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为了提高水稻小孢子培养效率,加速小孢子培养技术在水稻育种中的应用,本研究以上海本地早熟杂交粳稻品种‘申优17’为材料,对F_1小孢子培养中前期幼穗低温预处理时间(12和16 d)、诱导胚性愈伤转分化培养时间(4周、7周和13周)以及自发加倍频率等技术环节进行了研究。结果表明:低温(4°C)处理水稻幼穗12 d在诱导愈伤组织产量和分化绿苗产量上都要优于16 d处理,且在3个转分化培养时间上都显现出这一优势效果;比较3个转分化培养时间,其中7周的转分化培养时间可以获得较高的分化绿苗数;小孢子再生植株中自发加倍成二倍体频率约为50%,而单倍体约占40%。上述结果表明,选择合适的幼穗低温预处理时间(1～2周)和转分化培养时间(7周)有利于提高水稻小孢子培养中的绿苗分化频率。
In order to improve the efficiency of microspore culture in rice and accelerate the application of microspore culture technology in rice breeding, this study took ’Shanghai Youyou 17’, a japonica hybrid cultivar with early maturity in Shanghai as material, to study the effects of low temperature pretreatment time (12 and 16 d), induction of embryogenic callus differentiation and culture time (4 weeks, 7 weeks and 13 weeks) and frequency of spontaneous doubling and other technical aspects were studied. The results showed that the treatment of low temperature (4 ° C) for young rice panicle 12 d was better than that of 16 d in induction of callus production and differentiation of green shoot production, and showed the advantage in 3 transdifferentiation culture time The frequency of differentiated green shoots was higher in the 7-week transdifferentiation culture. The frequency of spontaneous doubling in microspore-regenerated plants was about 50%, while that in haploid 40%. The above results indicated that the selection of the appropriate pretreatment time (1-2 weeks) and transdifferentiation (7 weeks) for young spikelets were helpful to improve the frequency of green shoot differentiation in the microspore culture of rice.