scholarly journals Bioactive exometabolites drive maintenance competition in simple bacterial communities

2021 ◽  
Author(s):  
John L. Chodkowski ◽  
Ashley Shade
Keyword(s):  
Stationary Phase ◽  
Pseudomonas Syringae ◽  
Interference Competition ◽  
Resource Limitation ◽  
Local Environment ◽  
Gene Clusters ◽  
Alternative Measure ◽  
Bacterial Cells ◽  
Bacterial Strains ◽  
Traditional Concept

AbstractDuring prolonged resource limitation, bacterial cells can persist in metabolically active states of non-growth. These maintenance periods, such as those experienced by cells in stationary phase cultures, can, perhaps counterintuitively, include upregulation of cellular secondary metabolism and release of exometabolites into the local environment, at the cost of an energetic commitment to growth. As resource limitation is a characteristic feature of many habitats that harbor environmental microbial communities, we hypothesized that neighboring bacterial populations employ exometabolites to compete or cooperate during maintenance, and that these exometabolite-facilitated interactions can drive community outcomes. Here, we evaluated the consequences of exometabolite interactions over stationary phase among three well-known environmental bacterial strains: Burkholderia thailandensis E264 (ATCC 700388), Chromobacterium violaceum ATCC 31532, and Pseudomonas syringae pv.tomato DC3000 (ATCC BAA-871). We assembled these stains into laboratory-scale synthetic communities that only permitted chemical interactions among them. We compared the responses (transcripts) and behaviors (exometabolites) of each member with and without neighbors. We found that transcriptional dynamics were altered in the presence of different neighbors, and that these changes could be attributed to the production of or response to bioactive exometabolites employed for competition during maintenance. B. thailandensis was especially influential and competitive within its communities, as it consistently upregulated additional biosynthetic gene clusters involved in the production of bioactive exometabolites for both exploitative and interference competition. Additionally, some of these bioactive exometabolites were upregulated and produced in a non-additive manner in the 3-member community. These results demonstrate that the active investment in competition during maintenance can contribute to both bacterial population fitness and community-level outcomes. It also suggests that the traditional concept of defining competitiveness by growth outcomes may be too narrow, and that maintenance competition could be an alternative measure.

scholarly journals Competition sensing alters antibiotic production in Streptomyces

2020 ◽  
Author(s):  
Sanne Westhoff ◽  
Alexander Kloosterman ◽  
Stephan F. A. van Hoesel ◽  
Gilles P. van Wezel ◽  
Daniel E. Rozen
Keyword(s):  
Genetic Relatedness ◽  
Cell Damage ◽  
Antibiotic Production ◽  
Interference Competition ◽  
Resource Limitation ◽  
Gene Clusters ◽  
Regulatory Control ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters ◽  
The Social

ABSTRACTOne of the most important ways that bacteria compete for resources and space is by producing antibiotics that inhibit competitors. Because antibiotic production is costly, the biosynthetic gene clusters coordinating their synthesis are under strict regulatory control and often require “elicitors” to induce expression, including cues from competing strains. Although these cues are common, they are not produced by all competitors and so the phenotypes causing induction remain unknown. By studying interactions between 24 antibiotic-producing streptomycetes we show that inhibition between competitors is common and occurs more frequently if strains are closely related. Next, we show that antibiotic production is more likely to be induced by cues from strains that are closely related or that share secondary metabolite biosynthetic gene clusters (BGCs). Unexpectedly, antibiotic production is less likely to be induced by competitors that inhibit the growth of a focal strain, indicating that cell damage is not a general cue for induction. In addition to induction, antibiotic production often decreased in the presence of a competitor, although this response was not associated with genetic relatedness or overlap in BGCs. Finally, we show that resource limitation increases the chance that antibiotic production declines during competion. Our results reveal the importance of social cues and resource availability in the dynamics of interference competition in streptomycetes.SIGNIFICANCE STATEMENTBacteria secrete antibiotics to inhibit their competitors, but the presence of competitors can determine whether these toxins are produced. Here, we study the role of the competitive and resource environment on antibiotic production in Streptomyces, bacteria renowned for their production of antibiotics. We show that Streptomyces are more likely to produce antibiotics when grown with closely related competitors or that share biosynthetic pathways for secondary metabolites, but not when they are threatened by competitor’s toxins, in contrast to predictions of the competition sensing hypothesis. Streptomyces also often reduce their output of antibiotics when grown with competitors, especially under nutrient limitation. Our findings highlight that interactions between the social and resource environments strongly regulate antibiotic production in these medicinally important bacteria.

