Nematode assemblages in the rhizosphere of spring barley (Hordeum vulgare L.) depended on fertilisation and plant growth phase

Nematodes from rhizosphere soil of barley grown at three fertiliser treatments (control (0), NK and NPK) were studied in a field experiment. Sampling was done twice, during vegetative growth and flowering, respectively, to determine how fertiliser effects on nematode assemblages depended on plant growth phase. At the growth stage the proportion of fungal feeding nematodes (dominated by Aphelenchoides spp. and Aphelenchus sp.) was highest in NK. During flowering, the abundance and proportion of fungal feeders in the 0 and NPK plots had increased and reached a level similar to the NK plot. Overall densities of bacterial feeders (mainly Cephalobidae and Rhabditidae) were similar, but opportunistic bacterial feeders constituted a higher proportion in the fertilised plots compared to the unfertilised. Ectoparasitic plant feeders (Tylenchorhynchus sp.) were more numerous in NK and NPK than in the control at both sampling dates. Endoparasite (Pratylenchus spp.) numbers were lower in the NPK plot at the growth stage. Numbers of Tylenchidae increased between samplings. The classification of Tylenchidae as epidermal cell and root hair feeders as opposed to hyphal feeders is discussed. Results thus indicate that: (i) bacterial and especially fungal feeding nematodes are stimulated by unbalanced fertilisation; (ii) ectoparasitic plant feeders are stimulated by N-fertilisation, while migratory endoparasites are inhibited at high and balanced fertilisation; (iii) nutrient effects diminish after plants reach the flowering stage.     

The bioprotective effect of AM root colonization against the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici in barley depends on the barley variety

The systemic effect of root colonization by the arbuscular mycorrhizal fungus (AMF) Glomus mosseae on the susceptibility of old and modern barley varieties to the soil-borne fungal pathogen Gaeumannomyces graminis var. tritici (Ggt) was studied in a split-root system. Plants were precolonized on one side of the split-root system with the AMF and thereafter the other side of the split-root system was inoculated with the pathogen. At the end of the experiment the level of bioprotection was estimated by quantifying lesioned roots and the determination of the root fresh weight. AM root colonization provided protection in some of the barley genotypes tested, but not in others. This protective effect seemed to vary in the oldest and the most modern barley variety tested.     

Use of an artificial root to examine the influence of 8-hydroxyquinoline on soil microbial activity and bacterial community structure

Invasive plant species have been shown to alter the microbial community composition of the soils they invade and it is suggested that this below-ground perturbation of potential pathogens, decomposers or symbionts may feedback positively to allow invasive success. Whether these perturbations are mediated through specific components of root exudation are not understood. We focussed on 8-hydroxyquinoline, a putative allelochemical of Centaurea diffusa (diffuse knapweed) and used an artificial root system to differentiate the effects of 8-hydroxyquinoline against a background of total rhizodeposition as mimicked through supply of a synthetic exudate solution. In soil proximal (0-10 cm) to the artificial root, synthetic exudates had a highly significant (P < 0.001) influence on dehydrogenase, fluorescein diacetate hydrolysis and urease activity. In addition, 8-hydroxyquinoline was significant (p = 0.003) as a main effect on dehydrogenase activity and interacted with synthetic exudates to affect urease activity (p = 0.09). Hierarchical cluster analysis of 16S rDNA-based DGGE band patterns also identified a primary affect of synthetic exudates and a secondary affect of 8-hydroxyquinoline on bacterial community structure. Thus, we show that the artificial rhizosphere produced by the synthetic exudates was the predominant effect, but, that the influence of the 8-hydroxyquinoline signal on the activity and structure of soil microbial communities could also be detected.     

Distribution of microbial biomass across the rhizosphere of barley (Hordeum vulgare L.) in soils

Summary The microbial activity at the soil-root interface (rhizosphere) of barley was examined using a rhizobox system. In this system, the soil was placed in several compartments separated from each other by a 500-mesh nylon cloth. Plants were grown in the central compartment and after a 2-month growing period the roots were still confined to this compartment. The soil from each compartment was then analyzed for ATP, NO₃ ^/–, total N, total C and CO₂ production. The increase in ATP concentration was found in a range of 4 mm around the roots. The ATP content and CO₂ production across the rhizosphere were correlated in all the soils used, but changes in NO₃ ^– were not correlated with ATP changes. The range of NO₃ ^– change was wider than that of the ATP change, indicating that NO₃ ^– production is not influenced by the biological activity in the rhizosphere.     

