食品科学 ›› 2019, Vol. 40 ›› Issue (10): 134-140.doi: 10.7506/spkx1002-6630-20180529-402

• 生物工程 • 上一篇    下一篇

扩增区域对鲊广椒细菌MiSeq测序的影响

王玉荣,杨成聪,葛东颖,尚雪娇,张振东,郭 壮*   

  1. 湖北文理学院食品科学技术学院,鄂西北传统发酵食品研究所,湖北 襄阳 441053
  • 出版日期:2019-05-25 发布日期:2019-05-31
  • 基金资助:
    湖北文理学院教师科研能力培育基金项目(2017kypy051)

Influence of Different Amplified Regions on Results of Bacterial Diversity in Zhaguangjiao, a Chinese Traditional Fermented Chili Product, by MiSeq Sequencing

WANG Yurong, YANG Chengcong, GE Dongying, SHANG Xuejiao, ZHANG Zhendong, GUO Zhuang*   

  1. Northwest Hubei Research Institute of Traditional Fermented Food, College of Food Science and Technology, Hubei University of Arts and Science, Xiangyang 441053, China
  • Online:2019-05-25 Published:2019-05-31

摘要: 在使用引物27F/338R、338F/806R和515F/907R分别对细菌16S rRNA基因的V1~V2区、V3~V4区和V4~V5区进行扩增的基础上,采用MiSeq高通量测序评价扩增区域对鲊广椒细菌多样性分析结果的影响。结果发现,扩增16S rRNA基因V4~V5区的非特异性序列所占比例显著偏低(P<0.05),而扩增V1~V2区样品细菌的Chao1指数显著偏高(P<0.05)。虽然扩增16S rRNA基因的V3~V4区会导致Cyanobacteria(蓝细菌门)和Pediococcus(小球菌)相对含量显著偏高(P<0.05),但不同扩增区域扩增出的同一样品其微生物群落结构无显著差异(P>0.05)。在对鲊广椒细菌多样性进行解析时,建议选择引物515F/907R扩增16S rRNA基因的V4~V5区。

关键词: 鲊广椒, 引物, 扩增区域, MiSeq测序, 细菌多样性

Abstract: This study aimed to evaluate whether different amplified regions of 16S rRNA gene influence the results of bacterial diversity in Zhaguagnjiao by MiSeq sequencing. The V1–V2, V3–V4 and V4–V5 regions were amplified using bacterial genomic DNA extracted from Zhaguangjiao samples as template with three different primer pairs, 27F/338R, 338F/806R and 515F/907R. The results indicated that the percentage of non-specific amplification sequences were significantly lower in the amplified products of V4–V5 region (P < 0.05), and the Chao 1 index was significantly higher in the amplified products of V1–V2 regions (P < 0.05). Although the relative abundance of Cyanobacteria and Pediococcus were significantly higher in the amplified products of V3–V4 region (P < 0.05), there was no significant difference in the microbial community structure of the same samples with different amplified variable regions (P > 0.05). Thus, amplification of the V4–V5 region using primer pair 515F/907R is recommended for analysis of the bacterial diversity of Zhaguangjiao.

Key words: Zhaguangjiao, primer, amplified regions, MiSeq sequencing, bacterial diversity

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