Influence of growth-promoting bacteria on the growth of wheat in different soils and temperatures

Plant-growth-promoting bacteria isolated from the rhizosphere, phyllosphere and soil of the root zone in different climatic regions of Germany and Uzbekistan were analysed for plant-growth-promoting effects and nutrient uptake on winter wheat on different soils and under different temperature regimes. The investigations were carried out in pot experiments using loamy sand and sandy loam soils from Müncheberg, Germany and Calcisol soil from Tashkent, Uzbekistan. The temperature and soil types were found to influence growth-promoting effects. Inoculation with bacterial strains Pseudomonas fluorescens PsIA12, Pantoea agglomerans 050309 and Mycobacterium sp. 44 isolated from Müncheberg (semi-continental climate) was found to significantly increase the root and shoot growth of winter wheat at 16 °C compared to 26 °C in loamy sand. Mycobacterium phlei MbP18 and Mycoplana bullata MpB46 isolated from Tashkent (semi-arid climate) were found to significantly increase the root and shoot growth of winter wheat in nutrient-poor Calcisol at 38 °C as well as in nutrient-rich loamy sand at 16 °C. Bacterial inoculation also resulted in significantly higher N, P, and K contents of plant components. The bacteria isolates were able to survive in the rhizosphere and in the soil of winter wheat after root and shoot inoculation.     

Azospirillum strains isolated from roots and rhizosphere soil of wheat (Triticum aestivum L.) grown under different soil moisture conditions

The present study deals with the isolation and characterization of Azospirillum strains isolated from roots and rhizosphere soil of wheat (at tillering and anthesis stages) plants grown under different moisture regimes in the field and in pots. The survival of Azospirillum isolates from plants of irrigated field and those from well-watered pots was higher than that of Azospirillum strains isolated from roots and rhizosphere soils of plants grown under arid and semiarid (14–8% soil moisture) field conditions and under water-stressed (8% soil moisture) conditions in pots. On the basis of carbon/nitrogen source utilization, the Azospirillum strains isolated from wheat under field and pot conditions were grouped in three groups. The unweighted pair group method with arithmetic means cluster analysis based on random amplification of polymorphic DNA showed that two groups of Azospirillum were similar. The strains isolated from plants (at tillering stage) grown under low moisture conditions either in pots or in field were genetically similar to strains isolated from plants grown under well-watered conditions in both pots and field. Inoculation of wheat with isolates from water-stressed plants induced tolerance to water stress in inoculated plants. Isolates from water-stressed conditions exhibited lower production of indole acetic acid, gibberellic acid, and trans zeatin riboside but a higher production of abscisic acid.     

Effect of Azospirillum brasilense inoculation on rhizobacterial communities analyzed by denaturing gradient gel electrophoresis and automated ribosomal intergenic spacer analysis

Nucleic acid-based techniques allow the exploration of microbial communities in the environments such as the rhizosphere. Azospirillumbrasilense, a plant growth promoting rhizobacterium (PGPR), causes morphological changes in the plant root system. These changes in root physiology may indirectly affect the microbial diversity of the rhizosphere. In this study, the changes in the rhizobacterial structure following A. brasilense inoculation of maize (Zea mays) plants was examined by PCR-denaturating gradient gel electrophoresis (DGGE) and automated ribosomal intergenic spacer analysis (ARISA), using two universal primers sets for the 16S rRNA gene, and an intergenic 16S-23S rDNA primer set, respectively. Similar results were obtained when using either ARISA or DGGE performed with these different primer sets, and analyzed by different statistical methods: no prominent effect of A. brasilense inoculation was observed on the bacterial communities of plant roots grown in two different soils and in different growth systems. In contrast, plant age caused significant shifts in the bacterial populations.     

