MICROPROPAGATION OF CYMBIDIUM SPP. BY SOMATIC EMBRYOGENESIS TECHNIQUE

Abstract

Cymbidium spp. was regularly produced through micropropagation by protocorm like bodies (PLBs) and multiple shoots; micropropagation takes a lot of labor, and has high cost of seedlings, energy and material. The purpose of this paper was to study the new technique of using in vitro embryogenesis culturing for microprogation to resolve the above difficulties. The method was to use PLBs as planting materials to produce somatic callus cell and embryos. Results were followed as: PLBs were cut into slices and placed on the medium for callus initiated and used as materials for embryo formation study. A fresh weight of callus were used for the experiment of about 100 mg. The medium for the initiation of embryonic callus was composed of: MS + BA (0.1 mg/l) + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1mg/l) or 2.4D (1 mg/l) and was proliferated on the medium MS + pepton (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) or 2.4D (1 mg/l). Somatic cell suspensions were initiated and proliferated on the medium consisting of MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) + BA (0.1 mg/l). Somatic cell suspensions were differentiated to embryonic cell suspensions on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with BA (1 mg/l). Embryonic cell suspensions were plating and regeneration on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (0.1 mg/l) + BA (1 mg/l). Micropropagation of Cymbidium spp. via embryogenesis technique was set up to produce 3,800 plantlets per one liter of somatic embryogenesis suspension.