土壤宏基因组学技术及其应用

传统的基于培养的研究方法只能反映土壤中少数(0.1%～10 %)微生物的信息,而大部分微生物目前还不能培养,因而这部分微生物资源尚难以被有效地开发利用.宏基因组学是分子生物学技术应用于环境微生物生态学研究而形成的一个新概念,主要技术包括土壤DNA的提取、文库的构建和目标基因克隆的筛选.它可为揭示微生物生态功能及其分子基础提供更全面的遗传信息,并已在微生物新功能基因筛选、活性物质开发和微生物多样性研究等方面取得了显著成果.本文对土壤宏基因组学技术的方法和应用作了详细介绍.     

宏组学方法在污水处理系统中的应用进展

污水生物处理由微生物生理过程驱动,宏组学方法能够获得不同水平的分子信息,为认识污水处理系统中的微生物提供了新途径。本文对宏基因组学、宏转录组学、宏蛋白质组学与代谢组学等宏组学方法的发展进行综述,着重介绍各组学及整合宏组学在污水处理系统中的研究现状,并指出其应用前景。     

宏组学在发酵食品微生物群落研究中的应用进展

发酵食品是人类饮食的重要组成部分,深入研究其中的微生物群落结构、代谢及其活性,对于充分探索微生物资源、促进发酵食品的质量具有深远意义。目前,随着后基因组学时代的到来,测序技术的迅猛发展及其成本的降低,主要基于宏基因组学、宏转录组学,宏蛋白质组学和代谢组学在内的宏组学技术已经应用于微生物群落的多样性、结构及潜在基因功能等方面,为发酵食品潜在分子机制的研究奠定了基础。总结了在发酵食品中应用的宏组学(宏基因组学、宏转录组学,宏蛋白质组学和代谢组学)技术。随后,对奶酪、发酵酒精饮料、发酵蔬菜、发酵茶、发酵豆制品和食醋环境中微生物群落结构和功能方面的宏组学研究进行了综述,揭示了发酵食品中有益基因和生物活性物质的来源,并简单概述了这些技术的优势及其现阶段所存在的不足,旨为未来发酵食品的科学研究和工业生产提供一定参考。     

基于宏组学方法认识微生物群落及其功能

进入后基因组学时代,测序技术飞速发展,测序成本明显下降,形成了涵盖宏基因组学、宏转录组学和宏蛋白质组学的宏组学技术,推动了对微生物群落的多样性、结构及潜在基因功能方面的深入研究。最近随着整合的宏组学技术的提出及应用,全面系统分析微生物群落动态变化及其代谢功能已成为可能,这将成为微生物生态学研究的新趋势。本文综述了宏组学在研究海洋湖泊、深海热泉、人体肠道、牛瘤胃生境、森林土壤与堆肥生境等环境中微生物群落的结构和功能方面的最新进展与成功应用案例。     

宏蛋白质组学技术在废水生物处理工艺研究领域中的应用

随着后基因组时代的到来,宏蛋白质组学逐渐兴起并在生命科学基础领域和临床医药领域成功运用,宏蛋白质组学技术现已成为各研究领域炙手可热的方法之一.宏蛋白质组学技术在废水生物处理研究领域中的应用刚起步,但已展示其强大功能.本文主要综述近年来国内外宏蛋白质组学在废水生物处理研究领域的研究进展,回顾及总结了宏蛋白质组学的研究策略及应用,如鉴定功能性蛋白质/酶、揭示污染物的微生物降解途径、推断废水生物处理系统的关键代谢途径、及探讨不同污泥微生物群落微生态变化等.
     

宏蛋白质组学信息分析的基本策略及其挑战

宏蛋白质组学是一门新型科学,它运用质谱技术规模化地采集自然界微生物种群的蛋白质信息,并结合多种组学数据,开展微生物种群的遗传特征及其生物功能的研究.宏蛋白质组学的信息分析与传统蛋白质组学方法有较大的不同,亟需拓展新的分析思路.由于宏蛋白质组的研究对象是复杂度极高的微生物样品,因此,需要构建尽可能囊括样本中所含微生物的基因组信息的物种数据库.面对庞大的数据库,必须考虑到分析过程中所消耗的计算资源和鉴定结果的质控标准,因此,需要高度优化库容量、搜库、假阳性控制等参数.鉴于宏蛋白质组数据中广泛存在复杂的同源蛋白质序列,因此,需要充分利用NCBI数据库中的分类信息进行匹配,并运用LCA算法过滤处理才能将蛋白质有效地归组到物种.本文立足于宏蛋白质组学信息分析,从宏蛋白质组的数据库建立、蛋白质归并、生物学意义发掘等几个方面着手,对该领域的发展现状、面临挑战以及未来研究方向进行了评述.     

