Patterns of gastrointestinal bacterial exchange between chimpanzees and humans involved in research and tourism in western Uganda

Ecological overlap may increase the risks of microbial exchange between humans and wild non-human primates. Escherichia coli bacteria were collected from chimpanzees and humans in Kibale National Park, western Uganda, in May and June 2004, in order to examine whether interaction between humans and apes in the wild might affect gastrointestinal bacterial communities in the two species. Chimpanzees harbored bacteria genetically more similar to those of humans employed in chimpanzee-directed research and tourism than to those of humans from a local village. Most humans (81.6%) and 4.4% of chimpanzees harbored at least one isolate resistant to locally available antibiotics. In isolates from both humans and chimpanzees, resistance was higher to five of these antibiotics than to Ceftiofur, an antibiotic not available in the region. These data indicate that humans and apes interacting in the wild can share genetically and phenotypically similar gastrointestinal bacteria, presumably originating from common environmental sources. Strategies to limit transmission of pathogens between humans and primates, whether that transmission is direct or indirect, would benefit both human health and primate conservation.     

Soil colloids-bound plasmid DNA: Effect on transformation of E. coli and resistance to DNase I degradation

The adsorption and binding of plasmid p34S DNA on four different colloidal fractions from a Brown soil and clay minerals in the presence of various Ca²⁺ concentrations, the ability of bound DNA to transform competent cells of CaCl₂-treated Escherichia coli, and the resistance of bound DNA to degradation by DNase I were studied. DNA adsorption on soil colloids and clay minerals was promoted in the presence of Ca²⁺. Kaolinite exhibited the highest adsorption affinity for DNA among the examined soil colloids and clay minerals. In comparison with organo-mineral complexes (organic clays) and fine clays (<0.2 μm), DNA was tightly adsorbed by H₂O₂-treated clays (inorganic clays) and coarse clays (0.2-2 μm). The transformation efficiency of bound DNA increased with increasing concentrations of Ca²⁺ at which soil colloid or clay mineral-DNA complexes were formed. DNA bound by kaolinite showed the lowest transformation efficiency, and especially no transformants were observed with kaolinite-DNA complex prepared at 5-100 mM Ca²⁺. Compared to organic clays and fine clays, DNA bound on inorganic clays and coarse clays showed a lower capacity to transform E. coli at different Ca²⁺ concentrations. The presence of soil colloids and minerals provided protection to DNA against degradation by DNase I. Montmorillonite, organic clays and fine clays showed stronger protective effects for DNA than inorganic clays and coarse clays. The protection mechanisms as well as the differences in transforming efficiency of plasmid DNA molecules bound on various soil colloidal particles are discussed. The information obtained in this study is of fundamental significance for the understanding of the horizontal dissemination of recombinant DNA and the fate of extracellular DNA in soil environments.     

Fate of gram-negative bacterial biomass in soil—mineralization and contribution to SOM

Soil microorganisms contribute to the formation of non-living soil organic matter (SOM) by metabolic transformation of plant-derived material. After cell death, their biomass components with a specific molecular character become incorporated into SOM imprinting its chemical properties, although this process has not yet been quantified. In order to elucidate the contribution to SOM formation, we investigated the fate of gram-negative bacterial model biomass (Escherichia coli usually introduced into soil with manure or feces) during incubation of soil with isotopically (¹³C) and genetically (lux gene) labeled cells. The decline of living cells was monitored by the loss of bioluminescence. The carbon turnover and mineralization was balanced by bulk soil stable isotope analysis, and the persistence of nucleic acids was investigated by PCR amplification of the lux gene. During incubation, the number of viable E. coli cells decreased rapidly (99.9% within the first 42 d) serving as substrate for other microorganisms or for the formation of SOM, and bioluminescent cells could only be detected during the first 56 d. However, the lux gene was still detected after 224 d, which indicates stabilization of DNA in SOM. Although the survival of E. coli in soil is limited, only about 65% of the added labeled biomass carbon was mineralized to ¹³CO₂ and 51% remained in soil after 224 d with an average ¹³C recovery of 117%. The amount of ¹³C found in the PLFA representative of living cells had decreased to 25% of the initial value, suggesting a proportional decrease of the ¹³C in the soil microbial biomass. The extent of this decrease is higher than the mineralization of the bulk E. coli C and thus the difference of around 25% has to be stabilized as metabolites, or in non-living SOM. The data provide evidence that the genetic information and a considerable part of the carbon from dying bacterial biomass were retained in both the soil microbial food web and in non-living SOM.     

