三种室内饲养鱼类肠道微生物群落PCR-DGGE指纹分析
PCR-DGGE FINGERPRINTING ANALYSIS ON INTESTINAL MICROBIAL COMMUNITY OF THREE INDOOR REARING FISHES
查看参考文献29篇
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文摘
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以室内饲养的斑点叉尾鮰(Ictalurus punctatus)、银鲫和异育银鲫(中科三号)(Carassius auratus gibelio)幼鱼为对象,通过PCR-DGGE指纹技术对其肠道微生物群落进行了探索研究。在三种鱼的肠道中检测到不同的PCR-DGGE指纹谱带,其中斑点叉尾鮰的平均谱带数(7.5)相对于银鲫和异育银鲫的谱带数(分别为15和14)要少。基于PCR-DGGE指纹谱带及各谱带相对丰度的UPGMA聚类和MDS排序结果显示银鲫和异育银鲫肠道微生物相似性高,而与斑点叉尾鮰的差异比较大;rank-abundance散点图及回归分析也显示斑点叉尾鮰与银鲫、异育银鲫肠道微生物群落存在显著差异(P<0.05),而银鲫和异育银鲫之间无显著差异(P=0.383)。在斑点叉尾鮰肠道中检测到的菌群主要是变形杆菌,包括γ-变形杆菌和α-变形杆菌;而银鲫和异育银鲫肠道中菌群主要包括梭杆菌属中的类群,还包括变形杆菌门中的气单胞菌属,以及一些未知的类群。以上结果均表明在所研究的三种鱼的幼鱼阶段,其肠道微生物组成在不同种类鱼中存在差异,且该差异受基因型的影响可能更大。 |
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其他语种文摘
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PCR-DGGE fingerprinting was explored to study intestinal microbial community of three indoor rearing ju-venile fishes(channel catfish, silver prussian carp and hybridized prussian carp).Each fishes with three repetitions were reared in a semi-recirculating system in Institute of Hydrobiology.During the experiment, water temperature was 23℃—30℃, and fish were fed to satiation twice daily(9:00 and 15:00).After 30 days, 3 fishes of each treatment were randomly collected to extract microbial DNA.Gastrointestinal samples of all fishes were obtained by aseptically dis-secting the fish and carefully extracting the entire gastrointestinal tract.After dissecting, different segments of each gastrointestinal tracts from 3 fishes of the same treatment were pooled to analyse the bacterial communities.DNA from species samples were obtained according to the traditional Phenol-chloroform method.Bacterial domain-specific prim-ers F357(5’-CCTACGGGAGGCAGCAG-3’ with GC clamp in the 5’ end) and R518(5’-ATTACCGCGGCTGC TGG-3’) were used to amplify the variable V3 region of 16S rRNA genes.PCR products were separated on a 9% polyacrylamide gel(ratio of acrylamide to bisacrylamide was 37.5∶1) in a 1×TAE buffer and a denaturing gradient from 35% to 50% of urea and formamide.Unweighted pair-group method using arithmetic averages(UPGMA) and Multidimensional Scaling(MDS) were used to investigate community similarity of bacterial among different fish species.Moreover, rank– abun-dance plots were used to determine differences in bacterial community structure based on taxa relative abundance.For each plot, a linear regression model was fitted, and it was represented by the equation, log10y = a + bx, where a was the intercept and b was the slope of the plot.The slope(b) was subsequently used as a descriptive statistic for changes in community structure.Linear regressions, coefficients of determination(r) and significance(P) were calculated by using SPSS 13.0 software.Special DNA bands on DGGE profile were sequenced by Shanghai Sunny Biotechnology Co.As showed in DGGE profile, various number of 16S rDNA bands were detected in different samples, and the average bands detected in channe1 catfish intestine(7.5) were less than that of silver prussian carp and hybridized prussian carp(15 and 14, respectively).In addition, Both UPGMA clustering and MDS ordination based on 1/0 matrix showed that the similarity of intestinal microbe of silver prussian carp and hybridized prussian carp was higher when comparing to the similarity with channel catfish.Also, rank-abundance plots and linear regression on the basis of relative abundance of DGGE bands revealed that the microbial community structure of the three fishes were significantly different.Bacterial community structures in silver prussian carp and hybridized prussian carp showed no significant difference(P=0.383), while they showed difference(P<0.05) comparing with the intestinal bacteria in channe1 catfish.On the other hand, sequencing results showed that the bacteria in channel catfish intestine belonged to Proteobacteria, containing γ-Proteobacterium and α-Proteobacterium, while the bacteria in silver prussian carp and hybridized prussian carp intes-tine contained Fusobacterium, Aeromonas and some unclassified-bacteria.In conclusion, the intestinal bacterial compo-sition is not identical in different kinds of juvenile fishes reared in the same condition. |
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来源
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水生生物学报
,2011,35(3):423-429 【核心库】
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关键词
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斑点叉尾鮰
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银鲫
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异育银鲫
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PCR-DGGE
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肠道微生物群落
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地址
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中国科学院水生生物研究所, 中国科学院水生生物多样性与保护重点实验室, 武汉, 430072
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中国科学院水生生物研究所, 淡水生态与生物技术国家重点实验室, 武汉, 430072
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语种
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中文 |
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文献类型
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研究性论文 |
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ISSN
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1000-3207 |
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学科
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普通生物学;分子生物学 |
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基金
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国家973计划
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国家自然科学基金资助
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文献收藏号
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CSCD:4384281
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