scholarly journals Competition Sensing Changes Antibiotic Production in Streptomyces

mBio ◽  
2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Sanne Westhoff ◽  
Alexander M. Kloosterman ◽  
Stephan F. A. van Hoesel ◽  
Gilles P. van Wezel ◽  
Daniel E. Rozen
Keyword(s):  
Genetic Relatedness ◽  
Cell Damage ◽  
Antibiotic Production ◽  
Interference Competition ◽  
Resource Limitation ◽  
Gene Clusters ◽  
Regulatory Control ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters ◽  
The Social

ABSTRACT One of the most important ways that bacteria compete for resources and space is by producing antibiotics that inhibit competitors. Because antibiotic production is costly, the biosynthetic gene clusters coordinating their synthesis are under strict regulatory control and often require “elicitors” to induce expression, including cues from competing strains. Although these cues are common, they are not produced by all competitors, and so the phenotypes causing induction remain unknown. By studying interactions between 24 antibiotic-producing strains of streptomycetes, we show that strains commonly inhibit each other’s growth and that this occurs more frequently if strains are closely related. Next, we show that antibiotic production is more likely to be induced by cues from strains that are closely related or that share secondary metabolite biosynthetic gene clusters (BGCs). Unexpectedly, antibiotic production is less likely to be induced by competitors that inhibit the growth of a focal strain, indicating that cell damage is not a general cue for induction. In addition to induction, antibiotic production often decreases in the presence of a competitor, although this response was not associated with genetic relatedness or overlap in BGCs. Finally, we show that resource limitation increases the chance that antibiotic production declines during competition. Our results reveal the importance of social cues and resource availability in the dynamics of interference competition in streptomycetes. IMPORTANCE Bacteria secrete antibiotics to inhibit their competitors, but the presence of competitors can determine whether these toxins are produced. Here, we study the role of the competitive and resource environment on antibiotic production in Streptomyces, bacteria renowned for their production of antibiotics. We show that Streptomyces cells are more likely to produce antibiotics when grown with competitors that are closely related or that share biosynthetic pathways for secondary metabolites, but not when they are threatened by competitor’s toxins, in contrast to predictions of the competition sensing hypothesis. Streptomyces cells also often reduce their output of antibiotics when grown with competitors, especially under nutrient limitation. Our findings highlight that interactions between the social and resource environments strongly regulate antibiotic production in these medicinally important bacteria.

scholarly journals Exometabolite Dynamics over Stationary Phase Reveal Strain-Specific Responses

mSystems ◽  
2020 ◽  
Vol 5 (6) ◽  
pp. e00493-20
Author(s):  
John L. Chodkowski ◽  
Ashley Shade
Keyword(s):  
Stationary Phase ◽  
Pseudomonas Syringae ◽  
Stationary Phases ◽  
Interspecies Interactions ◽  
Bacterial Strains ◽  
Community Interactions ◽  
Succinate Production ◽  
Intact Cells ◽  
Content Type ◽  
Environmental Bacteria

ABSTRACTMicrobial exponential growth is expected to occur infrequently in environments that have long periods of nutrient starvation punctuated by short periods of high nutrient flux. These conditions likely impose nongrowth states for microbes. However, nongrowth states are uncharacterized for the majority of environmental bacteria, especially in regard to exometabolite production. We compared exometabolites produced over stationary phase across three environmental bacteria: Burkholderia thailandensis E264 (ATCC 700388), Chromobacterium violaceum ATCC 31532, and Pseudomonas syringae pv. tomato DC3000 (ATCC BAA-871). We grew each strain in monoculture and investigated exometabolite dynamics from mid-exponential to stationary phases. We focused on exometabolites that were released into the medium and accumulated over 45 h, including approximately 20 h of stationary phase. We also analyzed transcripts (transcriptome sequencing [RNA-seq]) to interpret exometabolite output. We found that the majority of exometabolites released were strain specific, with a subset of identified exometabolites involved in both central and secondary metabolism. Transcript analysis supported that exometabolites were released from intact cells, as various transporters had either increased or consistent transcripts through time. Interestingly, we found that succinate was one of the most abundant identifiable exometabolites for all strains and that each strain rerouted their metabolic pathways involved in succinate production during stationary phase. These results show that nongrowth states can be metabolically dynamic and that environmental bacteria can enrich a minimal environment with diverse chemical compounds as a consequence of growth and postgrowth maintenance in stationary phase. This work provides insights into microbial community interactions via exometabolites under conditions of growth cessation or limitation.IMPORTANCE Nongrowth states are common for bacteria that live in environments that are densely populated and predominantly nutrient exhausted, and yet these states remain largely uncharacterized in cellular metabolism and metabolite output. Here, we investigated and compared stationary-phase exometabolites and RNA transcripts for each of three environmental bacterial strains. We observed that diverse exometabolites were produced and provide evidence that these exometabolites accumulate over time through release by intact cells. Additionally, each bacterial strain had a characteristic exometabolite profile and exhibited dynamics in exometabolite composition. This work affirms that stationary phase is metabolically dynamic, with each strain tested creating a unique chemical signature in the extracellular space and altering metabolism in stationary phase. These findings set the stage for understanding how bacterial populations can support surrounding neighbors in environments with prolonged nutrient exhaustion through exometabolite-mediated interspecies interactions.