Characterization of brushite as a re-crystallization product formed during bacterial solubilization of hydroxyapatite in batch cultures

Two strains of bacteria (Burkholderia sp., strain FeGL01, and Burkholderia caribiensis, strain FeGL03) were isolated from a Brazilian high phosphorus iron ore. The capacity of both strains to solubilize hydroxyapatite, Ca₅(PO₄)₃(OH), was assessed in plate and batch cultures. In batch cultures, the concentration of solution-P showed two kinetics: an initial one, characterized by a continuously increasing kinetics and a second one, characterized by oscillatory kinetics. To understand the nature of these oscillations, phosphatic residues in the spent broth were collected before, during and after the oscillations, and characterized using scanning electron microscopy (SEM), energy-dispersive X-ray chemical microanalyses (EDX) and X-ray diffraction (XRD). From these studies, it was found that drops in P concentration were related to the formation of an intermediate phosphate in the residues, identified as brushite, CaHPO₄·2H₂O. Later increase of available P in the solution was found to be a consequence of re-dissolution of brushite crystals previously formed. Re-crystallization of brushite was also detected in plate cultures after 12-14 days of incubation     

Impact of heavy metal amended sewage sludge on forest soils as assessed by bacterial and fungal biosensors

Bacterial and fungal bioluminescence-based biosensors were used as indicators of potential heavy metal toxicity to microorganisms in the needle litter of a mature Pinus radiata forest under heavy metal contaminated sewage sludge. Sewage sludge was amended with increasing concentrations of Cu, Ni and Zn and applied to the surface of a mature P. radiata forest. The response of the bacterial and fungal biosensors to soluble Cu, Ni and Zn in needle litter extracts was investigated. The bioluminescence response of the bacterial biosensor Escherichia coli HB101 pUCD607 declined as water-soluble Zn concentrations increased. The effective concentrations that gave a 50% reduction in bioluminescence (EC₅₀ values) for water-soluble Zn and total litter Zn were 1.3 mg l⁻¹ and 3700 mg kg⁻¹, respectively. The bioluminescence response of the fungal biosensor Armillaria mellea declined as soluble Cu concentrations increased. The EC₅₀ values for water-soluble Cu and total litter Cu were 0.12 mg l⁻¹ and 540 mg kg⁻¹, respectively. No decline in bioluminescence was noted for either the bacterial or fungal biosensor on exposure to increasing concentrations of water-soluble Ni. The use of a combination of bacterial and fungal biosensors offers a rapid and sensitive tool for assessing toxicity of heavy metals to microorganisms and, thus, elucidating the environmental impact of contaminants in sewage sludge on litter dwelling microorganisms.     

柚木优良无性系根系养分吸收动力学研究

以4个柚木优良无性系1年生苗木为试材,采用营养液培养和离子消耗曲线模拟方法,测定了根系的Ca2+、Mg2+、K+、NO-3吸收动力学参数。结果表明,以根系总吸收面积和总干重计算的Ca2+、Mg2+、K+、NO3-最大吸收速率相接近。在Ca2+、Mg2+、K+、NO3-最大吸收速率(Vmax)及离子流入速率(α值)指标上,不同基因型之间差异较大,而在养分离子吸收亲合力(Km)指标上,则没有明显差异。缅甸种源无性系VI-23根系对Ca2+、Mg2+、NO3-的Vmax及α值均为最大,而印度种源无性系70-12根系则对K+的Vmax及α值为最大,表明缅甸种源无性系VI-23为Ca、Mg和NO3--N硝态氮高效吸收基因型,印度种源无性系70-12为钾高效吸收基因型。     

Suppression of root-knot disease by Pseudomonas fluorescens CHA0 in tomato: importance of bacterial secondary metabolite, 2,4-diacetylpholoroglucinol

