Genomic analysis of plant-associated bacteria and their potential in enhancing phytoremediation efficiency

Phytoremediation is an emerging technology that uses plants in order to cleanup pollutants including xenobiotics and heavy metals from soil, water and air. Inoculation of plants with plant growth promoting endophytic and rhizospheric bacteria can enhance efficiency of phytoremediation. Genomic analysis of four plant-associated strains belonging to the Stenotrophomonas maltophilia species revealed the presence of genes encoding proteins involved in plant growth promotion, biocontrol of phytopathogens, biodegradation of xenobiotics, heavy metals resistance and plant-bacteria-environment interaction. The results of this analysis suggest great potential of bacteria belonging to Stenotrophomonas maltophilia species in enhancing phytoremediation efficiency

Comparative genomics of Pantoea ananatis species reviles genes involved in plant-endophytic bacteria interactions

tekst w j. pol. i ang.Celem pracy było znalezienie genów kodujących białka zaangażowane w oddziaływania endofit–roślina poprzez analizę porównawczą genomów bakterii z gatunku Pantoea ananatis

Analiza udziału białek ściany komórkowej w odpowiedzi na stres temperaturowy u modelowego gatunku trawy Brachypodium distachyon

Praca zawiera artykuły w języku angielskim.Increasing climate changes increase the average temperature of the Earth's surface and a higher frequency of extreme weather phenomena, which are a threat to food production. Given the growing human population and declining farmland, there is an urgent need to develop new crops adapted to changing climate. It is possible by developing new plant varieties, either with the use of classical breeding methods or through the use of genetic engineering. The latter approach requires a detailed understanding of the molecular mechanisms underlying various stress responses, particularly temperature stress induced by low and high temperatures. Temperature stress changes photosynthetic efficiency and disturbs redox potential. It also leads to rearrangement of the cell wall cytoskeleton and cell wall remodelling. All these processes are relatively poorly understood. Therefore, this work focused on the response to the temperature stress of leaves of a model species for grasses in the temperate climate zone, Brachypodium distachyon. This study aimed to analyse the response of Brachypodium distachyon cell wall proteins in leaves subjected to low (4 °C) and high (40 °C) temperature. Another objective was to obtain mutants with inactivated genes encoding the Fasciclin arabinogalactan protein (Bradi3g39740) and pectin methylesterase (Bradi3g24750). To characterize the effect of cellular stress on the cell wall proteins, immunocytochemistry with antibodies binding specifically to arabinogalactan proteins and extensins epitopes, analysis of the expression profile of genes encoding arabinogalactan and extensin proteins, as well as analysis of the cell wall proteome were applied. In order to obtain mutants with inactivated genes, site-directed mutagenesis based on CRISPR/Cas9 system was used. In addition, the transformation of the embryogenic callus induced from immature embryos was performed using Agrobacterium tumefaciens bacterial cells. Studies on the distribution of AGP epitopes in leaves of B. distachyon revealed their presence mainly in the vascular bundle and extensins in the mesophyll. Differences in the distribution and intensity of signals were demonstrated for the four antibodies recognizing AGP: JIM8, JIM16, LM2, and LM6. At the same time, no differences in the distribution and intensity of signals were observed for antibodies recognizing extensin epitopes. Analysis of the expression profile of the genes encoding AGP and extensin revealed an increase in the expression level, significantly greater in the plants incubated at high temperatures. In turn, proteomic analysis of the cell wall allowed the identification of 46 proteins with the differentiated presence at high temperature compared to the control. These changes suggest lower protease activity, cell wall lignification and expansion, and changes in the architecture of cell wall polymers, especially pectins. Using the CRISPR/Cas9 system, four mutants with inactivated genes encoding the Fasciclin arabinogalactan protein and pectin methylesterase were obtained. Although the obtained mutants show no change in response to temperature stress, they show slower growth in response to salt stress

Selecting Bacteria Candidates for the Bioaugmentation of Activated Sludge to Improve the Aerobic Treatment of Landfill Leachate

