基于454高通量测序的黄土高原不同乔木林土壤细菌群落特征

选取黄土高原不同乔木林（辽东栎,LDL;侧柏,CB;刺槐,CH;油松,YS）表层土壤为研究对象,利用第二代高通量测序技术罗氏454平台对其进行16S rRNA基因V1~V3可变区测序,通过分析其Alpha多样性、物种组成和丰度、群落结构,结合土壤的理化性质研究其对细菌群落结构的影响.结果表明：所有土壤样品共检测到36个门,84个纲,187个目.不同乔木林土壤中优势菌来自变形菌门（Proteobacteria）、放线菌门（Actinobacteria）、酸杆菌门（Acidobacteria）、绿弯菌（Chloroflexi）和浮霉菌门（Planctomycetes）,主要的优势菌纲为放线杆菌纲（Actinobacteria）、α-变形菌纲（Alphaproteobacteria）、酸杆菌（Acidobacteria）、β-变形菌纲（Betaproteobacteria）、浮霉菌纲（Planctomycetacia）.不同乔木林土壤中绿弯菌门（Chloroflexi）与pH值呈极显著的正相关,总磷和蓝细菌（Cyanobacteria）呈极显著的正相关,微生物生物量碳与芽单胞菌门（Gemmatimonadetes）呈极显著的负相关,土壤总磷含量可能为CB样地区别于其他土样群落组成的主要因素.LDL样地细菌群落受环境影响较小.     

污水回用中主要病原菌解析及其紫外消毒效应

本研究以污水处理厂二级出水中的微生物为研究对象,通过454焦磷酸测序技术分析其群落结构组成,揭示了主要病源菌的种类和比例;通过培养法、q PCR、Q-RT-PCR这3种方法分析紫外剂量为60 m J·cm-2时对指示菌大肠杆菌和典型病原菌沙门氏菌及分枝杆菌的去除特性.结果表明,二级出水中共有11种病原菌,主要为梭菌属(2.96%)、弓形杆菌属(0.82%)和分枝杆菌(0.36%).60 m J·cm-2剂量的紫外消毒可以有效去除99.9%可培养的大肠杆菌和沙门氏菌,对可培养分枝杆菌的去除率不足90%.但是,该剂量紫外消毒对活性大肠杆菌、沙门氏菌和分枝杆菌的去除率较低,Q-RT-PCR检测方法可以较准确评价微生物的存活状态.60 m J·cm-2紫外剂量会导致大量病原菌进入具有活性但不可培养(VBNC)状态,需结合其他深度处理工艺进一步去除活性病原菌以保障污水回用的安全利用.     

活性污泥系统聚糖菌的富集与菌群多样性分析

在实验室规模续批式反应器(SBR)内,采用人工配水作为系统进水,以乙酸钠为唯一碳源,限制进水磷浓度,调整适宜的运行方式对活性污泥进行驯化,培养富集聚糖菌。90 d的培养过程中,系统的进水COD浓度始终维持在260mg/L左右,进水PO3-4-P浓度从20 mg/L逐渐降至2 mg/L,系统除磷能力逐渐丧失。系统内活性污泥的TP/TSS从40.5%降至10.4%,污泥中的糖原/TSS从14.5%增至38.2%,同时COD去除率能保持在90%以上,说明系统中成功富集聚糖菌。采用454高通量测序法分析该系统的活性污泥菌群结构,发现α-proteobacteria,β-proteobacteria,γ-proteobacteria 3类细菌占微生物总量的比例达83.78%。在属的等级上,Tepidicella占活性污泥菌群的比例最高,为20.60%。     

九龙江流域潜在病原菌污染分析

近年来,由于流域经济快速发展和城镇化进程加快等原因,九龙江水污染问题日趋严重.为全面认识九龙江流域病原菌的分布状况,应用16S rRNA基因-454焦磷酸测序技术测定九龙江支流西溪、北溪水体和沉积物中细菌16S rRNA基因V1~V3高变区,共获得204 216条高质量序列.通过与病原菌参考数据库对比分析发现,九龙江分布有68个潜在病原菌属,占序列总量的6.1%.其中梭菌属(Clostridium)、分支杆菌属(Mycobacterium)和鞘氨醇单胞菌属(Sphingomonas)在所有样品中都有检出,且丰度最高,分别占病原菌属序列总量的54.5%、5.9%和5.6%.在种水平上,九龙江分布有48种病原菌,占序列总量的0.76%.其中Afipia felis、Mycobacterium asiaticum、Clostridium baratii、Brucella melitensis和Delftia tsuruhatensis是丰度最高的5种病原菌,分别占病原菌种序列总量的48.9%、20.3%、8%、2.7%和1.7%.统计分析表明,九龙江水体中分布的病原菌种类数(属或种)显著高于沉积物中,且西溪水体检出的病原菌种类(属或种)和丰度最高,说明九龙江水体,尤其是西溪水体受病原菌污染的风险较高.此外,相关分析表明,九龙江水体中病原菌的种类数(属或种)和丰度与营养盐(氮、磷)有着显著的正相关关系,说明九龙江水体中分布的病原菌与沿岸的人类活动,如养殖业、污水排放等密切相关.因此,为保证流域的公共卫生安全,需进一步加强污染源清理整顿工作,开展水环境病原菌的实时监测.     