The antimicrobial metabolites 2,4-diacetylphloroglucinol (2,4-DAPG) and pyoluteorin contribute to the ability of Pseudomonas fluorescens strain CHA0 to control plant diseases caused by soil-borne pathogens. P. fluorescens strain CHA0 and its derivatives CHA89 (antibiotics-deficient) and CHA0/pME3424 (antibiotics overproducing) were investigated as potential biocontrol agents against Meloidogyne javanica the root-knot nematode. Exposure of root-knot nematode to culture filtrates of P. fluorescens under in vitro conditions significantly reduced egg hatch and caused substantial mortality of M. javanica juveniles. Nutrient broth yeast extract (NBY) medium amended with 2% (w/v) glucose or 1 mM EDTA markedly repressed hatch inhibition activity of the strain CHA0 but not that of CHA0/pME3424 or CHA89. On the other hand, NBY medium amended with glucose significantly enhanced nematicidal activity of the strain CHA0/pME3424. Neither glucose nor EDTA had an influence on the nematicidal activity of the strains CHA0 and CHA89. Under in vitro conditions, antibiotic overproducing strain CHA0/pME3424 and CHA0 expressed phl‘-’lacZ reporter gene but strain CHA89 did not. Expression of the reporter gene reflects actual production of DAPG. In general, CHA0/pME3424 expressed reporter gene to a greater extent compared to its wild type counterpart CHA0. Regardless of the bacterial strains, reporter gene expression was markedly enhanced when NBY medium was amended with glucose but EDTA had no such effect. A positive correlation between the degree of juvenile mortality and extent of phl‘-’lacZ reporter gene expression was also observed in vitro. Strain CHA0 produced zones of 4-6 mm on MM medium containing gelatin while strain CHA0/pME3424 and CHA89 did not. When MM medium containing gelatin was amended with 2% glucose of 1 mM EDTA size of haloes produced by the strain CHA0 reduced to 2 mm. Under glasshouse conditions aqueous cell suspension of the strains CHA0 or CHA0/pME3424 at various inoculum levels (10⁷, 10⁸ or 10⁹ cfu ml⁻¹) significantly reduced root-knot development. CHA89 caused significant reduction in galling when applied at 10⁹ cfu ml⁻¹. To better understand the mechanism of nematode suppression, split root bioassay was performed. Split-root experiments, that guarantee a spatial separation of inducing agent and a challenging pathogen, showed that soil treatment of one half of the root system with cell suspension of CHA0 or CHA0/pME3424 resulted in a significant systemic induced resistance leading to reduction of M. javanica infection of tomato roots in the non-baterized nematode treated half. The results clearly suggest that the antibiotic 2,4-DAPG from P. fluorescens CHA0 act as the inducing agents of systemic resistance in tomato roots. Populations of CHA0 and its derivatives declined progressively by 10-fold between first and fourth harvests (0-21 days after inoculation). However, bacterial populations increased at final harvest (28 days after application).     

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

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

Short-term effects of single or combined application of mineral N fertilizer and cattle slurry on the fluxes of radiatively active trace gases from grassland soil