生态毒理基因组学和生态毒理蛋白质组学研究进展

将基因组学和蛋白质组学知识整合到生态毒理学中形成了生态毒理基因组学和生态毒理蛋白质组学.通过生态毒理基因组学和生态毒理蛋白质组学的研究能够在基因组和蛋白质组水平更深入理解毒物的作用机制,寻找更敏感、有效的生物标记物,形成潜在的强有力的生态风险评价工具.介绍了生态毒理基因组学和生态毒理蛋白质组学的研究进展,以及DNA芯片技术和2D-凝胶电泳技术在持久性有毒污染物的生态毒理学研究中的应用.     

宏基因组学:土壤微生物研究的新策略

土壤中多数微生物不可培养,这限制了微生物资源的开发利用。宏基因组学方法在开发和利用不可培养微生物资源方面有巨大潜力,可以将其运用到土壤微生物学研究中。对土壤宏基因组DNA的提取、宏基因组文库的构建和筛选等方面的研究现状和进展进行了简要综述。     

微生物蛋白质组学的定量分析

越来越多的微生物基因组序列数据为系统地研究基因的调节和功能创造了有利条件.由于蛋白质是具有生物功能的分子,蛋白质组学在微生物基因组的功能研究中异军突起、蓬勃发展.微生物蛋白质组学的基本原则是,用比较研究来阐明和理解不同微生物之间或不同生长条件下基因的表达水平.显而易见,定量分析技术是比较蛋白质组学中急需发展的核心技术.对蛋白质组学定量分析技术在微生物蛋白质组研究中的进展进行了综述.     

环境微生物学的组学技术应用和突破（英文稿）

由于研究环境变化和微生物群落的需要,近年来高通量组学技术得到了迅猛开发和应用.其中,基于测序和芯片技术的宏基因组学是一个关键的、最成熟的组学技术,为大多数的其它组学技术提供了支撑.相比较而言,宏转录组学、宏蛋白质组学和宏代谢组学也取得了少数的有限成功,但已经显示出可喜的潜力.所有的组学技术都有赖于生物信息学,使得后者成为组学技术应用的一个主要的技术瓶颈.这些新的组学技术对环境微生物学领域产生了革命性的影响,极大地丰富了我们对于环境微生物基因资源和功能活性的了解.     

植物、土壤及土壤管理对土壤微生物群落结构的影响

土壤微生物是土壤生态系统的重要组成部分,对土壤微生物群落结构多样性的研究是近年来土壤生态学研究的热点。本文综述了有关植物、土壤类型以及土壤管理措施对土壤微生物群落结构影响的最新研究结果,指出植物的作用因植物群落结构多样性、植物种类、同种植物不同的基因型,甚至同一植物不同根的区域而异;而土壤的作用与土壤质地和有机质含量等因素有关;植物和土壤类型在对土壤微生物群落结构影响上的作用存在互作关系。不同的土壤管理措施对土壤微生物群落结构影响较大,长期连作、大量的外援化学物质的应用降低了土壤微生物的多样性;而施用有机肥、免耕可以增加土壤微生物群落结构多样性,有利于维持土壤生态系统的功能。     

Estimating respiration of roots in soil: Interactions with soil CO2, soil temperature and soil water content

Little information is available on the variability of the dynamics of the actual and observed root respiration rate in relation to abiotic factors. In this study, we describe I) interactions between soil CO₂ concentration, temperature, soil water content and root respiration, and II) the effect of short-term fluctuations of these three environmental factors on the relation between actual and observed root respiration rates. We designed an automated, open, gas-exchange system that allows continuous measurements on 12 chambers with intact roots in soil. By using three distinct chamber designs with each a different path for the air flow, we were able to measure root respiration over a 50-fold range of soil CO₂ concentrations (400 to 25000 ppm) and to separate the effect of irrigation on observed vs. actual root respiration rate. All respiration measurements were made on one-year-old citrus seedlings in sterilized sandy soil with minimal organic material.Root respiration was strongly affected by diurnal fluctuations in temperature (Q₁₀ = 2), which agrees well with the literature. In contrast to earlier findings for Douglas-fir (Qi et al., 1994), root respiration rates of citrus were not affected by soil CO₂ concentrations (400 to 25000 ppm CO₂; pH around 6). Soil CO₂ was strongly affected by soil water content but not by respiration measurements, unless the air flow for root respiration measurements was directed through the soil. The latter method of measuring root respiration reduced soil CO₂ concentration to that of incoming air. Irrigation caused a temporary reduction in CO₂ diffusion, decreasing the observed respiration rates obtained by techniques that depended on diffusion. This apparent drop in respiration rate did not occur if the air flow was directed through the soil. Our dynamic data are used to indicate the optimal method of measuring root respiration in soil, in relation to the objectives and limitations of the experimental conditions.     

