Burkholderia Strains Research Articles

Antifungal potential of endophytic <i>Burkholderia</i> spp., plant extracts, and curcumin silver nanoparticles against <i>aspergillus</i> spp. and <i>fusarium</i> sp.

Numerous members of Aspergillus spp. and Fusarium spp. are pathogens possessing a broad spectrum of their host, including plants, animals, and humans. In this study, five endophytic Burkholderia spp., five plant extracts, and curcumin silver nanoparticles (C-AgNPs) were used to screen their antifungal activities against Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, and Fusarium sp. ATCC 60289 (F. sp.). The results of dual assays showed that all five Burkholderia spp. strains NC119, NC148, NC160, NC166, and NC206 exhibited the antifungal activities with the percentage of inhibition (PI) ranging from 34.34% to 76.01%, in which the strain NC166 showed the strongest antifungal activity against all four studied fungi. Notably, Burkholderia spp. appeared to be effective against F. sp., with the PI greater than 50% in four out of five bacterial strains. In contrast, the results of well-diffusion assays with five plant extracts from Perilla frutescens L. (leaves, stems, roots), Piper betle L. (leaves), and Zingiber officinale Rosc. (rhizomes) had low probabilities of inhibiting F. sp. However, the bacterial strain Burkholderia metallica isolated from Perilla root showed the antifungal activities against F. sp with PI of 47.1%. In addition, the C-AgNPs performed considerable effectiveness in inhibiting the growth of all four fungi, with the highest PI of 71.17 ± 1.44% against Aspergillus terreus. These outcomes not only emphasize the potential of Burkholderia spp. and C-AgNPs as antimicrobial agents for the management of Aspergillus spp. and Fusarium spp., but also primarily rule out the antifungal possibility against F. sp. of some plant extracts, providing suggestions for future approaches in the research of these pathogenic fungi.

Employing Genomic Tools to Explore the Molecular Mechanisms behind the Enhancement of Plant Growth and Stress Resilience Facilitated by a Burkholderia Rhizobacterial Strain.

The rhizobacterial strain BJ3 showed 16S rDNA sequence similarity to species within the Burkholderia genus. Its complete genome sequence revealed a 97% match with Burkholderia contaminans and uncovered gene clusters essential for plant-growth-promoting traits (PGPTs). These clusters include genes responsible for producing indole acetic acid (IAA), osmolytes, non-ribosomal peptides (NRPS), volatile organic compounds (VOCs), siderophores, lipopolysaccharides, hydrolytic enzymes, and spermidine. Additionally, the genome contains genes for nitrogen fixation and phosphate solubilization, as well as a gene encoding 1-aminocyclopropane-1-carboxylate (ACC) deaminase. The treatment with BJ3 enhanced root architecture, boosted vegetative growth, and accelerated early flowering in Arabidopsis. Treated seedlings also showed increased lignin production and antioxidant capabilities, as well as notably increased tolerance to water deficit and high salinity. An RNA-seq transcriptome analysis indicated that BJ3 treatment significantly activated genes related to immunity induction, hormone signaling, and vegetative growth. It specifically activated genes involved in the production of auxin, ethylene, and salicylic acid (SA), as well as genes involved in the synthesis of defense compounds like glucosinolates, camalexin, and terpenoids. The expression of AP2/ERF transcription factors was markedly increased. These findings highlight BJ3's potential to produce various bioactive metabolites and its ability to activate auxin, ethylene, and SA signaling in Arabidopsis, positioning it as a new Burkholderia strain that could significantly improve plant growth, stress resilience, and immune function.

