A rapid in vitro protocol for callus production in Piper aduncum L. propagation of East Kalimantan supplemented with gradient sucrose solutions

Abstract

Piper aduncum L. is a medicinal plant which contains important active compounds such as phenyl propanoids, lignoids and flavonoids. Furthermore, it is one of larvicidal agent against mosquitoes Aedes aegypti L. Alternative use of in vitro callus P. aduncum L. are as a substitute for medicinal raw materials and becomes one way to maintain the sustainability of germ plasm utility, which is related to its difficulties in conventional cultivation. An experiment based on saline formulation of Murashige and Skoog (MS) and Wood Plant Medium (WPM) combined with different 2,4-D and kinetin concentration was evaluated. Several conditions without plant growth regulator hormones with nodule segment explants produced good multiplication process. Inoculated micro-cutting segments of nodules were grown in media with an additional 2 mg/L of 2,4-D, 2 mg/L of kinetin and 30 g sugar produced multiplication of clear shoots and callus formation and were developed after 4 weeks of planting. Additional treatment of 60, 90, 120 g sugar showed slow growth as indicated by callus formation after 6-8 weeks of planting. Treatment of 30 g sugar showed better growth than other treatments, which triggered the survival of callus growth until 12 weeks after planting. The other treatments showed cell death after callus initiation due to browning process.