scholarly journals Antimicrobial Activity Screening of Rhizosphere Soil Bacteria from Tomato and Genome-Based Analysis of their Antimicrobial Biosynthetic Potential

2020 ◽  
Author(s):  
Lu Zhou ◽  
Chunxu Song ◽  
Zhibo Li ◽  
Oscar P. Kuipers
Keyword(s):  
Antimicrobial Activity ◽  
Plant Growth ◽  
Disease Control ◽  
Pseudomonas Syringae ◽  
Rhizosphere Soil ◽  
Gene Clusters ◽  
Bacterial Strains ◽  
Vast Number ◽  
Tomato Disease

Abstract Background: Tomato plant growth is frequently hampered by a high susceptibility to pests and diseases. Traditional chemical control causes a serious impact on both the environment and human health. Therefore, seeking environment-friendly and cost-effective green methods in agricultural production becomes crucial nowadays. Plant Growth Promoting Rhizobacteria (PGPR) can promote plant growth through biological activity. Their use is considered to be a promising sustainable approach for crop growth. Moreover, a vast number of biosynthetic gene clusters (BGCs) for secondary metabolite production are being revealed in PGPR, which helps to find potential anti-microbial activities for tomato disease control.Results: We isolated 351 bacterial strains (181 of which are Bacillus sp.) from healthy tomato, rhizosphere soil, and tomato tissues. In vitro antagonistic assays revealed that 34 Bacillus strains have antimicrobial activity against Erwinia carotovora, Pseudomonas syringae; Rhizoctonia solani; Botrytis cinerea; Verticillium dahliae and Phytophthora infestans. The genomes of 10 Bacillus and Paenibacillus strains with good antagonistic activity were sequenced. Via genome mining approaches, we identified 120 BGCs encoding NRPs, PKs-NRPs, PKs, terpenes and bacteriocins, including known compounds such as fengycin, surfactin, bacillibactin, subtilin, etc. In addition, several novel BGCs were identified. We discovered that the NRPs and PKs-NRPs BGCs in Bacillus species are encoding highly conserved known compounds as well as various novel variants.Conclusions: This study highlights the great number of varieties of BGCs in Bacillus strains. These findings pave the road for future usage of Bacillus strains as biocontrol agents for tomato disease control and are a resource arsenal for novel antimicrobial discovery.

scholarly journals Plasma-Activated Water (PAW) as a Disinfection Technology for Bacterial Inactivation with a Focus on Fruit and Vegetables

Foods ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 166
Author(s):  
Aswathi Soni ◽  
Jonghyun Choi ◽  
Gale Brightwell
Keyword(s):  
Fruit And Vegetables ◽  
Atmospheric Plasma ◽  
Model Systems ◽  
Thermal Treatments ◽  
Bacterial Cells ◽  
Bacterial Inactivation ◽  
Gas Temperature ◽  
Bacterial Strains ◽  
Sensory Qualities ◽  
Plasma Activated Water

