Effect of 2, 4-D and BAP on Callus Induction of Piper retrofractum Vahl.

Abstract

Piper retrofractum Vahl. in Indonesia has potential to be one of the medicinal plants to treat infertility problems. Several secondary metabolite compounds contained in Piper retrofractum Vahl. are essential oils, alkaloids, piperins, saponins, flavonoids, triterpenoids, steroids and glycosides. Plant tissue culture method to induce callus media requires proper formulation to get the optimum result. The purpose of this research was to know the effect of growth regulators combination of 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and 6-Benzylaminopurine (BAP) on time of induction, percentage, fresh weight, dry weight, and morphology on callus culture of Piper retrofractum Vahl.'s leaf explant. Leaf explant of Piper retrofractum Vahl. cultured on Murashige and Skoog (MS) medium with combination of 2, 4-D and BAP with concentrations of 0.0mg/L, 0.5mg/L, 1mg/L, 1.5mg/L, 2mg/L. This research was a laboratory experimental research with the complete randomized design method. The data obtained were analyzed quantitatively and qualitatively. Quantitative data consisted of time of induction, percentage, fresh weight, dry weight of callus, while qualitative data was obtained from the morphology description of Piper retrofractum Vahl. The results showed that the combination of growth regulators 2, 4-D and BAP added in culture medium influenced the growth of Piper retrofractum Vahl. leaf explant. Both at the time of induction, percentage, fresh weight and dry weight of callus. The best combination of growth regulators 2, 4-D 1.0mg/L and BAP 1.0mg/L showed the average time in inducing callus 15 days after planting. The combination of growth regulators 2, 4-D 1.0mg/L and BAP 2.0mg/L was the best combination resulted in an average of fresh weight and dry weight of callus 31.20mg and 5.92mg. Morphology of callus was produced by explant mostly textured friable greenish-white which cultured in medium with combination of growth regulators 2, 4-D 1.0mg/L and BAP 2.0mg/L in all parts of edge and surface explant.