摘要
以内蒙古锡林郭勒镶黄旗呼日敦高勒嘎查牧场内牛、山羊、绵羊、马、人的粪便和土壤为研究对象,利用Pac Bio SMRT测序平台对细菌16S rRNA基因全长进行测序,运用生物信息学分析方法对人畜肠道微生物及土壤微生物多样性进行全面评估.结果发现,人畜肠道菌群均以厚壁菌门和拟杆菌门为优势菌门,而土壤以酸杆菌门和变形菌门为优势菌门.在属水平,人畜以梭菌属、拟杆菌属为优势菌属,而土壤以Blastocatella和芽孢杆菌属为优势菌属.在种水平人以Escherichia/Shigella dysenteriae和Streptococcus salivarius为优势菌种,而4种家畜的优势菌种均为Oscillibacter valericigenes和Eubacterium coprostanoligenes.土壤以Blastocatella fastidiosa和Bacillus longiquaesitum为优势菌种.比较各样品α多样性发现,人肠道微生物的多样性显著低于其他食草动物.通过主坐标和层次聚类分析发现可将6种样品分为:组1(人)、组2(马)、组3(绵羊、牛、山羊)和组4(土壤)4组.本研究对差异OTU进行比对分析探究了造成该分组的原因.因此本研究揭示了土壤、人和家畜肠道微生物组成和结构存在显著的差异,为进一步研究不同生境中微生物多样性提供数据参考.
As an important part of ecosystem, microbes are widely distributed in various habitats. In recent years, more and more attention has been paid to the study on gut microbiota. The gut microbiota and their metabolites influence human and animal nutrition processing, metabolic balance, immune function, gastrointestinal development and other physiological activities. With the deepening of studies on human and animal gut microbiota, it has been found that some factors, such as diet, age, gender, and living environment, impacting on the composition of gut microbiota, while the differences among animal species have more significant influence on the gut microbiota composition. In relatively primitive grassland ecosystems, soil microbes interact with human and animal activities. On the one hand, microbes in the soil environment are the driving forces for the transformation and circulation of organic matter and nutrients. The improvement of soil microbial community diversity is beneficial for the soil fertility. On the other hand, human and animal activities will affect the diversity of soil microbial community. Although there are a lot of researches on soil microbiota, animal and human gut microbiota, their differences in diversity and composition within the same environment have not been studied. The advent of sequencing technology provides an effective mean for the accurate and comprehensive understanding of microbes, especially for the study of uncultivable microorganisms. The PacBio single-molecule real-time(SMRT) technology is advantageous in producing long sequence reads with high accuracy. Based on sequencing the full length 16 S rRNA genes, the microbiota composition can be identified to the species level. Therefore, it is an effective approach for studying microbial diversity. We collected 56 stool and soil samples from Xilinguole, including 6, 10, 11, 9, and 10 stool samples from human, goat, cattle, horse, and sheep, respectively, as well as 10 soil samples. Genomic DNA was extracted from the samples. After DNA extraction and quality check, the 16 S rRNA genes of all samples were amplified from the genomic DNA. The PCR products were sequenced using the PacBio RS II instrument. The QIIME software(V1.7) was used to analyze the sequencing data, and the R software(version 3.5.0) was used to further analyze and visualize the results. Firstly, it was found that the gut microbiota diversity of human was significantly lower than other samples(P<0.01). The overall composition of the human and animal gut microbiota were dominated by the Firmicutes and Bacteroidetes phyla. However, the soil microbiota was dominated by Proteobacteria and Acidobacteria. At the genus level, the sheep, goat and cattle gut microbiota were dominated by Clostridium, Bacteroides, and Oscillibacter, while the horse gut microbiota was mainly composed of Clostridium, Eubacterium, and Treponema. The soil microbiota was composed mainly of Blastocatella and Bacillus. The human gut microbiota comprised much of Veillonella, Clostridium, Escherichia/Shigella. At the species level, the human gut microbiota mainly contained Escherichia/Shigella, dysenteriae, Streptococcus salivarius. The sheep, goat, cattle, and horse gut microbiota were dominated by Oscillibacter valericigenes and Eubacterium coprostanoligenes. The major species in soil were Blastocatella fastidiosa and Bacillus longiquaesitum. Moreover, principal coordinate analysis(PCoA) and hierarchical clustering analysis showed some differences in the microbiota structure among human gut, animal gut and soil samples. The gut microbiota structure was similar among cattle, goats and sheep. They were more different from the samples collected from human, horse and soil. We classified all samples into four clusters. Cluster 1 only included human samples;cluster 2 comprised the horse samples;cluster 3 was consisted of the sheep, goat, and cattle samples;while cluster 4 contained only the soil samples. Lastly, we identified the discriminatory OTUs and assigned them taxonomically to the species level. In conclusion, there were significant differences between animal gut microbiota and soil microbiota. The soil microbiota was more complex. As an omnivore, human gut microbiota diversity was significantly lower than other herbivorous animal(namely cattle, goat, horse and sheep). Although cattle, sheep, goat and horses are all herbivorous animals, the distinct features of the digestive systems could contribute to the difference in gut microbiota composition of horse from those of sheep, goat and cattle. This study revealed the differences of gut microbiota diversity between human and other animals, as well as from the soil microbial community. This work has laid a theoretical foundation for further studies on microbial diversity in different habitats.
作者
王佼
王彦杰
侯强川
徐海燕
孙志宏
张和平
孙天松
Jiao Wang;Yanjie Wang;Qiangchuan Hou;Haiyan Xu;Zhihong Sun;Heping Zhang;Tiansong Sun(Key Laboratory of Dairy Biotechnology and Engineering,Ministry of Education,Key Laboratory of Dairy Products Processing,Ministry of Agriculture and Rural Affairs,Inner Mongolia Agricultural University,Huhhot 010018,China)
出处
《科学通报》
EI
CAS
CSCD
北大核心
2019年第3期337-347,共11页
Chinese Science Bulletin
基金
国家自然科学基金(31720103911
31622043)
国家现代农业产业(奶牛)技术体系建设(CARS-36)
内蒙古自治区科技重大专项资助
关键词
肠道微生物
土壤微生物
单分子实时测序
多样性
16S
rRNA基因全长测序
gut microbiota
soil microbiota
single-molecule real-time (SMRT)sequencing technology
diversity
16S rRNA full-length sequencing
