Effect of gelling agent, carbon source and sterilization methods on initiation and establishment of embryogenic cultures in Khasi pine (Pinus kesiya Royle ex. Gord.)

Embryogenic cultures were initiated and established in Khasi pine (Pinus kesiya Royle ex. Gord.) using mature zygotic–embryos. Factors such as gelling agent, carbon source and sterilization methods affecting the initiation of embryogenic cultures were investigated. Phytagel at a concentration of 4 g L⁻¹ induced white mucilaginous embryogenic callus but various brands of agars failed to produce embryogenic callus. Phytagel was a promising gelling agent for the initiation of embryogenic cultures in Pinus kesiya. Among the carbon sources tested, sucrose and maltose were found as medium and best carbon sources, respectively. Maltose @ 30 g L⁻¹ produced highest embryonal masses and was used as the best carbon sources. The growth rate of embryogenic callus was increased when carbon sources were filter sterilized. Also, the number of proembyros on embryogenic callus generally increased by the use of sterilized carbon source.