水稻原生质体培养及植株再生的研究

由粳稻77-170品系及籼稻品种IR-50的细胞悬浮培养物游离的原生质体,用琼脂糖包埋于RY-2培养基中,发生了持续分裂。前者植板率达2.5％以上,二者最后都再生出植株。对游离和培养方法做了如下改进:1)采用两步法,即先用果胶酶,再用果胶酶和纤维素酶的混合酶进行游离,可避免原生质体发生融合并获得高质量的原生质体;2)悬浮细胞培养基中加入ABA有利于原生质体的存活和分裂;3)琼脂糖包埋培养可大大提高植板率;4)用较高渗透压的培养基培养原生质体再生的细胞团及愈伤组织,可提高植株再生频率。由于这两个品种(系)的培养物都已继代一年半之久,再生植株均为白化苗。这是迄今第一个由籼稻原生质体再生植株的报道。

Studies on Protoplast Culture of Rice (Oryza saliva L.) and Plant Regeneration from Protoplast-derived Calli

Protoplasts isolated from suspension cells of a japonica rice (77-170) and an indica variety (IR-50) were cultured in RY-2 medium.The plating efficiency of 77-170 was as high as 2.5%.Plant regenerations were achieved in both subspecies.Some methodological improvements are as follows:1) A two-step isolation procedure was used to avoid the spontaneous fusions and obtain high quality protoplasts.2) Protoplasts isolated from suspension cells cultured in AA medium supplemented with ABA showed a distinctly high leved of survival and division.3) Postponing the transfer of the embrdded protoplasts into liquid medium may be beneficial to their survival and divisions.4) Using media with relatively high osmotic pressure in cul-turing protoplast-derived calli increased their plant regeneration frequency on subsequent differentiation media.This is the first report of plant regeneration from the protoplast culture of an indica rice.All regenerated plants are albinos,probably due to their long-term culture history (one and a half years).