提取曲拉样品的DNA,选择合适的通用引物扩增ITS特定区域,采用Illumina高通量测序技术检测目标区域的序列变异和丰度,通过分析曲拉样品中真菌的Alpha多样性、物种分类组成及Beta多样性,以揭示甘南牦牛曲拉中真菌菌群的群落结构。结果表明,曲拉中具有高度的真菌多样性,并且不同来源样品的Shannon指数、Simpson指数、Chao1指数和ACE指数存在差异;在曲拉样品中共检测出6个门和123个属的真菌,所有样品中的共有优势门为子囊菌门(Ascomycota),共有优势属为毕赤酵母属(Pichia)和双足囊菌属(Dipodascus);采用非度量多维尺度分析和非加权组平均法聚类分析可以将曲拉样品分为3类,结果与样品来源一致;置换多元方差分析表明,不同来源的样品间真菌组成差异显著(P=0.001);Spearman关联网络图表明,样品中真菌群落之间正相关占主导地位;研究结果可为牦牛曲拉的品质控制奠定基础。
This study analysed the fungi microbiota structure of Qula, a kind of fermented Yak milk product, using high throughput sequending technology. After extraction the metagenome DNA of Qula sample, the universal primers of fungi ITS marker gene were amplied and sequenced using Illumina high throughput sequencing technique. And the variation of microbiota structure and abundance were conducted by analyzing the Alpha diversity, species composition and Beta diversity of fungi in yak Qula samples from Gannan. The results showed that there was a high degree of fungal diversity in different Qula samples indicated in Shannon index, Simpson index, Chao1 index and ACE index. A total of 6 phyla and 123 genera of fungi were detected in the Qula samples. The common dominant phylum in all samples was Ascomycota, and the common dominant genera were Pichia and Dipodascus. The non-metric multidimensional scaling analysis and the unweighted group averaging clustering analysis were used to classify the Qula samples into three categories which were consistent with the sample sources. Adonis/PERMANOVA multivariate analysis of variance and replacement test showed that there were significant differences in fungal composition between samples from different sources (P=0.001). The Spearman correlation network diagram showed that there were positive correlation between dominant fungal communities. This study laid the foundation for the quality control of yak Qula.
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