菌肥对青稞根际土壤理化性质以及微生物群落的影响

应用化学分析、聚合酶链反应-变性梯度凝胶电泳(PCR-DGGE)技术和DNA测序技术,研究了西藏棕色砂壤土中微生物肥料不同施用量和施用期对青稞根际土壤理化性质和细菌群落多样性的影响。结果表明,施用谷特菌肥能显著提高土壤全氮、全磷、全钾、有机质、碱解氮、有效磷和速效钾的水平,如播前施用菌肥浓度750 ml hm-2的处理较不施用菌肥的处理上述指标分别提高13.32%、28.42%、16.20%、9.81%、21.36%、39.35%和30.48%,拔节期施用菌肥浓度2 250 ml hm-2的处理较不施用菌肥的处理分别提高7.25%、29.35%、18.04%、12.86%、15.90%、43.27%和53.99%。DGGE分析表明,相同施用方式中不同施用量土样中微生物的DGGE图谱相似。非加权组平均法(UPGMA)聚类分析将DGGE图谱分为2大类群。Shannon-Wiener指数表明,施用菌肥的土壤细菌多样性先增加后逐渐降低,播前以喷施谷特菌浓度750 ml hm-2时的细菌多样性最高;拔节期则以喷施谷特菌浓度2 250 ml hm-2处理的细菌多样性最高,且两种施用方式土壤养分的释放与Shannon指数的变化规律均为播前﹥拔节期。测序结果表明,不同施肥浓度土样微生物种群分布较为广泛,其中Actinobacteria纲细菌种类略多,少数菌种为未经培养菌种(Uncultured bacterium)。典型对应分析(CCA)表明,DGGE图谱条带分布与土壤理化性质密切相关,碱解氮、全磷和全氮是影响微生物群落的主要环境因子。研究结果表明,施用谷特菌肥可明显改善青稞根际土壤理化性状,提高土壤细菌多样性。     

盐地碱蓬根际土壤细菌群落结构及其功能

盐地碱蓬作为生物改良盐碱地的理想材料,其根际土壤微生物对土壤改良发挥着重要作用。为了深入探索环渤海滨海盐碱地碱蓬根际土壤细菌群落结构组成及其功能,采用Illumina Misep高通量测序平台对环渤海地区滨海盐碱地盐地碱蓬根际土壤和裸地土壤进行测序。从16个样本中获得有效序列734 792条, 4 285个OTUs,归属于41门、100纲、282目、400科、892属、1 577种。盐地碱蓬根际土壤细菌群落由变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、绿弯曲门(Chloroflexi)、拟杆菌门(Bacteroidetes)、芽单胞菌门(Gemmatimonadetes)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicutes)、蓝藻细菌门(Cyanobacteria)、髌骨细菌门(Patescibacteria、浮霉菌门(Planctomycetes)组成。Alpha多样性计算结果表明,盐地碱蓬根际土壤细菌群落结构多样性高并与裸地土壤间差异显著;LEfSe(LDAEffectSize)分析发现,盐地碱蓬与裸地差异指示种明显不同。PCoA与相关性Heatmap表明,盐地碱蓬、速效氮、速效钾、速效磷、电导率是影响土壤细菌目类水平群落组成的主要因子。PICRUSt(Phylogenetic InvestigationofCommunitiesbyReconstructionofUnobserved States)分析表明微生物群落在新陈代谢等40个功能方面盐地碱蓬根际土壤比裸地土壤高。本研究表明盐地碱蓬覆盖能够降低土壤盐分,增加土壤养分,对土壤细菌群落多样性及其功能有积极作用。     

Protease and deaminase activities in wheat rhizosphere and their relation to bacterial and protozoan populations

Protease and deaminase activities and population dynamics of bacteria and protozoa were measured in the rhizosphere of wheat to study their interactions with the mineralization of nitrogen. The experimental design allowed the separation of roots and soil material by means of a gauze. The most pronounced rhizosphere effect was detected for all the measured variables in the soil closest to the gauze. The number of bacteria was significantly higher in the presence than in the absence of plants up to 4 mm away from the soil-root interface and the closer to this interface the higher the number. Protozoan and bacterial population dynamics were positively correlated; generally, populations of flagellates and amoebae were comparable and their sum accounted for the population of total protozoa. For both enzyme activities the rhizosphere effect extended up to 2 mm away from the soil-root interface. The histidinase activity was of bacterial origin, while it is likely that bacteria, protozoa and root hair all contributed to the overall caseinase activity. Decomposition of root exudates and native organic matter in the rhizosphere, reflected by a growing microbial population, is associated with nitrogen mineralization through increases in caseinhydrolysing and L-histidine-deaminating activities. The adopted soil-plant microcosm is suitable for the study of the rhizosphere effect over time of incubation and distance gradient from the soil-root interface.     