Effects of PAH-Contaminated Soil on Rhizosphere Microbial Communities

Bacterial associations with plant roots are thought to contribute to the success of phytoremediation. We tested the effect of addition of a polycyclic aromatic hydrocarbon contaminated soil on the structure of the rhizosphere microbial communities of wheat (Triticum aestivum), lettuce (Lactuca sativa var. Tango), zucchini (Cucurbita pepo spp. pepo var. Black Beauty), and pumpkin (C. pepo spp. pepo var. Howden) 16S rDNA terminal restriction fragment length polymorphism (T-RFLP) profiles of rhizosphere microbial communities from different soil/plant combinations were compared with a pairwise Pearson correlation coefficient. Rhizosphere microbial communities of zucchini and pumpkin grown in the media amended with highest degree of contaminated soil clustered separately, whereas communities of these plants grown in unamended or amended with lower concentrations of contaminated soil, grouped in a second cluster. Lettuce communities grouped similarly to cucurbits communities, whereas wheat communities did not display an obvious clustering. The variability of 16S rDNA T-RFLP profiles among the different plant/soil treatments were mostly due to the difference in relative abundance rather than presence/absence of T-RFLP fragments. Our results suggest that in highly contaminated soils, the rhizosphere microbial community structure is governed more by the degree of contamination rather than the plant host type.     

Interactions between a plant growth-promoting rhizobacterium,an AM fungus and a phosphate-solubilising fungus in the rhizosphere of Lactuca sativa

This study evaluated the interactions between the inoculation with an arbuscular mycorrhizal fungus, Glomus intraradices Schenck & Smith, a plant growth-promoting rhizobacterium, Bacillus subtilis, and a filamentous soil fungus, Aspergillus niger, with respect to their effects on growth of lettuce plants and on indicators of biological soil quality (microbial biomass C, water-soluble C and carbohydrates and dehydrogenase, urease, acid phosphatase and benzoyl argininamide hydrolyzing protease activities). Water-soluble carbohydrates and microbial biomass were increased only in the rhizosphere soil of G. intraradices-plants. Rhizosphere soil from all microbial inoculation treatments had significantly higher dehydrogenase activity than the control soil, particularly in the soil inoculated with B. subtilis (about 21% higher than control soil). Inoculation with A. niger or B. subtilis increased significantly the urease, protease and phosphatase activities of the rhizosphere soil of the lettuce plants. The foliar P and K contents increased significantly with the B. subtilis or G. intraradices inoculation, alone or in combination. The most effective co-inoculation was observed in the combined treatment of inoculation with G. intraradices and B. subtilis, which synergistically increased plant growth compared with singly inoculated (about 77% greater with respect to the control plants).     

Nematode community structure as a sensitive indicator of microbial perturbations induced by a genetically modified Pseudomonas fluorescens strain

The effects of seed inoculation with the Pseudomonas fluorescens strains F113lacZY [a genetically marked biocontrol agent producing the anti-fungal agent 2,4-diacetylphloroglucinol (DAPG)] and F113G22 [a genetically modified (GM) derivative strain of F113lacZY incapable of producing DAPG] on associated nematode communities were investigated over 17 days of plant growth. Plant growth measurements and colony forming unit counts (CFU) derived from rhizosphere soil indicated only small and transient perturbations as a result of introductions of the GM bacteria. Total nematode numbers were increased significantly in the rhizosphere of inoculated plants compared with the non-inoculated control treatments. These increases were mainly due to increases in bacterial feeding nematodes. This indicates that inoculation with the GM P. fluorescens strains induced high bacterial growth rates in the rhizosphere of plants inoculated with these strains. No indication of greater root colonisation by fluorescent Pseudomonas spp. could be found using CFU counts on Pseudomonas-selective media. Numbers of fungal feeding nematodes decreased initially, probably as a result of lack of intact hyphae in the soil. However, inoculation with the two different GM P. fluorescens strains resulted in a rapid recovery of fungal feeding nematode populations, whereas in the non-inoculated control populations of fungal feeding nematodes remained small. This result is surprising as one of the strains (F113lacZY) produces the anti-fungal agent DAPG and it would be expected that this agent would result in a decrease in fungal activity.     

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.     