In this study, a multifaceted approach for selecting the suitable candidates for bioaugmentation of activated sludge (AS) that supports leachate treatment was used. To determine the exploitation of 10 bacterial strains isolated from the various matrices for inoculating the AS contaminated with the Kalina pond leachate (KPL), their degradative potential was analyzed along with their aptitude to synthesize compounds improving remediation of pollutants in wastewater and ability to incorporate into the AS flocs. Based on their capability to degrade aromatic compounds (primarily catechol, phenol, and cresols) at a concentration of 1 mg/mL and survive in 12.5% of the KPL, Pseudomonas putida OR45a and P. putida KB3 can be considered to be the best candidates for bioaugmentation of the AS among all of the bacteria tested. Genomic analyses of these two strains revealed the presence of the genes encoding enzymes related to the metabolism of aromatic compounds. Additionally, both microorganisms exhibited a high hydrophobic propensity (above 50%) and an ability to produce biosurfactants as well as high resistance to ammonium (above 600 g/mL) and heavy metals (especially chromium). These properties enable the exploitation of both bacterial strains in the bioremediation of the AS contaminated with the KPL

Analysis of the Bioaugmentation Potential of Pseudomonas putida OR45a and Pseudomonas putida KB3 in the Sequencing Batch Reactors Fed with the Phenolic Landfill Leachate

The treatment of landfill leachate could be challenging for the biological wastewater treatment systems due to its high toxicity and the presence of poorly biodegradable contaminants. In this study, the bioaugmentation technology was successfully applied in sequencing batch reactors (SBRs) fed with the phenolic landfill leachate by inoculation of the activated sludge (AS) with two phenol-degrading Pseudomonas putida OR45a and Pseudomonas putida KB3 strains. According to the results, the SBRs bioaugmented with Pseudomonas strains withstood the increasing concentrations of the leachate. This resulted in the higher removal efficiency of the chemical oxygen demand (COD) of 79–86%, ammonia nitrogen of 87–88% and phenolic compounds of 85–96% as compared to 45%, 64%, and 50% for the noninoculated SBR. Simultaneously, the bioaugmentation of the AS allowed to maintain the high enzymatic activity of dehydrogenases, nonspecific esterases, and catalase in this ecosystem, which contributed to the higher functional capacity of indigenous microorganisms than in the noninoculated AS. Herein, the stress level experienced by the microorganisms in the SBRs fed with the leachate computed based on the cellular ATP measurements showed that the abundance of exogenous Pseudomonas strains in the bioreactors contributed to the reduction in effluent toxicity, which was reflected by a decrease in the stress biomass index to 32–45% as compared to the nonbioaugmented AS (76%)

Effects of Low Concentration of Selected Analgesics and Successive Bioaugmentation of the Activated Sludge on Its Activity and Metabolic Diversity

In this study, we evaluated the impact of the successive bioaugmentation of the activated sludge (AS) with the defined bacterial consortium on the activity and functional capacity of the AS microorganisms. In parallel, the removal of low concentrations of the selected non-steroidal anti-inflammatory drugs (ibuprofen, naproxen, diclofenac) and analgesic paracetamol was studied. We found that the addition of the bacterial consortium consisting of three pharmaceuticals-degrading strains Bacillus thuringiensis B1 (2015b), Stenotrophomonas maltophilia KB2, and Pseudomonas moorei KB4 into the AS did not cause any significant changes in the biomass abundance and metabolic activity of the AS microorganisms. Although, the successive bioaugmentation of the AS caused a slight increase in the metabolic diversity, the intensity of carbohydrates usage, and metabolic richness. Microorganisms in the bioaugmented and non-bioaugmented AS were able to degrade the mixture of the analyzed drugs with similar e ciency, however, diclofenac was removed more e ectively in the bioaugmented AS. Several metabolites were identified and e ciently utilized, with the exception of 4-OH diclofenac. Two new diclofenac-degrading strains assigned as Serratia proteamaculans AS4 and Rahnella bruchi AS7 were isolated from the diclofenac-treated AS

Genome Mining Revealed a High Biosynthetic Potential for Antifungal Streptomyces sp. S-2 Isolated from Black Soot

The increasing resistance of fungal pathogens has heightened the necessity of searching for new organisms and compounds to combat their spread. Streptomyces are bacteria that are well-known for the production of many antibiotics. To find novel antibiotic agents, researchers have turned to previously neglected and extreme environments. Here, we isolated a new strain, Streptomyces sp. S-2, for the first time, from black soot after hard coal combustion (collected from an in-use household chimney). We examined its antifungal properties against plant pathogens and against fungi that potentially pose threat to human health (Fusarium avenaceum, Aspergillus niger and the environmental isolates Trichoderma citrinoviridae Cin-9, Nigrospora oryzae sp. roseF7, and Curvularia coatesieae sp. junF9). Furthermore, we obtained the genome sequence of S-2 and examined its potential for secondary metabolites production using anti-SMASH software. The S-2 strain shows activity against all of the tested fungi. Genome mining elucidated a vast number of biosynthetic gene clusters (55), which distinguish this strain from closely related strains. The majority of the predicted clusters were assigned to non-ribosomal peptide synthetases or type 1 polyketide synthetases, groups known to produce compounds with antimicrobial activity

