DEVELOPMENT OF AN AGROBACTERIUM-MEDIATED TRANSFORMATION SYSTEM FOR BLACK PEPPER (PIPER NIGRUM L.)

Leaf, petiole and stem explants from axenic seedlings of black pepper (Piper nigrum L.) were scored or pricked and then inoculated with the Agrobacterium tumefaciens strain LBA 4404 containing vectors carrying the npt II and gus genes.
After inoculation, the explants were first cultured in callus-inducing medium in the dark at 28°C and then transferred to shoot regeneration medium.
This is a MS-based medium with antibiotics carbenicillin and cefotaxime incorporated at 500 and 100 mg/1 respectively to eliminate Agrobacterium. The shoot regeneration medium was also supplemented with kanamycin sulfate at 75 mg/l for selection of transformed shoots.

Protuberances which resembled embryos were observed at the basal end of petioles cultured in MS medium supplemented with BAP and NAA. Multiple shoots were readily induced on stem explants which were cut right across the node.

At ten days after inoculation with the Agrobacterium, explants were sampled for GUS expression by X-gluc staining.
This study showed that gene transfer to pepper could be achieved by using suitable strains of Agrobacterium tumefaciens since expression of the gus gene confirmed gene transfer to recipient tissue.
Transformation, if any, needs to be confirmed by further analysis using shoots regenerated from the inoculated explants.

International Symposium on Biotechnology of Tropical and Subtropical Species Part 2

S.L. Sim, R. Jafar, J.B. Power, M.R. Davey

Piper nigrum, Agrobacterium tumifaciens, transformation

https://doi.org/10.17660/ActaHortic.1998.461.39