Subtle changes in rhizosphere microbial community structure in response to increased boron and sodium chloride concentrations

Two of the major constraints to grain production in large areas of South-East Australia and cropping soils worldwide are high levels of subsoil boron (B) and excessive salinity (NaCl). Although the effect of these constraints is often studied in plants, the effect on microbially mediated plant-beneficial processes is unclear. To that end, we investigated the impact of B and NaCl on soil microbial community structure (MCS) in the wheat rhizosphere using BIOLOG ecoplates and terminal restriction fragment length polymorphism (T-RFLP). In addition, the effects of B and NaCl on the nitrogen (N) cycle processes of N fixation and ammonia oxidation were assessed by the construction of clone libraries of diazotrophic (nifH) and ammonia oxidising (amoA) rhizobacteria. Analysis of BIOLOG plates using non-metric multidimensional scaling (MDS) revealed addition of both B and NaCl significantly changed MCS, the latter of which was also significant through the analysis of T-RFLP data. Utilisation of several chemical groups of BIOLOG substrates significantly changed in NaCl-amended soil; both B and NaCl affected utilisation of several individual substrates indicative of plant stress including serine and malic acid. A significant decrease in diversity and species richness was observed in high B rhizosphere soil. The community structure of ammonia-oxidising bacteria (AOB), all of which clustered with Nitrosospira-like sequences, did not significantly change in response to addition of B or NaCl, but addition of the latter resulted in a significant increase of diazotroph clones within the α-proteobacteria similar to Azospirillum sp. It appeared that the addition of B and NaCl to soil changed rhizosphere MCS indirectly through increased soil moisture and subtle changes in root exudate patterns, the addition of the latter producing a more distinct change through increased osmotic pressure, leading to a greater increase in rhizodeposition of nutrients, especially carbohydrates. The implications for the current study are that B and NaCl are more likely to affect rhizosphere MCS indirectly through root exudate quantity and/or quality than directly through microbial toxicity, and that plant health is a major determinant in rhizosphere MCS and normal N cycling.     

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.     

Host-plant specificity in the infection of cereals with Azospirillum spp

The specificity of the infection of maize, wheat and rice roots by N₂-fixing Azospirillum spp was studied in four greenhouse experiments using pots with unsterilized soil and in two field experiments. In all experiments A. lipoferum was most frequently isolated from externally sterilized roots of maize, and A. brasilense nir^? (nitrite reductase negative) from wheat and rice. In pot experiments, A. brasilense nir⁺ was isolated with moderate frequency from within maize roots but rarely from within wheat or rice roots. Inoculation of the pots with a mixture of representative strains of the three Azospirillum groups had no effect on the proportion of strains recovered from each plant species. In the field experiments, inoculation with spontaneous streptomycin-resistant mutants of two of the representative strains confirmed the apparent specificity of A. lipoferum for maize roots and of A. brasilense for wheat but the results were partially obscured by the unexpectedly high proportion of streptomycin-resistant strains isolated from within the roots of uninoculated plants.     

Effects of inoculation with ectomycorrhizal fungi on microbial biomass and bacterial functional diversity in the rhizosphere of Pinus tabulaeformis seedlings

This study investigated the effects of inoculation with three individual ectomycorrhizal (ECM) fungal species on soil microbial biomass carbon and indigenous bacterial community functional diversity in the rhizosphere of Chinese pine (Pinus tabulaeformis Carr.) seedlings under field experimental conditions. The results showed that ECM fungal inoculation significantly increased the ectomycorrhizal colonization compared with non-inoculated seedlings. ECM fungal inoculations have higher soil microbial biomass carbon than that of control, ranging from 49.6 μg C g^?1 dry soil in control to 134.02 μg C g^?1 dry soil in treatment inoculated with Boletus luridus Schaeff ex Fr. Multivariate analyses (PCA) of BIOLOG data revealed that the application of ECM fungi significantly influenced bacterial functional diversity in the rhizosphere of P. tabulaeformis seedlings. The highest average well-color development (AWCD) and functional diversity indices were also observed in treatment inoculated with B. luridus. A wider range of sole carbon sources were utilized by the bacterial community in the rhizosphere of inoculated seedlings. The data gathered from this study provides important information for utilization of ECM fungi in forest restoration project in the Northwestern China. The present study will also significantly broaden our understanding of practical importance in the application of ECM fungal inoculum to promote soil microbial community diversity of soil.     

