Does the depletion of pentachlorophenol in root–soil interface follow a simple linear dependence on the distance to root surfaces?

To understand root–soil–microbe interactions in rhizo-depletion of xenobiotics, we conducted a glasshouse study using specially designed laminar rhizoboxes which allow intact layers of near- (1–5 mm) and far- (>5 mm) rhizosphere soil to be harvested separately from root surfaces without the removal of the root material itself. Plant (Lolium perenne L.) seedlings were grown for 90 days in a soil treated with PCP at 20 and 50 mg kg⁻¹. Changes in PCP depletion, soil microbial biomass and community structure (as indicated by phospholipid fatty acids (PLFAs) profiles) with increasing distance from the root surfaces were then assessed after harvesting. Surprisingly, depletion of PCP in the planted rhizoboxes exhibited a nonlinear dependence on the distance to root surfaces, with the most rapid loss in the 2 or 3 mm near-rhizosphere layers, contrasting to the well-known linear gradient of root exudates and mineral nutrients etc. (generally, the extent gradually decreased with increasing distance from the root surface). Soil microbial biomass carbon, however, decreased linearly as expected with increasing distance from the roots. The microbial community structures as indicated by PLFA profiles showed distance-dependent selective enrichment of competent species that may be responsible for efficient PCP depletion. The results suggest that root exudates induced modifications of microbial communities in the PCP contaminated rhizosphere and spatially modified the dominant species within these communities, resulting in the nonlinear PCP depletion pattern.     

Facilitation of pentachlorophenol degradation in the rhizosphere of ryegrass (Lolium perenne L.)

The phytoremediation of xenobiotics depends upon plant-microbe interactions in the rhizosphere, but the extent and intensity of these effects are currently unknown. To investigate rhizosphere effects on the biodegradation of xenobiotics, a glasshouse experiment was conducted using a specially designed rhizobox where ryegrass seedlings were grown for 53 days in a soil spiked with pentachlorophenol (PCP) at concentrations of 8.7±0.5 and 18±0.5 mg kg⁻¹ soil. The soil in the rhizobox was divided into six separate compartments at various distances from the root surface. Changes in PCP concentrations with increasing distance from the root compartment of the rhizobox were then assessed. The largest and most rapid loss of PCP in planted soil was at 3 mm from the root zone where total PCP decreased to 0.20 and 0.65 mg kg⁻¹, respectively with the two PCP treatments. The degradation gradient followed the order: near-rhizosphere>root compartment>far-rhizosphere soil zones for both concentrations where ryegrass was grown. In contrast, there was no difference in PCP concentration with distance in the unplanted soil. The increases in both soil microbial biomass carbon and the activities of soil urease and phosphatase were accompanied by the enhanced degradation of PCP, which was higher in the near-rhizosphere than far-rhizosphere soil. The results suggest that the effect of root proximity is important in the degradation of xenobiotics such as PCP in soil.     

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.     

Nitrogen fertiliser rate affects the frequency of nitrate-dissimilating Pseudomonas spp. in the rhizosphere of Lolium perenne grown under elevated pCO2 (Swiss FACE)

The effect of elevated pCO₂ (60 Pa) on the frequency of nitrate-dissimilating Pseudomonas (NDP) was investigated in the rhizosphere of fertilised Lolium perenne swards in the Swiss Free Air Carbon dioxide Enrichment (FACE) experiment. Numbers of cultivable root-associated Pseudomonas were greater under elevated (60 Pa) than under ambient (36 Pa) pCO₂ in both high and low N-fertilised swards. For both pCO₂ conditions, the NDP frequency decreased with closer root proximity to L. perenne roots in low fertilised swards. Anyway, in high N swards the NDP frequency was similar in root and soil fractions. Thus, N availability may be a major factor influencing NDP populations under elevated pCO₂, most likely due to increased competition for N between plant and nitrate-dissimilating bacteria.     

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.     