Impact of template denaturation prior to whole genome amplification on gene detection in high GC-content species, Burkholderia mallei and B. pseudomallei

ObjectiveIn this study, we sought to determine the types and prevalence of antimicrobial resistance determinants (ARDs) in Burkholderia spp. strains using the Antimicrobial Resistance Determinant Microarray (ARDM).ResultsWhole genome amplicons from 22 B. mallei (BM) and 37 B. pseudomallei (BP) isolates were tested for > 500 ARDs using ARDM v.3.1. ARDM detected the following Burkholderia spp.-derived genes, aac(6), blaBP/MBL-3, blaABPS, penA-BP, and qacE, in both BM and BP while blaBP/MBL-1, macB, blaOXA-42/43 and penA-BC were observed in BP only. The method of denaturing template for whole genome amplification greatly affected the numbers and types of genes detected by the ARDM. BlaTEM was detected in nearly a third of BM and BP amplicons derived from thermally, but not chemically denatured templates. BlaTEM results were confirmed by PCR, with 81% concordance between methods. Sequences from 414-nt PCR amplicons (13 preparations) were 100% identical to the Klebsiella pneumoniae reference gene. Although blaTEM sequences have been observed in B. glumae, B. cepacia, and other undefined Burkholderia strains, this is the first report of such sequences in BM/BP/B. thailandensis (BT) clade. These results highlight the importance of sample preparation in achieving adequate genome coverage in methods requiring untargeted amplification before analysis.

Ingested soil bacteria breach gut epithelia and prime systemic immunity in an insect

Insects lack acquired immunity and were thought to have no immune memory, but recent studies reported a phenomenon called immune priming, wherein sublethal dose of pathogens or nonpathogenic microbes stimulates immunity and prevents subsequential pathogen infection. Although the evidence for insect immune priming is accumulating, the underlying mechanisms are still unclear. The bean bug Riptortus pedestris acquires its gut microbiota from ambient soil and spatially structures them into a multispecies and variable community in the anterior midgut and a specific, monospecies Caballeronia symbiont population in the posterior region. We demonstrate that a particular Burkholderia strain colonizing the anterior midgut stimulates systemic immunity by penetrating gut epithelia and migrating into the hemolymph. The activated immunity, consisting of a humoral and a cellular response, had no negative effect on the host fitness, but on the contrary protected the insect from subsequent infection by pathogenic bacteria. Interruption of contact between the Burkholderia strain and epithelia of the gut weakened the host immunity back to preinfection levels and made the insects more vulnerable to microbial infection, demonstrating that persistent acquisition of environmental bacteria is important to maintain an efficient immunity.

Comparative Genome Analyses Provide Insight into the Antimicrobial Activity of Endophytic Burkholderia.

Endophytic bacteria are endosymbionts that colonize a portion of plants without harming the plant for at least a part of its life cycle. Bacterial endophytes play an essential role in promoting plant growth using multiple mechanisms. The genus Burkholderia is an important member among endophytes and encompasses bacterial species with high genetic versatility and adaptability. In this study, the endophytic characteristics of Burkholderia species are investigated via comparative genomic analyses of several endophytic Burkholderia strains with pathogenic Burkholderia strains. A group of bacterial genes was identified and predicted as the putative endophytic behavior genes of Burkholderia. Multiple antimicrobial biosynthesis genes were observed in these endophytic bacteria; however, certain important pathogenic and virulence genes were absent. The majority of resistome genes were distributed relatively evenly among the endophytic and pathogenic bacteria. All known types of secretion systems were found in the studied bacteria. This includes T3SS and T4SS, which were previously thought to be disproportionately represented in endophytes. Additionally, questionable CRISPR-Cas systems with an orphan CRISPR array were prevalent, suggesting that intact CRISPR-Cas systems may not exist in symbiotes of Burkholderia. This research not only sheds light on the antimicrobial activities that contribute to biocontrol but also expands our understanding of genomic variations in Burkholderia's endophytic and pathogenic bacteria.

Polysaccharides Produced by Plant Growth-Promoting Rhizobacteria Strain Burkholderia sp. BK01 Enhance Salt Stress Tolerance to Arabidopsis thaliana.