生物质炭对水稻土团聚体微生物多样性的影响

生物质炭施用对土壤微生物群落结构的影响已有报道,但土壤团聚体粒组中微生物群落对生物质炭施用的响应的研究还相对不足。以施用玉米秸秆生物质炭两年后的水稻土为对象,采用团聚体湿筛法,通过高通量测序对土壤团聚体的微生物群落结构与多样性进行分析,结果表明:(1)与对照相比,生物质炭施用显著促进了大团聚体(2000—250μm)的形成,并提高了团聚体的稳定性。(2)不同粒径团聚体间微生物相对丰度存在显著差异。在未施生物质炭的处理(C0)中,随着团聚体粒径增大,变形菌门、子囊菌门、β-变形杆菌目、格孢腔菌目的相对丰度逐渐降低,而酸杆菌门、担子菌门、粘球菌目、类球囊霉目的相对丰度逐渐升高。(3)生物质炭施用显著改变了团聚体间的微生物群落结构。与C0处理相比,生物质炭施用处理的大团聚体中变形菌门、鞭毛菌门和β-变形杆菌目的相对丰度分别显著提高了14.37%、33.28%和33.82%;微团聚体(250—53μm)中酸杆菌门、子囊菌门和粘球菌目的相对丰度分别显著降低了20.15%、19.93%和17.66%;粉、黏粒组分(53μm)中担子菌门的相对丰度升高90.25%,而子囊菌门和鞭毛菌门的相对丰度分别降低12.15%和12.58%。由此可见,生物质炭不仅改变土壤团聚体组成和分布,同时伴随着土壤微生物群落结构的改变。     

The porosity of soil aggregates from bulk soil and from soil adhering to roots

Summary Total porosity and pore-size distribution (p.s.d.) were determined in soil aggregates taken in plots planted with maize and treated with farmyard manure and three rates of compost. Soil aggregates were collected from the soil adherent to the maize roots (root soil aggregates) and from bulk soil (bulk soil aggregates). Mercury intrusion porosimetry was used to evaluate the total porosity and the p.s.d. Treatments did not affect the total porosity of the bulk soil aggregates. The same was observed for the root soil aggregates. However the total porosity of the root soil aggregates was always lower than that of the bulk soil aggregates. The loss of total porosity was found to be due to a decrease in the percentage of larger pores with respect to the total.     

Available soil nitrogen in relation to fractions of soil nitrogen and other soil properties

The available N of 27 soils from England and Wales was assessed from the amounts of N taken up over a 6-month period by perennial ryegrass grown in pots under uniform environmental conditions. Relationships between availability and the distribution of soil N amongst various fractions were then examined using multiple regression. The relationship: available soil N (mg kg^–1 dry soil)=(N_min×0.672)+(N_inc×0.840)+(N_mom×0.227)–5.12 was found to account for 91% of the variance in available soil N, where N_min=mineral N, N_inc=N mineralized on incubation and N_mom=N in macro-organic matter. The N mineralized on incubation appeared to be derived largely from sources other than the macro-organic matter because these two fractions were poorly correlated. When availability was expressed in terms of available organic N as % of soil organic N (N_ao) the closest relationship with other soil characteristics was: N_ao=[N_inc×(1.395–0.0347×CN_mom]+[N_mom×0.1416], where CN_mom=CN ratio of the macro-organic matter. This relationship accounted for 81% of the variance in the availability of the soil organic N.The conclusion that the macro-organic matter may contribute substantially to the available N was confirmed by a subsidiary experiment in which the macro-organic fraction was separated from about 20 kg of a grassland soil. The uptake of N by ryegrass was then assessed on two subsamples of this soil, one without the macro-organic matter and the other with this fraction returned: uptake was appreciably increased by the macro-organic matter.     

