Influence of grass species and soil type on rhizosphere microbial community structure in grassland soils

A number of studies have reported species specific selection of microbial communities in the rhizosphere by plants. It is hypothesised that plants influence microbial community structure in the rhizosphere through rhizodeposition. We examined to what extent the structure of bacterial and fungal communities in the rhizosphere of grasses is determined by the plant species and different soil types. Three grass species were planted in soil from one site, to identify plant-specific influences on rhizosphere microbial communities. To quantify the soil-specific effects on rhizosphere microbial community structure, we planted one grass species (Lolium perenne L.) into soils from three contrasting sites. Rhizosphere, non-rhizosphere (bulk) and control (non-planted) soil samples were collected at regular intervals, to examine the temporal changes in soil microbial communities. Rhizosphere soil samples were collected from both root bases and root tips, to investigate root associated spatial influences. Both fungal and bacterial communities were analysed by terminal restriction fragment length polymorphism (TRFLP). Both bacterial and fungal communities were influenced by the plant growth but there was no evidence for plant species selection of the soil microbial communities in the rhizosphere of the different grass species. For both fungal and bacterial communities, the major determinant of community structure in rhizospheres was soil type. This observation was confirmed by cloning and sequencing analysis of bacterial communities. In control soils, bacterial composition was dominated by Firmicutes and Actinobacteria but in the rhizosphere samples, the majority of bacteria belonged to Proteobacteria and Acidobacteria. Bacterial community compositions of rhizosphere soils from different plants were similar, indicating only a weak influence of plant species on rhizosphere microbial community structure.     

Continuous defoliation of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) and associated changes in the composition and activity of the microbial population of an upland grassland soil

A microcosm study was conducted to investigate the effect of continuons plant defoliation on the composition and activity of microbial populations in the rhizosphere of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens). Continuons defoliation of ryegrass and clover resulted in sigmficant (P <0.01) increases in soil microbial biomass, although whilst increases were measured from day 2 in soil sown with clover significant increases were only seen from day 21 in soil sown with ryegrass. These increases were paralleled, from day 10 onwards, by increases in the numbers of culturable bacteria. Numbers ofPsendomonas spp. also increased in the later stages of the study. No influence on culturable fungal populations was detected. Whilst shifts in the composition of the microbial populations were measured in response to defoliation there was little effect on microbial activity. No changes in either dehydrogenase activity or microbial respiration in the rhizosphere of ryegrass or clover were measured in response to defoliation, but both dehydrogenase activity and microbial respiration were greater in ryegrass than clover when values over the whole study were combined. Continuous defoliation resulted in significant (P <0.001) reductions in the root dry weight of ryegrass and clover, of the order 19% and 16%, respectively.     

Continuous defoliation of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) and associated changes in the composition and activity of the microbial population of an upland grassland soil

A microcosm study was conducted to investigate the effect of continuons plant defoliation on the composition and activity of microbial populations in the rhizosphere of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens). Continuons defoliation of ryegrass and clover resulted in sigmficant (P <0.01) increases in soil microbial biomass, although whilst increases were measured from day 2 in soil sown with clover significant increases were only seen from day 21 in soil sown with ryegrass. These increases were paralleled, from day 10 onwards, by increases in the numbers of culturable bacteria. Numbers ofPsendomonas spp. also increased in the later stages of the study. No influence on culturable fungal populations was detected. Whilst shifts in the composition of the microbial populations were measured in response to defoliation there was little effect on microbial activity. No changes in either dehydrogenase activity or microbial respiration in the rhizosphere of ryegrass or clover were measured in response to defoliation, but both dehydrogenase activity and microbial respiration were greater in ryegrass than clover when values over the whole study were combined. Continuous defoliation resulted in significant (P <0.001) reductions in the root dry weight of ryegrass and clover, of the order 19% and 16%, respectively.     

