马铃薯原生质体融合技术的研究

Studies on the Techniques of Protoplast Fusion in Potato

采用聚乙二醇（ＰＥＧ）、电融合和ＰＥＧ－电融合三种诱导融合的方法，对马铃薯双单倍体品系８２－３６和二倍体野生种Ｓｏｌａｎｕｍｍｉｃｒｏｄｏｎｔｕｍ无菌苗叶肉原生质体的融合技术进行了研究，建立了三种融合方法的适宜条件。结果表明，使用镀金平行多电极的电融合法融合效果最好，在交流场强１５０Ｖ／ｃｍ、正弦波频率１．０ＭＨＺ和直流脉冲场强１．２ＫＶ／ｃｍ、宽幅６３７．５μｓ下，施加１次脉冲，原生质体的融合频率可达３０％。在对融合细胞的培养中发现：经过异质融合处理后的原生质体形成的愈伤组织具有较强的分化能力，其中有少数愈伤组织再生出了绿色植株。

The in vitro shoots of a dihaplid strain 82～36 from anther culture of Solanum tuberosum and a diploid wiod species S.microdontum were used as the donors.Three protoplast fusion methods were studied including polyethylene glycol(PEG) fusion ,electro fusion and PEG electro fusion.The optimal fusion conditions were found.The results showed that the best method was electro fusion with a gold plating parallel multi electrode in all the experiments.Using the electrode and under the treatment with 1.0 MHz frequency and 150V/cm alternating current(AC) field intensity followed by 1.2 KV/cm,637.5 μs direct current (DC) sharp wave pulse once,the protoplast fusion frequency could reach 30%.It was observed in the culture of fusion products that calli derived from heterogeneous fused cells had higher frequency of shoot regeneration,which could form green plantlets.