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【目的】研究简单重复序列(Simple sequence repeat,SSR)分子标记方法用于香菇原生质体单核体、孢子单核体及其杂交后代的分离和鉴定。【方法】利用基于香菇全基因组序列信息开发的SSR标记,分析由香菇品种“L808”双核菌丝制备的原生质体单核体、孢子单核体及其杂交后代的SSR指纹。【结果】对制备的原生质体单核体的鉴定中,在不经过杂交配对的情况下,鉴定出“L808”的两种不同极性的原生质体单核体,其分离比例为191:1,该鉴定结果得到SSR标记、随机扩增多态性DNA(Random amplified polymorphismic DNA,RAPD)标记及传统方法的验证。另外,开发的香菇SSR标记还能以多位点组合的方式,用于对孢子单核体及其杂交后代的鉴定。【结论】应用SSR标记可加快香菇单核体的制备进程,并提高鉴定单核体及相关杂交菌株的准确性,促进香菇遗传育种研究。
【Objective】 The objective of this study was to investigate the isolation and identification of simple sequence repeat (SSR) markers for protoplast monokaryons, sporophyte monokaryons and their hybrid progeny. 【Method】 The SSR markers of protoplast, sporophyte mononuclear and their hybrid progenies prepared from the mushroom “L808” binuclear mycelium were analyzed by SSR markers based on the whole genome sequence of mushroom. 【Result】 The results showed that the protoplast monokaryons with different polarities of “L808” were identified in the identification of the protoplast mononuclear bodies without hybridization, with the separation ratio of 191: 1. The results of the identification were confirmed by SSR markers and random amplified polymorphismic DNA (RAPD) markers and the traditional methods. In addition, the developed SSR markers of shiitake mushrooms can also be used for the identification of sporophytic mononuclear bodies and their hybrid progeny in a multi-site combination manner. 【Conclusion】 The application of SSR markers can accelerate the preparation of monokaryons and improve the accuracy of identification of monokaryons and related hybrid strains, and promote the research of mushroom genetic breeding.