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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 has degradation of lignin, oxidative degradation of phenolic compounds, inhibit bacteria, improve fruiting quality and so on. The combination of enzymolysis and protoplast fusion technology was used to establish the transformation system of laccase in P. edulis breeding. The protoplasts of Pleurotus ostreatus which had higher laccase activity and faster growth speed were inactivated by high temperature, The two fusion strains Ⅲ18C and Ⅲ2A with higher laccase activity were screened by RB-PDA plate colorimetric technique. The antagonistic test, RAPD molecular marker and laccase gene amplification were also carried out. The results showed that the two confluent strains screened out antagonistic lines with the amphiphile, then the bands amplified by the primers were significantly different from those of the parents, and a fragment of the laccase gene was amplified. At the same time, it also showed that the selection of fusion strains by laccase transformation system has the advantages of clear, accurate and rapid target.