利用原生质体技术构建双孢蘑菇杂交菌株

Construction of Agarrcus bisporus Hybrids by Protoplast Techniques

利用原生质体技术，从异核菌丝出发构建了双孢蘑菇的杂交菌株。蘑菇原生质体制备与再生的最佳条件是：异核菌丝在ＰＳＬ培养液中培养５天，菌丝经１．５％溶壁酶、０．６ＭＫＣｌ酶解数小时，纯化后在０．０１％琼脂－ＲＳＰＭ中预培养２０—２４小时，涂布到０．１８％琼脂ＲＳＰＭ上再生。根据再生菌落的形态推定出同核体与异核体，经栽培验证，继而将不孕的推定同核体配对。根据亲和性确证同核体，并分离异核化杂交种。研究结果表明：原生质体杂交育种比常规杂交育种具一定优越性；提出同核化是蘑菇原生质体制备过程中的特征之一。

The routine hybridization of Agaricus bisporus (L.)Imbach is difficult for its secondary homothallism.The optimum conditions of preparation and regeneration of protoplasts of this fungus were studied in this research. Protoplasts were prepared and then regenerated from heterocaryotic mycelia.Some regenerates were presumed to be homocaryotic based upon the morphology and growth rate of the colonies, and then verified by cultivation fructification. They were further confirmed to be authentic homocaryons based upon their pairing behaviors, and nearly 30 hybrids were eventually obtained. It is suggested that the protoplast hybridization method is superior to the routine ones to some extent.