石灰性紫色土硝化作用及硝化微生物对不同氮源的响应

土壤中发生的硝化作用是对p H高度敏感的典型过程。本文采用室内恒温培养法,结合定量PCR和高通量测序,研究石灰性紫色土硝化作用以及氨氧化细菌(Ammonia-oxidizing bacteria,AOB)、氨氧化古菌(Ammonia-oxidizing archaea,AOA)、亚硝酸盐氧化细菌(Nitrite-oxidizing bacteria,NOB)的丰度与群落结构对不同氮源的响应。结果表明:不同氮源均刺激土壤硝化作用的发生,CO(NH2)2处理下的净硝化速率最大,约是CK处理的4.76倍,(NH_4)2SO4和NH_4Cl处理下的净硝化速率分别为N 3.88和3.34 mg kg-1d-1。相比于(NH_4)2SO4和CO(NH2)2处理,NH_4Cl处理降低了硝态氮的累积量,抑制了铵态氮的减少量。AOB amo A基因拷贝数在28 d培养过程中变化显著(p0.05),在(NH_4)2SO4和CO(NH2)2处理中呈先增长后降低趋势,在NH_4Cl处理中呈持续增长趋势;而AOA amo A基因拷贝数无显著变化(p0.05)。说明石灰性紫色土硝化作用的主要推动者是AOB,而不是AOA。在28 d培养过程中,亚硝酸盐氧化细菌占总微生物的比例高于氨氧化细菌和古菌,意味着石灰性紫色土中可能存在全程氨氧化微生物(Comammox)。高通量测序的结果表明:石灰性紫色土中AOB的优势种群为亚硝化螺菌Nitrosospira Cluster 3,AOA的优势种群是土壤古菌Group 1.1b,NOB的优势种群是硝化螺菌Nitrospira。     

土壤亚硝酸气体(HONO)排放过程及其驱动机制

气态亚硝酸(HONO)是大气中氢氧自由基(OH·)的重要来源,直接影响到大气氧化能力和空气质量。通过比较外场测定和模型计算的HONO浓度,发现白天时存在未知的大气HONO来源。研究表明,土壤可以向大气中排放HONO。其机理可能是土壤亚硝态氮和氢离子的化学平衡作用;或土壤夜间吸附和白天解吸附的动态物理化学过程;或氨氧化细菌等微生物的直接排放;也可能是硝化过程中产生的羟胺,在土壤颗粒物等表面的化学反应。因此,土壤HONO排放通量与土壤亚硝态氮浓度、pH、氨氧化细菌丰度、土壤矿物、土壤湿度及C/N值等相关。目前对于土壤HONO排放的研究尚在起步阶段,国内亦少见相关成果报道。本文综述了土壤HONO排放的研究背景、探讨了土壤HONO排放的机理及影响因素,以期为减少氮素损失、提高氮肥利用率、评估氮肥的环境效应及城市空气质量等提供理论依据和科学指导。     

