Molecular analysis of bacterial communities associated with the roots of Douglas fir (Pseudotsuga menziesii) colonized by different ectomycorrhizal fungi

We studied the effect of ectomycorrhizal fungi on bacterial communities colonizing roots of Douglas fir (Pseudotsuga menziesii). Mycorrhizal tips were cleaned of soil and separated based on gross morphological characteristics. Sequencing of the internal transcribed spacers of the nuclear rRNA gene cluster indicated that the majority of the tips were colonized by fungi in the Russulaceae, with the genera Russula and Lactarius comprising 70% of the tips. Because coamplification of organellar 16S rRNA genes can interfere with bacterial community analysis of root tips, we developed and tested a new primer pair that permits amplification of bacterial 16S rRNA genes but discriminates more effectively against organellar sequences than commonly used bacterial primer sets. We then used terminal restriction fragment length polymorphism (T-RFLP) and sequence analysis of the 16S rRNA gene to examine differences in bacterial communities associated with the mycorrhizal tips. Cluster analysis of T-RFLP profiles indicated that there were different bacterial communities among the root tips; however, the communities did not seem to be affected by the taxonomic identity of the ectomycorrhizal fungi. Terminal restriction fragment profiling and sequencing of cloned partial 16S rRNA genes indicated that most bacteria on the ectomycorrhizal tips were related to the Alphaproteobacteria and the Bacteroidetes group.     

生物发酵稻秸对湖羊消化道上皮形态及微生物区系的影响

【目的】本研究旨在通过显微观察和16SrRNA基因高通量测序技术来探究饲喂生物发酵稻秸对湖羊肠道上皮形态及微生物区系的影响。【方法】试验选择70日龄、体重相近(25.15±0.47)kg的湖羊公羔21只,根据饲粮中粗饲料的组成随机分为3组：稻秸组、生物发酵稻秸组和苜蓿干草组,饲粮精粗比为6:4,试验持续7周,其中适应期3周,正试期4周,结束后屠宰取样,采集瘤胃、空肠和结肠上皮组织进行观察测量,收集对应肠段内容物用于微生物区系和代谢产物测定。【结果】与稻秸组相比,饲喂生物发酵稻秸显著提高湖羊瘤胃中纤维杆菌(Fibrobacteres)的相对丰度和挥发性脂肪酸(volatile fatty acids, VFA)的含量,促进瘤胃上皮的发育;显著提高湖羊空肠中厚壁菌门(Firmicutes)和疣微菌门(Verrucomicrobia)的相对丰度,改变空肠微生物菌群结构,促进空肠上皮组织的发育;改变湖羊结肠微生物菌群结构。【结论】饲喂生物发酵稻秸有利于湖羊肠道上皮的发育,并增加消化道内的微生物多样性。     

组学视角下养殖动物消化道微生物组结构与功能研究进展北大核心

养殖动物消化道中含有大量的微生物,不仅参与动物对营养物质的消化和吸收,还对宿主生长发育及免疫起重要调节作用。动物消化道微生物组研究是目前国内外的热点领域,取得了一系列重要研究进展。深入了解养殖动物消化道微生物组的结构与功能,将为今后调控和应用消化道微生物、提高动物生产性能、改善动物胃肠道健康和实现绿色健康养殖奠定理论基础。本文以4种代表性养殖动物(牛、羊、猪和鸡)为主体,对组学视角下其消化道微生物群落结构、功能等研究进展进行总结和分析;并对未来研究方向进行展望。     

Microbial community changes during the bioremediation of creosote-contaminated soil