Density, structure, and diversity of the cultivable arylsulfatase-producing bacterial community in the rhizosphere of field-grown rape and barley

In this study we investigated the arylsulfatase-producing bacterial community (ARS-BC) in the rhizosphere soils of field-grown rape in comparison with that of barley. For this, the rhizosphere soils from both plant species were sampled four times during plant growth. Soil arylsulfatase (ARS) activity and the density and the structure of the cultivable ARS-BC on M9-Xsulf medium were then determined. ARS activity in rape rhizosphere was greater than in barley rhizosphere and evolved along the phenology in the two rhizosphere soils. In parallel, the average density of ARS-BC in the rape rhizosphere was higher than that in the barley rhizosphere. Moreover, ARS activity is correlated with ARS-BC density both in rape and barley rhizosphere soils. The structure of the ARS-BC in the rape rhizosphere was different from that in the barley rhizosphere. In the rape rhizosphere, the ARS-BC was substantially more structured than in the barley rhizosphere. Among the ARS-BC, Actinobacteria and Pseudomonads were significantly present in the both rhizosphere soils. Actinobacteria predominated in the barley rhizosphere while Pseudomonads were mostly represented under rape. It is possible that the differences in ARS activity observed between rape and barley can be attributed to a different ARS-BC size and/or a different ARS-BC structure under these two plant covers. This impact of rape may be connected to a selective effect of rhizodeposits released by rape roots to the functional bacterial community. Our findings suggest that plant species, via their rhizodeposits, may affect the functional bacterial community and thus influence the dynamic of S in soil.     

Effects of rhizosphere and long-term fertilisation practices on the activity and community structure of ammonia oxidisers under double-cropping rice field

The potential nitrification activity (PNA), population size and composition of AOB and AOA communities in both rhizosphere and bulk soil from a long-term (32 year) fertiliser field experiment conducted during early rice and late rice seasons were investigated by using the shaken slurry method and quantitative real-time polymerase chain reaction. The experiment begins in 1986, and including five treatments: without fertiliser input, chemical fertiliser alone, rice straw residue and chemical fertiliser, low organic manure rate and chemical fertiliser and high organic manure rate and chemical fertiliser. The results showed that the application of chemical fertiliser greatly enhanced PNA and AOB abundance, while application of rice straw residue and organic manure increased AOA abundance. Moreover, the results showed that the PNA and population sizes of AOB and AOA were higher in the rhizosphere than in the bulk soil. Cluster and redundancy analyses further indicated that the rhizosphere effect play a more important role in shaping AOA community structure than long-term fertilization. In summary, the results indicated that AOB rather than AOA functionally dominate ammonia oxidation in the double-cropping rice paddy soils, and that rhizosphere effect and fertiliser regime play different roles in the activity and community structures of AOB and AOA.     

Bacteria and fungi on roots of different barley varieties (Hordeum vulgare L.)

Summary We investigated the abundance of bacteria and fungi on roots of different barley varieties grown in soil and in a nutrient solution. Measurements were made on the rhizoplane and, for soil-grown plants, also in the rhizosphere soil. Further, the influence of plant age was investigated. Barley variety, had a significant influence both for plants grown in soil and in the nutrient solution, and the effects were most pronounced on the rhizoplane. There were no significant differences among varieties in fungal hyphal lengths on the roots. Bacterial abundance on the rhizoplane was significantly decreased with increasing plant age. Varietal differences were maintained over different plant ages.     

