Microbial introductions to apple leaves: Influences of altered immigration on fungal community dynamics

Fungal immigration to apple leaves in the field was altered by the introduction of populations ofChaetomium globosum orAureobasidium pullulans to surface-disinfested leaves either immediately following, or 6 days after, disinfestation. Total numbers of fungal individuals and numbers of filamentous fungal and yeast individuals were estimated and compared over time for 4–7 weeks on control leaves (leaves disinfested but no populations applied), onAureobasidium-treated, and onChaetomium-treated leaves. Fungal communities developing on leaves during three experiments in two different time frames (experiment 1: July 9–August 27; experiments 2 and 3: July 29–August 27), and thus under different immigration regimes, were also compared. Survival of introduced populations was not related to the presence of prior fungal immigrants. Rates of increase in total numbers of fungi and numbers of filamentous fungi and yeasts per leaf varied among experiments, apparently in relation to differences in immigration and environmental history. Differences among leaves in immigration had a short-term (days) influence on community size. However, no long-term effects of altered immigration on phylloplane fungal community size were evident.     

Utility of Microcosm Studies for Predicting Phylloplane Bacterium Population Sizes in the Field

Population sizes of two ice nucleation-active strains of Pseudomonas syringae were compared on leaves in controlled environments and in the field to determine the ability of microcosm studies to predict plant habitat preferences in the field. The P. syringae strains investigated were the parental strains of recombinant deletion mutant strains deficient in ice nucleation activity that had been field tested for their ability to control plant frost injury. The population size of the P. syringae strains was measured after inoculation at three field locations on up to 40 of the same plant species that were studied in the growth chamber. There was seldom a significant relationship between the mean population size of a given P. syringae strain incubated under either wet or dry conditions in microcosms and the mean population size which could be recovered from the same species when inoculated in the field. Specifically, on some plant species, the population size recovered from leaves in the field was substantially greater than from that species in a controlled environment, while for other plant species field populations were significantly smaller than those observed under controlled conditions. Population sizes of inoculated P. syringae strains, however, were frequently highly positively correlated with the indigenous bacterial population size on the same plant species in the field, suggesting that the ability of a particular plant species to support introduced bacterial strains is correlated with its ability to support large bacterial populations or that indigenous bacteria enhance the survival of introduced strains. Microcosm studies therefore seem most effective at assessing possible differences between parental and recombinant strains under a given environmental regime but are limited in their ability to predict the specific population sizes or plant habitat preferences of bacteria on leaves under field conditions.     

Effect of sampling scale on the assessment of epiphytic bacterial populations

Bacterial populations on above-ground plant surfaces were estimated at three different biological scales, including leaflet disks, entire leaflets, and whole plants. The influence of sample scale on the estimation of mean bacterial population size per unit and per gram and on the variability among sampling units was quantified at each scale. Populations were highly variable among sampling units at every scale examined, suggesting that there is no optimal scale at which sample variance is reduced. The distribution of population sizes among sample units was sometimes, but not consistently, described by the lognormal. Regardless of the sampling scale, expression of population sizes on a per gram basis may not reduce variance, because population size was not generally a function of sample unit weight within any single sampling scale. In addition, the data show that scaling populations on a per gram basis does not provide a useful means of comparing population estimates from samples taken at different scales. The implications of these results for designing sampling strategies to address specific issues in microbial ecology are discussed. Correspondence to: L.L. Kinkel     

Nutrient use preferences among soil Streptomyces suggest greater resource competition in monoculture than polyculture plant communities

Background an aims

Nutrient use overlap among sympatric Streptomyces populations is correlated with pathogen inhibitory capacity, yet there is little information on either the factors that influence nutrient use overlap among coexisting populations or the diversity of nutrient use among soil Streptomyces.

Methods

We examined the effects of plant host and plant species richness on nutrient use of Streptomyces isolated from the rhizosphere of Andropogon gerardii (Ag) and Lespedeza capitata (Lc) growing in communities of 1 (monoculture) or 16 (polyculture) plant species. Growth on 95 carbon sources was assessed over 5d.

Results

Cumulative growth was significantly greater for polyculture vs. monoculture isolates, and for Lc vs. Ag isolates. Isolates from monocultures, but not polycultures, exhibited a drop in growth rates between 24 h and 72 h post-inoculation, suggesting resource allocation to non-growth functions. Isolates from high-carbon (polyculture) or high-nitrogen (Lc) soils had larger niche widths than isolates from low-C (monocultures) or low-N (Ag) soils. Sympatric isolates from polycultures were significantly more differentiated from one another in preferred nutrients for growth than sympatric isolates from monocultures.

Conclusions

These results suggest that Streptomyces populations respond to selection imposed by plant host and plant community richness and that populations from polyculture but not from monoculture, mediate resource competition via niche differentiation.

