金针菇成熟期和伸长期菌盖的转录组与蛋白组比较分析

Comparison of transcriptomes and proteomes in pilei between maturation and stipe elongation stages of Flammulina filiformis

菌盖是大型真菌的重要组成部分,也是其产生有性孢子的部位,但是其发育机制仍不明确。本研究以金针菇Flammulina filiformis为材料,采用转录组和蛋白组联合分析的方法,比较分析了金针菇成熟期和伸长期菌盖的差异基因与蛋白,并对其进行GO(gene ontology)功能聚类分析、KEGG(Kyoto encyclopedia of genes and genomes)富集分析和蛋白互作网络分析。本研究筛选到差异表达基因有1391个,差异表达蛋白147个,均以上调表达为主。GO功能聚类分析结果表明,催化活性(catalytic activity)条目富集基因最多,其次是细胞组分(cell part)、细胞过程(cellular process)和细胞器(organelle)。KEGG富集分析结果表明,差异表达基因和蛋白主要富集在碳水化合物代谢通路(carbohydrate metabolism)和氨基酸代谢通路(amino acid metabolism)等。本研究选取了9个关键的差异表达基因,使用实时荧光定量PCR(real-time quantitative PCR,RT-q PCR)对其表达量进行了验证。RT-qPCR验证结果与转录组测序结果相一致。蛋白互作网络分析表明,水解酶类、结构域类和转录调节类蛋白为互作网络的主要结点。本研究联合转录组、蛋白组测序数据,通过分析差异基因与蛋白,为深入了解金针菇菌盖发育机制提供数据参考。

The pileus is an important part of macro-fungi where spores are produced,but the mechanism of its development remains unclear.The differentially expressed genes(DEGs)and differentially expressed proteins(DEPs)of the pileus at maturation and stipe elongation stages of Flammulina filiformis were compared and analyzed with the conjoint method of transcriptome and proteome on the basis of gene ontology(GO),Kyoto encyclopedia of genes and genomes(KEGG)database and search tool for the retrieval of interacting genes(STRING)database.1391DEGs and 147 DEPs were obtained,most of which were up-regulated.GO analysis showed that most DEGs were clustered in catalytic activity term,secondly cellular components,cellular processes and organelles.Most DEGs and DEPs were mainly clustered in carbohydrate metabolism and amino acid metabolism according to analysis based on KEGG database.Nine key DEGs were selected and the expression patterns were verified by using real-time quantitative PCR(RT-qPCR).The results of RT-qPCR are consistent with those of transcriptome sequencing.The protein-interaction analysis based on STRING database showed that hydrolase,domain-like proteins and transcriptional regulation proteins were the main nodes of the protein-interaction network diagram.DEGs and DEPs obtained and analyzed by transcriptome and proteome sequencing provide data reference for the further study of the developmental mechanism of F.filiformis.