秸秆直接还田与炭化还田对潮土硝化微生物的影响

为比较秸秆直接还田和炭化还田对黄淮海平原小麦/玉米轮作体系典型潮土硝化作用及硝化微生物群落的影响,设置4个处理:全量小麦秸秆还田(S)、全量秸秆炭化还田(B)、半量秸秆半量生物质炭还田(SB)和不进行秸秆或生物质炭还田的对照(CK),连续进行3 a田间试验。对小麦、玉米两个生长季土壤理化性质进行分析,用末端限制性片段长度多态性(Terminal-restriction length polymorphism,T-RFLP)技术和克隆文库技术对氨氧化古菌(Ammonia-oxidizing archaea,AOA)和氨氧化细菌(Ammonia-oxidizing bacteria,AOB)群落结构和多样性进行分析。结果表明,在小麦季,与S处理相比,B处理显著降低了土壤容重,提高了土壤pH、有机碳(SOC)和速效钾(AK)含量(P0.05),但并未显著影响土壤水分、铵态氮(NH_4~+-N)和硝态氮(NO_3~--N)含量;B和SB处理的硝化潜势(Potential nitrification rate,PNR)分别为0.58、0.49μg·h~(-1)·g~(-1)(以NO_2~-计,下同),显著高于CK,与S处理(0.40μg·h~(-1)·g~(-1))差异不显著。玉米季,B处理显著提高了土壤水分、SOC和AK(P0.05),各处理玉米季的PNR整体低于小麦季,B处理最高(0.27μg·h~(-1)·g~(-1)),显著高于CK和S处理(P0.05)。小麦季PNR分别与AK、NH_4~+浓度和土壤容重显著相关(P0.05),与AOA和AOB群落组成均无显著关系;玉米季PNR仅与理化因子SOC显著相关,但该季节PNR与AOB群落结构显著相关。冗余分析(RDA)表明,土壤SOC、容重、pH和AK是显著影响硝化微生物群落结构的主要因子,对AOA和AOB群落结构总变异的解释量分别为76.4%和75.5%。系统发育树分析表明,AOA大部分属于土壤古菌Group1.1b,AOB多属于亚硝化螺菌Nitrosospira簇3。综上,与秸秆直接还田相比,炭化还田提高土壤硝化活性,改善部分土壤理化性质,引起土壤硝化微生物群落结构变化。     

不同酸碱性紫色土的硝化活性及微生物群落组成

为研究紫色土中的硝化作用、总微生物和硝化微生物的群落结构,以重庆永川的酸性紫色土(p H=5.3)和中性紫色土(p H=7.2)以及四川盐亭的石灰性紫色土(p H=8.5)为研究对象,采用稳定性同位素标记技术进行培养实验,并通过Miseq测序对三种紫色土微生物群落结构进行分析。每种土样共设有三种处理,包括~(13)CO_2标记处理、~(12)CO_2对照处理和~(13)CO_2+C_2H_2对照处理。结果表明,中性紫色土(p H=7.2)和石灰性紫色土(p H=8.5)经过56 d的培养后发生了强烈的硝化作用,而酸性紫色土(p H=5.3)中未发生明显的硝化作用,并且硝化作用类型都以自养硝化为主。三种紫色土中都存在着变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicutes)、绿弯菌门(Chloroflexi)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)和芽单胞菌门(Gemmatimonadetes),其中变形菌门在三种紫色土中都大约占有20%的比例。中性紫色土和石灰性紫色土各处理中硝化螺旋菌门(Nitrospirae)百分比大于酸性紫色土各处理。三种紫色土各处理中的氨氧化细菌(Ammonia-oxidizing bacteria,AOB)主要以亚硝化螺菌属(Nitrosospira)为主,亚硝酸氧化细菌(Nitrite-oxidizing bacteria,NOB)主要以硝化螺菌属(Nitrospira)为主。且NOB/AOB的值在三种紫色土各处理中最高可达到13,这意味着全程氨氧化细菌(Comammox)可能在紫色土的硝化作用中占据重要贡献。     

两个水稻品种根际土壤细菌和氨氧化细菌的群落结构差异

通过根盒试验比较了籼稻汕优63和粳稻武运粳7号苗期不同采样期根际土和土体土壤的硝化强度以及氨氧化细菌数量的差异,并且采用16S rDNA PCR-DGGE(Denaturing gradient gel electrophoresis)指纹图谱技术比较分析了上述两种水稻苗期不同采样期根际和土体土壤中细菌及其氨氧化细菌的群落结构变化。结果表明,两个水稻品种根际土壤中硝化强度和氨氧化细菌的数量随着生育期的延长均表现出一定的正相关性,汕优63籼稻根部土壤中的细菌和氨氧化细菌的丰富度和群落变化特征随着水稻生育时期的延长较武运7号粳稻的变化更为多样,说明籼稻品种根系和根际硝化作用更强,在其根系附近会产生更多的硝态氮。这种差异性严重影响水稻植株对氮素的利用效率。     

