Protocol establishment of culturing doubled dihaploid plants in vitro on chilli (Capsicum sp.)

Abstract

Haploid shoots can be obtained by in vitro ovary and anther culturing, which is the first step in the process of breeding using plant tissue culturing. Anthers and unfertilized ovaries of 4 kinds of chilli peppers including hot red pepper, horn peppers, bell peppers and bullhorn peppers were cultured. The result showed that the death rate and the rate of callus formation from ovary and anther were improved through cold pretreatment. Callus from anthers and ovaries were obtained from all four types of peppers in suitable culture media. Callus when transferred to regenerating media, changed from white to yellow green and became suitable for shoot regeneration. Embryos and embryonic callus were induced from bell pepper anthers on MS + 2mg/l 2,4-D + 1mg/l kinetin or MS + 0.5mg/l TDZ (2.7 – 4.67% of anthers), of which only 42.86% regenerated shoots successfully on MS medium added 0.5mg/l BA. It was impossible to regenerate shoot from calluses of hot red peppers. calluses from 3 remaining kinds of peppers were capable of regenerating somatic embryo.