 27 days).
authors.affiliation: Department of Biotechnology and Genetic Engineering, School of Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India.
authors.forename: Kondeti
authors.initials: K
authors.lastname: Subramanyam
authors.forename: Manoharan
authors.initials: M
authors.lastname: Rajesh
authors.forename: Balusamy
authors.initials: B
authors.lastname: Jaganath
authors.forename: Amirthalingam
authors.initials: A
authors.lastname: Vasuki
authors.forename: Jeevaraj
authors.initials: J
authors.lastname: Theboral
authors.forename: Dhandapani
authors.initials: D
authors.lastname: Elayaraja
authors.forename: Sivabalan
authors.initials: S
authors.lastname: Karthik
authors.forename: Markandan
authors.initials: M
authors.lastname: Manickavasagam
authors.forename: Andy
authors.initials: A
authors.lastname: Ganapathi
mesh_terms: D000098:Acetophenones / Q000494:pharmacology; D060054:Agrobacterium / Q000235:genetics*; D018920:Coculture Techniques; D005818:Genetic Engineering / Q000379:methods*; D005838:Genotype; D018525:Germination; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000235:genetics* / Q000254:growth & development; D032323:Solanum melongena / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D013010:Sonication; D013501:Surface-Active Agents / Q000494:pharmacology; D013997:Time Factors; D014170:Transformation, Genetic / Q000187:drug effects; D019076:Transgenes / Q000235:genetics; D014618:Vacuum
chemical_list: D000098:Acetophenones; D013501:Surface-Active Agents; C051667:acetosyringone; D005966:Glucuronidase