Comparison on bacterial community of rhizosphere and bulk soil of poplar plantation based on pyrosequencing北大核心CSCD

By the 454 pyrosequencing technology, this research compared the bacterial communities in poplar plantation rhizosphere and bulk soil for an accurate understanding of bacterial community colonization in the two soil environments. The species annotation showed that rhizosphere soil contained 145 bacterial genera and bulk soil contained 141 bacterial genera, with 8 common genera shared by both at a relative abundance of more than 4%. The 8 genera in common were Acidobacterium GP1, Acidobacterium GP3, Acidobacterium GP6, Gemmatimonas, Bradyrhizobium, Burkholderia, Streptomyces and Acidobacterium GP4. The relative abundance of the same bacterial community was significantly different between rhizosphere and bulk soil environments. Alpha diversity analysis showed that the bacterial community diversity of rhizosphere soil was higher than that of bulk soil, but the difference was not significant. The results of bacterial communities sorting could reflect the variation of soil bacterial communities from rhizosphere to the bulk and the spatial variation among different sampling points, indicating a contribution of about 21.2% variance of bacterial communities by the effect of rhizosphere. Beta diversity analysis showed great difference between rhizosphere and bulk soil samples in bacterial community composition. There were 15 genera specific to rhizosphere soil and 11 to bulk soil. The abundance of 23 genera, mainly cellulose degrading bacteria and nitrogen-fixing bacteria, changed significantly. Selectivity of root to rhizosphere microorganisms is an important mechanism leading to significant differences in the rhizosphere microbial community composition and structure, which may significantly impact the carbon and nitrogen cycles of the root-soil interface.     

The molecular diversity of arbuscular mycorrhizal fungi in the arsenic mining impacted sites in Hunan Province of China

Arbuscular mycorrhizal fungi(AMF) can establish a mutualistic association with most terrestrial plants even in heavy metal contaminated environments. It has been documented that high concentrations of toxic metals, such as arsenic(As) in soil could adversely affect the diversity and function of AMF. However, there are still gaps in understanding the community composition of AMF under long-term As contaminations. In the present study, six sampling sites with different As concentrations were selected in the Realgar mining area in Hunan Province of China. The AMF biodiversity in the rhizosphere soils of the dominant plant species was investigated by sequencing the nuclear small subunit ribosomal RNA(SSU rR NA) gene fragments using 454-pyrosequencing technique. A total of 11 AMF genera were identified,namely Rhizophagus, Glomus, Funneliformis, Acaulospora, Diversispora, Claroideoglomus, Scutellopora,Gigaspora, Ambispora, Praglomus, and Archaeospora, among which Glomus, Rhizophagus, and Claroideoglomus clarodeum were detected in all sampling sites, and Glomus was the dominant AMF genus in the Realgar mining area. Redundancy analysis indicated that soil pH, total As and Cd concentrations were the main factors influencing AMF community structure. There was a negative correlation between the AMF species richness and the total As concentration in the soil,but no significant correlation between the Shannon–Wiener index of the AMF and plants. Our study showed that high As concentrations can exert a selective effect on the AMF populations.     

A comparative study of microbial communities in soils amended by manures from pigs fed with organic versus synthetic feeds

We applied pyrosequencing to determine bacterial communities in soil samples taken from test fields amended by manures of pigs fed with green feeds (GPMS), synthetic feeds (APMS), and the reference field without swine manure amendment (NPMS), respectively. For each sample, 7050 effective sequences were selected and utilized to do the bacterial diversity and abundance analysis, respectively. In total, 2986, 2873, and 1602 operational taxonomic units were obtained at a 3% distance cut-off in GPMS, APMS, and NPMS, respectively. Bacterial phylotype richness in GPMS was higher than the other samples. NPMS had the least richness. The most dominant class in both GPMS and APMS is Betaproteobacteria, whereas Alphaproteobacteria is dominant in NPMS. Circa 7.5% sequences in GPMS were found to be affiliated with Burkholderiales order. Bacterial diversity and abundance in the soil varied with swine manure amendment, due to nutrient elements as well as toxic metals in swine manures. Compared with the soil amendment by manures from pigs bred with synthetic feeds, more bacterial diversity but less potentially pathogenic bacterial genera in the soil amended by manures of pigs fed with green feeds were found, which indicated that these manures were better land-applied organic fertilizers.     