After implementation of legislative measures for the reduction of environmental hazards from nitrate leaching and ammonia volatilisation when using organic manures and fertilizers in Europe, much attention is now paid to the specific effects of these fertilizers on the dynamics of global warming-relevant trace gases in soil. Particularly nitrogen fertilizers and slurry from animal husbandry are known to play a key role for the CH₄ and N₂O fluxes from soils. Here we report on a short-term evaluation of trace gas fluxes in grassland as affected by single or combined application of mineral fertilizer and organic manure in early spring. Methane fluxes were characterised by a short methane emission event immediately after application of cattle slurry. Within the same day methane fluxes returned to negative, and on average over the 4-day period after slurry application, only a small but insignificant trend to reduced methane oxidation was found. Nitrous oxide emissions showed a pronounced effect of combined slurry and mineral fertilizer application. In particular fresh cattle slurry combined with calcium ammonium nitrate (CAN) mineral fertilizer induced an increase in mean N₂O flux during the first 4 days after application from 10 to 300 μg N₂O-N m⁻² h⁻¹. ¹⁵N analysis of emitted N₂O from ¹⁵N-labelled fertilizer or manure indicated that easily decomposable slurry C compounds induced a pronounced promotion of N₂O-N emission derived from mineral CAN fertilizer. Fluxes after application of either mineral fertilizer or slurry alone showed an increase of less than 5-fold. The NO_x sink strength of the soil was in the range of −6 to −10 μg NO_x-N m⁻² h⁻¹ and after fertilization it showed a tendency to be reduced by no more than 2 μg NO_x-N m⁻² h⁻¹, which was a result of both, increased NO emission and slightly increased NO₂ deposition. Associated determination of the N₂O:N₂ emission ratio revealed that after mineral N application (CAN) a large proportion (c. 50%) was emitted as N₂O, while after application of slurry with easily decomposable C and predominantly -N serving as N-source, the N₂O:N₂ emission ratio was 1:14, i.e. was changed in favour of N₂. Our work provides evidence that particularly the combination of slurry and nitrate-containing N fertilizers gives rise to considerable N₂O emissions from mineral fertilizer N pool.     

Rhizosphere priming of soil organic matter by bacterial groups in a grassland soil

Plants often impact the rate of native soil organic matter turnover through root interactions with soil organisms; however the role of root-microbial interactions in mediation of the “priming effect” is not well understood. We examined the effects of living plant roots and N fertilization on belowground C dynamics in a California annual grassland soil (Haploxeralf) during a two-year greenhouse study. The fate of ¹³C-labeled belowground C (roots and organic matter) was followed under planted (Avena barbata) and unplanted conditions, and with and without supplemental N (20 kg N ha⁻¹ season⁻¹) over two periods of plant growth, each followed by a dry, fallow period of 120 d. Turnover of belowground ¹³C SOM was followed using ¹³C-phospholipid fatty acid (PLFA) biomarkers. Living roots increased the turnover and loss of belowground ¹³C compared with unplanted soils. Planted soils had 20% less belowground ¹³C present than in unplanted soils after 2 cycles of planting and fallow. After 2 treatment cycles, unlabeled soil C was 4.8% higher in planted soils than unplanted. The addition of N to soils decreased the turnover of enriched belowground ¹³C during the first treatment season in both planted and unplanted soils, however no effect of N was observed thereafter. Our findings suggest that A. barbata may increase soil C levels over time because root and exudate C inputs are significant, but that increase will be moderated by an overall faster C mineralization rate of belowground C. N addition may slow soil C losses; however, the effect was minor and transient in this system. The labeled root-derived ¹³C was initially recovered in gram negative (highest enrichment), gram positive, and fungal biomarkers. With successive growing seasons, the labeled C in the gram negative and fungal markers declined, while gram positive markers continued to accumulate labeled belowground C. The rhizosphere of A. barbata shifted the microbial community composition, resulting in greater abundances of gram negative markers and lower abundances of gram positive, actinobacteria and cyclopropyl PLFA markers compared to unplanted soil. However, the longer-term utilization of labeled belowground C by gram positive bacteria was enhanced in the rhizosphere microbial community compared with unplanted soils. We suggest that the activities of gram positive bacteria may be major controllers of multi-year rhizosphere-related priming of SOM decomposition.     

Biological control of damping-off caused by Rhizoctonia solani using chitinase-producing Paenibacillus illinoisensis KJA-424

A bacterium having strong chitinolytic activity was isolated from a coastal soil in Korea and identified as Paenibacillus illinoisensis KJA-424 on the basis of the nucleotide sequence of a 16S rRNA gene. By activity staining after SDS-PAGE, three major chitinase bands with chitinolytic activity, approximate molecular weight of 63, 54 and 38 kDa were detected. On co-culture Rhizoctonia solani with KJA-424, abnormal swelling and deformation of R. solani hyphae were observed, where the release of N-acetyl-d-glucosamine was detected. The bacterium suppressed the symptom of damping-off cucumber seedlings caused by R. solani, in greenhouse trial.     