氮肥减施对玉米幼苗根系分泌物影响的根际代谢组学分析

植物根系分泌物的根际代谢组学分析有助于更好地理解土壤根际微域内植物根系与土壤和土壤生物之间化学信号交流的根际过程。本文采用核磁共振氢谱(~1H nuclear magnetic resonance,~1H NMR)技术,对基于不同施氮处理下[常规施氮(180 kg×hm~(-2))、80%常规氮量和55%常规氮量]土壤培养收集的玉米幼苗的根系土壤沥出液(soil leachate,SL)、根鞘土浸提液(rhizosheath soil,RS)和根系水培液(distilled water cultivation,DWC)内的根系分泌物进行检测,并结合多维统计分析对比了不同收集方法以及不同施氮量下玉米幼苗根系分泌物的不同。结果表明:3种不同方法收集的玉米幼苗根系分泌物的核磁共振氢谱谱图轮廓及主要标志物明显不同。其中SL法的谱图峰信号及检测到的根系分泌物数目少,而RS和DWC法的谱图峰信号较多且可检测到玉米幼苗根系分泌物中的糖、有机酸和氨基酸等组分。与常规施氮量比较,在85%施氮量下,玉米幼苗根系分泌物中的α-葡萄糖、苹果酸、亮氨酸、缬氨酸水平显著增加;而当施氮量减少到55%时,玉米幼苗根系分泌物水平不再显著增加并呈现下降趋势。上述根系分泌物的变化可能和玉米根系对土壤氮营养供应水平的适应性调节有关。采用~1H NMR技术,结合RS和DWC收集方法进行根际代谢组学分析,可为根际生态及根际氮素营养研究提供重要理论依据。     

Increased microbial activity and nitrogen mineralization coupled to changes in microbial community structure in the rhizosphere of Bt corn

The interactions between plant roots and soil microorganisms are essential for the function and stability of ecosystems, primary agricultural production and plant health. Despite the importance of soil microbes the response of these microbes to large-scale cultivation of genetically modified (GM) crops is still poorly understood. This study evaluated the potential impact of two lines of transgenic Bt maize on rhizosphere microorganisms. A time-course field experiment was conducted over a period of two years in two fields in Guadalajara (Spain) with monthly sampling from April to September. Rhizosphere soil was collected from transgenic (TG) and unmodified (WT) maize plants from each field and sampling time for the analysis of several important functional and structural soil quality parameters. Total microbial activity, as determined by H³-Thymidine and C¹⁴-Leucine incorporation, was found to be higher in the rhizospheres of the transgenic plants. Similarly, differences in potential ammonification and nitrification were observed in the second year of the study. In contrast, bacterial and fungal microbial catabolic abilities, as determined by Biolog ECO and FF plate analyses, respectively, were more influenced by sampling time than the transgenic nature of the plants. Microbial community structure was also studied by bacterial and phylum-specific PCR-DGGE and PCR cloning approaches. In general, differences were again more pronounced between sampling times, as opposed to between TG versus WT plants, although marked differences were observed within the Betaproteobacteria between plant lines. For the first time it describes the presence of Iamiaceae family in soil, specifically to TG plant rhizosphere. To summarize, the study showed that some important properties of rhizopshere microbes may be impacted by Bt maize cultivation and highlighted the fact that such potential effects need to be viewed within the context of seasonal and spatial variability.     

根际增氧对水稻根系形态和生理影响的研究进展

根际氧是影响稻田土壤环境和水稻根系生理代谢的重要环境因子,已有的关于水稻根际氧的综述多从低氧或缺氧胁迫的角度展开,随着技术的进步,越来越多的学者在水稻栽培中实施了主动的根际增氧措施,取得了一定的研究成果。根际增氧显著影响了水培水稻根系形态和结构,使其呈现细而长的特征,增氧条件下水稻根系形态、结构及其功能需求间存在内在的一致性;根际增氧对不同生育时期水稻的根系活力均有明显的促进作用,其增幅从10%到150%不等,并存在明显的品种间差异;从水稻根系形态、生理活性以及根部氮素形态转化等多个方面来看,增氧处理有利于水稻根系吸收氮素,但其对水稻氮素积累量的影响则与增氧处理方式和程度有关,过度增氧抑制了水稻植株对氮的利用,从而限制了其生物量的增加,反过来抑制了对氮的吸收。水稻对根际增氧的响应规律并非其对低氧和缺氧胁迫响应规律的简单倒转,饱和氧处理下水稻生物量和产量的剧烈降低表明了水稻对富氧响应的复杂性。探索根际增氧对三叶期前水稻幼苗的影响,完善根际增氧对水稻氮代谢的影响研究,并量化水稻田间需氧量,探索简单易行的苗期增氧措施,对进一步完善水稻育秧理论,改进水稻育秧技术具有重要意义。     

Rapid changes in the rhizosphere bacterial community structure during re-colonization of sterilized soil