     

Stability of grassland production is robust to changes in the consumer food web

Theory predicts that consumers may stabilise or destabilise plant production depending on model assumptions, and tests in aquatic food webs suggest that trophic interactions are stabilising. We quantified the effects of trophic interactions on temporal variability (standard deviation) and temporal stability (mean/standard deviation) of grassland biomass production and the plant diversity–stability relationship by experimentally removing heterotrophs (large vertebrates, arthropods, foliar and soil fungi) from naturally and experimentally assembled grasslands of varying diversity. In both grassland types, trophic interactions proportionately decreased plant community biomass mean and variability over the course of 6 years, leading to no net change in temporal stability or the plant diversity–stability relationship. Heterotrophs also mediated plant coexistence; their removal reduced diversity in naturally assembled grasslands. Thus, herbivores and fungi reduce biomass production, concurrently reducing the temporal variability of energy and material fluxes. Because of this coupling, grassland stability is robust to large food web perturbations.     

Oil red-O stains non-adipogenic cells: a precautionary note

Bovine adipofibroblasts, 3T3-L1 cells, L-6 myogenic cells, and sheep satellite cells were allowed to proliferate for 48 h. Oil red-O (ORO) was dissolved in three different solvents isopropanol, propylene glycol and triethyl phosphate. At 48 h, the proliferative cultures were stained with the three stains. ORO stain prepared in both propylene glycol and triethyl phosphate resulted in bright red droplets appearing in all cultures, whereas ORO dissolved in isopropanol was not taken up by any of the cells. These data suggest that certain preparations of ORO may stain cells in non-adipogenic lineages as well as undifferentiated pre-adipocytes. Caution must be exercised when choosing solvents for ORO in differentiation studies using cells of the fat/adipose lineage.     

Spatial variation in frequency and intensity of antibiotic interactions among Streptomycetes from prairie soil

Antibiotic interactions are believed to be significant to microbial fitness in soil, yet little is known of the frequency, intensity, and diversity of antibiotic inhibition and resistance among indigenous microbes. To begin to address these issues, we studied the abilities of streptomycete isolates from prairie soil to inhibit growth and display resistance to antibiotics produced by a test collection of 10 streptomycete isolates. Wide variations in antibiotic inhibition and resistance for prairie isolates among three locations and four soil depths within a 1-m2 plot were revealed. Fewer than 10% of 153 prairie isolates inhibited all 10 test isolates, while more than 40% of the isolates did not inhibit any of the test isolates. No field isolate was resistant to all of the test isolates, nor was any isolate susceptible to all of the test isolates. No correlation between inhibition and resistance phenotypes was found, suggesting that inhibition and resistance are under independent selection. The significant spatial variation in the frequency and intensity of antibiotic inhibition implies that the fitness benefits of antibiotic production are not the same among locations in soil. In contrast, the consistency of resistance over space indicates that its significance to fitness across locations is stable or the costs of maintaining resistance in the absence of selection are small or nonexistent. The spatial clustering of antibiotic inhibitory activity suggests a variable matrix of selection pressures and microbial responses across the soil landscape.     

Characterization of Kir1.1 channels with the use of a radiolabeled derivative of tertiapin

Inward rectifier potassium channels (Kir) play critical roles in cell physiology. Despite representing the simplest tetrameric potassium channel structures, the pharmacology of this channel family remains largely undeveloped. In this respect, tertiapin (TPN), a 21 amino acid peptide isolated from bee venom, has been reported to inhibit Kir1.1 and Kir3.1/3.4 channels with high affinity by binding to the M1-M2 linker region of these channels. The features of the peptide-channel interaction have been explored electrophysiologically, and these studies have identified ways by which to alter the composition of the peptide without affecting its biological activity. In the present study, the TPN derivative, TPN-Y1/K12/Q13, has been synthesized and radiolabeled to high specific activity with (125)I. TPN-Y1/K12/Q13 and mono-iodo-TPN-Y1/K12/Q13 ([(127)I]TPN-Y1/K12/Q13) inhibit with high affinity rat but not human Kir1.1 channels stably expressed in HEK293 cells. [(125)I]TPN-Y1/K12/Q13 binds in a saturable, time-dependent, and reversible manner to HEK293 cells expressing rat Kir1.1, as well as to membranes derived from these cells, and the pharmacology of the binding reaction is consistent with peptide binding to Kir1.1 channels. Studies using chimeric channels indicate that the differences in TPN sensitivity between rat and human Kir1.1 channels are due to the presence of two nonconserved residues within the M1-M2 linker region. When these results are taken together, they demonstrate that [(125)I]TPN-Y1/K12/Q13 represents the first high specific activity radioligand for studying rat Kir1.1 channels and suggest its utility for identifying other Kir channel modulators.