分子营养 Molecular Nutrition

利用高通量测序技术分析川中黑山羊瘤胃纤毛虫种群结构

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  • 四川农业大学动物医学院, 动物疫病与人类健康四川省重点实验室, 环境公害与动物疾病四川省高校重点实验室, 成都 611130
刘旗(1992-),男,河南漯河人,硕士研究生,研究方向为中西兽医与临床。E-mail:398772948@qq.com

收稿日期: 2016-11-04

  网络出版日期: 2017-05-13

基金资助

长江学者和创新团队发展计划创新团队项目(IRT0848);四川农业大学双支计划(03572070)

Ruminal Ciliate Community Structure of Chuanzhong Black Goats: An Analysis Using High-Throughput Sequencing Technology

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  • Key Laboratory of Animal Disease and Human Health of Sichuan Province, Key Laboratory of Environmental Hazard and Animal Disease of Sichuan Province, College of Veterinary Medicine, Sichuang Agricultural University, Chengdu 611130, China

Received date: 2016-11-04

  Online published: 2017-05-13

摘要

本试验旨在应用高通量测序技术研究川中黑山羊瘤胃纤毛虫种群结构。选取3只4月龄川中黑山羊[体重(15.53±0.21)kg],正常饲喂20 d后采集瘤胃液(A)样品,间隔40 d后再次采集瘤胃液(F)样品,提取样品总DNA后,扩增真核生物18S rRNA V4区,扩增产物使用Illumina MiSeq平台测序。结果表明:1)共获得高质量有效序列242 321条,聚类后得1 650个操作分类单位(OTU)。2)A样品、F样品在alpha多样性Chao、ACE、Shannon和Simpson指数上,差异不显著(P>0.05)。3)在纲水平分类上,2个时间点样品相对丰度最高的均为纤毛门,侧口纲(A样品为46.0%、F样品为44.7%),2个样品的相对丰度差异不显著(P>0.05)。4)在科水平分类上,A样品优势科为头毛科(31.8%),其次为均毛科(14.2%);F样品优势科为头毛科(42.8%);并且F样品头毛科相对丰度显著高于A样品(P<0.05),A样品均毛科相对丰度显著高于F样品(P<0.05)。5)在属水平分类上,A样品与F样品相对丰度最高的属均为多加多泡双毛属(A样品为20.9%、F样品为25.4%),无显著差异(P>0.05);存在显著性差异的属是均毛属、头毛属、腹甲双毛属、刺甲双毛属相对丰度,其中A样品均毛属(14.1% vs. 1.9%)、腹甲双毛属相对丰度(2.8% vs. 1.5%)显著高于F样品(P<0.05),而头毛属(6.7% vs. 12.5%)、刺甲双毛属相对丰度(0.3% vs. 2.5%)显著低于F样品(P<0.05)。本试验结果表明,幼龄川中黑山羊瘤胃液纤毛虫相对丰度最高的种属为多加多泡双毛属,瘤胃中还有许多未被分类鉴定且相对丰度较高的真核生物,需要进一步研究。

本文引用格式

刘旗, 陈芸, 邓俊良, 任志华, 杨颜铱, 高爽, 陈憧 . 利用高通量测序技术分析川中黑山羊瘤胃纤毛虫种群结构[J]. 动物营养学报, 2017 , 29(5) : 1574 -1581 . DOI: 10.3969/j.issn.1006-267x.2017.05.016

Abstract

This study was designed to reveal the ruminal ciliate community structure of Chuanzhong black goats using MiSeq sequencing technology. Three 4-month-old Chuanzhong black goats weighted (15.53±0.21) kg were used to collect rumen fluid, and the samples were collected after normal feeding of 20 (sample A) and 60 d (sample F), respectively. Total DNA was extracted for amplification V4 area of 18S rRNA, and the products were sequenced by Illumina MiSeq sequencing system. The results showed as follows: 1) a total of 242 321 high quality valid sequences and 1 650 operational taxonomic units were obtained. 2) There were no significant difference on alpha diversity indexes (Chao, ACE, Shannon and Simpson) between samples A and F (P>0.05). 3) At class level, the most abundant class in samples A and F were Ciliophora, Litostomatea (sample A was 46.0%, sample F was 44.7%), and there was no significant difference between samples A and F (P>0.05). 4) At family level, the most abundant family in sample A was Ophryoscolecidae (31.8%) followed by Isotrichidae (14.2%); the most abundant family in sample F was Ophryoscolecidae (42.8%); the relative abundance of Ophryoscolecidae in sample F was significantly higher than that in sample A (P<0.05), and the relative abundance of Isotrichidae in sample A was significantly higher than that in sample F (P<0.05). 5) At genus level, the most abundant ciliate genus was Polyplastron(sample A was 20.9%, sample F was 25.4%), and there are no significant differences between samples A and F (P>0.05); there were significant differences of the relative abundances of Isotricha, Ophryoscolex, Diploplastron and Enoploplastron, the relative abundances of Isotricha (14.1% vs. 1.9%) and Diploplastron (2.8% vs. 1.5%) in sample A were significantly higher than those in sample F (P<0.05), and relative abundances of Ophryoscolex (6.7% vs. 12.5%) and Enoploplastron (0.3% vs. 2.5%) in sample A were significantly lower than those in sample F (P<0.05). In conclusion, Polyplastron is the most abundant genus in the rumen ciliate communities of youth Chuanzhong black goats, and there are still a lot of unclassified and unidentified eukaryotic microorganisms with high abundance worth further study in rumen.

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