丝状蓝藻原生质球的释放及再生研究

Investigation on the Spheroplast Liberation and Regeneration of Filament in Cyanobacteria Phormidium luridum

以丝状蓝藻 Phormidium luridum 为材料,pH6.8磷酸缓冲液中加入溶菌醉(0.05％,W/V),于 35℃保温2 h,获得了有再生力的原生质球,新生的原生质球 24 h内能重建胞壁和片层结构;48 h细胞开始分裂;3～6 d后在平板上出现有厚胶质鞘细胞群;10 d再生成丝状藻体;15 d后形成肉眼可见的藻落,实验证明,BG-11培养基内附加维生素B1、维生素B12、维生素B6、吲哚丁酸、甘露醇(或山梨醇)

Conditions for liberating spheroplast and regenerating new filament in Plurmiditm lstidttmwere investigated. It has been shown that spheroplast can be liberated easily from mother filament by 2h digestion with 0. 05% (w/v) fyaozyme m phosphate buffer pH 6. 8 at 35℃. On a suitable culture plafe depicted below, (he fresh sphetoplast would resume its cell wall and internal lamella structure in 24h, launch into cell divisionin 48 h, reform a jelly sheath around the newiy formed cell colony in 72 h, regenerate a new filament and grow up a filamentous mass on the following days. It was found that rate of algal multiplication is promoted by addition of vitamin B,, B12, B6, indole 3-butyric acid to BG-ll in the dose of 100. 5, 100 and 500 μg/l6,respecively. Besides these, addition of manniitol or sorbitol in 45 g/1, promotes the rate conspicuously while cane sugar is a powerful retardant.