北方黑土土壤细菌16S rDNA扩增体系的优化

采用降落PCR方法扩增目的基因,设计两次正交试验对PCR反应体系中的各组分浓度进行筛选,同时对退火时间、延伸时间及循环次数进行摸索。结果表明,适合北方黑钙土土壤细菌16S rDNA扩增体系为:在25μl体系中,10×buffer2.5μl,DNA模板20ng,Mg2+2.5mmolL-1,dNTPs0.25mmolL-1,引物0.3μmolL-1,TaqDNA聚合酶1.5U。降落PCR扩增程序为:95℃,5min;95℃,45s;65~56℃,60s;72℃,90s;每两个循环降低1℃。95℃,45s;5.5℃,60s;72℃,90s;10个循环。最后72℃,10min。     

Short-term population dynamics of ammonia oxidizing bacteria in an agricultural soil

Ammonia oxidizing bacteria (AOB) control the rate limiting step of nitrification, the conversion of ammonia (NH₄⁺) to nitrite (NO₂⁻). The AOB therefore have an important role to play in regulating soil nitrogen cycling. Tillage aerates the soil, stimulating rapid changes in soil N cycling and microbial communities. Here we report results of a study of the short term responses of AOB and net nitrification to simulated tillage and NH₄⁺ addition to soil. The intensively farmed vegetable soils of the Salinas Valley, California, provide the context for this study. These soils are cultivated frequently, receive large N fertilizer inputs and there are regional concerns about groundwater N concentrations. An understanding of N dynamics in these systems is therefore important. AOB population sizes were quantified using a real-time PCR approach. In a 15 day experiment AOB populations, increased rapidly following tillage and NH₄⁺ addition and persisted after the depletion of soil NH₄⁺. AOB population sizes increased to a similar degree, over a 1.5-day period, irrespective of the amount of NH₄⁺ supplied. These data suggest selection of an AOB community in this intensively farmed and C-limited soil, that rapidly uses NH₄⁺ that becomes available. These data also suggest that mineralization may play an especially important role in regulating AOB populations where NH₄⁺ pool sizes are very low. Methodological considerations in the study of soil AOB communities are also discussed.     

Characterization of nitrifying bacteria communities of soils from different ecological regions of China by molecular and conventional methods

Soil nitrification rate is very different among soil types, as a result of differences in physical and chemical properties. Little is known about the composition of the nitrifying bacteria community. In this investigation, three soils (fluvo-aquic soil, permeable paddy soil and red earth) from different geo-ecological regions in China were characterized for their nitrification activities and their nitrifying bacteria communities determined either by molecular approaches or by conventional culture methods. A 28-day long-term soil incubation showed that the maximum nitrification potential was found in the fluvo-aquic soil with almost 100% of inorganic N present as NO₃^–-N, while the minimum nitrification potential was in red earth with only a 4.9% conversion rate from ammonium into nitrate. There was no relationship between nitrification potential and numbers of nitrifiers in the soil. The conventional most probable number (MPN) method could enumerate ammonia oxidizers, but failed in enumerating nitrite oxidizers. Therefore, we used an MPN-PCR procedure which gave a convincing nitrite oxidizer count result, instead of MPN-diphylamine. Soils were characterized by denaturing gradient gel electrophoresis (DGGE) of DNA extracted from soils and amplified using a primer specific for the 16S rRNA gene and/or for the amoA gene. The DGGE columns of the three soils differed from each other. There were two similar bands present in DGGE columns of the fluvo-aquic and permeable paddy soils, but no similar band was found in DGGE columns of the red earth. The sequence of amoA indicated that all ammonia oxidizers in these soils were grouped into Nitrosospira clusters 1 and 3, and each soil had a common band similar to the other soils and a special band which differed from the other soils.     