Diversity has been shown to be pivotal in ecosystem stability and resilience. It is therefore important to increase our knowledge about the development of diversity. The aim of this study was to investigate the temporal dynamics of the bacterial community structure in the rhizosphere of wheat plants growing in a soil in which the initial conditions for bacterial re-colonization were modified by mixing different amounts of sterilized with native soil at ratios of 19:1, 9:1, 4:1 and 1:1. Additional treatments comprised sterilized soil or native soil. Plant dry weight at day 20 decreased with increasing percentage of native soil in the mix. The bacterial community structure in the rhizosphere was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (DGGE) at days 3, 14 and 20 after planting. The bacterial community in the sterilized soil had a lower diversity and evenness than the native soil. Both diversity and evenness increased with time in the sterilized soil. Community structure in the different mixes changed over time and the changes were mix-specific. Principal component analyses of the DGGE banding patterns showed clear differences between the treatments particularly at day 3 and day 14 and revealed changes in community structure within a few days in a given treatment. The results of the present study show that bacterial communities rapidly re-colonize sterilized soil. During re-colonization, the community structure changes rapidly with a general trend towards higher diversity and evenness. The changes in community structure over time are also affected by the amount of sterile substrate to be re-colonized.     

Impact of artificial root exudates on the bacterial community structure in bulk soil and maize rhizosphere

Bacterial densities, metabolic signatures and genetic structures were evaluated to measure the impact of soil enrichment of soluble organic carbon on the bacterial community structures. The exudates chosen were detected in natural maize exudates (glucose, fructose, saccharose, citric acid, lactic acid, succinic acid, alanine, serine and glutamic acid) and were used at a rate of 100 μg C g⁻¹ day⁻¹ for 14 days. Moreover two synthetic solutions with distinct carbon/nitrogen ratios (20.5 and 40.1), obtained by varying carboxylic and amino acids concentrations, were compared in order to evaluate the potential role of organic N availability. The in vitro experiment consisted of applying exudate solutions to bulk soil. In the case of the control, only distilled water was added. Both solutions significantly increased bacterial densities and modified the oxidation pattern of Biolog® GN2 plates with no effect of the C/N ratio on these two parameters. Genetic structure, measured by means of ribosomal intergenic spacer analysis (RISA), was also consistently modified by the organic amendments. N availability levels led to distinct genetic structures. In a second experiment, one of the previous exudate solutions (C/N 20.5) was applied to 15-day-old maize plants to determine the structural influence of exudates on the rhizosphere microbial community (in situ experiment). Bacterial densities were significantly increased, but to a lesser extent than had been found in the in vitro experiment. Metabolic potentials and RISA profiles were also significantly modified by the organic enrichment.     

Field survival of the phytostimulator Azospirillum lipoferum CRT1 and functional impact on maize crop, biodegradation of crop residues, and soil faunal indicators in a context of decreasing nitrogen fertilisation

Heavy nitrogen fertilisation is often implemented in maize cropping systems, but it can have negative environmental effects. Nitrogen-fixing, phytohormone-producing Azospirillum plant growth-promoting rhizobacteria (PGPR) have been proposed as crop inoculants to maintain high yield when decreasing nitrogen fertilisation. In this context, agronomic and ecological effects of the inoculation of maize seeds with the PGPR Azospirillum lipoferum CRT1 were studied in two consecutive years. The inoculant was recovered from maize at 10⁵ CFU g⁻¹ root or higher. Inoculation enhanced root growth and development based on results of root biomass, rooting depth and/or parameters describing root system architecture, and a transient positive effect on shoot height was observed in the first year. Inoculation did not increase yield, but reducing mineral nitrogen fertilisation had only a minor effect on yield. This suggests that the lack of positive effect of the PGPR on yield was due to the fact that the whole field was heavily fertilised in years prior to the start of the experiment. Soil nitrogen levels decreased during the 2 years of the study, and the inoculant had no effect on residual soil nitrogen levels at harvest. Inoculation had no impact on Fusarium symptoms and concentration of the mycotoxin deoxynivalenol in maize kernels, but both were influenced by the interaction between inoculation and nitrogen fertilisation level. Inoculation did not influence meso/macrofaunal soil populations, but had a small but significant effect (smaller than the effect of added nitrogen) on decomposition, nitrogen mineralisation and mesofaunal colonisation of maize leaves (in litter bags). Overall, the ecological impact of seed inoculation with the PGPR A. lipoferum CRT1 was small, and its magnitude was smaller than that of chemical nitrogen fertilisation.     

