An experimental slow sand filter (SSF) was constructed to study the spatial and temporal structure of a bacterial community suppressive to an oomycete plant pathogen, Phytophthora cryptogea. Passage of water through the mature sand column resulted in complete removal of zoospores of the plant pathogen. To monitor global changes in the microbial community, bacterial and fungal numbers were estimated on selective media, direct viable counts of fungal spores were made, and the ATP content was measured. PCR amplification of 16S rRNA genes and denaturing gradient gel electrophoresis (DGGE) were used to study the dynamics of the bacterial community in detail. The top layer (1 cm) of the SSF column was dominated by a variable and active microbial population, whereas the middle (50 cm) and bottom (80 cm) layers were dominated by less active and diverse bacterial populations. The major changes in the microbial populations occurred during the first week of filter operation, and these populations then remained to the end of the study. Spatial and temporal nonlinear mapping of the DGGE bands provided a useful visual representation of the similarities between SSF samples. According to the DGGE profile, less than 2% of the dominating bands present in the SSF column were represented in the culturable population. Sequence analysis of DGGE bands from all depths of the SSF column indicated that a range of bacteria were present, with 16S rRNA gene sequences similar to groups such as Bacillus megaterium, Cytophaga, Desulfovibrio, Legionella, Rhodococcus rhodochrous, Sphingomonas, and an uncharacterized environmental clone. This study describes the characterization of the performance, and microbial composition, of SSFs used for the treatment of water for use in the horticultural industry. Utilization of naturally suppressive population of microorganisms either directly or by manipulation of the environment in an SSF may provide a more reproducible control method for the future.Fungal plant diseases are a major problem within the horticultural industry, resulting in reduced yields and occasionally major crop damage. Contaminated irrigation water has long been recognized as an important source of fungal plant pathogens and is an important factor in disease spread on commercial horticultural nurseries (4). Water from many commonly used sources, such as rainwater from glasshouse roofs and, in particular, water stored in open reservoirs and ponds, can often contain large numbers of infective propagules of fungal plant pathogens, such as Pythium and Phytophthora spp. (1,4,34,46,48,53). A particularly high risk of disease spread is associated with the collection, recycling, and reuse of irrigation water, often referred to as recirculation (45,57,58), which is becoming more popular as attempts are increasingly being made to conserve valuable water supplies. Rapid dispersal of the pathogen in water is often achieved by asexual flagellate zoospores, and a key element for pathogen control is the removal of zoospores from water supplies. A wide...
The microbiological characteristics associated with disease-suppressive peats are unclear. We used a bioassay for Pythium sylvaticum-induced damping-off of cress seedlings to identify conducive and suppressive peats. Microbial activity in unconditioned peats was negatively correlated with the counts of P. sylvaticum at the end of the bioassay. Denaturing gradient gel electrophoresis (DGGE) profiling and clone library analyses of small-subunit rRNA gene sequences from two suppressive and two conducive peats differed in the bacterial profiles generated and the diversity of sequence populations. There were also significant differences between bacterial sequence populations from suppressive and conducive peats. The frequencies of a number of microbial groups, including the RhizobiumAgrobacterium group (specifically sequences similar to those for the genera Ochrobactrum and Zoogloea) and the Acidobacteria, increased specifically in the suppressive peats, although no single bacterial group was associated with disease suppression. Fungal DGGE profiles varied little over the course of the bioassay; however, two bands associated specifically with suppressive samples were detected. Sequences from these bands corresponded to Basidiomycete yeast genera. Although the DGGE profiles were similar, fungal sequence diversity also increased during the bioassay. Sequences highly similar to those of Cryptococcus increased in relative abundance during the bioassay, particularly in the suppressive samples. This study highlights the importance of using complementary approaches to molecular profiling of complex populations and provides the first report that basidiomycetous yeasts may be associated with the suppression of Pythium-induced diseases in peats.
SUMMARY A total of 144 isolates of Microdochium nivale from stem bases of winter wheat were taken from 30 sites and 91 isolates from grain were taken from seven sites throughout the UK. Identification by polymerase chain reaction (PCR) amplification of the internal transcribed spacer (ITS) region followed by restriction enzyme digestion of the PCR product revealed that 70% of stem base isolates were M. nivale var. majus and 30% var. nivale while 93% grain isolates were var. majus and 7% var. nivale. Almost all isolates were resistant to the benzimidazole fungicide benomyl. Perithecial production in vitro was more common in var. majus isolates and occurred in almost all grain isolates, but was less common in stem base isolates. The implications of the findings in terms of epidemiology and chemical control of this important cereal pathogen are discussed.