Defoliation and fertiliser influences on the soil microbial community associated with two contrasting Lolium perenne cultivars

The influence of repeated defoliation on soil microbial community (SMC) structure and root turnover was assessed in two contrasting Lolium perenne cultivars (AberDove and S23) grown in fertilised (+F) and non-fertilised (NF) soil. BiOLOG sole carbon source utilisation profiles (SCSUPs) indicated consistently greater potential carbon utilisation in defoliated (+D) compared to non-defoliated (ND) soils regardless of cultivar and fertiliser, and was accounted for in a variety of substrate groups (sugars, carboxylic, amino and phenolic acids). Potential carbon utilisation was also stimulated in +F compared to NF soils, primarily through increased potential utilisation of carboxylic acids. PLFA indicators for the bacterial biomass did not significantly differ between cultivar, soil fertilisation, or defoliation. Defoliated swards grown in fertilised soil (+F+D) had a higher fungal:bacterial ratio and a greater bacterial stress index (cy19:0/18:1w7c), compared to that of +F ND, NF ND and NF+D, and regardless of cultivar. Overall SMC structure (canonical variate (CV) analysis of PLFAs) discriminated based on cultivar, defoliation and soil fertilisation. Primary discrimination of the SMCs could be related to differences in root density and total plant biomass, and in the case of NF soils, secondary community shifts, evident with defoliation, related to root disappearance over the growing season. Despite the strong common effects of defoliation, and to a lesser extent soil fertilisation, cultivar specific drivers of the soil microbial community were maintained, resulting in consistent, but subtle, discrimination of the SMC associated with the contrasting L. perenne cultivars.     

Short-term effects of defoliation on the soil microbial community associated with two contrasting Lolium perenne cultivars

Intra-species variation in response to defoliation and soil amendment has been largely neglected in terms of the soil microbial community (SMC). The influence of defoliation and soil fertiliser amendment on the structure of the SMC was assessed with two Lolium perenne cultivars contrasting in ability to accumulate storage reserves. Plant response to defoliation was cultivar specific and depended on the nutrient amendment of the soil. Results suggested a greater ability to alter plant biomass allocation in the low carbohydrate accumulating cultivar (S23) compared to the high carbohydrate cultivar (AberDove) when grown in improved (IMP), but not in unimproved (UNI), soil. Although differences in plant growth parameters were evident, no treatment effects were detected in the size of the active microbial biomass (total phospholipid fatty acid (PLFA) 313.8 nmol g⁻¹ soil±33.9) or proportions of PLFA signature groups. A lower average well colour development (AWCD) of Biolog sole carbon source utilisation profiles (SCSUPs) in defoliated (D) compared to non-defoliated (ND) treatments may be indicative of lower root exudation 1 week following defoliation, as a consequence of lower root non-structural carbohydrate (NSC) concentrations. Within the bacterial community the lower cyclopropyl-to-precursor ratio of PLFAs, and the trans/cis ratio of 16:1w7, in UNI relative to IMP soil treatments indicates lower physiological stress in UNI soils regardless of L. perenne cultivar. Discrimination of broad scale SMC structure, measured by PLFA analysis, revealed that soil treatment interacted strongly with cultivar and defoliation. In IMP soils the SMCs discriminated between cultivars while defoliation had little effect. Conversely, in UNI soils defoliation caused a common shift in the SMC associated with both cultivars, causing convergence of overall community structure. Separation of SMC structure along the primary canonical axis correlated most strongly (P<0.001) with root:shoot ratio (47.6%), confirming that differences in cultivar C-partitioning between treatments were influential in defining the rhizosphere microbial community.     

Decomposer animals (Lumbricidae, Collembola) and organic matter distribution affect the performance of Lolium perenne (Poaceae) and Trifolium repens (Fabaceae)

Decomposer animals stimulate plant growth by indirect effects such as increasing nutrient availability or by modifying microbial communities in the rhizosphere. In grasslands, the spatial distribution of organic matter (OM) rich in nutrients depends on agricultural practice and the bioturbation activities of large detritivores, such as earthworms. We hypothesized that plants of different functional groups with contrasting nutrient uptake and resource allocation strategies differentially benefit from sites in soil with OM accumulation and the presence of decomposer animals. In a greenhouse experiment we investigated effects of spatial distribution of ¹⁵N-labelled grass litter, earthworms and collembola on a simple grassland community consisting of Lolium perenne (grass) and Trifolium repens (legume). Litter aggregates (compared to homogeneous litter distribution) increased total shoot biomass, root biomass and ¹⁵N uptake by the plants. Earthworms and collembola did not affect total N uptake of T. repens; however, the presence of both increased ¹⁵N uptake by T. repens and L. perenne. Earthworms increased shoot biomass of T. repens 1.11-fold and that of L. perenne 2.50 fold. Biomass of L. perenne was at a maximum in the presence of earthworms, collembola and with litter concentrated in a single aggregate. Shoot biomass of T. repens increased in the presence of collembola, with L. perenne generally responding opposingly. The results indicate that the composition of the decomposer community and the distribution of OM in soil affect plant competition and therefore plant community composition.     

