An Efficient Anther Culture on Double-Layered Media to Produce Doubled Haploid Plants of Pepper (Capsicum annuum).

An efficient anther culture on double-layered media to produce doubled haploid (DH) plants in pepper (Capsicum annuum) was clearly shown to outperformed other techniques such as anther culture on solid medium and direct isolated microspore culture on liquid medium. It was even used for DH production in a cayenne type of hot pepper which was previously known as less responsive or even more recalcitrant to androgenesis than sweet bell pepper. Indeed, anther culture on double-layered media has been routinely used to produce DH plants on broad genotypes of C. annuum as parental candidates to develop hybrid varieties. The step-by-step protocol of pepper anther culture on double-layered media, we hereby present in detail, includes the growth of donor plants, the use of flower buds as anther source, flower bud disinfection, anther isolation, anther culture and incubation processes, embryo germination and plant acclimatization process, and transplanting of plants to the soil-compost medium in pots.