A research on the reactions of some pepper (Capsicum annuum L.) genotypes on different androgenesis methods

In this study, anther culture and shed microspore culture methods were used to obtain haploid and DH plants from 2 commercial pepper cultivars, Lumbard RZ F1 and Uc burun F1. In order to determine the appropriate anther development stage of the pepper varieties, the classification process was made considering the morphological appearance of the buds among themselves. Then, in order to determine the appropriate microspore stage, the buds classified were stained using ethidium bromide (EtBr) staining technique and observed under fluorescence microscope. Anthers of the appropriate stage were separated from the pepper bud and transferred to MS nutrient media, supplemented with 0,1 mg / l BAP +4 mg / l NAA + 15 mg / l AgNO3 and 2,5 g / l activated charcoal with pH 5,7-5,8 for the anther culture technique. Cultured anthers were first kept in incubator for 48 hours in the dark at +35 0C and then kept in the dark for 33 days in the growth chamber of +25 0C. 33 days after cultivation of pepper buds were transferred to hormone-free MS medium and taken to growth room with 3000 lux illumination at +25 0C for 16 hours light and 8 hours dark. Pepper buds were transferred to hormone-free MS medium 33 days after culturing and kept in growth chamber having 3000 lux illumination at 25 0C with 16 hours light and 8 hours dark Another method used in the research is the shed microspore method. In this method, pepper buds were collected at the appropriate stag and subjected to pretreatment at 4 0C for 24 hours. Anthers separated from pre-treated buds were placed in nutrient medium which was consisted of NN medium + %2 maltose +2,5 μM zeatin, 5 μM IAA + %1 activated charcoal and 6 g / l agar. The anther was then covered with liquid layer formed by adding + %2 maltose to NN medium. The pH of semi-solid and semi-liquid nutrient medium used in the culture medium was adjusted to pH 5, 7 – 5,8. Cultured anthers were kept in darkness at +4 0C duruing one week for cold pretreatment. After cold pre-treatment, anthers were kept at +27 0C until microspores callus, embryo or embryo-like structures were formed. The resulting structures were transferred to hormone-free MS medium and kept in a growth chamber with 16 hours light, 8 hours dark photoperiod conditions under 3000 lux illumination at +25 0C. As evaluation of results, embryo development was observed as %8,94 of Lumbard RZ F1 cultivars and %23,63 of the developing embryos were transformed into plants. The embryo formation rate of the Uç burun F1 cultivar used in this study was found to be %22,18, while their conversion rate to plants was %22,22. While both Lumbard RZ F1 and Ucburun F1 varieties responded to anther culture studies, in the shed microspore method, embryo formation was observed only from Lumbard RZ F1. Embryo formation rate of Lumbard RZ F1 was %11,11, while the rate of transformation of these embryos into plants was %7,87.