Salt stress is one of the most serious abiotic stresses leading to reduced agricultural productivity. Polysaccharides from seaweed have been used as biostimulants to promote crop growth and improve plant resistance to abiotic stress. In this study, PGPR strain Burkholderia sp. BK01 was isolated from the rhizosphere of wheat, and it was characterized for phosphorus (Pi) dissolution, indole-3-acetic acid (IAA) production, ammonia (NH3) and exopolysaccharides (EPS). In particular, strain BK01 can efficiently produce extracellular polysaccharide with a yield of 12.86 g/L, using sorbitol as carbon source. BK01 EPS was identified as an heteropolysaccharide with Mw 3.559 × 106 Da, composed of (D)-galactose (75.3%), (D)-glucose (5.5%), (L)-rhamnose (5.5%), (D)-galactouronic acid (4.9%) and (D)-glucuronic acid (8.8%). The present work aims to highlight the effect of the BK01 EPS on growth and biochemical changes in Arabidopsis thaliana under salt stress (100 mM). The purified BK01 EPS at a concentration of 100 mg/L efficiently promoted the growth of plants in pot assays, improved the chlorophyll content, enhanced the activities of SOD, POD and CAT, and decreased the content of MDA. This results suggested that the polysaccharides produced by PGPR strain Burkholderia sp. BK01 can be used as biostimulants to promote plant growth and improve plant resistance to salt stress.

Cloaking antibodies are prevalent in Burkholderia cepacia complex infection and their removal restores serum killing.

The Burkholderia cepacia complex encompasses a group of gram-negative opportunistic pathogens that cause chronic lung infections in people with cystic fibrosis. Distinct from other respiratory pathogens, Burkholderia causes a unique clinical disease in a subset of patients known as 'cepacia syndrome', fulminant pneumonia accompanied by bacteraemia and sepsis with a mortality rate of up to 75%. Due to the bacteraemia associated with this disease, the mechanisms that allow Burkholderia to resist the bactericidal effects of serum complement-depending killing are vital. Antibodies usually promote serum killing; however, we have described 'cloaking antibodies', specific for lipopolysaccharides that paradoxically protect serum-sensitive bacteria from complement-mediated lysis. Cloaking antibodies that protect Pseudomonas aeruginosa have been found in 24%-41% of patients with chronic lung diseases. The presence of these antibodies is also associated with worse clinical outcomes. Here, we sought to determine the relevance of cloaking antibodies in patients with Burkholderia infection. Twelve Burkholderia spp. were isolated from nine pwCF and characterised for susceptibility to healthy control serum. Patient serum was analysed for the titre of the cloaking antibody. The ability of the patient serum to prevent healthy control serum (HCS) killing of its cognate isolates was determined. We found that several of the Burkholderia strains were shared between patients. Ten of the 12 isolates were highly susceptible to HCS killing. Four of nine (44%) patients had cloaking antibodies that protected their cognate strain from serum killing. Depleting cloaking antibodies from patient serum restored HCS killing of Burkholderia isolates. Cloaking antibodies are prevalent in patients with Burkholderia pulmonary infection and protect these strains from serum killing. Removal of cloaking antibodies via plasmapheresis, as previously described for individuals with life-threatening Pseudomonas infection, may be a useful new strategy for those with serious and life-threatening Burkholderia infection.

Production of biohydrogen and green platform compound 2, 5-furandicarboxylic acid using rice straw hydrolysate

Lignocellulosic hydrolysate contains several inhibitors that hinder downstream biological processes. The immobilized strain Burkholderia sp. H-2 was used in this study to biodetoxify real rice straw hydrolysate and biotransform 5-hydroxymethylfurfural (5-HMF), an inhibitor, into 2, 5-furandicarboxylic acid (FDCA), a valuable platform compound. An electrodialysis with bipolar membranes (EDBM) system was applied to recover FDCA and perform secondary detoxification of the biodetoxified real rice straw hydrolysate. The rice straw hydrolysate after biodetoxification and EDBM treatment was used for anaerobic dark fermentative hydrogen production. The immobilized Burkholderia sp. H-2 could completely biotransform 1843 mg/L 5-HMF into 1532 mg/L FDCA within 24 h when the 2-fold diluted rice straw hydrolysate was used as the feedstock. The optimal EDBM system parameters to recover FDCA were a hydrolysate pH of 6, operated at a current density of 17.86 mA/cm2. The FDCA recovery efficiency was 44.6 %, when using the biodetoxified real rice straw hydrolysate as the EDBM feedstock. The rice straw hydrolysate after biodetoxification and EDBM treatment had better biohydrogen production performance than the undetoxified and biodetoxified rice straw hydrolysates.