AIMS: To investigate the effects of aeration on the ex situ biodegradation of polycyclic aromatic hydrocarbons (PAHs) in creosote-contaminated soil and its effect on the microbial community present. METHODS AND RESULTS: Aerated and nonaerated microcosms of soil excavated from a former timber treatment yard were maintained and sampled for PAH concentration and microbial community changes by terminal restriction fragment length polymorphism (TRFLP) analysis. After an experimental period of just 13 days, degradation was observed with all the PAHs monitored. Abiotic controls showed no loss of PAH. Results unexpectedly showed greater loss of the higher molecular weight PAHs in the nonaerated control. This may have been due to the soil excavation causing initial decompaction and aeration and the resulting changes caused in the microbial community composition, indicated by TRFLP analysis showing several ribotypes greatly increasing in relative abundance. Similar changes in both microcosms were observed but with several possible key differences. The species of micro-organisms putatively identified included Bacilli, pseudomonad, aeromonad, Vibrio and Clostridia species. CONCLUSIONS: Excavation of the contaminated soil leads to decompaction, aeration and increased nutrient availability, which in turn allow microbial biodegradation of the PAHs and a change in the microbial community structure. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the changes occurring in the microbial community during biodegradation of all PAHs is essential for the development of improved site remediation protocols. TRFLP allows useful monitoring of the total microbial community.     

Differences in the DNA restriction patterns between sheep with HbA and HbB

DNA samples obtained from sheep homozygous for HbA, for HbB and heterozygous were subjected to Southern blot analysis using a goat beta F and a rabbit beta 1-globin gene as probes. Sheep homozygous for HbA show a different restriction pattern from that of sheep homozygous for HbB with each of the used endonucleases. The DNAs from moufflon and sheep homozygous for HbA show indistinguishable restriction patterns with some endonucleases. By means of double digestions it has been possible to construct restriction maps of the beta B and beta C genes and to confirm the absence of the beta C gene in HbB sheep.     

Assessing genetic diversity in indigenous Veneto chicken breeds using AFLP markers

Genetic variation in four indigenous chicken breeds from the Veneto region of Italy was assessed using amplified fragment length polymorphism (AFLP) markers. A total of 99 individuals were analysed using three AFLP primer combinations that produced 70 polymorphisms. Four indigenous Veneto chicken breeds (Ermellinata, Padovana, Pépoi and Robusta) and a reference broiler line were included in the analysis. Breed-specific markers were identified in each breed. The expected heterozygosity did not differ significantly among the indigenous Veneto chicken breeds and the broiler line. The coefficient of gene variation (Gst) value across loci indicated that almost half of the total variability was observed among breeds. Nei's standard genetic distance between pairs of breeds showed that the distance between the broiler line and the Pépoi breed was greater than the distances between the broiler line and the other three chicken breeds. Cluster analysis based on standard genetic distances between breeds indicated that the Padovana and Pépoi breeds were closely related. Factorial analysis based on a binary matrix of the AFLP data showed a clear distinction of all breeds.     

DNA polymorphism at the casein loci in sheep

By using seven endonucleases and four bovine cDNA probes specific for alpha S1-, alpha S2-, beta-, and kappa-casein genes, nine restriction fragment length polymorphisms (RFLPs) have been found in the sheep orthologous DNA regions. In contrast to the low level of variation observed at the protein level, these DNA polymorphisms determine a high level of heterozygosity and, therefore, represent useful tools for genetic analyses since they can also be obtained without the need for gene expression. In fact, informative matings suggest that in sheep, as in cattle, the four loci are linked.     

Genetic variability between two breeds based on restriction fragment length polymorphisms (RFLPs) of major histocompatibility complex class I genes in the pig

Summary Restriction fragment length polymorphism analyses of SLA class I genes were performed on 55 Duroc and 24 Hampshire boars from the 1986–87 national performance tests of each breed. Few boars were inbred. Southern blotting and hybridization procedures were performed on genomic DNA isolated from white blood cells by using Pvu II, Bam HI, and Eco RI endonucleases and a swine MHC class I probe. Genetic variability within and between the two breeds was estimated in terms of nucleotide diversity, by using a mathematical analysis based on the different RFLP patterns. The nucleotide diversity calculated within each breed was less than that between the two breeds. The results from the nucleotide diversity analysis suggested that genetic variability was greater in the Duroc breed than in the Hampshire breed. A relatively high level of genetic variability was shown in the class I major histocompatibility complex genes in the pig.     