Impact of fresh root material and mature crop residues of oilseed rape (Brassica napus) on microbial communities associated with subsequent oilseed rape

Glasshouse bioassays were conducted to assess the impact of different inputs of oilseed rape plant material on soil and rhizosphere microbial diversity associated with subsequently grown oilseed rape (Brassica napus) plants. The first bioassay focussed on the effect of oilseed rape rhizodeposits and fresh detached root material on microbial communities, in a rapid-cycling experiment in which oilseed rape plants were grown successively in pots of field soil for 4 weeks at a time, with six cycles of repeated vegetative planting in the same pot. Molecular analyses of the microbial communities after each cycle showed that the obligate parasite Olpidium brassicae infected the roots of oilseed rape within 4 weeks after the first planting (irrespective of the influence of rhizodeposits alone or in the presence of fresh detached root material), and consistently dominated the rhizosphere fungal community, ranging in relative abundance from 43 to 88 % when oilseed rape was grown more than once in the same soil. Fresh detached root material also led to a reduction in diversity within the soil fungal community, due to the increased relative abundance of O. brassicae. In addition, rhizosphere bacterial communities were found to have a reduced diversity over time when fresh root material was retained in the soil. In the second glasshouse experiment, the effect of incorporating mature, field-derived oilseed rape crop residues (shoots and root material) on microbial communities associated with subsequently grown oilseed rape was investigated. As before, molecular analyses revealed that O. brassicae dominated the rhizosphere fungal community, despite not being prevalent in either the residue material or soil fungal communities.     

Comparison of root system architecture and rhizosphere microbial communities of Balsas teosinte and domesticated corn cultivars

The progenitor of maize is Balsas teosinte (Zea mays subsp. parviglumis) which grows as a wild plant in the valley of the Balsas river in Mexico. Domestication, primarily targeting above-ground traits, has led to substantial changes in the plant's morphology and modern maize cultivars poorly resemble their wild ancestor. We examined the hypotheses that Balsas teosinte (accession PI 384071) has a) a different root system architecture and b) a structurally and functionally different rhizosphere microbial community than domesticated cultivars sweet corn (Zea mays subsp. mays accession PI 494083) and popping corn (Zea mays subsp. mays accession PI 542713). In a greenhouse experiment, five plants from each corn variety were grown in individual pots containing a Maury silt loam – perlite (2:1) mixture and grown to the V8 growth stage at which rhizosphere bacterial and fungal community structure was assessed using terminal restriction fragment length polymorphism and fatty acid methyl ester analysis. Functional characteristics of the rhizosphere were assayed by examining the potential activity of seven extracellular enzymes involved in carbon, nitrogen and phosphorus cycling. Root system architecture was characterized by root scans of sand grown plants at the V5 growth stage. Compared to the control the sweet corn rhizosphere had different bacterial and fungal community structure, decreased fungal diversity and increased bacterial abundance. Teosinte caused a significant change in the rhizosphere bacterial and fungal community structure and increased bacterial abundance, but no significant decrease in bacterial or fungal diversity where the former was found to be significantly greater than in the sweet corn rhizosphere. Popping corn did not trigger significant changes in the bacterial or fungal diversity and bacterial abundance in the soil. The individual popping corn plants changed the bacterial and fungal communities in different directions and the overall effect on community structure was significant, but small. Of the enzymes analyzed, potential N-acetylglucosaminidase (NAG) activity was found to contributed most to the differentiation of teosinte rhizosphere samples from the other corn varieties. The teosinte root system had proportionally more very fine (diameter < 0.03 mm) roots than popping corn and sweet corn and it developed the highest root to shoot dry weight ratio, followed by popping corn. Sweet corn had significantly lower average root diameter than popping corn and teosinte and grew proportionally the least below-ground dry mass. The results allude to functional and structural differences in the rhizosphere microbial communities of the corn varieties that, with additional research, could lead to useful discoveries on how corn domestication has altered rhizosphere processes and how plant genotype influences nutrient cycling.     