Whole genome sequencing of Penicillium and Burkholderia strains antagonistic to the causal agent of kauri dieback disease (Phytophthora agathidicida) reveals biosynthetic gene clusters related to antimicrobial secondary metabolites.

Phytophthora agathidicida is a virulent soil pathogen of Aotearoa New Zealand's iconic kauri tree species (Agathis australis (D. Don) Lindl.) and the primary causal agent of kauri dieback disease. To date, only a few control options are available to treat infected kauri that are expressing symptoms of dieback disease. Previous research has identified strains of Penicillium and Burkholderia that inhibited the mycelial growth of P. agathidicida in vitro. However, the mechanisms of inhibition remain unknown. By performing whole genome sequencing, we screened the genomes of four Penicillium and five Burkholderia strains to identify secondary metabolite encoding biosynthetic gene clusters (SM-BGCs) that may be implicated in the production of antimicrobial compounds. We identified various types of SM-BGCs in the genome of each strain, including polyketide synthases (PKSs), non-ribosomal peptide synthetases (NRPSs), and terpenes. Across all four of the Penicillium strains, five SM-BGCs were detected that encoded the biosynthesis of napthopyrone, clavaric acid, pyranonigrin E, dimethyl coprogen and asperlactone. Across all five of the Burkholderia strains, three SM-BGCs were detected that encoded the biosynthesis of ornibactin, pyochelin and pyrrolnitin. Our analysis detected numerous SM-BGCs which could not be characterised. Further efforts should be made to identify the compounds encoded by these SM-BGCs so that we can explore their antimicrobial potential. The potential inhibitory effects of the compounds encoded by the SM-BGCs identified in this study may be worthy of further investigation for their effect on the growth and virulence of P. agathidicida.

Rhizobacteria control damping-off and promote growth of lima bean with and without co-inoculation with Rhizobium tropici CIAT899.

Rhizoctonia solani compromises the production of lima bean, an alternative and low-input food source in many tropical regions. Inoculation of bacterial strains has been used, but research on their biocontrol and growth promotion potential on lima bean is scarce. The objective of this study was to evaluate the effects of inoculation with rhizobacterial strains of the genera Bacillus, Brevibacillus, Paenibacillus, Burkholderia, Pseudomonas, and Rhizobium in combination or not with N2-fixing Rhizobium tropici on the control of damping-off disease and growth promotion in lima bean plants. Greenhouse experiments were conducted to evaluate the inoculation with bacterial strains with biocontrol potential in combination or not with R. tropici in substrate infected with R. solani CML 1846. Growth promotion of these strains was also assessed. Strains of Brevibacillus (UFLA 02-286), Pseudomonas (UFLA 02-281 and UFLA 04-885), Rhizobium (UFLA 04-195), and Burkholderia (UFLA 04-227) co-inoculated with the strain CIAT 899 (Rhizobium tropici) were the most effective in controlling R. solani, reducing the disease incidence in 47-60% on lima bean. The promising strains used in the biocontrol assays were also responsive in promoting growth of lima bean under disease and sterile conditions. A positive synergistic effect of co-inoculation of different genera contributed to plant growth, and these outcomes are important first steps to improve lima bean production.

Rapid Biodegradation of the Organophosphorus Insecticide Acephate by a Novel Strain Burkholderia sp. A11 and Its Impact on the Structure of the Indigenous Microbial Community.

The acephate-degrading microbes that are currently available are not optimal. In this study, Burkholderia sp. A11, an efficient degrader of acephate, presented an acephate-removal efficiency of 83.36% within 56 h (100 mg·L-1). The A11 strain has a broad substrate tolerance and presents a good removal effect in the concentration range 10-1600 mg·L-1. Six metabolites from the degradation of acephate were identified, among which the main products were methamidophos, acetamide, acetic acid, methanethiol, and dimethyl disulfide. The main degradation pathways involved include amide bond breaking and phosphate bond hydrolysis. Moreover, strain A11 successfully colonized and substantially accelerated acephate degradation in different soils, degrading over 90% of acephate (50-200 mg·kg-1) within 120 h. 16S rDNA sequencing results further confirmed that the strain A11 gradually occupied a dominant position in the soil microbial communities, causing slight changes in the diversity and composition of the indigenous soil microbial community structure.

