水稻(Oryza sativa)原生质体经钝化后诱导融合再生可育体细胞杂种

本研究在初步实现水稻原生质体培养的程序化后,选用普通栽培稻P339和特种稻苏御糯选的原生质体为融合亲本,利用碘乙酸(IA)和罗丹明-6G(R-6G)这两个代谢互补抑制剂钝化处理亲本原生质体,确定了合适的抑制条件。P339用0.25mmol/L IA,苏御糯选用50μg/ml R-6G分别经30min钝化处理,通过PEG和高Ca~(2 )、高pH法诱导融合,异源融合体具有代谢互补效应,经培养得到愈伤组织17块,并进一步分化获得不同形态的再生植株12棵。移栽存活的再生植株成熟后可育,通过对这些植株的形态以及酯酶和过氧化物酶同工酶电泳的分析表明是融合后的体细胞杂种植株。

REGENERATION OF RICE (Oryaz Sativa) FERTILE SOMATIC HYBRIDS AFTER PROTOPLAST INACTIVATION AND FUSION

After the schedulization of rice protoplast culture was primarily achieved, the P339 and ShuYuNuoXuan were selected as fusion partners. When rice protoplasts were treated with either IA (Iodoacetic acid ) or R-6G (Rhodamine 6-G) and then cultured in vitro, their growth were inhibited, but the heterofused protoplasts were metabolically complemented. The P339 protoplasts were treated with 0. 25 mmol/L IA, the ShuYuNuoXuan protoplasts with 50 fig/ml R-6G, for 30min before fusion. Using PEG6000 and high Ca2-,high pH solution to induce proto- plast fusion, we obtained 17 regenerated cell colonies in liquid thin layer culture and 12 regenerated plants in differentiation media. The genetic characters of the plants regenerated from fused protoplasts and parent protoplasts were compared. It showed that the plants regenerated from fused protoplasts were different from parent plants in morphological characters. The isozyme zymograms of peroxidase of the regenerated plants and esterase of seeds of the hybrid plants were different from those of the control plants. The fertile somatic hybrids of rice were obtained.