Relationships between microbial indices in roots and silt loam soils forming a gradient in soil organic matter

Perennial rye grass (Lolium perenne) was grown in a greenhouse pot experiment on seven soils to answer the question whether the microbial colonisation of roots is related to existing differences in soil microbial indices. The soils were similar in texture, but differed considerably in soil organic matter, microbial biomass, and microbial community structure. Ergosterol and fungal glucosamine were significantly interrelated in the root material. This ergosterol was also significantly correlated with the average ergosterol content of bulk and rhizosphere soil. In addition, the sum of fungal C and bacterial C in the root material revealed a significant linear relationship with microbial biomass C in soil. The colonisation of roots with microorganisms increased apparently with an increase in soil microbial biomass. In the root material, microbial tissue consisted of 77% fungi and 23% bacteria. In soil, the fungal dominance was slightly, but significantly lower, with 70% fungi and 30% bacteria. Fungal glucosamine in the root material was significantly correlated with that in soil (r=0.65). This indicates a close relationship between the composition of dead microbial remains in soil and the living fraction in soil and root material for unknown reasons.     

Interactions between mycorrhizal fungi and Collembola: effects on root structure of competing plant species

Mycorrhizal fungi influence plant nutrition and therefore likely modify competition between plants. By affecting mycorrhiza formation and nutrient availability of plants, Collembola may influence competitive interactions of plant roots. We investigated the effect of Collembola (Protaphorura fimata Gisin), a mycorrhizal fungus (Glomus intraradices Schenck and Smith), and their interaction on plant growth and root structure of two plant species, Lolium perenne L. (perennial ryegrass) and Trifolium repens L. (white clover). In a laboratory experiment, two individuals of each plant species were grown either in monoculture or in competition to the respective other plant species. Overall, L. perenne built up more biomass than T. repens. The clover competed poorly with grass, whereas the L. perenne grew less in presence of conspecifics. In particular, presence of conspecifics in the grass and presence of grass in clover reduced shoot and root biomass, root length, number of root tips, and root volume. Collembola reduced shoot biomass in L. perenne, enhanced root length and number of root tips, but reduced root diameter and volume. The effects of Collembola on T. repens were less pronounced, but Collembola enhanced root length and number of root tips. In contrast to our hypothesis, changes in plant biomass and root structure in the presence of Collembola were not associated with a reduction in mycorrhizal formation. Presumably, Collembola affected root structure via changes in the amount of nutrients available and their spatial distribution.     

间套作玉米对线辣椒根际土壤微生物生态特征的影响

采用常规稀释平板法、氯仿熏蒸法、BIOLOG GN微平板反应系统及种间根系分隔技术, 以线辣椒单作(SC)为试验对照, 研究了间套作处理[玉米/线辣椒套作+根部塑料膜分隔(ICP)、玉米/线辣椒套作+根部尼龙网分隔(ICM)、玉米/线辣椒套作根部无分隔(ICN)]对线辣椒根际土壤微生物生态特征的影响。结果表明: 整个线辣椒生育期内, 各套作处理线辣椒根际土壤微生物总数与细菌总数具有相同的变化趋势, ICN和ICM处理的真菌、细菌、放线菌数量和细菌/真菌(B/F)、放线菌/真菌(A/F)比值均大于ICP与SC处理。盛果期, ICN处理根际土壤微生物量碳和微生物量氮比同处理其他生育期增加14.2%~54.0%和10.6%~54.7%。各处理土壤微生物群落AWCD的变化随培养时间呈现明显的"S"型曲线。间套作玉米显著提高了线辣椒根际土壤微生物群落的Shannon-Wiener指数(P<0.05)、Simpson指数、种间相遇几率和McIntosh指数(P<0.05), 并改变了土壤微生物对单一碳源的利用能力。线辣椒根际土壤微生物的不同多样性指数分别与其生物学产量之间存在显著或极显著正相关。说明间套作改善了土壤微生态环境。     