Überleben inokulierter Rhizosphärenbakterien und deren Einfluß auf native Bakterienpopulationen im Wurzelraum von Luzerne

Survival of inoculated rhizosphere bacteria and their influence on native bacterial populations in the rhizosphere of alfalfa The survival of inoculated bacteria and their influence on native bacterial populations in the rhizosphere of alfalfa were investigated in a greenhouse experiment. The plant growth promoting strains Rhizobium meliloti me18 and Pseudomonas fluorescens PsIA12 were reisolated from the rhizosphere about 7 weeks after single and mixed strain inoculation. They did not induce lasting changes in the diversity of the native bacterial communities of the rhizosphere. Only within the first week after inoculation was an increase in total bacterial abundance observed. In general, the diversity of bacterial communities increased with plant age and with proximity to the root tip.     

Influence of introduced potential biocontrol agents on maize seedling growth and bacterial community structure in the rhizosphere

Two species of Pseudomonas chromosomally tagged with gfp, which had shown antagonistic activity against the tomato pathogen Ralstonia solanacearum in a previous study, were assessed for their impact in the rhizosphere of maize. Plant growth characteristics, numbers of indigenous heterotrophic bacteria, changes in the bacterial community structure according to the r/K strategy concept, and shifts in MIDI-FAME profiles of culturable bacterial fractions as well as total rhizosphere microbial communities were determined in relation to seed and soil treatment with the exogenous pseudomonads. The maize rhizosphere proved to be a suitable habitat for the introduced P. chlororaphis IDV1 and P. putida RA2, which showed good survival after introduction. However, both inoculants showed a small growth-reducing effect towards maize, which might have been caused by the high densities of inoculants used (i.e. competition for nutrients and action of metabolites produced) and/or changes in microbial community structure (both culturable bacterial fraction and the total microflora). Probably, an altered balance among the indigenous maize rhizosphere populations occurred. Thus, the culturable bacteria, as well as the total microflora in the rhizosphere, changed in response to the introduced pseudomonads, and their development was dependent on the growth stage of the plant. The FAME analyses showed that these microbial communities comprised different populations, and were separated according to, first, the method used (direct versus cultivation-based), second, sampling time, and, finally, inoculation level.     

Interactions of arbuscular-mycorrhizal fungi and Bacillus strains and their effects on plant growth,microbial rhizosphere activity (thymidine and leucine incorporation) and fungal biomass (ergosterol and chitin)

The effects of two Bacillus strains (Bacillus pumillus and B. licheniformis) on Medicago sativa plants were determined in single or dual inoculation with three arbuscular-mycorrhizal (AM) fungi and compared to P-fertilization. Shoot and root plant biomass, values of thymidine and leucine incorporation as well as ergosterol and chitin in rhizosphere soil were evaluated to estimate metabolic activity and fungal biomass, respectively, according to inoculation treatments. For most of the plant parameters determined, the effectiveness of AM fungal species was influenced by the bacterial strain associated. Dual inoculation of Bacillus spp. and AM fungi did not always significantly increase shoot biomass compared to single AM-colonized plants. The most efficient treatment in terms of dry matter production was the dual Glomus deserticola plus B. pumillus inoculation, which produced similar shoot biomass and longer roots than P-fertilization and a 715% (shoot) and 190% (root length) increase over uninoculated control. The mycorrhizas were more important for N use-efficiency than for P use-efficiency, which suggests a direct mycorrhizal effect on N nutrition not mediated by P uptake. Both chemical and biological treatments affected thymidine and leucine incorporation in the rhizosphere soil differently. Thymidine was greater in inoculated than in control rhizospheres and B. licheniformis was more effective than B. pumillus in increasing thymidine. Non-inoculated rhizospheres showed the lowest thymidine and leucine values, which shows that indigenous rhizosphere bacteria increased with introduced inocula. The highest thymidine and leucine values found in P-fertilized soils indicate that AM plants are better adapted to compete with saprophytic soil bacteria for nutrients than P-amended plants. Chitin was only increased by coinoculation of B. licheniformis and G. intraradices. B. pumillus increased ergosterol (indicative of active saprophyte fungal populations) in the rhizosphere of AM plants and particularly when colonized by G. mosseae. The different AM fungi have different effects on bacterial and/or fungal saprophytic populations and for each AM fungus, this effect was specifically stimulated or reduced by the same bacterium. This is an indication of ecological compatibilities between microorganisms. Particular Glomus–bacterium interactions (in terms of effect on plant growth responses or rhizosphere population) do not seem to be related to the percentage of AM colonization. The effect on plant growth and stimulation of rhizosphere populations, as a consequence of selected microbial groups, may be decisive for the plant establishment under limiting soil conditions.     