Soil bacterial diversity in a loblolly pine plantation: influence of ectomycorrhizas and fertilization

We studied the effect of ectomycorrhizas and fertilization on soil microbial communities associated with roots of 10-year-old loblolly pine. Ectomycorrhizas were identified using a combination of community terminal restriction fragment profiling and matching of individual terminal restriction fragments to those produced from ectomycorrhizal clones and sequences recovered from roots and sporocarps. Differences between bacterial communities were initially determined using cluster analysis on community terminal restriction fragment profiles and through subsequent recovery of 16S rDNA clones. Analysis of bacterial clones revealed that terminal restriction fragment length was often shared between taxonomically dissimilar bacterial types. Consequently, we could not reliably infer the identity of peaks in the bacterial community profile with some exceptions, notably chloroplast rDNA that generated an approximate peak size of 80.2 bp. Fertilization increased the frequency of a Piloderma-like ectomycorrhiza. However, we did not detect clear effects of fertilization or the presence of viable ectomycorrhizas on bacterial communities. Bacterial communities seemed to be determined largely by the carbon and nitrogen content of soil. These results suggest that important soil microbial groups respond differently to soil conditions and management practices, with ectomycorrhizal communities reflecting past nutrient conditions and bacterial communities reflecting current environmental conditions of soil microsites.     

Genetic diversity and subdivision of 57 European and Middle-Eastern sheep breeds

The population structure and genetic diversity of 57 European and Middle Eastern marginal and cosmopolitan sheep breeds from 15 countries were analysed by typing 31 microsatellite markers. Mean unbiased expected heterozygosities ranged from 0.63 in British Exmoor Horn to 0.77 in Albanian Ruda. South-eastern European and Middle-Eastern sheep breeds were significantly more variable than northwestern and western European breeds. An overall heterozygote deficiency (f) across all loci was observed (P < 0.001), while genetic differentiation (theta) was 5.7%. Principal component analysis and Bayesian model-based clustering indicate a south-east to north-west cline, but also revealed distinct groups of Middle-Eastern fat-tailed sheep, south-eastern European sheep and north-western/western European sheep. Within the last group, two less-distinct clusters comprised the Merino-type and Alpine breeds respectively. The incomplete demarcations of most clusters probably reflects cross-breeding and/or upgrading.     

采用PCR-RFLP技术在不同水平上鉴定大豆根瘤菌

采用16S rRNA基因PCR扩增与限制性酶切片段多态性分析(RFLP)技术对选自弗氏中华根瘤菌(S.fredii)、大豆慢生根瘤菌(B.japonicum)和埃氏慢生根瘤菌(B.elkanii)的19株代表菌进行了比较分析,根据用3种限制性内切酶的RFLP分析结果,可将供试菌株分为S.fredii,B.japonicum, B.elkanii Ⅱ和B.elkanii Ⅱa等4种基因型。各类菌株之间没有交叉,因此本研究采用的PCR-RFLP技术不失为一种快速鉴别大豆根瘤菌的新方法。采用本技术已将分离自中国的22株快生菌和19株慢生菌分别鉴定为S.fredii和B.japonicum。对供试参比菌株和野生型菌株进行的16S～23S基因间隔DNA(IGS)的PCR-RFLP分析结果表明：S.fredii和B.japonicum菌株的IGS长度不同,所有供试S.fredii菌株的IGS为2.1 kb,而供试B.japonicum菌株则为2.0 kb。依据RFLP的差异,可将来自中国两个不同地区的S.fredii株区分为2个基因型,而来自中国东北黑龙江地区的19株B.japonicum菌株则可分为11个基因型。对上述野生型菌株还进行了REP-PCR和ERIC-PCR分析并确定其具有菌株水平的特异性。     

中国7个地方绵羊品遗传多样性的微卫星分析

利用26个微卫星标记分析了中国7个地方绵羊（Ovis aries）品种的遗传多样性。通过计算基因频率,平均杂合度（H）、多态信息含量（PIC）及有效等位基因数（Ne）,并根据Nei氏标准遗传距离,利用UPGMA法进行了聚类分析,评估其种内遗传变异和品种间遗传关系。结果表明：26个微卫星位点共检测到278个等位基因,Ne在2.1288-13.3924之间;26个位点均为高度多态位点,PIC在0.6628-0.8712之间,聚类分析表明哈萨克羊、阿勒泰羊和巴音布鲁克羊遗传关系最近;然后与白藏羊,黑藏羊聚为一类,湖羊和晋中羊聚为一类,各绵羊各种的聚类关系与其来源,育成史及地理分布基本一致。     