pH regulates key players of nitrification in paddy soils

Increasing lines of evidence have suggested the functional importance of ammonia-oxidizing archaea (AOA) rather than bacteria (AOB) for nitrification in upland soils with low pH. However, it remains unclear whether niche specialization of AOA and AOB occurs in rice paddy wetlands constrained by oxygen availability. Using DNA-based stable isotope probing, we conclude that AOA dominated nitrification activity in acidic paddy soils (pH 5.6) while AOB dominated in alkaline soils (pH 8.2). Nitrification activity was stimulated by urea fertilization and accompanied by a significant increase of AOA in acid soils and AOB in alkaline soils. DNA-based stable isotope probing indicated significant assimilation of ¹³CO₂ for AOA only in acidic paddy soil, while AOB was the solely responsible for ammonia oxidation in the alkaline paddy soil. Phylogenetic analysis further indicated that AOA members within the soil group 1.1b lineage dominated nitrification in acid soils. Ammonia oxidation in the alkaline soil was catalyzed by Nitrosospira cluster 3-like AOB, suggesting that the physiological diversity of AOA is more complicated than previously thought, and soil pH plays important roles in shaping the community structures of ammonia oxidizers in paddy field.     

Contributions of ammonia-oxidizing archaea and bacteria to nitrification in Oregon forest soils

Ammonia oxidation, the first step of nitrification, is mediated by both ammonia-oxidizing archaea (AOA) and bacteria (AOB); however, the relative contributions of AOA and AOB to soil nitrification are not well understood. In this study we used 1-octyne to discriminate between AOA- and AOB-supported nitrification determined both in soil-water slurries and in unsaturated whole soil at field moisture. Soils were collected from stands of red alder (Alnus rubra Bong.) and Douglas-fir (Pseudotsuga menziesii Mirb. Franco) at three sites (Cascade Head, the H.J. Andrews, and McDonald Forest) on acidic soils (pH 3.9–5.7) in Oregon, USA. The abundances of AOA and AOB were measured using quantitative PCR by targeting the amoA gene, which encodes subunit A of ammonia monooxygenase. Total and AOA-specific (octyne-resistant) nitrification activities in soil slurries were significantly higher at Cascade Head (the most acidic soils, pH < 5) than at either the H.J. Andrews or McDonald Forest, and greater in red alder compared with Douglas-fir soils. The fraction of octyne-resistant nitrification varied among sites (21–74%) and was highest at Cascade Head than at the other two locations. Net nitrification rates of whole soil without NH₄⁺ amendment ranged from 0.4 to 3.3 mg N kg⁻¹ soil d⁻¹. Overall, net nitrification rates of whole soil were stimulated 2- to 8-fold by addition of 140 mg NH₄⁺-N kg⁻¹ soil; this was significant for red alder at Cascade Head and the H.J. Andrews. Red alder at Cascade Head was unique in that the majority of NH₄⁺-stimulated nitrifying activity was octyne-resistant (73%). At all other sites, NH₄⁺-stimulated nitrification was octyne-sensitive (68–90%). The octyne-sensitive activity—presumably AOB—was affected more by soil pH whereas the octyne-resistant (AOA) activity was more strongly related to N availability.     

Community composition of ammonia-oxidizing bacteria in the rhizosphere of intercropped wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.), maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.), and faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.)

Cereal/cereal and cereal/legume intercropping systems are popular in the north, northwest, and southwest of China and often result in yield increases compared to monocropping. Rhizosphere interactions may play a significant role in the yield increases, particularly with respect to nutrient availability. The aim of this study was to investigate the effects of intercropping on N availability and community composition of ammonia-oxidizing bacteria in the rhizosphere of wheat, maize, and faba bean at different growth stages. Denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes was used to analyze the community composition of bacterial ammonia oxidizers belonging to β-proteobacteria. The results showed that intercropping with faba bean significantly increased nitrate concentrations in the rhizosphere of wheat and maize at the second sampling time (20 June) compared to monocropping or intercropping between maize and wheat. Intercropping significantly affected the community composition of ammonia-oxidizing bacteria in the rhizosphere compared to monocropping, and the effects were most pronounced in the maize/faba bean and wheat/maize intercropping systems when faba bean and wheat were at anthesis and maize was in seedling stage. In wheat/faba bean intercropping, the effects of intercropping on community composition of ammonia-oxidizing bacteria were less pronounced at the seedling stage of the two species but were significant at anthesis.     

