Ploidy of broccoli regenerated from microspore culture versus anther culture

Abstract

The use of microspore or anther culture to generate doubled-haploids (DH) is an important adjunct to broccoli breeding. Regenerated populations from broccoli anther culture are usually mixtures of ploidy. However, ploidy composition of populations derived from microspore culture has not been reported. The purpose of the present study was to characterize regenerants derived from microspore culture, to evaluate factors influencing these characteristics and to compare results with those from anther culture. Eight populations, four from each culture method, were generated simultaneously using the same four F1 hybrids as donor parents. The ploidy level of all regenerants was determined by DNA flow cytometry: the majority of them were diploid. As in anther culture, a mixture of ploidy was observed in all populations derived from microspore culture. Ploidy variation was more frequent among clonal families from anther culture (10%) than microspore culture (5%). 'Everest' was the most productive donor parent with both methods, while 'Greenbelt' and 'Major' were least productive in anther and microspore culture, respectively. Genotype specificity for the total number of regenerated plants and ploidy composition occurred in both culture methods.