Effect of Piriformospora indica and Azospirillum strains from saline or non-saline soil on mitigation of the effects of NaCl

Salinity toxicity is a worldwide agricultural and eco-environmental problem. The intent of this study was to determine the salt tolerance of Piriformospora indica and strains of Azospirillum, isolated from non-saline and saline soil, as well as to determine their affect on the tolerance of wheat to soil salinity. In this study, an experiment was conducted to investigate the salt stress tolerance abilities of the endophytic fungi, P. indica, and Azospirillum strains, isolated from non-saline and saline soil, at five NaCl levels (0, 0.1, 0.2, 0.3, 0.4, 0.5 mol L^?1). Additionally, a greenhouse experiment was conducted to test the effects of these selected microorganisms under increasing salinity levels on seedling growth, solute accumulation (proline and sugars), and photosynthetic pigments (Chl a, b, ab) of seedling wheat. Azospirillum strains were isolated in Iran from the root of field-grown maize from non-saline soil with an EC = 0.7 dS m^?1 and from saline soil with an EC = 4.7 dS m^?1. Plants were irrigated with non-saline water–tap water with an electrical conductivity water (ECw) value of 0.2 dS m^?1, as well as low, moderate and severe saline water-irrigation with saline water with an ECw of 4 dS m^?1, 8 dS m^?1 and 12 dS m^?1, respectively. The upper threshold of P. indica salinity tolerance was 0.4 mol L^?1 NaCl in both liquid and solid broth medium. The upper thresholds of the salt adapted and non-adapted Azospirillum strains were 0.2 and 0.4 mol L^?1 NaCl, respectively. The results indicated a positive influence of the organisms on salinity tolerance, more with the saline-adapted Azospirillum strains than the non-adapted strains. P. indica was more effective than the Azospirillum strains. These results could be related to a better water status, higher photosynthetic pigment contents and proline accumulation in wheat seedlings inoculated with P. indica. The benefits of both isolates and P. indica depended on two factors: water salinity and growth stage of the host plant. Inoculation with the two isolates increased salinity tolerance of wheat plants; the saline-adapted Azospirillum strains showed better performance with respect to improved fresh and dry weights at 80 and 100 days after sowing under both non-saline and saline conditions. When compared to plants inoculated with non-saline-adapted Azospirillum strains, those inoculated with adapted Azospirillum strains had much better performance with respect to the presence of photosynthetic pigment (Chl a, b and ab) and proline accumulation. Overall, these results indicate that the symbiotic association between P. indica fungus and wheat plants improved wheat growth, regardless of the salinity. It is concluded that the mechanisms for protecting plants from the detrimental effects of salinity by P. indica fungus and Azospirillum strains may differ in their salinity tolerance and influence the uptake of water, photosynthetic pigment contents and proline accumulation in wheat seedlings.     

