野生稻原生质体培养与植株再生

Protoplast Culture and Plant Regeneration in Wild Oryza Species with Promotion by Nurse Cell Liquid and Suitable Proliferation Media

三个野生稻（Ｏ．ｒｕｆｉｐｏｇｏｎ，Ｏ．ｇｌｕｍａｅｐａｔｕｌａ和Ｏ．ｌａｔｉｆｏｌｉａ）成熟胚，经愈伤组织诱导和悬浮细胞培养，分离原生质体，利用简化原生质体培养基（ＳＰＣＭ），结合琼脂糖包埋，并附加看护细胞培养液，这三个野生稻原生质体均得到了成功培养，其中两个（Ｏ．ｒｕｆｉｐｏｇｏｎ和Ｏ．ｇｌｕｍａｅｐａｔｕｌａ）分化了绿色植株。看护细胞液在野生稻原生质体培养中，可促进细胞分裂，提早细胞分裂始期，细胞分裂频率（３．２％）和植板率（１９．０％）分别比未加看护细胞液的培养方法高出１３．３％和６．２％。看护细胞液可大大减少野生稻愈伤组织在液体中培养始期的死亡率，提高建立其悬浮细胞的成功率。原生质体克隆在适当的后培养基上进行培养，可以显著改良其结构，明显促进绿苗分化。尤其是Ｌ３培养基诱导原生质体克隆形成胚性愈伤组织或类似胚状体结构的频率达３２．１％，显著高于ＭＳ和Ｎ６培养基，从而导致较高绿苗分化频率。

The protoplasts of three Oryza species (O. rufipogon, O. glumaepatula and O. latifolia) were successfully cultured by using agrose-bead method in simplified protoplast culture medium (SPCM) with or without the nurse cell liquid through cell suspensions from mature embryos. Two of them (O.rufipogon, O.glumaepatula) regenerated green plantlets from protoplast-derived calli. The nurse cell liquid covering agrosebead protoplasts could promote cell division and significantly increase the frequency of cell division (33.2%) andcolony formation (19.0%). The hardening for protoplast-derived calli on the suitable proliferation media greatlybenefited green plantlet regeneration. In the course of callus hardening, L3 medium showed the highest efficiency (32.1%) in the formation of embryogenic calli and embryo-like structure, thus resulting in producing moregreen plants.