Phenotypic structure of Pseudomonas populations is altered under elevated pCO2 in the rhizosphere of perennial grasses

The increasing atmospheric CO₂ content (pCO₂) is likely to modify the ecosystem functioning including rhizosphere bacteria that are directly dependent on rhizodeposition. This may include alteration of Pseudomonas populations that display phenotypic traits in relation with plant fitness. In the present study, 1228 Pseudomonas strains were isolated from the non-rhizosphere soil, rhizosphere soil and root fractions of perennial grassland systems: Lolium perenne and Molinia coerulea. Both plants were grown under ambient (36 Pa) and elevated (60 Pa) pCO₂ in the Swiss Free Air CO₂ Enrichment (FACE) system. Pseudomonas spp. were tested for their ability to produce auxin, siderophores and hydrogen cyanide, and to dissimilate nitrate. No effect of root proximity and elevated pCO₂ was observed on the proportions of auxin producers. For L. perenne and M. coerulea, siderophore and hydrogen cyanide Pseudomonas producers were stimulated in the root fraction. In contrast lower frequencies of nitrate reducers were observed in the root fraction compared to non-rhizosphere soil. The frequencies of siderophore producers and nitrate dissimilating strains were higher, and those of hydrogen cyanide producers lower, under elevated pCO₂ for L. perenne. This alteration of the phenotypic structure of Pseudomonas guild in the root fraction is discussed in relation with the physico-biochemical modifications of the rhizosphere condition via rhizodeposition and environmental changes occurring under elevated pCO₂.     

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.     

Determination of the impact of continuous defoliation of Lolium perenne and Trifolium repens on bacterial and fungal community structure in rhizosphere soil

To determine the effects of defoliation on microbial community structure, rhizosphere soil samples were taken pre-, and post-defoliation from the root tip and mature root regions of Trifolium repens L. and Lolium perenne L. Microbial DNA isolated from samples was used to generate polymerase chain reaction–denaturing gradient gel electrophoresis molecular profiles of bacterial and fungal communities. Bacterial plate counts were also obtained. Neither plant species nor defoliation affected the bacterial and fungal community structures in both the root tip and mature root regions, but there were significant differences in the bacterial and fungal community profiles between the two root regions for each plant. Prior to defoliation, there was no difference between plants for bacterial plate counts of soils from the root tip regions; however, counts were greater in the mature root region of L. perenne than T. repens. Bacterial plate counts for T. repens were higher in the root tip than the mature root region. After defoliation, there was no effect of plant type, position along the root or defoliation status on bacterial plate counts, although there were significant increases in bacterial plate counts with time. The results indicate that a general effect existed during maturation in the root regions of each plant, which had a greater impact on microbial community structure than either plant type or the effect of defoliation. In addition there were no generic consequences with regard to microbial populations in the rhizosphere as a response to plant defoliation.     

Atmospheric CO2 enrichment and nutrient additions to planted soil increase mineralisation of soil organic matter, but do not alter microbial utilisation of plant- and soil C-sources

Plants link atmospheric and soil carbon pools through CO₂ fixation, carbon translocation, respiration and rhizodeposition. Within soil, microbial communities both mediate carbon-sequestration and return to the atmosphere through respiration. The balance of microbial use of plant-derived and soil organic matter (SOM) carbon sources and the influence of plant-derived inputs on microbial activity are key determinants of soil carbon-balance, but are difficult to quantify. In this study we applied continuous ¹³C-labelling to soil-grown Lolium perenne, imposing atmospheric CO₂ concentrations and nutrient additions as experimental treatments. The relative use of plant- and SOM-carbon by microbial communities was quantified by compound-specific ¹³C-analysis of phospholipid fatty acids (PLFAs). An isotopic mass-balance approach was applied to partition the substrate sources to soil respiration (i.e. plant- and SOM-derived), allowing direct quantification of SOM-mineralisation. Increased CO₂ concentration and nutrient amendment each increased plant growth and rhizodeposition, but did not greatly alter microbial substrate use in soil. However, the increased root growth and rhizosphere volume with elevated CO₂ and nutrient amendment resulted in increased rates of SOM-mineralisation per experimental unit. As rhizosphere microbial communities utilise both plant- and SOM C-sources, the results demonstrate that plant-induced priming of SOM-mineralisation can be driven by factors increasing plant growth. That the balance of microbial C-use was not affected on a specific basis may suggest that the treatments did not affect soil C-balance in this study.     