Interactions between the arbuscular mycorrhizal fungus Glomus intraradices and the plant growth promoting rhizobacteria Paenibacillus polymyxa and P. macerans in the mycorrhizosphere of Cucumis sativus

Interactions between the arbuscular mycorrhizal (AM) fungus Glomus intraradices and bacteria from the genus Paenibacillus (P. macerans and P. polymyxa) were examined in a greenhouse pot experiment with Cucumis sativus with and without organic matter amendment (wheat bran). P. polymyxa markedly suppressed AM fungus root colonization irrespective of wheat bran amendment, whereas P. macerans only suppressed AM fungus root colonization in combination with wheat bran amendment. Dual inoculation with P. macerans and G. intraradices in combination with wheat bran amendment also caused severe plant growth suppression. Inoculation with G. intraradices was associated with increased levels of dehydrogenase activity and available P in the growth substrate suggesting that mycorrhiza formation accelerated the decomposition of organic matter resulting in mobilization of phosphorus. Inoculation with both Paenibacillus species increased all measured microbial fatty acid biomarkers in the cucumber rhizosphere, except for the AM fungus biomarker 16:1ω5, which was reduced, though not significantly. Similarly, inoculation with G. intraradices increased all measured microbial fatty acid biomarkers in the cucumber rhizosphere, except for the Gram-positive bacteria biomarker 15:0 anteiso, which was overall decreased by G. intraradices inoculation. In combination with wheat bran amendment G. intraradices inoculation caused a 39% reduction in the amount of 15:0 anteiso in the treatment with P. polymyxa, suggesting that G. intraradices suppressed P. polymyxa in this treatment. In conclusion, plant growth promoting species of Paenibacillus may have suppressive effects of AM fungi and plant growth, especially in combination with organic matter amendment. The use of an inert plant growth media in the present study allowed us to study rhizosphere microbial interactions in a relative simple substrate with limited interference from other soil biota. However, the results obtained in the present work mainly show potential interactions and should not be directly extrapolated to a soil situation.     

Elevated CO2 increases the effect of an arbuscular mycorrhizal fungus and a plant-growth-promoting rhizobacterium on structural stability of a semiarid agricultural soil under drought conditions

In arid and semiarid Mediterranean regions, an increase in the severity of drought events could be caused by rising atmospheric CO₂ concentrations. We studied the effects of the interaction of CO₂, water supply and inoculation with a plant-growth-promoting rhizobacterium (PGPR), Pseudomonas mendocina Palleroni, or inoculation with an arbuscular mycorrhizal (AM) fungus, Glomus intraradices (Schenk & Smith), on aggregate stabilisation of the rhizosphere soil of Lactuca sativa L. cv. Tafalla. The influence of such structural improvements on the growth of lettuce was evaluated. We hypothesised that elevated atmospheric CO₂ concentration would increase the beneficial effects of inoculation with a PGPR or an AM fungus on the aggregate stability of the rhizosphere soil of lettuce plants. Leaf hydration, shoot dry biomass and mycorrhizal colonisation were decreased significantly under water-stress conditions, but this decrease was more pronounced under ambient vs elevated CO₂. The root biomass decreased under elevated CO₂ but only in non-stressed plants. Under elevated CO₂, the microbial biomass C of the rhizosphere of the G. intraradices-colonised plants increased with water stress. Bacterial and mycorrhizal inoculation and CO₂ had no significant effect on the easily-extractable glomalin concentration. Plants grown under elevated CO₂ had a significantly higher percentage of stable aggregates under drought stress than under well-watered conditions, particularly the plants inoculated with either of the assayed microbial inocula (about 20% higher than the control soil). We conclude that the contribution of mycorrhizal fungi and PGPR to soil aggregate stability under elevated atmospheric CO₂ is largely enhanced by soil drying.     

Quantification of the biocontrol agent Pseudomonas fluorescens Pf153 in soil using a quantitative competitive PCR assay unaffected by variability in cell lysis- and DNA-extraction efficiency