Techniques for observing phosphorus mobilization in the rhizosphere

Summary The techniques described here were developed to visualize the dissolution of sparingly soluble calcium phosphate and the presence of root-borne phosphatase in the rhizosphere. Newly formed calcium phosphate precipitate was suspended in agar containing other essential nutrients. The agar was poured into Petri dishes and acrylglass boxes and was used as a growth medium for seedlings of wheat, rape, buckwheat, and rice. With NH ₄ ⁺ applied as the N source, the precipitate dissolved in the root vicinity and this was attributed to acidification. No dissolution occurred with NO ₃ ^– as the N source. The release of a neutral phosphatase from roots was verified by embedding the roots of young seedlings in agar at pH 7 containing phenolphthalein phosphate. After pH was raised to the alkaline range by adding sodium hydroxide, the agar around the roots turned purple, especially around the roots of P-deprived plants. The most intensive phosphatase activity was found in apical root regions.     

Nitrogen acquisition by pea and barley and the effect of their crop residues on available nitrogen for subsequent crops

Nitrogen acquisition by field pea (Pisum sativum L.) and spring barley (Hordeum vulgare L.) grown on a sandy loam soil and availability of N in three subsequent sequences of a cropping system were studied in an outdoor pot experiment. The effect of crop residues on the N availability was evaluated using ¹⁵N-labelled residues. Field pea fixed 75% of its N requirement and the N₂ fixation almost balanced the N removed with the seeds. The barley crop recovered 80% of the ¹⁵N-labelled fertilizer N supplied and the N in the barley grain corresponded to 80% of the fertilizer N taken up by the crop. The uptake of soil-derived N by a test crop (N catch crop) of white mustard (Sinapis alba L.) grown in the autumn was higher after pea than after barley. The N uptake in the test crop was reduced by 27% and 34% after pea and barley residue incorporation, respectively, probably due to N immobilization. The dry matter production and total N uptake of a spring barley crop following pea or barley, with a period of unplanted soil in the autumn/winter, were significantly higher after pea than after barley. The barley crop following pea and barley recovered 11% of the pea and 8% of the barley residue N. The pea and barley residue N recovered constituted only 2.5% and <1%, respectively, of total N in the N-fertilized barley. The total N uptake in a test crop of mustard grown in the second autumn following pea and barley cultivation was not significantly influenced by pre-precrop and residue treatment. In the short term, the incorporation of crop residues was not important in terms of contributing N to the subsequent crop compared to soil and fertilizer N sources, but residues improved the conservation of soil N in the autumn. In the long-term, crop residues are an important factor in maintaining soil fertility and supplying plant-available N via mineralization.     

Distribution of phosphatase activity and various bacterial phyla in the rhizosphere of Hordeum vulgare L. depending on P availability

Despite the importance of the rhizosphere for nutrient turnover, little is known about the spatial patterns of organic phosphorus mineralization by plants and by microorganisms in the rhizosphere. Therefore, the distribution of acid and alkaline phosphatase activity and the abundance of bacteria belonging to various bacterial phyla were investigated in the rhizosphere of barley (Hordeum vulgare L.) as dependent on the availability of inorganic P. For this purpose, we conducted a greenhouse experiment with barley growing in inclined boxes that can be opened to the bottom side (rhizoboxes), and applied soil zymography and fluorescence-in situ-hybridization (FISH). Acid phosphatase activity was strongly associated with the root and was highest at the root tips. Due to P fertilization, acid phosphatase activity decreased in the bulk soil, and less strongly in the rhizosphere. Alkaline phosphatase activity, i.e., microbial phosphatase activity was high throughout the soil in the control treatment and was reduced due to inorganic P fertilization especially in the rhizosphere and less strongly in the bulk soil. P-fertilization slightly increased the total number of bacteria in the rhizosphere. Moreover, P-fertilization decreased the abundance of Firmicutes and increased the abundances of Beta- and Gamma-Proteobacteria. The total number of bacterial cells was significantly higher at the root surface than at the root tip and at a distance of 30 μm from the root surface. Our results show that alkaline phosphatase activity decreased more strongly in the rhizosphere than in the bulk soil due to P fertilization, which might be because of greater C deficiency in the bulk soil compared to the rhizosphere. Furthermore, the results indicate a spatial separation between hotspots of acid phosphatase activity and hotspots of bacteria in the rhizosphere of H. vulgare. Taken together, our study shows that bacteria and phosphatase activity were very heterogeneously distributed in soil, and that the effects of P fertilization on phosphatase activity differed strongly between bulk soil and rhizosphere as well as between various zones of the rhizosphere.