The effect of root hairs on rhizosphere phosphatase activity

Background: Phosphatases in soil are of great importance for plant P acquisition. It is hypothesized that root hairs increase rhizosphere phosphatase activity as they release enzymes into soil and stimulate microbial activity. Methods: To test the effect of root hairs on soil phosphatase activity, we grew barley (Hordeum vulgare ‘Pallas') wild type and its root‐hairless mutant in rhizoboxes and determined phosphatase activity using soil zymography. Measurements were done at three moisture levels (30, 15, and 5% VWC). Rhizosphere phosphatase activity was estimated for the two genotypes and two locations along the root [root tip region (0–4 cm behind tip) and mature roots (> 7 cm behind tip)]. Results: Rhizosphere phosphatase activity was similar in the two locations along the root (root tip region vs. mature root parts). In contrast, rhizosphere phosphatase extension was two times larger for the root tip region of the wild type than for the mutant at 30% and 15% VWC. However, as phosphatase activities at the root surface of tips and mature root parts were slightly higher for the mutant than for the wild type, average enzyme activities were unaffected by the genotype. Conclusions: We conclude that the mutant seems to compensate for the lack of root hairs by increased phosphatase activity close to the root surface. However, the increased rhizosphere phosphatase extension for the wild type may be equally efficient as it allows P mobilization and uptake from large soil volumes.     

Soil bacterial community response to sulfadiazine in the soil–root zone

Sulfonamide antibiotics reach soil via manure and adversely affect microbial diversity. Clear effects of these bacteriostatic, growth‐inhibiting antibiotics occur in the presence of a parallel input of microbial activity stimulating manure. Natural hot spots with already increased soil microbial activity are located in the rhizosphere, comprising microorganism such as Pseudomonas with plant growth promoting and pathogenic strains. The hypothesis was therefore that the antibiotic activity of sulfonamides is promoted in the rhizosphere even in the absence of manure, followed by shifts of the natural plant‐specific microbial community structure. This was evaluated by a laboratory experiment with Salix fragilis L. and Zea mays L. After 40 d of incubation, sub‐areas such as non‐rhizosphere soil, rhizosphere soil and plant roots were sampled. Effects on microbial community structure were analyzed using 16S rRNA gene fragment patterns of total bacteria community and Pseudomonas. Selected exoenzymes of N‐, P‐, and C‐cycling were used to test effects on microbial functions. Compared to the factors soil sub‐area and sulfadiazine (SDZ) content, plant species had the largest influence on the bacterial community structure and soil exoenzyme activity pattern. This was also reflected by an up to 1.5‐fold higher acid phosphatase activity in samples from maize‐ compared to willow‐planted soil. We conclude that antibiotic effects on the bacterial community structures are influenced by the antibiotic concentration and root influence.     

Bacterial community structures in rhizosphere microsites of ryegrass (Lolium perenne var. Nui) as revealed by pyrosequencing

Management of soils to facilitate plant beneficial microbial interactions requires basic knowledge of the species composition and microbial community structures in the plant rhizosphere. Here, we examined composition of bacterial communities associated with rhizosphere microsites located at the root tips and mature root zones of Lolium perenne when grown in Chilean ash-derived volcanic soils (Andisols: Freire and Piedras Negras soil series). Community structures were analyzed by pyrosequencing of 16S ribosomal RNA (rRNA) genes followed by in silico analysis for phylogenetic assignments (MOTHUR and Visualization tool for Taxonomic Compositions of Microbial Community (VITCOMIC)). Analysis of the community structure revealed significant differences in community structures in relation to the soil series, which differed particularly in the relative abundance of Cyanobacteria and Firmicutes. However, no significant differences were observed with respect to root microsite location in the same Andisol series. Predominant taxa included members of the Proteobacteria, Actinobacteria, and Acidobacteria. Analysis by VITCOMIC showed that dominant bacterial groups comprised only 5 to 10 % of the total bacterial community and the remaining majority of bacteria included low-abundant taxa (Fusobacteria, Thermotogae, Lentisphaerae, Tenericutes, Deferribacteres Spirochaetes, Planctomycetes, Thermotogae, and Deinococcus-Thermus), most of which have not been previously reported or associated with the plant rhizosphere according to GenBank database. The results indicate that most of bacteria in the Chilean Andisols have not been described to the rhizosphere plants and their functional traits are still largely unknown.     