Comparison of the colonization ability of Burkholderia strain B23 in the citrus rhizoplane and rhizosphere and assessment of the underlying mechanisms using full-length 16S rDNA amplicon and metatranscriptomic analyses.

The characterization of bacterial strains with efficient root colonization ability and the mechanisms responsible for their efficient colonization is critical for the identification and application of beneficial bacteria. In this study, we found that Burkholderia strain B23 exhibited a strong niche differentiation between the rhizosphere and rhizoplane (a niche with more abundant easy-to-use nutrients but stronger selective pressures compared with the tightly adjacent rhizosphere) when inoculated into the field-grown citrus trees. Full-length 16S rDNA amplicon analysis demonstrated that the relative abundance of B23 in the rhizoplane microbiome at 3, 5, and 9 days post-inoculation (dpi) was always higher than that at 1 dpi, whereas its relative abundance in the rhizosphere microbiome was decreased continuously, as demonstrated by a 3.18-fold decrease at 9 dpi compared to 1 dpi. Time-series comparative expression profiling of B23 between the rhizoplane and rhizosphere was performed at representative time points (1, 3, and 9 dpi) through metatranscriptomic analysis, and the results demonstrated that multiple genes involved in the uptake and utilization of easy-to-use carbohydrates and amino acids and those involved in metabolism, energy production, replication, and translation were upregulated in the rhizoplane compared with the rhizosphere at 1 dpi and 3 dpi. Several genes involved in resistance to plant- and microbial competitor-derived stresses exhibited higher expression activities in the rhizoplane compared with the rhizosphere. Furthermore, gene loci responsible for the biosynthesis of the key antifungal and antibacterial metabolites occidiofungin and ornibactin were induced, and their expression levels remained relatively stable from 3 dpi to 9 dpi in the rhizoplane but not in the rhizosphere. Collectively, our findings provide novel lights into the mechanisms underlying the root colonization of the inoculated bacterial strains and serve as a basis for the identification of strains with efficient colonization ability, thus contributing to the development of beneficial bacteria applications.

BysR, a LysR-Type Pleiotropic Regulator, Controls Production of Occidiofungin by Activating the LuxR-Type Transcriptional Regulator AmbR1 in Burkholderia sp. Strain JP2-270.

Occidiofungin is a highly effective antifungal glycopeptide produced by certain Burkholderia strains. The ocf gene cluster, responsible for occidiofungin biosynthesis, is regulated by the cluster-specific regulators encoded by an ambR homolog(s) within the same gene cluster, while the extent to which occidiofungin biosynthesis is connected with the core regulation network remains unknown. Here, we report that the LysR-type regulator BysR acts as a pleiotropic regulator and is essential for occidiofungin biosynthesis. Magnaporthe oryzae was used as an antifungal target in this study, and deletion of bysR and ocfE abolished the antagonistic activity against M. oryzae in Burkholderia sp. strain JP2-270. The ΔbysR defect can be recovered by constitutively expressing bysR or ambR1, but not ambR2. Electrophoretic mobility shift assays (EMSAs) collectively showed that BysR regulates ambR1 by directly binding to its promoter region. In addition, transcriptomic analysis revealed altered expression of 350 genes in response to bysR deletion, and the genes engaged in flagellar assembly and bacterial chemotaxis constitute the most enriched pathways. Also, 400 putative BysR-targeted loci were identified by DNA affinity purification sequencing (DAP-seq) in JP2-270. These loci include not only genes engaged in key metabolic pathways but also those involved in secondary metabolic pathways. To conclude, the occidiofungin produced by JP2-270 is the main substance inhibiting M. oryzae, and BysR controls occidiofungin production by directly targeting ambR1, an intracluster transcriptional regulatory gene that further activates the transcription of the ocf gene cluster. IMPORTANCE We report for the first time that occidiofungin production is regulated by the global transcriptional factor BysR, by directly targeting the specific regulator ambR1, which further promotes the transcription of ocf genes. BysR also acts as a pleiotropic regulator that controls various cellular processes in Burkholderia sp. strain JP2-270. This study provides insight into the regulatory mechanism of occidiofungin synthesis and enhances our understanding of the regulatory patterns of the LysR-type regulator.

