Breeding of edible mushroom using cell fusion technique

在为电灌注技术提供原型适合物，并具有良好的高可用性，在3小时内观察到氢化反应时，在30 ° C在3 - 5天内使用酶解决方案，用于微波培养3 - 5天。该溶液含有2%的纤维素酶、0.6%的氧化酶和6个单位的钛酶，因此，适当地。原塑料的产量超过10/毫升，平均平板的效率接近20%。对原塑料的超结构的调查显示，那些与细胞塑料有一个密集的集中的原塑料，具有较高的平板效率。在聚变条件下，获得了最佳结果，将0.5mM氯化镁和0.1mM氯化钙转化为0.5M氯化钠溶液，脉冲长度和电压设置为40秒，4 kV/cm，响应。而且这给了大约60%的融合率。然而，进一步的研究仍将通过监管来进行降解，以防止聚合的原塑料在一个高频率电场中被添加的electrolyte引起，以及关于在高频率再生中增加的高频率电压的影响，等等。同位素分析的研究展示了一个特定的同位素模式可以被观察到esterase和malate脱氢酶，这是一个有效的选择指数，以筛选出体杂交瘤。“有约束力的标记者的发展，就像一种辅助性突变体一样，也在某种程度上受到怀疑。筛选测试现在已经被运送出来了，尽管还没有新的异源性混合体出现过。

Optimum result in obtaining protoplast suitable for an electrofusion technique and having a high viability as well,was observed when hydrolysis was conducted for 3 hours at 30゜C in an enzyme solution on mycelia cultured for 3 to 5 days in soyonion medium. The solution contained 2% of cellulase, 0.6% zymolyase, and 6 units of chitinase, respectively. The yield of protoplast was over 10/ml, the average plating efficiency being approximately 20%. Investigation of the ultrastructure of the protoplast showed that those dikaryon protoplasts with a dense concentration of cytoplasm, had a higher plating efficiency. In terms of fusion conditions,the best result was obtained by adding 0.5mM magnesium chloride and 0.1mM calcium chloride to a 0.5M mannitolsolution, with the pulse length and voltage set at 40 sec, and 4kV/cm, respectively. And this gave a fusion rate of approximately 60%. However, further investigations have yet to be conducted with regard to the decrease in aggregation of protoplasts in a high frequency electric field caused by the addition of electrolyte, and on influences of increased high frequency voltage on the hyphae regeneration, etc. Study of isozyme analysis exhibited that a specific isozyme pattern could be observed for esterase and malate dehydrogenase, thus constituting an efficient selection index for the screening of somatic hybridoma. The development of strains having markers, such as auxotrophic mutants is also under consideration. Screening tests are now being carried out although as yet no new somatic hybridoma have been obtained.