Quantitative assessment of the rhizoplane microflora by direct microscopy

The bacterial populations on roots of several grassland species were estimated by combining the techniques of direct counting of stained bacteria in situ with standard sampling techniques of plant ecology. The eight plant species examined had bacterial cover of the root surface within the range 4–10 per cent. In a more detailed experiment with two plant species, the method was sufficiently precise to demonstrate that a 7.7 per cent cover of roots of Lolium perenne L. by bacteria was significantly different (P < 0-05) from a 6.3 per cent cover on Plantago lanceolata L. The length of fungal mycelium per unit root surface area was estimated by a modification of the line intercept method, which was originally developed for measuring root lengths. When hyphal diameters were also measured, fungal cover per cent and volume could be estimated.Estimates of the numbers of rhizoplane bacteria and fungi made by the spread-plate counting method on a non-selective agar showed that the mean number of bacteria from Lolium was greater (but not significantly different) than the counts on Plantago: however. Lolium did have significantly more Gram-negative bacteria as estimated by crystal violet agar than did Plantago.The numbers of bacteria estimated by direct microscopy were about 10-fold greater than estimates by plate counts. For a given period of time, direct microscopy gave a more precise estimate of total bacterial and fungal abundance than did plate counts hut the technique does require continuous concentration by the observer over long periods.     

Hydrocarbon degradation potential and activity of endophytic bacteria associated with prairie plants

Phytoremediation systems for organic compounds such as petroleum hydrocarbons rely on a synergistic relationship between plants and their root-associated microbial communities. To determine the probable role of endophytic bacterial communities in these systems, this study examined both rhizosphere and endophytic communities of five different plant species at a long-term phytoremediation field site. Hydrocarbon degradation potential and activity were assessed using MPN assays, PCR analysis of catabolic genes associated with hydrocarbon degradation, and mineralization assays with C-14 labeled hydrocarbons. Microbial community structure in each niche was assessed by DGGE analysis of 16S rRNA gene fragments and subsequent band sequencing. Both endophytic degrader populations and endophytic degrader activity showed substantial inter-species variation, largely independent of that shown by the respective rhizosphere populations. Endophytic hydrocarbon degradation was linked to dominant bacterial endophytes. Pseudomonas spp. dominated endophytic communities exhibited increased alkane hydrocarbon degradation potential and activity, while Brevundimonas and Pseudomonas rhodesiae dominated endophytic communities were associated with increased PAH degradation potential and activity. In one plant species, Lolium perenne, increased endophytic alkane hydrocarbon degradation was associated with increased rhizosphere alkane degradation and decreased rhizosphere PAH degradation. Our results show that diverse plant species growing in weathered-hydrocarbon contaminated soil maintain distinct, heterogeneously distributed endophytic microbial populations, which may impact upon the ability of plants to promote the degradation of specific types of hydrocarbons.     

Effects of pioneering plants on microbial structures and functions in a glacier forefield

This study investigates the small-scale spatial impact of the pioneering plant Leucanthemopsis alpina (L.) Heywood (L. alpina) on biological and chemical–physical parameters in an early successional stage of a glacier forefield. Considering the frequent occurrence of isolated patches of this pioneer plant in the forefield of the Dammaglacier (Switzerland), we hypothesized that the impact of the plant would establish gradients in nutrients, and microbial community structure and activity that may be of importance for the successional processes occurring in the forefield. Our results indicated that, in young successional soils, the rhizosphere effect of L. alpina plant patches can influence bacterial cell numbers and activities not only within the root zone, but even at 20 cm distance from the plant. Microbial cell counts, active cells, and saccharase, glucosidase, and acid phosphatase activities revealed significant distance effects, decreasing from soil directly underneath the plant to soils at 20 and 40 cm distance. Soil chemical and physical parameters did not exhibit significant trends. Fingerprinting analysis of amplified 16S rDNA fragments was used to characterize the microbial community. A selective effect of the plant on the microbial community could not be shown because the bacterial communities were similar regardless of distance to the plant.     

