The effects of plant use at different genetic levels on haploid plant production in pepper (capsicum annuum l.) anther and microspore culture

This study was conducted to reveal the effects using pepper lines at four different genetic levels on haploid embryo formation in anther and microspore culture. For this purpose, pepper genotypes buds were classified by staining ethidium bromide dye technique in order to determinate the suitable anther and microspore development stage. In this study, MS basic media, supplemented with 4 mg/l NAA + 1 mg/l BA, was used for anther culture whereas B5 basic media with the combination of 0.1 mg/l 2,4-D + 0.2 mg/l kinetin was used for microspore culture. At anther culture of study, anthers were subjected to heat pretreatment during for 8 days at +35°C temperature. At microspore culture of study, for first 7 days heat pretreatment and carbohydrate starvation treatment at 32°C were applied to promote the haploid embryo formation by stimulating microspores. Petri dishes belong to anther culture were transferred to growth chamber having 25°C, 16 h light/8 h dark photoperiod and 3000 lux illumination whereas petri dishes belong to microspore culture were maintained in 25°C in dark conditions. No positive effects of heat pretreatments and carbohydrate starvation treatment on embryo formation was observed in the study. On the other hand callus, embryo likes structures and embryos were obtained and recorded. In terms of their response to anther culture among genotypes studied, anthers of F4 plants gave thetorpedo embryos at the rate of 15.07% and determined as the most suitable genetic level. According to the assessment made in the microspore culture, F4 plants had the best results as in anther culture with 13.89% embryo formation.