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从黑木耳(Auricularia auricula)种内杂交子H2J3的子实体上单孢分离培养得到F1代52个单核体菌株,将F1子代及亲本(H2,J3)等所有供试单核体菌株的菌丝体两两配对培养,采用核荧光染色观察核相,和光学显微镜观察锁状联合,鉴别菌丝交配反应是否形成双核体菌丝,同时测定了亲本菌株所有F1子代的交配型。根据交配型表型将F1子代菌株分成两群(F1-A1,F1-A2),将每群内各自菌株的DNA等量混合,构建交配型基因的等基因池,通过64个随机引物的RAPD分析,发现引物S126在2个等基因池间扩增出差异带S1261021,且在属于同一交配型的亲本及F1子代之间扩增结果基本一致,表明S1261021是与交配型基因连锁的分子标记。
Fifty-two mononuclear strains of F1 generation were isolated from the fruiting body of hybrid H2J3 in Auricularia auricula. F1 monokaryotic strains of F1 hybrids and their parents (H2, J3) The mycelia were pairwise cultured. The nuclear phase was observed by nuclear fluorescence staining and the lock-like combination was observed with optical microscopy to identify whether the mycelium mating reaction formed dinuclear mycelia. The mating type of all the F1 progenies of the parental strain was also determined. According to the mating phenotype, the F1 progeny strains were divided into two groups (F1-A1 and F1-A2), and the DNAs of the respective strains in each group were mixed in the same amount to construct the isogenic pool of mating genes. Sixty-four random primers The result of RAPD analysis showed that S126 was amplified in S1261021 between two isogenic pools, and the result of amplification between parents and F1 progenies belonging to the same mating type was basically the same, indicating that S1261021 is a molecule linked to the mating type gene mark.