Impact of microbial consortia on organic maize in a temperate climate varies with environment but not with fertilization

Inoculating crops with specifically designed microbial consortia (MC) is increasingly discussed as an instrument for sustainably promoting arable crop growth. While some promising results were achieved in laboratory, climate chamber, and greenhouse experiments, field applications of microbial inoculants under temperate climate conditions have been much less investigated. Here, we assess the effect of three novel (MC_B, MC_C, MC_C_AMF) and one commercial (Micosat F) MC in a factorial combination with three fertilization levels (unfertilized, 110 kg nitrogen ha-1, and 200 kg nitrogen ha-1) on organic maize (Zea mays L.) productivity. Split-plot experiments were performed in two consecutive years (2020 and 2021) at different locations in the Rhineland, Germany, resulting in three trial environments. The novel MC consisted of well-studied single strains of Azotobacter, Bacillus, Burkholderia, Pseudomonas, Rahnella, and Glomus. At stem elongation in 2020, crop growth (+17%), shoot nitrogen (+34%), and phosphorus uptake (+25%) were significantly enhanced upon inoculation with MC_C, possibly due to atmospheric N2-fixation of A. chroococcum LS132 and phosphorus solubilization of P. fluorescens DR54. However, MC_C failed to impact maize productivity in 2021. This difference between years was likely due to distinct weather and pre-crop effects. Significant increases in grain yield were observed only for MC_C_AMF (+10%) in 2021. Interactions between MC inoculation and organic fertilization were not observed. Therefore, the assumption that MC are more effective in low-input systems cannot be confirmed by this study. However, the effect of fertilization on crop growth and nutrition was more common and consistent than the impact of MC inoculation in both years. As revealed by the varying performance of MC across trial environments, achieving significant, agronomically relevant, and reproducible benefits from MC inoculation still remains a considerable challenge in field-grown arable crops in temperate climate.

143. Genomics analysis of Carbapenem resistance in BCC Identify PenR E151V substitution and novel Burkholderia cepacia complex specific OXA-1043 subgroup

Abstract Background Treatment options for Burkholderia cepacia complex (Bcc) remain limited, and previous study revealed 26% of strains was susceptible to meropenem. In B. cenocepacia and many Bcc species, ampC and its regulator, ampR, named penR in Burkholderia genomes are not associated, sharing a divergent promoter, as in other previously studied species. Recently, Josselin Bodilis et el (2018) found that there was no different in genetic organization between imipenem-sensitive and imipenem-resistant strains. Our study is to provide an answer to the different of genetic expression pattern between imipenem resistance and imipenem sensitive Burkholderia cepacia complex species. Methods The Bcc species genome was deeply sequenced using Nanopore technology. Protein-coding genes, and coding and non-coding RNAs in the chromosomes and plasmids were annotated via the NCBI PGAP pipeline. Antibiotic-resistant genes (ARGs) were predicted by aligning protein-coding genes against the Comprehensive Antibiotic Resistance Database (CARD) using Diamond. Results In our study, drug susceptibility test of different antibiotic showed only Burkholderia cenocepacia strain MC0-3 revealed sensitive to imipenem (Table 1). Further genetic organization between imipenem-resistance and imipenem-sensitive strains showed similar β-lactamase-encoding genes (Figure 1). In the multiple sequence alignment of PenR, nine imipenem-resistance Bcc stains, which amino acid sequence at position 151 is glutamic acid, however, in imipenem-sensitive strain amino acid substitute glutamic acid with valine (Figure 2b). PROVEAN also showed amino acid substitutions at position E151 in PenR is deleterious (Table 2). We also analyzed 95 strains of Burkholderia, and drew the phylogenetic tree which showed PenR E151V subsititution consisted in other imipenem-sensitive strains of Burkholderia. In addition, a novel Burkholderia cepacia complex specific OXA-1043 subgroup is found in stain toggle 2, toggle 3 and toggle 4 (Figure 3). Figure 1.Genetic organization of imipenem resistance and imipenem sensitive in different strain of Burkholderia cepacia complex. Conclusion The results of this study indicate that antibiotic resistance related to the amino acid substitutions, which may explain the imipenem-sensitive strain of Burkholderia cenocepacia. Bedsides, a new OXA family is found in Burkholderia cenocepacia strain, and is named OXA 1403. Figure 2.Multiple sequence alignment PenR in different strains of Burkholderia cepacia complex and phylogenetic tree of Burkholderia spp. Disclosures All Authors: No reported disclosures.

