An improved in vitro technique for isolated microspore culture of barley

A detailed procedure for isolated microspore culture of barley is presented along with examples of response across genotypes. Over 30 genotypes, including winter and spring growth habit and 2-row and 6-row genotypes, have shown an essentially genotype independent response, averaging about 10,000 embryos per 5 cm petri culture plate. The regeneration frequency, checked on samples of 500 embryos per plate ranged from 36 to 97% with most genotypes being in the range of 70 to 90%. About 70 to 80% of the plants regenerated have been completely fertile doubled haploids, thus eliminating the need to double the chromosome number of plants. Many little details are critical to success of the microspore procedure and while it saves much time compared to anther culture, greater attention to details and cleanliness is essential.