Haploid plant production in Zantedeschia aethiopica ‘Hong Gan’ using anther culture

► A high yielding protocol for the anther culture of Zantedeschia aethiopica was established. ► Regenerants were obtained via a callus phase. ► Both haploid and diploid plants originated from the microspores. ► 25% of the DHs equaled the original cultivar in growth vigor, 37.5% showed fertility. This report describes advances in the anther culture of Zantedeschia aethiopica. Important factors for improvement as compared to the earlier procedure were: (1) using flowers from inflorescences developed at relatively low temperature during winter, (2) high temperature stress treatment at 32 °C for 2 days in the beginning of the culture, (3) use of Gamborg B5 as anther culture medium, and (4) addition of sucrose at high concentration of 8% in the culture medium. Plants were obtained via a callus phase. Frequency of anthers producing calli was around 4–5%. About 87% of the calli gave regenerants, of which 52% were haploid, 36% were diploid and the rest had other ploidy levels. In addition to chromosome counting, cytological examination of the microspore development and amplified fragment length polymorphism (AFLP) analysis of the regenerants showed that haploid as well as diploid plants originated from the microspores. Finally, 12 doubled haploid (DH) plants could be produced from each inflorescence. One quarter of the DHs equaled the original cultivar in growth vigor, while more than one third showed good fertility, indicating that inbreeding depression was not so severe in this heterozygous species. The improved protocol now enables production of sufficient number of DHs for application of haploid technology in genetic improvement and breeding of Z. aethiopica.