A novel bacterial strain Burkholderia sp. F25 capable of degrading diffusible signal factor signal shows strong biocontrol potential.

Vast quantities of synthetic pesticides have been widely applied in various fields to kill plant pathogens, resulting in increased pathogen resistance and decreased effectiveness of such chemicals. In addition, the increased presence of pesticide residues affects living organisms and the environment largely on a global scale. To mitigate the impact of crop diseases more sustainably on plant health and productivity, there is a need for more safe and more eco-friendly strategies as compared to chemical prevention. Quorum sensing (QS) is an intercellular communication mechanism in a bacterial population, through which bacteria adjust their population density and behavior upon sensing the levels of signaling molecules in the environment. As an alternative, quorum quenching (QQ) is a promising new strategy for disease control, which interferes with QS by blocking intercellular communication between pathogenic bacteria to suppress the expression of disease-causing genes. Black rot caused by Xanthomonas campestris pv. campestris (Xcc) is associated with the diffusible signal factor (DSF). As detailed in this study, a new QQ strain F25, identified as Burkholderia sp., displayed a superior ability to completely degrade 2 mM of DSF within 72h. The main intermediate product in the biodegradation of DSF was identified as n-decanoic acid, based on gas chromatography-mass spectrometry (GC-MS). A metabolic pathway for DSF by strain F25 is proposed, based on the chemical structure of DSF and its intermediates, demonstrating the possible degradation of DSF via oxidation-reduction. The application of strain F25 and its crude enzyme as biocontrol agents significantly attenuated black rot caused by Xcc, and inhibited tissue maceration in the host plant Raphanus sativus L., without affecting the host plant. This suggests that agents produced from strain F25 and its crude enzyme have promising applications in controlling infectious diseases caused by DSF-dependent bacterial pathogens. These findings are expected to provide a new therapeutic strategy for controlling QS-mediated plant diseases.

Microplastic accelerate the phosphorus-related metabolism of bacteria to promote the decomposition of methylphosphonate to methane

Microplastic (MP) contaminants in marine water have become a global public health concern because of their persistence and potentially adverse effects on organisms. MP can affect the growth and metabolism of marine microorganisms and further impact the microbial environmental functions. The molecular impact mechanisms of MP on specific functional microbes with the capability of decomposing methylphosphonate (MPn) to release methane (CH4) in oxygenated water have rarely been reported upon. Herein, we investigated the effects of MP on microbes and concomitant methanogenesis via the microbial degradation of MPn. Furthermore, the specific perturbation was revealed at the molecular level combined with transcriptomics and metabolomics. The results showed that intracellular phosphorus utilization by MPn-degrading strain Burkholderia sp. HQL1813 was enhanced by accelerating the catabolism of MPn. Phosphorus transport-related genes (phnG-M, pstSCAB, phnCDE) were upregulated in the MP exposure groups. Amino acid metabolism, the phosphotransferase system and nucleotide metabolism were also perturbed after MP exposure. Notably, released CH4 increased by 24 %, 29 % and 14 % in the exposure group. In addition, the responses of the strain were dose-independent with increasing MP doses. These findings are beneficial for clarifying the effect of MP on specific functional microbes at the molecular level and their degradation of CH4 by MPn.

Characterization of sulfur-oxidizing bacteria isolated from mustard (Brassica juncea L.) rhizosphere having the capability of improving sulfur and nitrogen uptake.