Genetic diversity of Fusarium oxysporum populations isolated from different soils in France

The genetic diversity of soil-borne populations of Fusarium oxysporum was assessed using 350 isolates collected from six different French soils. All isolates were characterised by restriction fragment analysis of the PCR-amplified ribosomal intergenic spacer (IGS). Twenty-six IGS types were identified among the 350 isolates analysed. Five to nine different IGS types were detected in each soil. None of the IGS types was common to all of the soils. An analysis of the molecular variance based on IGS type relationships and frequency revealed that the genetic structure of the populations of F. oxysporum varied widely among the soils. Some populations were both highly diverse within the soils and differentiated between the soils. A possible relationship between the intrapopulation or interpopulation level of diversity and some external factors such as the soil type or the crop history was evaluated. A subsample representative of the diversity of the six populations was further characterised by analysing the genomic distribution of two transposable elements, impala and Fot1. One to 10 copies of the impala element were present in most of the isolates, irrespective of their soil of origin. The Fot1 element was only detected in 40% of the isolates originating from the three populations less diverse in terms of IGS types, but in 82.6% of the isolates originating from the three more diverse populations.     

中国绵羊起源、进化和遗传多样性研究进展

中国地方绵羊品种资源丰富,部分品种具有繁殖力高、毛皮品质好、多角、多乳头、大尾脂、抗逆性强等独特性状,这些遗传资源引起了学者们对其进行深入研究的兴趣,但目前仍然存在绵羊起源问题的争议,缺乏对我国绵羊的遗传多样性进行全面系统研究等问题。本文综述了绵羊起源、品种分化等方面的研究进展,并从父系、母系、常染色体分子标记等不同层面介绍了中国绵羊遗传多样性的研究概况,为中国绵羊遗传资源的保护和利用、绵羊新品种(系)的培育以及我国绵羊产业良性发展提供参考。     

Genetic structure and admixture in sheep from terminal breeds in the United States

Selection for performance in diverse production settings has resulted in variation across sheep breeds worldwide. Although sheep are an important species to the United States, the current genetic relationship among many terminal sire breeds is not well characterized. Suffolk, Hampshire, Shropshire and Oxford (terminal) and Rambouillet (dual purpose) sheep (n = 248) sampled from different flocks were genotyped using the Applied Biosystems Axiom Ovine Genotyping Array (50K), and additional Shropshire sheep (n = 26) using the Illumina Ovine SNP50 BeadChip. Relationships were investigated by calculating observed heterozygosity, inbreeding coefficients, eigenvalues, pairwise Wright’s F_ST estimates and an identity by state matrix. The mean observed heterozygosity for each breed ranged from 0.30 to 0.35 and was consistent with data reported in other US and Australian sheep. Suffolk from two different regions of the United States (Midwest and West) clustered separately in eigenvalue plots and the rectangular cladogram. Further, divergence was detected between Suffolk from different regions with Wright’s F_ST estimate. Shropshire animals showed the greatest divergence from other terminal breeds in this study. Admixture between breeds was examined using admixture , and based on cross-validation estimates, the best fit number of populations (clusters) was K = 6. The greatest admixture was observed within Hampshire, Suffolk, and Shropshire breeds. When plotting eigenvalues, US terminal breeds clustered separately in comparison with sheep from other locations of the world. Understanding the genetic relationships between terminal sire breeds in sheep will inform us about the potential applicability of markers derived in one breed to other breeds based on relatedness.     

Succession of bacterial community composition over two consecutive years in two aquatic systems: a natural lake and a lake-reservoir

The succession in bacterial community composition was studied over two years in the epilimnion and hypolimnion of two freshwater systems: a natural lake (Pavin Lake) and a lake-reservoir (Sep Reservoir). The bacterial community composition was determined by cloning-sequencing of 16S rRNA and by terminal restriction fragment length polymorphism. Despite large hydrogeological differences, in the Sep Reservoir and Pavin Lake the dominant bacteria were from the same taxonomic divisions, particularly Actinobacteria and Betaproteobacteria. In both ecosystems, these major bacterial divisions showed temporal fluctuations that were much less marked than those occurring at a finer phylogenetic scale. Nutrient availability and mortality factors, the nature of which differed from one lake to another, covaried with the temporal variations in the bacterial community composition at all sampling depths, whereas factors related to seasonal forces (temperature and outflow for Sep Reservoir) seemed to account only for the variation of the hypolimnion bacterial community composition. No seasonal reproducibility in temporal evolution of bacterial community from one year to the next was observed.     