Biotransformation of pentachlorophenol by Chinese chive and a recombinant derivative of its rhizosphere-competent microorganism, Pseudomonas gladioli M-2196

The use of plants or microorganisms to detoxify contaminated soil or groundwater is a potentially cost-effective alternative to traditional remediation technologies. This study investigated the effects of a rhizosphere microbe on the biotransformation of pentachlorophenol (PCP). Chinese chive (Allium tuberosum Rottler) and its rhizosphere-competent bacterium, Pseudomonas gladioli M-2196, were used as a plant-bacterium pair. The genes encoding PCP-degrading enzymes from Sphingobium chlorophenolicum ATCC39723 were introduced into the chromosome of P. gladioli M-2196. The resultant transformants were able to degrade PCP almost completely in liquid medium within 4 d in culture. PCP degradation experiments showed that the amount of PCP in soil (3.3 μg g⁻¹) planted with the P. gladioli transformant (T-9) and Chinese chive decreased by 40% as compared with untreated soil (control) by day 28. Strain T-9, which was used in the PCP degradation experiments, retained the ability to colonize the Chinese chive rhizosphere after 28 d. Tetrachlorocatechol (TCC) was detected as a metabolite of PCP in Chinese chive extract. The amount of PCP in soil treated only with Chinese chive decreased by 30% as compared with the control, but the total amount of PCP plus TCC detected in the plant was less than 10% of the amount of PCP removed from soil. This might be due to the enhancement of a soil microflora population capable of degrading PCP by root exudates from Chinese chive. Therefore, Chinese chive itself, in addition to the rhizosphere-competent bacterium, seemed to play an important role in reducing the PCP level in the soil.     

根际修复中五氯酚耐性草种的筛选及其应用效果研究

以8种国内外应用较为广泛的草坪草种为材料,进行了根际修复中五氯酚(PCP)耐性草种的筛选及其应用效果研究,结果表明,多年生黑麦草顶峰(Pinnacle)为PCP耐性品种,在PCP污染土壤的根际修复中具有通过优化土壤环境而促进PCP在根际微域中快速降解的能力。研究结果为PCP污染土壤根际修复中目标植物的合理选取提供了科学试验依据。     

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.     

Culture-based and culture-independent assessment of the impact of mixed and single plant treatments on rhizosphere microbial communities in hydrocarbon contaminated flare-pit soil

Phytoremediation is a novel treatment option for weathered, hydrocarbon contaminated, flare-pit soil in prairie ecosystems. The remediation potential of six different naturalized prairie plants was assessed by examining their impact on the degradation potential of indigenous bacterial communities. Culture-based and culture-independent microbiological methods were used to determine if mixed plant treatments stimulate different microbial communities and catabolic genotypes in comparison to individual plant species that comprise the mix. DGGE analysis of PCR-amplified 16S rRNA genes revealed that alfalfa (Medicago sativa) had a dominant effect on the structure of rhizosphere microbial communities in mixed plant treatments, stimulating relative increases in specific Bacteroidetes and Proteobacteria populations. Alfalfa and mixes containing alfalfa, while supporting 100 times more culturable PAH degraders than other treatments, exhibited only 10% TPH reduction, less than all planted treatments except perennial rye grass (Lolium perenne). Total petroleum hydrocarbon (TPH) reduction was greatest in single-species grass treatments, with creeping red fescue (Festuca rubra) reducing the TPH concentration by 50% after 4.5 months. Overall TPH reduction throughout the study was positively correlated (p<0.001) to culturable n-hexadecane degraders.     

