李婷, 姜丹, 胡小松, 常晓茜, 许贞, 巩颖, 华国栋, 刘春生. 基于高通量测序技术对市售柏子仁表面真菌多样性的研究J. 药学学报, 2019,54(11): 2100-2105. doi: 10.16438/j.0513-4870.2019-0512
引用本文: 李婷, 姜丹, 胡小松, 常晓茜, 许贞, 巩颖, 华国栋, 刘春生. 基于高通量测序技术对市售柏子仁表面真菌多样性的研究J. 药学学报, 2019,54(11): 2100-2105. doi: 10.16438/j.0513-4870.2019-0512
LI Ting, JIANG Dan, HU Xiao-song, CHANG Xiao-xi, XU Zhen, GONG Ying, HUA Guo-dong, LIU Chun-sheng. Study on the surface fungus diversity of commercially Platycladi semen based on high-throughput sequencing technologyJ. Acta Pharmaceutica Sinica, 2019,54(11): 2100-2105. doi: 10.16438/j.0513-4870.2019-0512
Citation: LI Ting, JIANG Dan, HU Xiao-song, CHANG Xiao-xi, XU Zhen, GONG Ying, HUA Guo-dong, LIU Chun-sheng. Study on the surface fungus diversity of commercially Platycladi semen based on high-throughput sequencing technologyJ. Acta Pharmaceutica Sinica, 2019,54(11): 2100-2105. doi: 10.16438/j.0513-4870.2019-0512

基于高通量测序技术对市售柏子仁表面真菌多样性的研究

Study on the surface fungus diversity of commercially Platycladi semen based on high-throughput sequencing technology

  • 摘要: 为了分析市售柏子仁表面真菌群落的结构组成,揭示其表面真菌生物多样性和结构差异。收集了河南、山东、香港3个地区的柏子仁药材,对其进行DNA抽提,并对DNA中ITS片段进行PCR扩增,PCR产物经质检合格后进行Illumina Hiseq 2500平台上Miseq测序,得到序列OTU聚类后进行生物信息学分析。三个地区的柏子仁样品肉眼均观察不到微生物群落,测序结果显示其表面微生物群落均具有较高的生物多样性,但物种组成上存在明显差异。7份柏子仁样品得到345 947条有效序列,划分为267个OTU聚类,分3门,18纲,40目,82科,120属,191种真菌。在属水平上,曲霉属丰度占比最高,达到(93.36±6.01)%,为优势菌属。7份样品均被黄曲霉菌污染,污染程度分别为14.58%、15.98%、17.64%、16.44%、0.97%、23.39%、18.86%。除5号样品筛曲霉丰度占比较高,其余6份样品均以黑曲霉、烟曲霉以及黄曲霉为核心菌群。通过分析不同产地柏子仁真菌分布的多样性,充分了解了柏子仁表面真菌的情况,为黄曲霉菌的污染及其毒素污染进行早期的风险预警奠定基础,为柏子仁质量安全提供理论依据。

     

    Abstract: The structural composition of the surface fungal community of commercially Platycladi semen was analyzed to reveal the surface fungal biodiversity and structural differences. Platycladi semen was collected from Henan, Shandong and Hong Kong, their DNA was extracted, ITS fragments in DNA were amplified by PCR. Miseq was sequenced on Illumina Hiseq 2500 platform after the PCR products were qualified for quality inspection. The sequence OTU cluster was obtained and bioinformatics analysis was carried out. Microbial communities were not observed in the eyes of the Platycladi semen in the three regions. Sequencing results showed that the surface microbial community had high biodiversity, but there were significant differences in species composition. Seven samples o Platycladi semen obtained 345 947 valid sequences, which were divided into 267 OTUs, 3 phylums. 18 classes, 40 orders, 82 families, 120 genus, 191 species fungi. At the genus level, Aspergillus is the dominant species, accounting for the highest proportion, reaching (93.36 ±6.01)%. Seven samples were contaminated by Aspergillus flavus, and the pollution levels were 14.58%, 15.98%, 17.64%, 16.44%, 0.97%, 23.39% and 18.86%. Except sample No. 5, Aspergillus cibarius was the most abundant, the other six samples were Aspergillus niger, Aspergillus fumigatus and Aspergillus flavus as the core microflora. By analyzing the diversity of fungi distribution in different habitats, we can fully understand the fungi on the surface of Platycladi semen, lay a foundation for early risk warning of Aspergillus flavus contamination and its aflatoxin contamination, and provide a theoretical basis for the quality and safety of Platycladi semen.

     

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