摘要
选择"桂味"和"妃子笑"两个荔枝(Litchi chinensis Sonn.)品种的花药胚性细胞悬浮系来源的原生质体作融合亲本,在优化电融合参数(排列电压5 V,脉冲电压80 V,脉冲持续时间45～65μs,原生质体密度5.0×105～7.5×105个/mL,融合液Ca2+浓度0.2 mmol/L,甘露醇浓度10%～11%)条件下,可获得20%以上双元异核融合率。在亲本原生质体混合前,以0.02%中性红10 min染色"桂味"原生质体,有助于双元异核融合体的有效确认。
The protoplasts of lychee （Litchi chinensis Sonn.） cv. Guiwei and cv. Feizixiao isolated respectively from cell suspension cultures from anther-derived calli were used for the fusion. Above 20% binuclear heterokaryons frequency was achieved under following conditions： alternate current field strength of 5 V, direct current pulse strength of 80 V, pulse duration of 45～65 μs, protoplast density at 5.0×10^5-7.5×10^5 per mL, 0.2 mmoFL Ca2＋ and 10%-11% mannitol in electrical fusion solution. Binuclear heterokaryons frequency could be easily determined by detaining patent protoplast of lychee cv. Guiwei with 0.02% netural red for 10min before fusing. The results could be a reference for large-scale electrofusing and subsequent hybrid screening of litchi protoplast.
出处
《热带作物学报》
CSCD
北大核心
2012年第6期1030-1033,共4页
Chinese Journal of Tropical Crops
关键词
荔枝
原生质体融合
中性红
Lychee （Litchi chinensis Sonn.）
Netural red
Protoplast electrofusion
