大豆幼荚子叶原生质体培养及植株再生

Protoplast Culture and Plant Regeneration of Immature Cotyledons of Soybean (Glycine max L.)

本文研究了13个栽培大豆(Glvcine max L.)品种原生质体培养的再生能力。从大豆幼荚子叶酶解游离原生质体,用Gellan Gum进行珠状包埋,悬浮在含2,4-D 0.1-0.2mg/L,BA0.5-1.0mg/L的改良MS液体培养基中,原生质体培养3天后开始第一次分裂,以后持续分裂。供试基因型间的10天植板率差异显著,变幅为33-67％。30天内形成大量的细胞团,各供试基因型50-60天内都能形成1-2mm大小的愈伤组织。把这种小愈伤组织转到含2,4-D 0.3mg/L,BA0.5mg/L的MSB固体培养基上,促使其进一步生长。再转入附加NAA 5.0mg/L,BA0.5mg/L,KT0.5mg/L和3％蔗糖的MSB分化培养基上,在黄色脆硬的瘤状愈伤组织表面可分化出胚状体,胚状体在含NAA 1.0mg/L和KT0.5mg/L的MSB培养基上可发育成再生植株。供试品种泗豆11号和铁丰8号都已得到再生植株。

Thirteen soybean (Gjiycme max L.) genotypes were evaluated for their regenerability from protoplasts. Protoplasts were isolated from immature cotyledons, buried in beads solidified with Gellan Gum, and cultured in modified MS liquid medium containing asparagine, glutamine, allantoin, and allantoic acid, supplemented with 0.1-0.2 mg/1 2, 4-D and 0.5-1.0 mg/1 BA. The protoplasts started to divide after 3 days of culture. Significant differences were observed in plating efficiency, which were varied from 33-67%. Sustained divisions resulted in mass produc-tion of cell colonies after 30 days of culture. 1-2 mm diameter colonies were formed in 50-60 days with all the genotypes tested. The colonies were then transferred onto MSB (MS salts+B5 or-ganics) medium with 0.3mg /l 2, 4-D and 0.5 mg / l BA for further growth. Once the callus had become compact and nodular, they were transferred to MSB regeneration medium with NAA 5.0 mg /l, 0.5 mg /l each of BA and KT, and 3% sucrose, then embryo could be differentiated from the callus upon regular subculturing, Embryo developed into plantlet on MSB medium with 1.0 mg /l NAA and 0.5mg /l KT. Plantlets have been regenerated from cultivars of Sidou No. ll and Tiefeng No.8.