The present investigation was carried out to isolate, screen and characterize potential sulfur-oxidizing bacteria (SOB) isolated from mustard field's soil. A total of 130 bacteria were isolated and after screening five maximum sulfate-producing isolates were optimized for culture conditions. The incubation time of 48 h was found optimum for all bacterial isolates and 30°C was the best temperature for the growth of SSD11, SSR1 and SSG8 whereas 35°C for SSF17. The pH8 was found best for all four isolates except SSF17 (6 pH). Media having glucose as a carbon source and ammonium sulphate as an N-source were producing maximum sulphate. The isolates SSF17, SSR1 and SSG8 were identified as Burkholderia cepacia (accession no. MT559819), Enterobacter cloacae (accession no. MT559820) and Klebsiella oxytoca (accession no. MT372097), respectively, on the basis of morphological, biochemical and molecular characterization. The isolates were also found to increase N and S uptake efficiently in both wheat and mustard crops. This study strongly concludes that SOB isolated from the mustard field can oxidize sulfur in vitro and in vivo conditions. The three best isolates come out of the study were identified as Burkholderia, Enterobacter and Klebsiella strains. Also, inoculation of SOB increased the uptake of S and N nutrient in mustard and wheat crops and thus may be proved as an important plant growth-promoting bacteria having the biofertilization capability. As we know, our soil is continuously deteriorating day by day due to excessive utilization and immoderate use of chemical fertilizers. The SOB could minimize the application of chemical fertilizers thus reducing environmental deterioration by improving soil health in sustainable agricultural practices.

A Wild Rice Rhizobacterium Burkholderiacepacia BRDJ Enhances Nitrogen Use Efficiency in Rice.

Rice domestication has dramatically improved its agronomic traits, albeit with unavoidable significantly reduced genetic diversity. Dongxiang common wild rice, the wild rice species distributed in northernmost China, exhibits excellent resistance against stress and diseases and provides a rich genetic resource for rice breeding. Most of the studies focus on the function of the plant genes, often disregarding the role of the root microbes associated with the plants. In this work, we isolated a Burkholderia strain from the root of Dongxiang wild rice, which we identified as Burkholderia cepacia BRDJ, based on a phylogenetic analysis. This strain promoted the rice growth under greenhouse conditions. The grain yield was higher in a rice line containing a small genomic fragment derived from the Dongxiang wild rice, compared to the indica rice cultivar Zhongzao 35. This new strain also increased the plant biomass under limiting nitrogen conditions. Interestingly, this strain had a differential effect on indica and japonica rice varieties under full nitrogen supply conditions. By genome sequencing and comparison with another two B. cepacia strains, we observed enriched genes related with nitrogen fixation and phytohormone and volatiles biosynthesis that may account for the growth-promoting effects of the BRDJ. BRDJ has the potential to be used as a biofertilizer in promoting nitrogen use efficiency and overall growth in rice.

Bacterial ectosymbionts in cuticular organs chemically protect a beetle during molting stages

In invertebrates, the cuticle is the first and major protective barrier against predators and pathogen infections. While immune responses and behavioral defenses are also known to be important for insect protection, the potential of cuticle-associated microbial symbionts to aid in preventing pathogen entry during molting and throughout larval development remains unexplored. Here, we show that bacterial symbionts of the beetle Lagria villosa inhabit unusual dorsal invaginations of the insect cuticle, which remain open to the outer surface and persist throughout larval development. This specialized location enables the release of several symbiont cells and the associated protective compounds during molting. This facilitates ectosymbiont maintenance and extended defense during larval development against antagonistic fungi. One Burkholderia strain, which produces the antifungal compound lagriamide, dominates the community across all life stages, and removal of the community significantly impairs the survival probability of young larvae when exposed to different pathogenic fungi. We localize both the dominant bacterial strain and lagriamide on the surface of eggs, larvae, pupae, and on the inner surface of the molted cuticle (exuvia), supporting extended protection. These results highlight adaptations for effective defense of immature insects by cuticle-associated ectosymbionts, a potentially key advantage for a ground-dwelling insect when confronting pathogenic microbes.