벼의 小胞子 培養에서 裸出前 高溫處理와 裸出方法이 callus形性 및 植物體 分化에 미치는 影響

Effects of preculture temperatures and isolation methods on callus formation and plant regeneration in rice microspore culture

To improve the rate of green plant regeneration in isolated microspore culture of Oryza sativa L., panicles of middle or late-uninucleate microspore stages were collected from cultivar Daecheongbyeo and were pretreated for 10 days at 10℃. Anthers were precultured in N6 liquid medium supplemented with 2mg/ℓ NAA and 1mg/ℓ kinetin for 2 or 7 days at 30 and 35℃ after preculture for 8 days at 25℃. Microspores were isolated with spontaneously shedded method of mechanically squashed method. The number of isolated microspore and survived microspore was remarkably higher by mechanically homogenized method than by spontaneously shedded method. The effect of high temerature culture in the late period of preculture on callus induction was the highest at 35℃ by mechanically homogenized isolation and that was mostly lower by spontaneously shedded isolation than by mechanically homogenized method. The plants were the most regenerated as 54 plants by mechanically homogenized isolation at 35℃, 36 were green and 18 were albino. Frequency of haploid of regenerated green plant was 29% and diploid was 69%.