Rhizosphere bacterial community composition in natural stands of Carex arenaria (sand sedge) is determined by bulk soil community composition

The relative importance of specific plant properties versus soil characteristics in shaping the bacterial community structure of the rhizosphere is a topic of considerable debate. Here, we report the results of a study on the bacterial composition of the rhizosphere of the wild plant Carex arenaria (sand sedge) growing at 10 natural sites in The Netherlands. The soil properties of the sandy soils at these sites were highly disparate, most notably in pH, chloride and organic matter content. Rhizosphere and bulk soil bacterial communities were examined by culture-independent means, namely, 16S rDNA-directed PCR-DGGE profiling. Large differences were observed between the bacterial communities of the different sites for both bulk and rhizosphere soil. Cluster analysis of bacterial profiles revealed that the rhizosphere community of each site was generally more closely related to the bulk soil community of that site rather than to rhizosphere communities of other sites. Hence, bacterial community structure within the rhizosphere of C. arenaria appeared to be determined to a large extent by the bulk soil community composition. This conclusion was supported by a reciprocal planting experiment, where C. arenaria shoots of different sites yielded highly similar rhizosphere communities when planted in the same soil.     

Phosphate solubilization by a wild type strain and UV-induced mutants of Aspergillus tubingensis

Phenotypic mutants of Aspergillus tubingensis were obtained by UV irradiation and phosphate solubilization ability of these mutants were studied and compared with the wild type strain. Low phosphate solubilizing mutant was also selected in this study. Among the different mutants, AtM-5 and AtM-2 showed highest P solubilization when tri-calcium phosphate and rock phosphate were used as P source compared to the wild type strain and other mutants. These mutants also showed maximum acid phosphatase and phytase activity. These results suggest that P solubilization by these isolates is due to lowering of pH of the culture filtrate and also the activity of acid phosphatase and phytase.     

The bacterial community of tomato rhizosphere is modified by inoculation with arbuscular mycorrhizal fungi but unaffected by soil enrichment with mycorrhizal root exudates or inoculation with Phytophthora nicotianae

Arbuscular mycorrhizal (AM) fungi have been shown to induce the biocontrol of soilborne diseases, to change the composition of root exudates and to modify the bacterial community structure of the rhizosphere, leading to the formation of the mycorrhizosphere. Tomato plants were grown in a compartmentalized soil system and were either submitted to direct mycorrhizal colonization or to enrichment of the soil with exudates collected from mycorrhizal tomato plants, with the corresponding negative controls. Three weeks after planting, the plants were inoculated or not with the soilborne pathogen Phytophthora nicotianae growing through a membrane from an adjacent infected compartment. At harvest, a PCR-Denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments amplified from the total DNA extracted from each plant rhizosphere was performed. Root colonization with the AM fungi Glomus intraradices or Glomus mosseae induced significant changes in the bacterial community structure of tomato rhizosphere, compared to non-mycorrhizal plants, while enrichment with root exudates collected from mycorrhizal or non-mycorrhizal plants had no effect. Our results support that the effect of AM fungi on rhizosphere bacteria would not be mediated by compounds present in root exudates of mycorrhizal plants but rather by physical or chemical factors associated with the mycelium, volatiles and/or root surface bound substrates. Moreover, infection of mycorrhizal or non-mycorrhizal plants with P. nicotianae did not significantly affect the bacterial community structure suggesting that rhizosphere bacteria would be less sensitive to the pathogen invasion than to mycorrhizal colonization. Of 96 unique sequences detected in the tomato rhizosphere, eight were specific to mycorrhizal fungi, including two Pseudomonas, a Bacillus simplex, an Herbaspirilium and an Acidobacterium. One Verrucomicrobium was common to rhizospheres of mycorrhizal plants and of plants watered with mycorrhizal root exudates.     