2-Phenylethylisothiocyanate concentration and microbial community composition in the rhizosphere of canola

Canola crops have been shown to inhibit soil-borne pathogens in following crops. This effect is mainly attributed to the release of low molecular S-containing compounds, such as isothiocyanates, during microbial degradation of the crop residues. We have assessed the effect of low concentrations of phenylethylisothiocyanate (PEITC) on soil microbial communities as well as its rate of degradation in soil and determined the concentration of PEITC and the microbial community structure in the rhizosphere of canola. PEITC was degraded within 96 h by soil microorganisms. PEITC added to the soil daily for 5 d affected both bacterial and eukaryotic community structure, determined by PCR-DGGE. Community structures of bacteria and eukaryotes changed at PEITC concentrations between 1300 and 3790 pmol g⁻¹ soil fresh weight but was unaffected at lower concentrations. The PEITC concentration in the rhizosphere of living canola roots was greater in first order laterals than in second order laterals. The maximal PEITC concentration detected in the rhizosphere was 1827 pmol g⁻¹. Redundancy analysis of the DGGE banding patterns indicated a significant correlation between the PEITC concentration in the rhizosphere and the community structure of the active fraction of eukaryotes and bacteria in the rhizosphere. Other important factors influencing the microbial community structure were soil moisture and plant dry matter. It is concluded that canola may affect the soil microbial community structure not only after incorporation of canola residues but also during active growth of the plants.     

Effects of glyphosate on rhizosphere soil microbial communities under two different plant compositions by cultivation-dependent and -independent methodologies

We studied, under two different plant compositions, the short-term effects of glyphosate on rhizosphere soil microbial communities through the utilization of cultivation-dependent and -independent techniques. A short-term pot study was carried out using factorial treatments that included two different compositions of forage plant species (triticale versus a mixture of triticale and pea) and two concentrations of glyphosate (50 and 500 mg active ingredient kg⁻¹ soil, as a commercial formulation, Roundup Plus) arranged in a completely randomized design experiment with four replicates. Control plants (no glyphosate added) were clipped in an attempt to compare two methods of weed control (manual = clipping; chemical = herbicide treatment). Rhizosphere soil was sampled 15 and 30 days after glyphosate treatment and the following soil components were determined: potentially mineralizable nitrogen, ammonium content, community-level physiological profiles using Biolog Ecoplates™, DNA microbial biomass and genotype diversity by means of PCR-DGGE. Fifteen days after herbicide treatment, a glyphosate-induced stimulation of the activity and functional diversity of the cultivable portion of the heterotrophic soil microbial community was observed, most likely due to glyphosate acting as an available source of C, N and P. On the other hand, 30 days after herbicide treatment, both the activity and diversity of the rhizosphere soil microbial communities showed an inconsistent response to glyphosate addition. Apart from its intended effect on plants, glyphosate had non-target effects on the rhizosphere soil microbial community which were, interestingly, more enhanced in triticale than in “triticale + pea” pots. Biolog™ was more sensitive than PCR-DGGE to detect changes in soil microbial communities induced by glyphosate and plant composition.     

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.     

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.     

Enhanced dissipation of chrysene in planted soil: the impact of a rhizobial inoculum

Results from an innovative approach to improve remediation in the rhizosphere by encouraging healthy plant growth and thus enhancing microbial activity are reported. Mixed grass-legume systems, together with microbial inoculants, were used to remediate a polycyclic aromatic hydrocarbon (chrysene) spiked agricultural soil. Inoculants were symbiotic rhizobia, which may play an important role in rhizoremediation by increasing plant and root growth. An inoculum of an isolate of Rhizobium leguminosarum bv. trifolii, selected for PAH tolerance, was produced using a peat carrier. The inoculum and white clover (Trifolium repens L.), were planted into soils with ryegrass (Lolium perenne L.). The soils spiked with chrysene (500 mg kg⁻¹) then aged for 4 weeks. Shoot- and root-biomass of plants, and the amount of root nodulation, were determined. Rhizobial populations, soil pH and soil nitrogen were also monitored throughout the trial. In addition, the ability of the inoculated rhizobial strain to utilise chrysene as a sole carbon source was assessed. Direct uptake and/or degradation of chrysene by the clover and ryegrass did not occur to a significant degree. Enhanced losses of chrysene were seen in planted, non-sterile soils that contained a rhizobial inoculum. No direct degradation of chrysene by R. leguminosarum bv. trifolii was observed and no enhanced losses of PAHs were detected in sterile soils after inoculation with rhizobia. Results suggest that the enhanced dissipation of chrysene, observed in the non-sterile planted inoculated pots, was not a result of degradation of chrysene by R. leguminosarum bv. trifolii. The symbiotic association with R. leguminosarum bv. trifolii improved plant vigour and growth in inoculated planted treatments. This may have stimulated the rhizospheric microflora to degrade chrysene.     