Plasma-activated water (PAW) is generated by treating water with cold atmospheric plasma (CAP) using controllable parameters, such as plasma-forming voltage, carrier gas, temperature, pulses, or frequency as required. PAW is reported to have lower pH, higher conductivity, and higher oxygen reduction potential when compared with untreated water due to the presence of reactive species. PAW has received significant attention from researchers over the last decade due to its non-thermal and non-toxic mode of action especially for bacterial inactivation. The objective of the current review is to develop a summary of the effect of PAW on bacterial strains in foods as well as model systems such as buffers, with a specific focus on fruit and vegetables. The review elaborated the properties of PAW, the effect of various treatment parameters on its efficiency in bacterial inactivation along with its usage as a standalone technology as well as a hurdle approach with mild thermal treatments. A section highlighting different models that can be employed to generate PAW alongside a direct comparison of the PAW characteristics on the inactivation potential and the existing research gaps are also included. The mechanism of action of PAW on the bacterial cells and any reported effects on the sensory qualities and shelf life of food has been evaluated. Based on the literature, it can be concluded that PAW offers a significant potential as a non-chemical and non-thermal intervention for bacterial inactivation, especially on food. However, the applicability and usage of PAW depend on the effect of environmental and bacterial strain-based conditions and cost-effectiveness.

scholarly journals Phenotypically and Genotypically Heterogeneous Strains of Pseudomonas syringae Associated With Alfalfa Leaf Spot Disease in Iran

Plant Disease ◽  
2019 ◽  
Vol 103 (12) ◽  
pp. 3199-3208 ◽  
Author(s):  
Maryam Ansari ◽  
S. Mohsen Taghavi ◽  
Sadegh Zarei ◽  
Soraya Mehrb-Moghadam ◽  
Hamzeh Mafakheri ◽  
...  
Keyword(s):  
Host Range ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Phylogenetic Analyses ◽  
Housekeeping Genes ◽  
Pcr Primers ◽  
Phylogenetic Position ◽  
Leaf Spot Disease ◽  
Bacterial Strains ◽  
Alfalfa Leaf

In this study, we provide a polyphasic characterization of 18 Pseudomonas spp. strains associated with alfalfa leaf spot symptoms in Iran. All of the strains were pathogenic on alfalfa, although the aggressiveness and symptomology varied among the strains. All strains but one were pathogenic on broad bean, cucumber, honeydew, and zucchini, whereas only a fraction of the strains were pathogenic on sugar beet, tomato, and wheat. Syringomycin biosynthesis genes (syrB1 and syrP) were detected using the corresponding PCR primers in all of the strains isolated from alfalfa. Phylogenetic analyses using the sequences of four housekeeping genes (gapA, gltA, gyrB, and rpoD) revealed that all of the strains except one (Als34) belong to phylogroup 2b of P. syringae sensu lato, whereas strain Als34 placed within phylogroup 1 close to the type strain of P. syringae pv. apii. Among the phylogroup 2b strains, nine strains were phylogenetically close to the P. syringae pv. aptata clade, whereas the remainder were scattered among P. syringae pv. atrofaciens and P. syringae pv. syringae strains. Pathogenicity and host range assays of the bacterial strains evaluated in this study on a set of taxonomically diverse plant species did not allow us to assign a “pathovar” status to the alfalfa strains. However, these results provide novel insight into the host range and phylogenetic position of the alfalfa-pathogenic members of P. syringae sensu lato, and they reveal that phenotypically and genotypically heterogeneous strains of the pathogen cause bacterial leaf spot of alfalfa.

Viability and respiratory activity ofPseudomonas syringaecells starved in buffer

1995 ◽  
Vol 41 (4-5) ◽  
pp. 372-377 ◽  
Author(s):  
João P. S. Cabral
Keyword(s):  
Oxygen Consumption ◽  
Electron Transport ◽  
Pseudomonas Syringae ◽  
Respiratory Activity ◽  
Bacterial Cells ◽  
Intact Cells ◽  
Rate Of Oxygen Consumption ◽  
Metabolic Activities ◽  
Different Levels ◽  
Number Of Cells

Pseudomonas syringae cells starved in buffer released orcinol-reactive molecules and materials that absorbed ultraviolet light. The number of cells culturable in nutrient medium decreased more rapidly than the number of intact particles determined by microscopy. The results suggested that starvation resulted in the lysis of an increasing number of cells, and that a fraction of the intact particles were not culturable. Starvation also resulted in a decrease in the rate of oxygen consumption with acetate, glycerol, and succinate, but at different levels. Whereas the respiration of acetate and glycerol decreased concomitantly with culturability, the respiration of succinate decreased to levels similar to the concentration of intact cells, suggesting that all intact particles respired the succinate, but only the culturable cells respired the acetate and glycerol. The results suggest that measuring the activity of the electron-transport system can overestimate the viability of starved bacterial cells, and that complex metabolic activities such as the respiration of acetate and glycerol are probably better suited for the evaluation of this parameter.Key words: Pseudomonas syringae, starvation, culturability, viability, respiration.

