摘要
采用电激细胞融合的方法 ,对紫罗兰MatthiolaincanaR Br与桂竹香CheiranthuscheiriL 原生质体培养及电激细胞融合效率导入最适条件作了探讨。紫罗兰的子叶原生质体分裂初期培养最适条件 :培养基为 1 2MS。培养基组成中 ,蔗糖质量分数为 4%、甘露糖醇浓度为 0 6mol L。加入谷酰胺酶无机态氮源时 ,效果明显。次之 ,加入植物生长调节物质 1mg LBA也达到高分裂率。再者 ,添加玉米素时效果也甚好。根据原生质分离前的子叶在 5℃预处理分裂率最高 ,培养温度 2 4～ 30℃为宜。电激细胞融合 (singlepairs)产生效率高的条件为 :紫罗兰原生质体密度 1× 10个 mL ,高周波电强度 1MHz ,15 0V cm ,脉冲幅 70 μs;桂竹香原生质体密度 1× 10个 mL高周波电强度 1MHz,10 0V cm ,脉冲幅 30 μs。
Through the electrical cell fusion of the stock and the wallflower, t he optimum conditions of the early-period cultivation of protoplast and the opti mum conditions which lead effectively to cellular fusion. The results were as fo llows: As a condition of early-period cultivation that increases the separati on of seedling protoplast of the stock, by cultivation in a basic culture medium that made up 1/2 MS, a sucrose concentration of 4% and a mannitol concentration of 0.6%, it was the most appropriate. In addition, when adding inorganic nitr ogen to glutamine, it got an excellent result. And when adding plant-growth regu lators BA as 1 mg/L it achieved a high rate of separation. Plus, the effects of th e addition of zeatin (maize factor) was good. Due to pre-treatment of seedlings prior to protoplast separation, the rate of separation achieved was high. The su itable temperature ranged from 24 ℃ to 30 ℃. The conditions which gave rise to a high proportion of single pairs through the medium of electrical cell fusi on were stock protoplast density of 1×105, high electrical field intensity of 1 MHz, the pulse voltage in 1.5 kV/cm and pulse width in 70 s;wallflower prot oplast density of 1×105 units/mL, high electrical field intensity of 1 MHz, 1 00 V/cm the pulse voltage in 2.0 kV/cm and pulse width in 30 μs.
出处
《中山大学学报（自然科学版）》
CAS
CSCD
北大核心
2003年第3期64-68,共5页
Acta Scientiarum Naturalium Universitatis Sunyatseni
基金
广东省自然科学基金资助项目 ( 980 737)