From the micro-scale to the habitat: Assessment of soil bacterial community structure as shown by soil structure directed sampling

Natural structural units of a luvisol under maize crop were studied to assess if soil structure directed sampling could improve the understanding of arrangements of bacteria in spatially constraint location. Three habitats were defined: (i) soil around fine lateral roots (rhizo-aggregates), (ii) soil close to basal roots (core clods) and (iii) unplanted soil between rows (bare soil clods). These habitats were also investigated with maize plants resulting from Azospirillum lipoferum CRT1 inoculated seeds as a model of enhanced fine root system. Rhizo-aggregates were clearly separated from each other (disconnected habitat) in contrast to micro-samples (fragments) from clods, which belong to cohesive macro-structures. Genetic fingerprints on metagenomic extracts were used to characterize the structure of bacterial communities on 95 micro-samples from the three habitats. For eubacteria, automated RISA (Ribosomal Intergenic Spacer Analysis) of ITS (Internal Transcribed Spacer) profiles were performed. PCR-RFLP on nifH gene were used to describe the N-fixer guilds. Exploratory multivariate analyses (PCA and MDS) revealed bacterial community patterns in the sampled habitats. On the basis of ITS profiles, rhizo-aggregates harboured closely related communities, distant from those of the unplanted soil, and each sampled rhizo-aggregate could therefore be considered as a sub-unit of the whole macro-habitat, comprising all the fine roots. The observed low dissimilarity of disconnected rhizo-aggregates is likely to result from the direct influence of maize root tips on the recruitment of rhizosphere bacteria. Molecular fingerprints of nifH from basal root clods (core) were more similar to bare soil than to rhizo-aggregates, indicating similar ecological conditions without, or with, at least, poor maize exudating root influence. Although our study was performed on a limited number of situations, the distribution of bacteria was revealed to be patterned by soil structure units, which is a first step to improve the modelling of microbial ecology in soils.     

Nurse shrubs increased the early growth of Cupressus seedlings by enhancing belowground mutualism and soil microbial activity

The influence of shrubs used as nurse plants was tested on the growth of Cupressus atlantica, on microbial activity and on arbuscular mycorrhizal (AM) soil potential in a Mediterranean environment. An experimental plantation was conducted combining uninoculated, arbuscular mycorrhized Cypress seedlings and an association between Lavandula stoechas planted close to newly planted C. atlantica seedlings. After three years plantation, this association between C. atlantica and L. stoechas lead to a higher growth of C. atlantica and better soil microbial characteristics compared to the control treatment. AM mycelium network, total microbial activity, dehydrogenase activity, phosphate-solubilizing fluorescent pseudomonads and N, P nutrient uptake by C. atlantica, were significantly higher in the presence of L. stoechas than those recorded in the other treatments. This pioneer shrubs facilitates the early establishment of Cypress seedlings by improving soil microbial characteristics and AM fungus community development. Given that the facilitative effect of one plant species to another increases with abiotic stress, the benefits of this technique would be useful in reforestation programs undertaken to rehabilitate degraded areas in Mediterranean region.     

Control of apothecia of Sclerotinia sclerotiorum by soil amendment with S-H mixture or Perlka in bean, canola and wheat fields

Ascospores of Sclerotinia sclerotiorum produced from apothecia are the primary source of inoculum for causing diseases such as white mold of common bean, pod rot of pea, stem blight of canola and head rot of sunflower and safflower in the Canadian prairies. A field study was conducted for 4 years to determine efficacy of control of production of apothecia from carpogenically germinated sclerotia of S. sclerotiorum by soil amendment with Perlka^® (calcium cyanamide) and S-H mixture (a formulated compound). Results of the 4-year experiments showed that amendment of soil with Perlka^® at low (30 g/m²) or high (60 g/m²) rate was effective in reducing carpogenic germination of sclerotia and production of apothecia under the canopy of host crops (common bean and canola) and a non-host crop (wheat). In the experiments of 1988, for example, the numbers of apothecia produced in the treatments of Perlka^®-low rate (30 g/m²), Perlka^®-high rate (60 g/m²) and untreated control were 42, 46, and 182 apothecia/plot (m²), respectively, for bean; 89, 42, and 318 apothecia/plot (m²), respectively, for canola; and 146, 143, and 412 apothecia/plot (m²), respectively, for wheat. However, soil amendment of S-H mixture at low (30 g/m²) or high (60 g/m²) rate was ineffective in reducing carpogenic germination of sclerotia and production of apothecia for all the 4 years of testing in all three crops. The ineffectiveness of S-H mixture and the practicality of Perlka^® for control of Sclerotinia diseases of crops grown under Canadian prairie conditions are discussed.