Table_6_De novo Sequencing and Comparative Transcriptome Analyses Provide First Insights Into Polysaccharide Biosynthesis During Fruiting Body Development of Lentinula edodes.XLSX

Polysaccharides separated from Lentinula edodes are well known for their medicinal properties. However, the precise molecular mechanisms of polysaccharide biosynthesis in L. edodes remain unclear. In this study, the fruiting bodies of L. edodes in four developmental stages with significant differences in polysaccharide yield were collected, and the characteristics of polysaccharides were studied. De novo sequencing and comparative transcriptomic analysis were performed by using high-throughput Illumina RNA-sequencing. KS1P30, KS2P30, KS3P30, and KS4P30 were obtained from the four developmental stages, respectively, by hot water extraction and 30% ethanol precipitation. These four polysaccharides had good immune activity in vitro; all of them were β-glucopyranose with a high molecular weight. Glucose was the main monosaccharide component of these polysaccharides. High-quality clean reads (57.88, 53.17, 53.28, and 47.56 million for different growth stages) and mapping ratios ranging from 84.75 to 90.11% were obtained. In total, 11,493 (96.56%) unigenes and 18,924 (97.46%) transcripts were successfully annotated in five public databases. The biosynthetic pathway and related genes of LEFP30 were mined. The molecular mechanism of LEFP30 yield change in the different developmental stages was predicted. The results provide some insights into the possible mechanisms involved in the biosynthetic pathway of this kind of polysaccharide in L. edodes fruiting bodies. They also indicate that candidate genes can be used as important resources for biotechnology and molecular breeding to regulate L. edodes fruiting body polysaccharide biosynthesis.

从香菇（Lentinula edodes）中分离出的多糖因其药用特性而广为人知。然而，香菇中多糖生物合成的精确分子机制尚不明确。在本研究中，收集了四个发育阶段香菇子实体，这些阶段在多糖产量上存在显著差异，并研究了多糖的特性。通过高通量Illumina RNA测序技术，对这四个发育阶段的香菇子实体进行了从头测序和比较转录组分析。分别从四个发育阶段中通过热水提取和30%乙醇沉淀获得了KS1P30、KS2P30、KS3P30和KS4P30。这四种多糖在体外表现出良好的免疫活性；它们均为高分子量的β-吡喃葡萄糖。葡萄糖是这些多糖的主要单糖组分。获得了高质量的清洁读段（不同生长阶段的数量分别为5788万、5317万、5328万和4756万），以及从84.75%到90.11%的映射率。在五个公共数据库中，共成功注释了11,493（96.56%）的unigenes和18,924（97.46%）的transcripts。挖掘了LEFP30的生物合成途径和相关基因。预测了LEFP30在不同发育阶段产量变化的分子机制。研究结果为揭示香菇子实体中此类多糖生物合成途径中涉及的潜在机制提供了一定的见解。此外，候选基因可作为生物技术和分子育种中调节香菇子实体多糖生物合成的重要资源。