Two immunodiagnostic detection assay procedures were compared with two conventional assays for their sensitivity in detecting propagules of Pythium ultimum var. sporangiiferum , Pythium Group F, Phytophthora cactorum and P. cryptogea in dilution series in sterile distilled water. The most sensitive assay for all four species was the zoospore trapping immunoassay (ZTI). Conventional membrane filtration-dilution plating gave similar results to ZTI with the two Phytophthora spp., but was less sensitive in Pythium detection. Immunodiagnostic dipstick assays and conventional bait tests showed similar sensitivities in the dilution series, and were generally about two orders of magnitude less sensitive than ZTI. The four techniques were also compared for their detection efficacy with water samples collected from horticultural nurseries and in in situ tests of infected root zones of Chamaecyparis , tomato and Chrysanthemum . In these comparisons, ZTI was again the most sensitive test for water samples, although membrane filtration-dilution plating proved to be a more consistent test. Dipstick and baiting assays were the best techniques for in situ testing, and dipsticks provided epidemiologically valuable, quantitative data on pathogen propagule numbers.
The total bacterial community of an experimental slow sand filter (SSF) was analyzed by denaturing gradient gel electrophoresis (DGGE) of partial 16S rRNA gene PCR products. One dominant band had sequence homology to Legionella species, indicating that these bacteria were a large component of the SSF bacterial community. Populations within experimental and commercial SSF units were studied by using Legionella-specific PCR primers, and products were studied by DGGE and quantitative PCR analyses. In the experimental SSF unit, the DGGE profiles for sand column, reservoir, storage tank, and headwater tank samples each contained at least one intense band, indicating that a single Legionella strain was predominant in each sample. Greater numbers of DGGE bands of equal intensity were detected in the outflow water sample. Sequence analysis of these PCR products showed that several Legionella species were present and that the organisms exhibited similarity to strains isolated from environmental and clinical samples. Quantitative PCR analysis of the SSF samples showed that from the headwater sample through the sand column, the number of Legionella cells decreased, resulting in a lower number of cells in the outflow water. In the commercial SSF, legionellae were also detected in the sand column samples. Storing prefilter water or locating SSF units within greenhouses, which are often maintained at temperatures that are higher than the ambient temperature, increases the risk of growth of Legionella and should be avoided. Care should also be taken when used filter sand is handled or replaced, and regular monitoring of outflow water would be useful, especially if the water is used for misting or overhead irrigation.
Abstract. Invasive ants commonly reach abnormally high abundances and have severe impacts on the ecosystems they invade. Current invasion theory recognises that not only negative interactions, such as natural enemy release, but positive interactions, such as facilitation, are important in causing this increased abundance. For invasive ants, facilitation can occur through mutualism with exudate‐producing plants and insects. To obtain such partnerships, however, invaders must first displace native ants, whose communities are highly structured around such resources. By manipulating the abundance of an invasive ant relative to a native, we show that a minimum threshold abundance exists for invasive ants to monopolise exudate‐producing resources. In addition, we show that behavioural dominance is context dependent and varies with spatial location and numerical abundance. Thus, we suggest a ‘facilitation‐threshold’ hypothesis of ant invasion, whereby a minimum abundance of invasive ants is required before facilitation and behavioural dominance can drive abundance rapidly upwards through positive feedback.
Artificial soils made from waste materials offer an alternative to imported natural top-soils, notably in large-scale groundworks and reclamation projects. Benefits include diversion of waste from landfill and recycling. Nonetheless, there is limited information on the characteristics needed to support plant growth in the long-term, particularly the existence of a sustainable nitrogen reservoir. Therefore we assessed the efficacy of nitrogen cycling and retention within an artificial soil composed of 25 % sand, 10 % clay, 32.5 % composted bark and 32.5 % composted green waste over 52 weeks. Leachate was analysed for nitrogen species and nitrogen concentrations and two of the soil columns had fertiliser added after 26 and 48 weeks.Results show that nitrate concentrations decreased from 6.73 to 0.36 mg N L -1 after 2 weeks, due to poor retention of this anion in soil, and remained low for 6 months, before increasing up to 5.87 mg N L -1 after week 26, in unfertilised soils. The sharp increase in dissolved nitrate was preceded by a decrease of the ratio of dissolved organic carbon over dissolved organic nitrogen in the soil leachate. This finding indicates that the soil had become carbonlimited, leading to mineralisation of organic nitrogen by soil organisms and excretion of nitrogen.We also found that fertilisation of the soil with nitrogen-rich substrate did not alleviate carbon-limitation and nitrogen-loss was greater in fertilised soils, indicating nitrogensaturation. After the onset of carbon-limitation, the dissolved nitrate concentrations in both the fertilised and unfertilised soils were close to exceeding the European Union threshold of concern for nitrate groundwater and river pollution. Thus while the deployment of artificial soils is a viable option for landscaping projects, loss of nitrogen may be environmentally significant and soil management protocols must take account of both the carbon and nitrogen status of the substrate.
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