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通过比较cDNA和gDNA探索瘤胃中活性尿素分解菌的多样性

Exploring the Diversity of Active Ureolytic Bacteria in the Rumen by Comparison of cDNA and gDNA.

作者信息

Liu Sijia, Zheng Nan, Zhao Shengguo, Wang Jiaqi

机构信息

State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, No.2 Yuanmingyuan West Road, Haidian, Beijing 100193, China.

出版信息

Animals (Basel). 2020 Nov 20;10(11):2162. doi: 10.3390/ani10112162.

Abstract

In this study we revealed the diversity of active ureolytic bacteria in the rumen by compared amplicons between gDNA and cDNA. Rumen fluid was collected from four Holstein dairy cows with rumen fistulas at 0, 2, and 6 h after morning feeding. Total microbial gDNA and RNA were isolated, and the RNA was reverse-transcribed into cDNA. The gene amplicons of gDNA and cDNA were produced and sequenced by MiSeq. These results revealed that the sampling time had no significant difference on the alphssa and beta diversity indices of the ureolytic bacteria. The Shannon diversity of the gene for cDNA was greater than that for gDNA ( < 0.05). There were significant difference in the beta diversity of ureolytic bacteria between gDNA and cDNA ( < 0.01), which indicates a shift in the community of active ureolytic bacteria. Approximately 67% of sequences from cDNA could not be confidently classified at the genus level. The active ureolytic bacteria were mainly from , , , , , and sp. Changes in the operational taxonomic units revealed that the top abundant genes were mostly consistent between gDNA and cDNA, and most differences occurred in the genes with lower abundances. These results revealed distinct ureolytic bacteria community profiles based on gDNA and cDNA. The dominant ureolytic bacteria had high transcriptional activity, and the differential were mainly distributed in the genus of low abundance.

摘要

在本研究中,我们通过比较基因组DNA(gDNA)和互补DNA(cDNA)的扩增子,揭示了瘤胃中活性尿素分解菌的多样性。在早晨喂食后0、2和6小时,从四头装有瘤胃瘘管的荷斯坦奶牛采集瘤胃液。分离总微生物gDNA和RNA,并将RNA反转录成cDNA。通过MiSeq对gDNA和cDNA的基因扩增子进行生成和测序。这些结果表明,采样时间对尿素分解菌的α多样性和β多样性指数没有显著差异。cDNA基因的香农多样性大于gDNA基因的香农多样性(P<0.05)。gDNA和cDNA之间尿素分解菌的β多样性存在显著差异(P<0.01),这表明活性尿素分解菌群落发生了转变。来自cDNA的约67%序列在属水平上无法可靠分类。活性尿素分解菌主要来自[具体菌属1]、[具体菌属2]、[具体菌属3]、[具体菌属4]、[具体菌属5]和[具体菌属6]种。操作分类单元的变化表明,gDNA和cDNA之间丰度最高的基因大多一致,大多数差异发生在丰度较低的基因中。这些结果揭示了基于gDNA和cDNA的不同尿素分解菌群落概况。优势尿素分解菌具有较高的转录活性,差异主要分布在低丰度属中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b9e/7699693/36f6bd0b6d7a/animals-10-02162-g001.jpg

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