Linking soil bacterial biodiversity and soil carbon stability

Native soil carbon (C) can be lost in response to fresh C inputs, a phenomenon observed for decades yet still not understood. Using dual-stable isotope probing, we show that changes in the diversity and composition of two functional bacterial groups occur with this ‘priming'' effect. A single-substrate pulse suppressed native soil C loss and reduced bacterial diversity, whereas repeated substrate pulses stimulated native soil C loss and increased diversity. Increased diversity after repeated C amendments contrasts with resource competition theory, and may be explained by increased predation as evidenced by a decrease in bacterial 16S rRNA gene copies. Our results suggest that biodiversity and composition of the soil microbial community change in concert with its functioning, with consequences for native soil C stability.Substrate inputs can stimulate decomposition of native soil organic carbon (SOC; Kuzyakov et al., 2000), a phenomenon known as the ‘priming effect'' (Kuzyakov, 2010), and is considered large enough to influence ecosystem C balance (Wieder et al., 2013). Two functionally distinct groups of microorganisms are postulated to mediate priming: one that grows rapidly utilizing labile C, and one that grows slowly, breaking down recalcitrant SOC (Fontaine et al., 2003; Blagodatskaya et al., 2007). However, distinguishing these groups is technically challenging. Here, we used dual-stable isotope probing with ¹³C-glucose and ¹⁸O-water to identify bacteria in these two groups growing in response to single and repeated pulses of glucose. Organisms that utilize labile C for growth assimilate both ¹³C-glucose and ¹⁸O-water into their DNA, whereas organisms that grow using SOC incorporate only ¹⁸O-water. Differential isotope incorporation leads to a range of DNA densities separable through isopycnic centrifugation, which can then be characterized by sequencing (Radajewski et al., 2000).We sequenced fragments of bacterial 16S rRNA genes following single and repeated glucose pulses. We hypothesized that the single pulse of labile C would stimulate growth of opportunistic organisms, thus immobilizing nutrients and suppressing growth and diversity of the SOC-utilizing community, decreasing SOC decomposition (negative priming), a response analogous to that observed in plant communities in response to chronic N additions (Tilman, 1987; Clark and Tilman, 2008). We hypothesized that multiple glucose additions would stimulate growth of a more diverse bacterial community, including more native SOC-utilizing organisms that possess enzymes to decompose recalcitrant compounds, causing positive priming (Fontaine et al., 2003; Kuzyakov, 2010).Soil from a ponderosa pine ecosystem was amended weekly for 7 weeks with 500 μg C-glucose per gram soil (2.65 atom % ¹³C) in 100 μl deionized water or with 100 μl deionized water (n=5). Measurements of δ¹³C–CO₂ and [CO₂] enabled the partitioning of CO₂ into that derived from added glucose or from native SOC (C_SOC):where C_total is CO₂–C from glucose-amended samples, δ_total is the δ¹³C–CO₂ from glucose-amended samples, δ_glucose is the δ¹³C of the added glucose and δ_SOC is the δ¹³C–CO₂ evolved from the non-amended samples. Priming was calculated as the difference between SOC oxidation of the amended and non-amended samples. With this approach, any evolved CO₂ carrying the ¹³C signature of the added glucose is considered respiration of glucose, including ¹³C-labeled biomass and metabolites derived from prior glucose additions. Thus, this approach quantifies priming as the oxidation of SOC present at the beginning of the experiment, consistent with many other studies of priming (Cheng et al., 2003; De Graaff et al., 2010).In a parallel incubation for dual-stable isotope probing, the repeated-pulse samples received unlabeled glucose (500 μg C-glucose per gram soil) for 6 weeks while the non-amended and single-pulse samples received sterile deionized water. In week 7, samples received one of four isotope treatments (n=3): 97 atom % H₂ ¹⁸O (non-amended soil), 99 atom % ¹³C-glucose and 97 atom % H₂ ¹⁸O (single- and repeated-pulse soil), ¹²C-glucose and 97 atom % H₂ ¹⁸O (repeated-pulse soil) or ¹²C-glucose and H₂ ¹⁶O (repeated-pulse soil). After incubating for 7 days, soil was frozen at −40 °C. DNA was extracted, separated through isopycnic centrifugation, and two density ranges were sequenced for the bacterial 16S rRNA gene (Supplementary Figure 1): 1.731–1.746 g ml⁻¹ (hereafter called the SOC-utilizing community) and 1.759–1.774 g ml⁻¹ (hereafter called the glucose-utilizing community).Amplicons of the V3–V6 16S rRNA region were bar coded with broad-coverage fusion PCR primers and pooled before sequencing on a Genome Sequencer FLX instrument. These sequence data have been submitted to the GenBank database under accession number SRP043371. Data were checked for chimeras (Edgar et al., 2011), demultiplexed and quality checked (Caporaso et al., 2010). Taxonomy was assigned to genus at the ⩾80% bootstrap confidence level (Cole et al., 2009).We used the Shannon''s diversity index (H′), commonly used in microbial systems (Fierer and Jackson, 2006), to assess changes in microbial diversity. Analysis of variance was used to compare the amount of DNA within densities between isotope treatments (Supplementary Figure 2) and to test the effects of the treatments on the Shannon''s diversity (Figure 2) and Pielou''s evenness (Supplementary Figure 3) of the active bacterial communities, with post hoc Student''s t-tests, α=0.05. PRIMER 6 and