Plant genotype strongly modifies the structure and growth of maize rhizosphere microbial communities

We studied the microbial communities in maize (Zea mays) rhizosphere to determine the extent to which their structure, biomass, activity and growth were influenced by plant genotype (su1 and sh2 genes) and the addition of standard and high doses of different types of fertilizer (inorganic, raw manure and vermicompost). For this purpose, we sampled the rhizosphere of maize plants at harvest, and analyzed the microbial community structure (PLFA analysis) and activity (basal respiration and bacterial and fungal growth rates). Discriminant analysis clearly differentiated rhizosphere microbial communities in relation to plant genotype. Although microorganisms clearly responded to dose of fertilization, the three fertilizers also contributed to differentiate rhizosphere microbial communities. Moreover, larger plants did not promoted higher biomass or microbial growth rates suggesting complex interactions between plants and fertilizers, probably as a result of the different performance of plant genotypes within fertilizer treatments, i.e. differences in the quality and/or composition of root exudates.     

Investigating microbial community structure in soils by physiological, biochemical and molecular fingerprinting methods

Several biochemical and molecular methods are used to investigate the microbial diversity and changes in microbial community structure in rhizospheres and bulk soils resulting from changes in management. We have compared the effects of plants on the microbial community, using several methods, in three different types of soils. Pots containing soil from three contrasting sites were planted with Lolium perenne (rye grass). Physiological (Biolog), biochemical (PLFA) and molecular (DGGE and TRFLP) fingerprinting methods were employed to study the change in soil microbial communities caused by the growth of rye grass. Different methods of DNA extraction and nested PCR on TRFLP profiles were examined to investigate whether they gave different views of community structure. Molecular methods were used for both fungal and bacterial diversity. Principal component analysis of Biolog data suggested a significant effect of the plants on the microbial community structure. We found significant effects of both soil type and plants on microbial communities in PLFA data. Data from TRFLP of soil bacterial communities showed large effects of soil type and smaller but significant effects of plants. Effects of plant growth on soil fungal communities were measured by TRFLP and DGGE. Multiple Procrustes analysis suggested that both methods gave similar results, with only soil types having a significant effect on fungal communities. However, TRFLP was more discriminatory as it generated more ribotype fragments for each sample than the number of bands detected by DGGE. Neither methods of DNA extraction nor the nested PCR had any effect on the evaluation of soil microbial community structure. In conclusion, the different methods of microbial fingerprinting gave qualitatively similar results when samples were processed consistently and compatible statistical methods used. However, the molecular methods were more discriminatory than the physiological and biochemical approaches. We believe results obtained from this experiment will have a major impact on soil microbial ecology in general and rhizosphere–microbial interaction studies in particular, as we showed that the different fingerprinting methods for microbial communities gave qualitatively similar results.     

Effects of Arbuscular Mycorrhizal Colonization on Microbial Community in Rhizosphere Soil and Fusarium Wilt Disease in Tomato

Fusarium wilt is caused by soil-borne pathogen Fusarium oxysporum. Tomato (Lycopersicon esculentum Mill.) is susceptible to Fusarium oxysporum f. sp. lycopersici race 1 and was infected with wilt disease. A pot experiment was conducted to investigate effects of inoculating arbuscular mycorrhizal (AM) fungus (Glomus etunicatium) on the microbial community in the rhizosphere soil and Fusarium wilt in tomato (cv. Oogatafukuju). The results indicated that AM fungal inoculation suppressed the Fusarium number in the rhizosphere soil of tomato and decreased the Fusarium wilt disease index. Compared to the control, AM fungal inoculation increased the actinomycete number but increased bacterial number. Bacterial and fungal numbers were high but actinomycetes number was low when tomato basal stems became discolored brown. Fusarium inoculation significantly suppressed development of AM colonization and decreased polyphenol oxidase (PPO) activity in leaves and roots of tomato. Inoculation with AM fungi and Fusarium maintained high PPO activity in leaves and roots. The AM colonization increased root growth of tomato, whereas Fusarium inoculation had no significant effect on tomato growth. These findings suggest that because AM fungal inoculation changes microbial communities and enhances PPO activity, it should suppress occurrence of Fusarium wilt in tomato.