黑木耳种内杂交子的鉴定技术*

采用原生质体技术,获得黑木耳(Auricularia auricula)栽培菌株He-1的单核化菌株H1、H2、H3和栽培菌株Ju-1的单核化菌株J1、J2、J3,将H1、H2、H3分别与J1、J2、J3配对杂交,核相观察确认H2J1、H2J2和H2J3均为双核体。酯酶同工酶分析表明,H2J1、H2J2和H2J3不仅具有相应的亲本单核体共有的酶带,而且具有两个亲本各自的特异性标记酶带。RAPD分析表明,引物S30和S62对杂交子H2J1、H2J2和H2J3的扩增图谱中不仅包含相应的亲本单核体所共有的DNA带,而且包含亲本单核体各自的特异性DNA带。拮抗和栽培试验表明,杂交子H2J1、H2J2、H2J3与双核体亲本He-1和Ju-1的菌落之间有窄细的黑色拮抗线,子实体形态上有较明显差异。

IDENTIFICATING TECHNIQUE OF INTRASPECIFIC HYBRIDS IN AURICULARIA AURICULA

Protoplasted monocaryons H1,H2 ,H3, J1, J2 and J3 were obtained from cultivated strain He-1 and Ju-1 of Auricularia auricula respectively by protoplasting technique.Hybridazation tests were carried out by mycelial mating between H1,H2 ,H3 and J1, J2 , J3 respectively. H2J1, H2J2 and H2J3 were confirmed as dicaryons by means of nuclear fluorescence staining. The result of esterase isozyme analysis showed that hybrids H2J1, H2J2 and H2J3 had not only the common isozyme bands of their corresponding monocaryotic parents, but also the specific ones from its parents. The RAPD analysis using 20 arbitrary primers demonstrated that the amplified patterns of hybrids H2J1, H2J2 and H2J3 included not only the common DNA bands of their corresponding monocaryotic parents, but also the specific ones from its parents.Antibiosis and cultivation tests showed that there were obvious antibiotic reaction and fruitbody morphological difference between every hybrid and its dicaryon parent. In a word, hybrids H2J1, H2J2 and H2J3 are genuine hybrids.