Partitioning root and microbial contributions to soil respiration in Leymus chinensis populations

Based on the enclosed chamber method, soil respiration measurements of Leymus chinensis populations with four planting densities (30, 60, 90 and 120 plants/0.25 m²) and blank control were made from July 31 to November 24, 2003. In terms of soil respiration rates of L. chinensis populations with four planting densities and their corresponding root biomass, linear regressive equations between soil respiration rates and dry root weights were obtained at different observation times. Thus, soil respiration rates attributed to soil microbial activity could be estimated by extrapolating the regressive equations to zero root biomass. The soil microbial respiration rates of L. chinensis populations during the growing season ranged from 52.08 to 256.35 mg CO₂ m⁻² h⁻¹. Soil microbial respiration rates in blank control plots were also observed directly, ranging from 65.00 to 267.40 mg CO₂ m⁻² h⁻¹. The difference of soil microbial respiration rates between the inferred and the observed methods ranged from −26.09 to 9.35 mg CO₂ m⁻² h⁻¹. Some assumptions associated with these two approaches were not completely valid, which might result in this discrepancy. However, these two methods' application could provide new insights into separating root respiration from soil microbial respiration. The root respiration rates of L. chinensis populations with four planting densities could be estimated based on measured soil respiration rates, soil microbial respiration rates and corresponding mean dry root weight, and the highest values appeared at the early stage, then dropped off rapidly and tended to be constant after September 10. The mean proportions of soil respiration rates of L. chinensis populations attributable to the inferred and the observed root respiration rates were 36.8% (ranging from 9.7 to 52.9%) and 30.0% (ranging from 5.8 to 41.2%), respectively. Although root respiration rates of L. chinensis populations declined rapidly, the proportion of root respiration to soil respiration still increased gradually with the increase of root biomass.     

Changes in microbial communities and carbohydrate profiles induced by the mycorrhizal fungus (Glomus intraradices) in rhizosphere of olive trees (Olea europaea L.)

The influence of inoculation of olive trees with arbuscular mycorrhizal (AM) fungi, Glomus (G) intraradices, on microbial communities and sugar concentrations, were examined in rhizosphere of olive trees (Olea europaea L.). Analyses of phospholipid and neutral lipid fatty acids (PLFA and NLFA, respectively) were then used to detect changes in microbial community structure in response to inoculation of plantlets with G. intraradices.Microscopic observations studies revealed that the extraradical mycelium of the fungus showed formation of branched absorbing structures (BAS) in rhizosphere of olive tree. Root colonization with the AM fungi G. intraradices induced significant changes in the bacterial community structure of olive tree rhizosphere compared to non-mycorrhizal plants. The largest proportional increase was found for the fatty acid 10Me18:0, which indicated an increase in the number of actinomycetes in mycorrhizal rhizosphere soil, whereas the PLFAs i15:0, a15:0, i16:0, 16:1ω7 and cy17:0 which were used as indicators of bacteria decreased in mycorrhizal treatment compared to non-mycorrhizal control treatment. A highest concentration of glucose and trehalose and a lowest concentration of fructose, galactose, sucrose, raffinose and mannitol were detected in mycorrhizal rhizosphere soil. This mycorrhizal effect on rhizosphere communities may be a consequence of changes in characteristics in the environment close to mycorrhizal roots.