斑玉蕈育种中漆酶转化体系建立的初步研究

漆酶具有降解木质素,氧化降解酚类物质,抑制杂菌,改善出菇品质等作用。采用酶学与原生质体融合相结合的技术,建立斑玉蕈育种中漆酶转化体系,将漆酶活性较高、生长速度较快的凤尾菇原生质体经高温灭活与斑玉蕈原生质体融合,通过RB-PDA平板显色技术筛选出具有漆酶活性较高的两融合菌株Ⅲ18C、Ⅲ2A,并对其进行了拮抗试验、RAPD分子标记、漆酶基因扩增等研究。结果表明,筛选出的两融合菌株与两亲本具有明显的拮抗线,随即引物扩增的条带与两亲本有明显的差异,并扩增出漆酶基因的一个片段。同时也表明利用漆酶转化体系筛选融合菌株具有目标明确、准确、快速的优点。

Laccase transformation system established in Hypsizigus marmoreus breeding

The laccase transformation system in Hypsizigus marmoreus breeding was established by enzymological technique in combination with protoplast fusion.Protoplast of Hypsizigus marmoreus and heat-treated protoplast of Pleurotus sajor-caju with faster growth and higher activity of laccase were fused by fusogen.Two fusion strains Ⅲ18C,Ⅲ2A were screened by the RB-PDA plate.Antagonistic test,RAPD-PCR and amplification of laccase gene were used to study the genetic stability of fusion strains and two parents.The results showed that there was an obvious antagonistic line between fusion strain and two parents.The fusion strain's amplification patterns of RAPD-PCR were different from parents,and a partial gene of laccase was amplified.Using laccase transformation system to screen fusion strains has shown an effective advantage.