新疆富蕴地震断裂带植被恢复对土壤古菌群落的影响

选取新疆富蕴地震断裂带8种次生植物根际土壤,以同土层裸地为对照,测量土壤化学性质,利用末端限制性片段长度多态性技术研究塌陷区次生植物对土壤古菌群落的影响.结果表明,多数植物根际土壤养分显著(P＜0.05)高于对照,其主要古菌类群为泉古菌门(Crenarchaeota)和广古菌门(Euryarchaeota),群落间多样性差异大,相似性低.不同植物根际土壤古菌的优势类群数量差异较大,鼠掌老鹳草高达18种,西北绢蒿少为2种.典范对应分析表明,土壤有机质含量对古菌优势类群分布影响最大(Hha Ⅰ酶切:r=0.94;Rsa l酶切:r=0.74),速效磷含量与古菌群落各多样性指数呈显著正相关(P＜0.05),总氮含量与均匀度指数E呈显著正相关(P＜0.05).新疆地震断裂带植被的恢复可影响根际土壤古菌群落的分布、组成和结构,其原因与植物根际土壤化学性质有关,同时在改善土壤肥力方面也有显著效果.     

宽体金线蛭消化道的组织学观察

运用解剖学和组织学方法对宽体金线蛭消化道的结构进行了组织学研究。结果表明,宽体金线蛭的嗉囊向两侧伸出11对侧盲囊,第6对侧盲囊狭长并延伸到直肠两侧;咽主要由黏膜层、肌层和外膜构成,外膜几乎不可见;食道、嗉囊、肠和直肠管壁由黏膜层、黏膜下层、肌层和浆膜构成;咽和直肠的上皮具纹状缘。除肠外,其他消化道的上皮细胞均无发达的纤毛,且黏膜上皮皆为单层柱状上皮;除肠和直肠外,腺体及导管较少;直肠的黏膜肌层为内环外纵两层,其他各部均为纵行肌一层;消化道各部黏膜下层较发达;外膜为浆膜,与黏膜下层分界不明显。     

Assessment of DNA fingerprinting for determining genetic diversity in sheep populations

Genomic DNA, prepared from 12 animals from four sheep flocks, was digested with either HaeIII or Hin fI and probed with three DNA fingerprinting probes. Mean DNA fingerprint band sharing and band frequency calculated for each flock were used to estimate genetic diversity. Each of the DNA fingerprinting systems showed the same trend in diversity within the sampled flocks, and greater diversity between the flocks than within the flocks. DNA fingerprinting therefore provides a useful measure of genetic diversity in sheep.     

Inferring the population structure of the Maghreb sheep breeds using a medium‐density SNP chip

North Africa has a great diversity of indigenous sheep breeds whose origin is linked to its environmental characteristics and to certain historical events that took place in the region. To date, few genome‐wide studies have been conducted to investigate the population structure of North African indigenous sheep. The objective of the present study was to provide a detailed assessment of the genetic structure and admixture patterns of six Maghreb sheep populations using the Illumina 50K Ovine BeadChip and comparisons with 22 global populations of sheep and mouflon. Regardless of the method of analysis used, patterns of multiple hybridization events were observed within all North African populations, leading to a heterogeneous genetic architecture that varies according to the breed. The Barbarine population showed the lowest genetic heterogeneity and major southwest Asian ancestry, providing additional support to the Asian origin of the North African fat‐tailed sheep. All other breeds presented substantial Merino introgression ranging from 15% for D'man to 31% for Black Thibar. We highlighted several signals of ancestral introgression between North African and southern European sheep. In addition, we identified two opposite gradients of ancestry, southwest Asian and central European, occurring between North Africa and central Europe. Our results provide further evidence of the weak global population structure of sheep resulting from high levels of gene flow among breeds occurring worldwide. At the regional level, signs of recent admixture among North African populations, resulting in a change of the original genomic architecture of minority breeds, were also detected.