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.

Mini-grafting of adult Passiflora edulis Sims f. flavicarpa Deg. scions onto vegetatively propagated adult rootstocks of P. mucronata Lam.

“…According to these authors, the repetitive pruning of the graft produced new stem tissues which suggests a regular flow of nutrients from the rootstock and consequently the success of the graft. The use of Parafilm ® in top wedge grafting in transgenic Jatropha curcas scions onto wild rootstock of the same species was rather effective with 100% genetic stability between the mother plant and the grafted plant (Jaganath et al, 2014). This reinforces our data on the usefulness of the fastening materials in improving the grafting process.…”

Agrobacterium-mediated in planta genetic transformation of sugarcane setts

“…Mayavan et al (2013) selected the transformed sugarcane seedlings by germinating in the sterile MS liquid medium supplemented with 2.0 mg l -1 BASTA Ò and then 2-month-old plants by spraying with 2.0 g l -1 BASTA Ò . In a similar fashion, the transformed Arabidopsis, Medicago truncatula, Brassica rapa, and Jatropha curcas were selected by spraying BASTA Ò (Mengiste et al 1997;Trieu et al 2000;Qing et al 2000;Jaganath et al 2014). …”

Effects of Media Composition and Auxins on Adventitious Rooting of Bienertia sinuspersici Cuttings

“…Moreover, vegetative propagation is efficient because it reduces the amount of time required for seedling establishment and a large number of healthy living stock plants can be produced rapidly from a small amount of available original plant material. Rooted cuttings can also serve as potential rootstocks for grafting experiments for plant species that are recalcitrant to adventitious root induction process or for multiplication of transgenic shoots [8] [9]. In addition, findings on nutrient requirements and effects of auxins on rooting of B. sinuspersici cuttings may provide valuable insight into factors that influence the in vitro regeneration process.…”