scholarly journals First Report of Pseudomonas syringae pv. aptata Causing Bacterial Leaf Spot on Sugar Beet in Serbia

Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 281-281 ◽  
Author(s):  
V. Stojšin ◽  
J. Balaž ◽  
D. Budakov ◽  
Slaviša Stanković ◽  
I. Nikolić ◽  
...  
Keyword(s):  
Sugar Beet ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Leaf Surface ◽  
Negative Control ◽  
Gyrb Gene ◽  
Bacterial Suspension ◽  
Bacterial Strains ◽  
Gene Sequences ◽  
Bacterial Leaf Spot

A severe bacterial leaf spot was observed during June and July 2013 on commercial cultivars of sugar beet (Beta vulgaris var. saccharifera) in the Vojvodina Province of Serbia. Serbia is a major sugar beet production area in southeastern Europe, with 62,895 ha and 3 million tons of sugar beet yield in 2013. A foliar leaf spot observed in 25 commercial sugar beet fields surveyed ranged from 0.1 to 40% severity. Symptoms were characterized as circular or irregular, 5- to 20-mm diameter, white to light brown necrotic spots, each with a dark margin. Diseased leaves were rinsed in sterilized, distilled water (SDW) and dried at room temperature, and leaf sections taken from the margin of necrotic tissue were macerated in SDW. Isolations from 48 symptomatic leaves onto nutrient agar with 5% (w/v) sucrose (NAS) produced bacterial colonies that were whitish, circular, dome-shaped, and Levan-positive. Representative isolates (n = 105) were Gram negative; aerobic; positive for catalase, fluorescence on King's medium B, and tobacco hypersensitivity; and negative for oxidase, potato rot, and arginine dehydrolase. These reactions corresponded to LOPAT group Ia, which includes Pseudomonas syringae pathovars (2). Repetitive extragenic palindromic sequence (rep)-PCR was used for genetic fingerprinting the isolates using the REP, ERIC, and BOX primers. Twenty-five different profiles were obtained among the strains. From each profile group, one representative strain was sequenced for the gyrB gene (1). Four heterogenic groups were observed, and representative gyrB gene sequences of each group were deposited in the NCBI GenBank (Accession Nos. KJ950024 to KJ950027). The sequences were compared with those of pathotype strain P. syringae pv. aptata CFBP 1617 deposited in the PAMDB database; one strain was 100% homologous, and the other three were 99% homologous. To fulfill identification of the Serbian sugar beet isolates, gltA and rpoD partial gene sequences were determined (1), and the sequences were deposited as Accession Nos. KM386838 to KM386841 for gltA and KM386830 to KM38683033 for rpoD. The sequences were 100% homologous with those of pathotype strain CFBP 1617. Pathogenicity of each of four representative bacterial strains was tested on 3-week-old plants of the sugar beet cultivars Marinela, Serenada, and Jasmina (KWS, Belgrade, Serbia) and Lara (NS Seme, Novi Sad, Serbia) by atomizing a bacterial suspension of ~106 CFU/ml of the appropriate isolate onto the abaxial leaf surface of three plants per cultivar until water-soaking of the leaf surface was observed. Three plants of each cultivar atomized similarly with P. syringae pv. aptata CFBP 2473 and SDW served as positive and negative control treatments, respectively. Inoculated plants were kept in a clear plastic box at 80 to 100% RH and 17 ± 1°C and examined for symptom development over 3 weeks. For all test isolates and the control strain, inoculated leaves first developed water-soaked lesions 7 days after inoculation (DAI). By 10 to 14 DAI, lesions were necrotic and infection had spread to the petioles. By 21 DAI, wilting was observed on more than 50% of inoculated plants. Negative control plants were symptomless. Bacteria re-isolated onto NAS from inoculated leaves had the same colony morphology, LOPAT results, and gyrB partial gene sequences as described for the test strains. No bacteria were re-isolated from negative control plants. Based on these tests, the pathogen causing leaf spot on sugar beet in Serbia was identified as P. syringae pv. aptata. References: (1) P. Ferrente and M. Scortichini. Plant Pathol. 59:954, 2010. (2) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966.