反刍动物饲料中乳源性成分与牛羊肉骨粉的区分

利用奶粉的溶解性,通过水和0.5%次氯酸钠溶液洗涤混有乳源性成分和牛、羊肉骨粉的饲料样品,去除饲料中的乳粉成分后,再使用16S rDNA PCR方法进行动物源性检测。结果表明,实验所建立的方法能够完全区分饲料中的乳粉与肉骨粉。当乳粉含量分别为25%、50%和75%时,混合饲料中牛、羊肉骨粉的检出限分别为2%、0.5%和0.1%。此方法操作简单,容易掌握,可用于鉴别反刍动物饲料中非乳源性成分的牛、羊源性成分。     

Responses of ammonia-oxidizing bacteria and archaea to nitrogen fertilization and precipitation increment in a typical temperate steppe in Inner Mongolia

As the first and rate-limiting step of nitrification, ammonia oxidation can be realized either by ammonia-oxidizing bacteria (AOB) or archaea (AOA). However, the key factors driving the abundance, community structure and activity of ammonia oxidizers are still unclear, and the relative importance of AOA and AOB in ammonia oxidation is unresolved. In the present study, we examined the effects of long-term (6 years) nitrogen (N) addition and simulated precipitation increment on the abundance and community composition of AOA and AOB based on a field trial in a typical temperate steppe of northern China. We used combined approaches of quantitative PCR, terminal-restriction fragment length polymorphism (T-RFLP) and clone library analyses of amoA genes. The study objective was to determine (1) AOA and AOB diversity and activity in response to N addition and increased precipitation and (2) the relative contributions of AOA and AOB to soil ammonia oxidation in the typical temperate steppe. The results showed that the potential nitrification rate (PNR) increased with N addition, but decreased with increased precipitation. Both N addition and increased precipitation significantly increased AOB but not AOA abundance, and a significant correlation was only observed between PNR and AOB amoA gene copies. The T-RFLP analysis showed that both N and precipitation were key factors in shaping the composition of AOB, while AOA were only marginally influenced. Phylogenetic analysis indicated that all AOA clones fell within the soil and sediment lineage while all AOB clones fell within the Nitrosospira. The study suggested that AOA and AOB had distinct physiological characteristics and ecological niches. AOB were shown to be more sensitive to N and precipitation than AOA, and the ammonia oxidation process was therefore supposed to be mainly driven by AOB in this temperate steppe.     

桂西北不同植被恢复阶段土壤氨氧化细菌遗传多样性研究

以桂西北喀斯特不同植被恢复阶段(草丛、灌木林、次生林、原生林)生态系统为研究对象,运用分子生物学技术分析了土壤氨氧化细菌amoA功能基因多样性,探讨了其与脲酶活性和土壤理化性质的关系.结果显示,随着植被的恢复,土壤氨氧化细菌多样性指数与均匀度指数呈增大趋势(灌木林例外),且土壤中氨氧化细菌群落结构发生了改变:主要表现在因Nitrosospira3簇种群对铵态氮浓度敏感度差异导致其在3a、3b簇中分布不一致;相关分析表明;土壤脲酶活性与铵态氮浓度呈正相关关系,土壤脲酶可能通过影响铵态氮浓度改变氨氧化细菌多样性,但植被恢复后期土壤铵态氮浓度减少并未降低土壤氨氧化细菌多样性.LIBSHUFF和RDA分析揭示,植被类型和土壤脲酶活性及pH与氨氧化细菌群落结构紧密相关,说明植被和土壤氮素有效性以及pH可能是决定土壤氨氧化细菌多样性的主要因子,为深入理解喀斯特地区土壤氮素循环提供了一定的科学依据.     