Although often neglected, variability in cell lysis efficiency and DNA extraction yield represents the major hurdles of any polymerase chain reaction (PCR)-based quantification protocol in soil and other natural environments. In this study we developed a technique that minimizes the effects of these constraints, providing at the same time a reliable internal control to distinguish between PCR-inhibition and negative results. We used Pseudomonas fluorescens Pf153, a root-colonizing bacterium that shows biocontrol activity against tobacco and cucumber black root rot, as the target organism for PCR quantification. Prior to DNA extraction, the genetically engineered, cognate reference strain P. fluorescens CHA0/c2 was inoculated in a reference soil. CHA0/c2 in the reference soil and Pf153 in the soil sample were lysed in parallel and afterward the lysates were mixed in known proportions. CHA0/c2 carries the plasmid pME6031-cmp2 that contains an allelic variant (competitor) of the Pf153 specific sequence Pf153_2. In a quantitative competitive PCR (QC-PCR) assay the competitor allows the quantification of the target strain down to 0.66 Pf153 CFU/mg soil. Processing the reference strain in the same way as Pf153 enables the exact quantification of the target strain in biocontrol assays performed in natural soil, overcoming differences in DNA extraction efficiency and PCR amplification from different soil environments. This technique is easily adaptable to other Pseudomonas strains simply by replacing the competitor used here with one derived from a SCAR-marker which is specific for the strain of choice.     

Effects of microbial inoculations on soil chemical,biochemical and microbial biomass carbon of cassava (Manihot esculenta Crantz) growing Vertisols

Cassava is an important subsidiary food in the tropics. In Tamil Nadu, India, microbial cultures were used to eradicate the tuberous root rot of cassava. Hence, an experiment was conducted for two consecutive years to test the effects of coinoculation of microbes on soil properties. The surface soil from the experimental site was analysed for soil available nutrients, soil enzyme activities and microbial biomass carbon. The treatment of Azospirillum with Trichoderma at the 50% recommended N:P₂O₅:K₂O (NPK) rate (50:25:50 kg ha^?1) significantly increased soil available nitrogen (142.81 kg ha^?1) by 72.66% over uninoculated control. There was a significant increase in available phosphorus in soil by the inoculation of AM (arbuscular mycorrhizal) fungi with Trichoderma at the 50% recommended NPK rate (41.04 kg ha^?1) compared to other treatments. The application of Pseudomonas fluorescens with Trichoderma at the 50% recommended NPK rate significantly increased available iron (19.34 µg g^?1) in soil. The treatment of Azospirillum with Trichoderma increased urease enzyme activity at the recommended NPK rate (816.32 μg urea hydrolyzed g^?1 soil h^?1). Soil application of all cultures at the 50% recommended NPK rate significantly increased dehydrogenase activity (88.63 μg TPF g^?1 soil) and β-glucosidase activity (48.82 μg PNP g^?1 soil) in soil. Inoculation of Trichoderma alone at the 50% recommended NPK rate significantly increased microbial biomass carbon (3748.85 μg g^?1 soil). Thus, the microbial inoculations significantly increased soil available nutrient contents, enzyme activities such as urease, dehydrogenase and β-glucosidase activity and microbial biomass carbon by reducing the amount of the required fertilizer.     

Nitrogen and phosphorus transformations in the rhizospheres of three tree species in a nutrient-poor sandy soil

We examined acid phosphatase activity (APA), N mineralization and nitrification rates, available N and P, and microbial biomass C, N and P in rhizosphere and bulk soils of 18-year-old Siberian elm (Ulmus pumila), Simon poplar (Populus simonii) and Mongolian pine (Pinus sylvestris var. mongolica) plantations on a nutrient-poor sandy soil in Northeast China. The main objective was to compare the rhizosphere effects of different tree species on N and P cycling under nutrient-deficient conditions. All tree species had the similar pattern but considerably different magnitude of rhizosphere effects. The APA, potential net N mineralization and nitrification rates increased significantly (by 27–60%, 110–188% and 106–142% respectively across the three species) in rhizosphere soil compared to bulk soil. This led to significantly higher Olsen-P and NH₄⁺-N concentrations in rhizosphere soil, whereas NO₃⁻-N concentration was significantly lower in rhizosphere soil owing to increased microbial immobilization and root uptake. Microbial biomass C and N generally increased while microbial biomass P remained constant in rhizosphere soil relative to bulk soil, indicating the N-limited rather than P-limited microbial growth. Rhizosphere effects on P transformation were most pronounced for Siberian elm, while rhizosphere effects on N transformation were most pronounced for Mongolian pine, implying the different capacities of these species to acquire nutrients.