水稻游离花粉培养的高频率再生植株

HIGH-EFFICENCY PLANT REGENERATION FROM IN VITRO CULTURED MICROSPORES OF RICE (ORYZA SATIV A L.SUB.JAPONICA)

用二个水稻栽培品种(Oryza sativa L Sub.japonica.)中花11号和盐粳的花粉处于单核靠边期的花药,经低温处理10—20天,在无糖培养基中预培养2—4天后游离花粉进行培养。培养基为KM8P,附加1mg/L 2,4-D,100mg/L脯氨酸,500mg/L水解酪蛋白,9%蔗糖。培养5天,花粉进行一次分裂,10天后分裂频率为21.3%,21天可见小愈伤组织形成。随即将直径为0.5—1.0mm的愈伤组织移至分化培养基上,2—4星期后得到绿苗,频率为70%。并从许多花粉诱导的愈伤组织克隆中筛选出高频率再生绿苗的花粉细胞悬浮系。

After 10-20 day's cold-treatment of 4℃, anthers of two japonica rice cultivars were precultured for 2-4 days in modified N6 medium without any sugar. Then the isolated uninucleate microspores were cultured in KM 8 P medium supplemented with 1 mg/L 2,4-D, 100mg/L proline, 500 mg/L casamino acids and 9% sucrose. The first cell divisions were observed 5 days later and the highest cell division frequency of 21.3% could be obtained at the 10th day of culture. Microcalli appea- red after 21 day's culture and were trans-fered to the media for plant regeneration when they grew to the diameter about 0.5-1 mm, green seedlings regenerated from these calli after 2-4 weeks. Among the numerious microspore-derived clones, the highest regeneration efficiency of green plant was 70%. Thus a suspension cell line with fast growth rate and high-regeneration efficiency was obtained and it is useful for genetic manipulation.