Ammonia-oxidizing communities in agricultural soil incubated with organic waste residues

The impact of organic compounds present in different kinds of organic fertilizers, i.e., anaerobically digested household waste, composted organic household waste, swine manure, and cow manure, on microbial communities in arable soil was investigated using microcosms. Soil was amended with dried residues or organic extracts of the residues and incubated for 12 weeks at 25°C. The microbial community composition was investigated by phospholipid fatty acid (PLFA) analysis, and the community of ammonia-oxidizing bacteria (AOB) was assessed by denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments, followed by sequencing. All dried residues increased the AOB activity, determined as potential ammonia oxidation, whereas the organic extracts from the thermophilically digested waste and the swine manure caused a decreased potential activity. However, no differences in the DGGE banding patterns were detected, and the same AOB sequences were present in all samples treated with the residue extracts. Moreover, the PLFA composition showed that none of the residue additions affected the overall microbial community structure in the soil. We conclude that the AOB community composition was not affected by the organic compounds in the fertilizers, although the activity in some cases was.     

Bacterial diversity and occurrence of ammonia-oxidizing bacteria in the Atacama Desert soil during a “desert bloom” event

The Atacama Desert, located in northern Chile, is one of the driest deserts on the Earth. However, in some years, short and sporadic rainfall in the southern end of this desert has increased the soil moisture that produces ephemeral “desert bloom”. Our goal was to assess the composition of the bacterial community and determine variations in the ammonia-oxidizing bacteria guild diversity from soils collected during the course of the “desert bloom” event. The bacterial composition from this arid soil was determined by cloning and sequencing the 16S rRNA gene. A relatively high diversity of clones belonging to 14 bacterial groups was detected. The ammonia-oxidizers showed a significantly higher diversity of amoA gene clones after the “desert bloom” than during or at the beginning of this event. All sequences obtained were related to Nitrosospira genera and environmental clones. These results suggest that the diversity of ammonia-oxidizing bacteria in this arid soil can be affected by the occurrence of “desert bloom”.     

Survival of bacterial DNA and culturable bacteria in archived soils from the Rothamsted Broadbalk experiment

Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 10⁵ genomes g⁻¹ soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.     

黑土区大豆基因型的根际细菌群落结构时空动态变化

The dynamics of rhizosphere microbial communities is important for plant health and productivity, and can be influenced by soil type, plant species or genotype, and plant growth stage. A pot experiment was carried out to examine the dynamics of microbial communities in the rhizosphere of two soybean genotypes grown in a black soil in Northeast China with a long history of soybean cultivation. The two soybean genotypes, Beifeng 11 and Hai 9731, differing in productivity were grown in a mixture of black soil and siliceous sand. The bacterial communities were compared at three zone locations including rhizoplane, rhizosphere, and bulk soil at the third node (V3), early flowering (R1), and early pod (R3) stages using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rDNA. The results of principal component analyses (PCA) showed that the bacterial community structure changed with growth stage. Spatially, the bacterial communities in the rhizoplane and rhizosphere were significantly different from those in the bulk soil. Nevertheless, the bacterial communities in the rhizoplane were distinct from those in the rhizosphere at the V3 stage, while no obvious differences were found at the R1 and R3 stages. For the two genotypes, the bacterial community structure was similar at the V3 stage, but differed at the R1 and R3 stages. In other words, some bacterial populations became dominant and some others recessive at the two later stages, which contributed to the variation of the bacterial community between the two genotypes. These results suggest that soybean plants can modify the rhizosphere bacterial communities in the black soil, and there existed genotype-specific bacterial populations in the rhizosphere, which may be related to soybean productivity.     

一种可用于PCR扩增的直接提取土壤细菌DNA的方法

本文以澳大利亚桉树林和松树林的土壤为例 ,采用Napp提取液和SDS直接溶解土壤细菌 ,并配合温浴 -玻璃珠震荡、苯酚 -氯仿萃取和异丙醇提取以及纯化DNA等步骤 ,直接从土壤样品中提取了土壤细菌DNA。所得DNA完全适用于酶解和PCR扩增的要求。该方法高效简单 ,费用低 ,在土壤微生物研究中具有重要的应用价值