scholarly journals Seasonal quantitative dynamics and ecology of pelagic rotifers in an acidified boreal lake

2017 ◽  
Vol 77 (1) ◽  
Author(s):  
Svein Birger Wærvågen ◽  
Tom Andersen
Keyword(s):  
Community Structure ◽  
Environmental Gradients ◽  
Seasonal Succession ◽  
Biotic Interactions ◽  
Resource Limitation ◽  
Local Environment ◽  
Food Competition ◽  
Variance Partitioning ◽  
Boreal Lake ◽  
Rotifer Community

Lake Gjerstadvann is a dimictic, oligotrophic, slightly acidified boreal lake in southern Norway (northwest Europe). The planktonic rotifer community of this lake was studied quantitatively during one year in order to investigate the impacts of the local environment and biotic interactions on seasonal succession and habitat selection. Pure suspension feeders (mainly Keratella spp., Conochilus spp., and Kellicottia longispina) together with raptorial graspers or specialised feeders (mainly Polyarthra spp. and Collotheca spp.) dominated the rotifer community over prolonged periods, whereas carnivorous/omnivorous species (mainly Asplanchna priodonta) were extremely uncommon. Low bicarbonate buffering capacity resulted in a distinctive seasonal oscillating pH between 5.0 and 5.6, defining a special acid-transition lake category. The pH values were highest in the productive period during summer, and lowest during ice break-up coinciding with the peak reactive aluminium concentrations of 250-300 mg L-1. As in typical Norwegian boreal perch lakes, the most abundant cladoceran was Bosmina longispina due to perch predation on the genus Daphnia. Rotifer community structure was significantly related to temperature and oxygen (P=0.001 and P=0.022), illustrating the important effects of the seasonal cycle and vertical density stratification. The most significant competition indicator species were B. longispina and Eudiaptomus gracilis (both with P=0.001). A variance partitioning indicated that 14% of the total community composition variance could only be explained by biotic interactions, while 19% of the variance could be attributed to environmental gradients. Of the variance, 23% could not be resolved between biotic interactions and environmental gradients, while a residual of 44% was not explainable by any of the variables. Acid conditions alone cannot account for all the observed changes in the rotifer community of this lake with low humic content, since resource limitation and food competition are also important factors shaping rotifer population dynamics and the community structure.

scholarly journals Effects of atmospheric conditions on ice nucleation activity of <i>Pseudomonas</i>

2012 ◽  
Vol 12 (22) ◽  
pp. 10667-10677 ◽  
Author(s):  
E. Attard ◽  
H. Yang ◽  
A.-M. Delort ◽  
P. Amato ◽  
U. Pöschl ◽  
...  
Keyword(s):  
Environmental Factors ◽  
Pseudomonas Syringae ◽  
Ice Nucleation ◽  
Acidic Ph ◽  
Bacterial Cells ◽  
Atmospheric Conditions ◽  
Ice Nucleation Protein ◽  
Ice Melt ◽  
Nucleation Activity ◽  
Ice Nucleation Activity

Abstract. Although ice nuclei from bacterial origin are known to be efficient at the highest temperatures known for ice catalysts, quantitative data are still needed to assess their role in cloud processes. Here we studied the effects of three typical cloud conditions (i) acidic pH (ii) NO2 and O3 exposure and (iii) UV-A exposure on the ice nucleation activity (INA) of four Pseudomonas strains. Three of the Pseudomonas syringae strains were isolated from cloud water and the phyllosphere and Pseudomonas fluorescens strain CGina-01 was isolated from Antarctic glacier ice melt. Among the three conditions tested, acidic pH caused the most significant effects on INA likely due to denaturation of the ice nucleation protein complex. Exposure to NO2 and O3 gases had no significant or only weak effects on the INA of two P. syringae strains whereas the INA of P. fluorescens CGina-01 was significantly affected. The INA of the third P. syringae strain showed variable responses to NO2 and O3 exposure. These differences in the INA of different Pseudomonas suggest that the response to atmospheric conditions could be strain-specific. After UV-A exposure, a substantial loss of viability of all four strains was observed whereas their INA decreased only slightly. This corroborates the notion that under certain conditions dead bacterial cells can maintain their INA. Overall, the negative effects of the three environmental factors on INA were more significant at the warmer temperatures. Our results suggest that in clouds where temperatures are near 0 °C, the importance of bacterial ice nucleation in precipitation processes could be reduced by some environmental factors.

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