Stability and instability processes in the calli of Fagopyrum tataricum that have different morphogenic potentials

The morphogenic callus (MC) of Fagopyrum tataricum contains a large amount of flavonoids, especially rutin, and exhibits a high level of antioxidant activity. A non-morphogenic callus (NC) may appear on the surface of the MC after two to three years of cultivation and is then subjected to a consistently high level of oxidative stress. The elucidation of the molecular background of this instability is essential for gaining a better understanding of the somaclonal variation mechanisms in tissue cultures that have different morphogenic potentials. Thus, in this study we show that continuous oxidative stress in a NC might be connected with a rapid senescence process and as a result, in the upregulation of the genes that are connected with the telomere complexity, ethylene biosynthesis and the expression of DNA methyltransferases. Moreover, we analysed the presence of the hydroxyproline-rich glycoproteins in the calli and demonstrated the differences between the MC and NC. The LM2 antibody can be useful as a marker of the cells in the MC that are embryogenically determined, while the MAC207 antibody seems to be a positive marker of a MC as its signal was absent in the NC. This study also provides the first report on the effect of trichostatin A on the DNA methyltransferases and demethylases in a MC

Organic micropollutants paracetamol and ibuprofen - toxicity, biodegradation, and genetic background of their utilization by bacteria

Currently, analgesics and nonsteroidal anti-inflammatory drugs (NSAIDs) are classified as one of the most emerging group of xenobiotics and have been detected in various natural matrices. Among them, monocyclic paracetamol and ibuprofen, widely used to treat mild and moderate pain are the most popular. Since long-term adverse effects of these xenobiotics and their biological and pharmacokinetic activity especially at environmentally relevant concentrations are better understood, degradation of such contaminants has become a major concern. Moreover, to date, conventional wastewater treatment plants (WWTPs) are not fully adapted to remove that kind of micropollutants. Bioremediation processes, which utilize bacterial strains with increased degradation abilities, seem to be a promising alternative to the chemical methods used so far. Nevertheless, despite the wide prevalence of paracetamol and ibuprofen in the environment, toxicity and mechanism of their microbial degradation as well as genetic background of these processes remain not fully characterized. In this review, we described the current state of knowledge about toxicity and biodegradation mechanisms of paracetamol and ibuprofen and provided bioinformatics analysis concerning the genetic bases of these xenobiotics decomposition

Hydroxyproline-Rich Glycoproteins as Markers of Temperature Stress in the Leaves of Brachypodium distachyon

Plants frequently encounter diverse abiotic stresses, one of which is environmental thermal stress. To cope with these stresses, plants have developed a range of mechanisms, including altering the cell wall architecture, which is facilitated by the arabinogalactan proteins (AGP) and extensins (EXT). In order to characterise the localisation of the epitopes of the AGP and EXT, which are induced by the stress connected with a low (4 C) or a high (40 C) temperature, in the leaves of Brachypodium distachyon, we performed immunohistochemical analyses using the antibodies that bind to selected AGP (JIM8, JIM13, JIM16, LM2 and MAC207), pectin/AGP (LM6) as well as EXT (JIM11, JIM12 and JIM20). The analyses of the epitopes of the AGP indicated their presence in the phloem and in the inner bundle sheath (JIM8, JIM13, JIM16 and LM2). The JIM16 epitope was less abundant in the leaves from the low or high temperature compared to the control leaves. The LM2 epitope was more abundant in the leaves that had been subjected to the high temperatures. In the case of JIM13 and MAC207, no changes were observed at the di erent temperatures. The epitopes of the EXT were primarily observed in the mesophyll and xylem cells of the major vascular bundle (JIM11, JIM12 and JIM20) and no correlation was observed between the presence of the epitopes and the temperature stress. We also analysed changes in the level of transcript accumulation of some of the genes encoding EXT, EXT-like receptor kinases and AGP in the response to the temperature stress. In both cases, although we observed the upregulation of the genes encoding AGP in stressed plants, the changes were more pronounced at the high temperature. Similar changes were observed in the expression profiles of the EXT and EXT-like receptor kinase genes. Our findings may be relevant for genetic engineering of plants with increased resistance to the temperature stress