In vitro androgenic response of minipaprika (Capsicum annuum L.) genotypes in different culture media

Abstract

The in vitro androgenic response of minipaprika F1 hybrid cv. Vine sweet (red, yellow, and orange form) was investigated using Dumas de Vaulx (CP) and Murashige and Skoog (MS) culture media to ascertain the effectiveness of androgenesis in haploid production. The frequency of callus and embryo formation varied in anther cultures of all minipaprika forms in both culture media. Anthers in both culture media responded to form the callus without regeneration in all minipaprika forms. The anthers of ‘Vine sweet-yellow’ produced 62.5% and 46.7% normal embryo in CP and MS medium, respectively. ‘Vine sweet-yellow’ produced almost four fold more normal-looking embryos than ‘Vine sweet-red’ in both culture media, and ‘Vine-sweet orange’ showed low androgenic response to anther culture. The anthers cultured in CP medium gave more normal embryo in all minipaprika forms as compared to MS medium. Among a total of 51 embryos transferred to growth regulator free MS medium for regeneration, 45 plants were regenerated. Flow cytometry analysis revealed that 44.5%, 42.4%, and 33.3% plants were haploids, and 55.5%, 57.6%, and 66.7% plants were spontaneous diploids in red, yellow and orange forms, respectively. The chromosome number of haploid plant was 12 and that of spontaneous diploid was 24. Stomata characters and chloroplast count in the guard cell of leaf stomata were also found to be reliable and simple method to analyze the ploidy level of regenerated plants. The spontaneous diploids confirmed as doubled haploids (DHs) following the self-pollination. The obtained DH and haploid plants from anther culture would be the valuable breeding materials for heterosis breeding.