微氧水解酸化处理石化废水的生物降解特性

本研究采用微氧水解酸化技术处理石化废水,以抑制硫酸盐的还原,减少硫化氢的产生.同时,通过与厌氧水解酸化的对比试验,研究了微氧水解酸化的生物降解特性.微氧反应器的ORP控制在(-290±71)m V,厌氧反应器的ORP为(-398±31)m V.反应器运行近7个月的结果表明,在进水COD为202~514 mg·L-1、硫酸根浓度为350~650 mg·L-1及HRT为12 h时,微氧水解酸化反应器COD的平均去除率为31.2%,高于厌氧水解酸化的26.4%.厌氧出水的VFA浓度((2.34±0.60)mmol·L-1)高于微氧出水((1.89±0.48)mmol·L-1).微氧出水的平均比紫外吸收值(UV254/DOC)为0.017,显著低于厌氧出水(0.025),表明微氧环境可以提高兼性水解酸化菌的生理代谢功能,强化难降解芳香有机物和含共轭双键大分子化合物的去除.微氧水解酸化出水的硫离子浓度((0.11±0.04)mg·L-1)显著低于厌氧出水((1.27±1.22)mg·L-1).454焦磷酸测序结果表明:微氧水解酸化菌群中,变形菌门、绿弯菌门和放线菌门菌群丰度(所占比例分别为39.7%、20.3%、1.9%)高于厌氧水解酸化菌群(分别为36.9%、17.5%、1.3%),对难降解大分子有机物的去除效果好;厌氧水解酸化菌群中拟杆菌门和酸杆菌门所占比例较大,酸化效果更好.在属的水平上,微氧水解酸化污泥中鉴定出的硫酸盐还原菌的种群多样性和丰度均低于厌氧污泥,这与其出水较低的硫离子浓度一致,表明微氧环境能够有效抑制硫酸盐还原菌的活性.上述研究结果表明,微氧水解酸化是一种很有前途的石化废水预处理技术.     

模拟氮沉降对干旱半干旱温带草原土壤细菌群落结构的影响

随着社会经济发展,活性氮源随人为活动逐年递增,导致氮沉降相应增加,由此带来的生态环境问题也日益严重.本研究基于细菌16S rRNA基因高通量测序技术,分析长期人工模拟氮沉降氮输入量为0、1、4、8、32 g·(m~2·a)~(-1)对干旱半干旱区草原土壤微生物群落结构的影响.结果表明,模拟氮沉降7 a增加了土壤中可利用氮含量,引起草原土壤酸化.高通量测序结果表明,细菌OTU(operational taxonomy unit)数量随氮输入量的增加呈现增加的趋势,高氮处理[8 g·(m~2·a)~(-1)和32g·(m~2·a)~(-1)]显著高于低氮和对照处理;细菌α多样性指数呈现先增加后降低的趋势.不同氮输入水平下土壤细菌群落结构差异显著,且与土壤p H和氮的有效性显著相关.不同细菌相对丰度在纲的水平对氮输入的响应趋势不同.氮的长期大量输入可直接或间接通过改变土壤性状而影响草地土壤细菌群落结构和多样性.     

Bacterial community analysis of Tatsoi cultivated by hydroponics

Tatsoi (Brassica narinosa) is a popular Asian salad green that is mostly consumed as a source of fresh produce. The purpose of this study was to assess the microbial diversity of Tatsoi cultivated in a hydroponic system and of its ecosystem. Tatsoi leaves, nutrient solution, and perlite/earth samples from a trickle feed system (TFS) and an ebb-and-flow system (EFS) were collected and their microbial communities were analyzed by pyrosequencing analysis. The results showed that most bacteria in the leaves from the TFS contained genus Sporosarcina (99.6%), while Rhizobium (60.4%) was dominant in the leaves from the EFS. Genus Paucibacter (18.21%) and Pelomonas (12.37%) were the most abundant microbiota in the nutrient solution samples of the TFS. In the EFS, the nutrient solution samples contained mostly genus Rhodococcus and Acinetobacter. Potential microbial transfer between the leaves and the ecosystem was observed in the EFS, while samples in the TFS were found to share only one species between the leaves, nutrient solution, and earth. Together, these results show that the bacterial populations in Tatsoi and in its ecosystem are highly diverse based on the cultivation system.