title: An efficient in planta transformation of Jatropha curcas (L.) and multiplication of transformed plants through in vivo grafting.
abstract: An efficient and reproducible Agrobacterium-mediated in planta transformation was developed in Jatropha curcas. The various factors affecting J. curcas in planta transformation were optimized, including decapitation, Agrobacterium strain, pin-pricking, vacuum infiltration duration and vacuum pressure. Simple vegetative in vivo cleft grafting method was adopted in the multiplication of transformants without the aid of tissue culture. Among the various parameters evaluated, decapitated plants on pin-pricking and vacuum infiltrated at 250 mmHg for 3 min with the Agrobacterium strain EHA 105 harbouring the binary vector pGA 492 was proved to be efficient in all terms with a transformation efficiency of 62.66%. Transgene integration was evinced by the GUS histochemical analysis, and the GUS positive plants were subjected to grafting. Putatively transformed J. curcas served as "Scion" and the wild type J. curcas plant severed as "Stock". There was no occurrence of graft rejection and the plants were then confirmed by GUS histochemical analysis, polymerase chain reaction (PCR) and Southern hybridization. Genetic stability of the grafted plants was evaluated by using randomly amplified polymorphic DNA (RAPD), marker which showed 100% genetic stability between mother and grafted plants. Thus, an efficient in planta transformation and grafting based multiplication of J. curcas was established.
authors.affiliation: Department of Biotechnology and Genetic Engineering, School of Biotechnology, Bharathidasan University, Tiruchirappalli, 620024, Tamil Nadu, India.
authors.forename: Balusamy
authors.initials: B
authors.lastname: Jaganath
authors.forename: Kondeti
authors.initials: K
authors.lastname: Subramanyam
authors.forename: Subramanian
authors.initials: S
authors.lastname: Mayavan
authors.forename: Sivabalan
authors.initials: S
authors.lastname: Karthik
authors.forename: Dhandapani
authors.initials: D
authors.lastname: Elayaraja
authors.forename: Rajangam
authors.initials: R
authors.lastname: Udayakumar
authors.forename: Markandan
authors.initials: M
authors.lastname: Manickavasagam
authors.forename: Andy
authors.initials: A
authors.lastname: Ganapathi
mesh_terms: D018744:DNA, Plant / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D031286:Jatropha / Q000235:genetics* / Q000254:growth & development; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic
chemical_list: D018744:DNA, Plant