PERMANOVA were used to create the nonmetric multidimensional scaling ordination and to compare bacterial communities between glucose treatments and the two sequenced density ranges.The single pulse of glucose suppressed SOC oxidation, whereas repeated pulses increased SOC oxidation (Figure 1). Few experiments to date have examined priming in response to repeated substrate amendments (Hamer and Marschner, 2005; Qiao et al., 2014), even though in nature soil receives repeated substrate pulses from litterfall and rhizodeposition. Our results demonstrate the dynamic response of SOC decomposition to repeated labile C inputs.Open in a separate windowFigure 1Weekly priming rates calculated as the difference in SOC respired between glucose-amended and non-amended soil (n=5).Dual-stable isotope probing was able to separate the growing bacteria into two groups with distinct DNA densities (P<0.001, PERMANOVA; Figure 3a), indicating differential uptake of ¹³C-glucose and ¹⁸O-water. In response to the initial glucose addition, the diversity of the growing glucose- and SOC-utilizing bacterial communities declined compared with the non-amended community (P<0.001, t-tests; Figure 2), driven by a strong decrease in evenness (Supplementary Figure 3). In the SOC-utilizing community, where DNA was labeled with ¹⁸O only, the relative abundance of Bacillus increased 4.9-fold compared with the non-amended control to constitute 31.6% of the community (Figure 3b). Bacillus survives well under low-nutrient conditions (Panikov, 1995), and is able to synthesize a suite of extracellular enzymes capable of degrading complex substrates (Priest, 1977), traits that are conducive for using SOC for growth. In the glucose-utilizing community, where DNA was labeled with both ¹³C and ¹⁸O, Arthrobacter increased 67.7-fold relative to the non-amended control to constitute 75.5% of the growing bacteria (Figure 3b). In culture experiments, Arthrobacter can rapidly take up and store glucose for later use (Panikov, 1995) and here we find it dominating the high-density DNA fractions, signifying that it is using the labeled glucose to grow. The increased biomass of Arthrobacter may have resulted in greater resource competition, thus reducing the diversity of the growing community, as is frequently found in plant communities (Bakelaar and Odum, 1978; Clark and Tilman, 2008).Open in a separate windowFigure 2Shannon''s diversity index (H′) of the non-amended, single-pulse, and repeated-pulse treatments (n=3) in the SOC- (mid-density) and glucose-utilizing (high-density) communities. Treatments with the same letter are not significantly different from each other (Student''s t, α=0.05).Open in a separate windowFigure 3(a) Nonmetric multidimensional scaling ordination showing differences in growing bacterial communities at the genus taxonomic level in the SOC-utilizing (mid-density; open symbols) and glucose-utilizing (high-density; closed symbols) groups of non-amended (Δ), single-pulse (○) and repeated-pulse (□) treatments (n=3). (b) Pie charts of genera in the SOC- and glucose-utilizing communities of the single- and repeated-pulse treatments (n=3). Genera with relative abundances >5% are listed in the figure legend.After repeated glucose amendments, the diversity of the growing community recovered to non-amendment levels (Figure 2) without strongly dominant organisms (Figure 3b and Supplementary Figure 3). The higher diversity found after repeated glucose pulses may be explained by trophic interactions where predators graze on prey populations that have been enlarged by resource addition, suppressing competition between prey species and causing secondary mobilization of nutrients (Clarholm, 1985). The decrease in total bacterial 16S rRNA gene copies in the repeated-pulse—compared with the single-pulse—treatment (Supplementary Figure 4) supports predation as a potential mechanism explaining the observed diversity increase after repeated glucose pulses.The recovery of diversity after repeated glucose pulses contrasts with resource competition theory (Tilman, 1987). When chronic additions of a limiting resource are applied, species diversity and evenness typically decrease (Bakelaar and Odum, 1978; Clark and Tilman, 2008) because competitive organisms become dominant. We observed this after the single glucose pulse, but not after repeated pulses. This diversity response may be the result of community shifts facilitated by short bacterial life cycles and the tens to hundreds of generations expected during the 7-week incubation (Behera and Wagner, 1974). In contrast, systems on which most ecological theory is based (for example, plants) might achieve perhaps 20 generations in a multi-decadal field experiment (Bakelaar and Odum, 1978; Clark and Tilman, 2008). With more generations, more community dynamics can occur, including increased resource cascades in which extracellular enzymes, metabolites or lysed cells of one functional group increase substrates for another (Blagodatskaya and Kuzyakov, 2008). Our results highlight the opportunity to test ecological theories in microbial ecosystems (Prosser et al., 2007), particularly as the short life cycles of microbes makes them well suited for pursuing ecological questions in an evolutionary framework (Jessup et al., 2004).The priming effect is ubiquitous, yet its drivers remain elusive. Our results suggest that changes in the diversity and composition of the growing bacterial community contribute to priming, and thus that ecosystem properties such as soil C storage may be sensitive to soil microbial biodiversity.     

