Breeding and Gene Manipulation for the Sawdust-Based Cultivation of Lentinula edodes

以木屑为基质培养香菇，研究其出菇过程中漆酶和纤维素酶mRNA的转录调控。（材料与方法）1。本研究共检测了12个菌种。KS-3、-4、-16、-23、-46、-56、-58、-60、-67和-76（宽气候品系），KS-12和-24（寒冷气候品系）。锯末基基质培养按照标准方法进行。在7个不同生长阶段测定了漆酶和纤维素酶的活性。生长期分为菌丝生长期（90 d）、聚集期（94 d）、针形期（98 d）、扣眼期（100 d）、花被期（102 d）、衰老期（104 d）和二次开花期（140 d）.通过浸泡处理调整木屑基质的水分状况。水分状况通过水势（φ）值进行评价.用RT-PCR方法检测漆酶和纤维素酶基因的转录调控。（结果）胞外酶 关于我们 以香菇（Lentinulaedodes）为试材，研究了不同生长阶段漆酶和纤维素酶的活性及其转录调控。漆酶和纤维素酶活性的变化与子实体的发育阶段有明显的关系。漆酶和纤维素酶活性在基因转录水平上受到调控。漆酶mRNA的表达水平在完全定殖阶段达到最高，在子实体发育过程中逐渐下降。纤维素酶mRNA在子实体形成期开始积累。纤维素酶mRNA转录本在子实体发育的幕裂阶段表达最多。这种趋势是明确的，在与广泛的气候菌株的出菇文化比非出菇寒冷的天气菌株。漆酶和纤维素酶基因的转录也受到木屑基基质的水分条件的影响。原基起始发生时，培养基的水势高，快速mRNA转录的菌丝体。少

Transcriptional regulation of laccase and cellulase mRNA were investigated for fruiting on the sawdust-based cultivation of Lentinula edodes.(Materials and Methods)1. Twelve spawn strains were examined in this study. KS-3, -4, -16, -23, -46, -56, -58, -60, -67, and -76 (wide-weather strains), KS-12 and -24 (cold weather strains).2. The sawdust-based substrate cultivation was done according to the standard methods. Activities of laccase and cellulase were measured at the 7 different growth stages. The growth stages were defined to mycelial growth (90-days old), aggregate (94-days old), pin (98-days old), button (100-days old), veil break (102-days old), senescent (104-days old), and second flush (140-days old).3. Moisture condition of the sawdust-based substrate was adjusted soaking treatment. The moisture condition was evaluated by the water potential (φ) values.4. Transcriptional regulation of laccase and cellulase genes was measured by the RT-PCR method.(Results)Extracellular enzyme … More activities of laccase and cellulase and their transcriptional regulation were investigated at various growth stages in a sawdust-based substrate for Lentinula edodes. Changes of laccase and cellulase activities revealed a clear relationship with fruit body development stages. Laccase and cellulase activities were regulated at the level of gene transcription. The level of laccase mRNA was maximal at the fully colonized stage and declined during fruit body development. Cellulase mRNA began to accumulate at the pin (miniature fruit bodies) formation stage. Cellulase mRNA transcripts were maximally expressed at the veil-break stage of fruit body development. This tendency was clearer in the fruiting cultures with the wide-range-weather strains than in non-fruiting cold-weather strains. Transcription of laccase and cellulase genes was also affected by the water conditions of the sawdust-based substrate. Primordia initiation occurred when the water potential of the medium was high for rapid mRNA transcription by the mycelium. Less