耕作方式对潮土土壤团聚体微生物群落结构的影响

为探究不同耕作方式对潮土土壤团聚体微生物群落结构和多样性的影响,采用磷脂脂肪酸(PLFA)法测定了土壤团聚体中微生物群落。试验设置4个耕作处理,分别为旋耕+秸秆还田(RT)、深耕+秸秆还田(DP)、深松+秸秆还田(SS)和免耕+秸秆还田(NT)。结果表明:与RT相比,DP处理显著提高了原状土壤和>5 mm粒级土壤团聚体中真菌PLFAs量和真菌/细菌,为真菌的繁殖提供了有利条件,有助于土壤有机质的贮存,提高了土壤生态系统的缓冲能力;提高了5～2 mm粒级土壤团聚体中细菌PLFAs量,降低了土壤革兰氏阳性菌/革兰氏阴性菌,改善了土壤营养状况;提高了<0.25 mm粒级土壤团聚体中微生物丰富度指数。总的来说,深耕+秸秆还田(DP)对土壤团聚体细菌和真菌生物量有一定的提高作用,并且在一定程度上改善了土壤团聚体微生物群落结构,有利于增加土壤固碳能力和保持土壤微生物多样性。冗余分析结果表明,土壤团聚体总PLFAs量、细菌、革兰氏阴性菌和放线菌PLFAs量与土壤有机碳相关性较强,革兰氏阳性菌PLFAs量与总氮相关性较强。各处理较大粒级土壤团聚体微生物群落主要受碳氮比、含水量、pH值和团聚体质量分数的影响,较小粒级土壤团聚体微生物群落则主要受土壤有机碳和总氮的影响。     

酸性硫酸盐土水改旱后土壤化学性状的变异初报

探讨了酸性硫酸盐水稻土改为旱作后土壤化学性状的变异以及比较不同利用方式之间的经济效益．结果表明,酸性硫酸盐水稻土改种甜玉米和蔬菜后,土壤化学性状发生显著变化．耕层土壤酸度、水溶性硫酸根含量、土壤活性铝和活性铁含量均显著降低．经济效益得到显著提高．建议对水改旱后的环境效应进行深入研究以及进行定位观测,以便合理利用这一特殊的土壤资源     

Temporal dynamics of soil bacterial network regulate soil resistomes

Soil bacteria are diverse and form complicated ecological networks through various microbial interactions, which play important roles in soil multi-functionality. However, the seasonal effects on the bacterial network, especially the relationship between bacterial network topological features and soil resistomes remains underexplored, which impedes our ability to unveil the mechanisms of the temporal-dynamics of antibiotic resistance genes (ARGs). Here, a field investigation was conducted across four seasons at the watershed scale. We observed significant seasonal variation in bacterial networks, with lower complexity and stability in autumn, and a wider bacterial community niche in summer. Similar to bacterial communities, the co-occurrence networks among ARGs also shift with seasonal change, particularly with respect to the topological features of the node degree, which on average was higher in summer than in the other seasons. Furthermore, the nodes with higher betweenness, stress, degree, and closeness centrality in the bacterial network showed strong relationships with the 10 major classes of ARGs. These findings highlighted the changes in the topological properties of bacterial networks that could further alter antibiotic resistance in soil. Together, our results reveal the temporal dynamics of bacterial ecological networks at the watershed scale, and provide new insights into antibiotic resistance management under environmental changes.