Amyloliquefaciens FZB42 Research Articles

Microbial Consortia Versus Single-Strain Inoculants as Drought Stress Protectants in Potato Affected by the Form of N Supply

This study investigated the drought protection effects of six fungal and bacterial inoculants and ten consortia thereof on vegetative growth, nutritional status, and tuberization of potato under controlled and field conditions. It was hypothesized that microbial consortia offer improved drought protection as compared with single strains, due to complementary or synergistic effects, with differential impacts also of N fertilization management. Under NO3− fertilization, a 70% reduction in water supply over six weeks reduced shoot and tuber biomass of non-inoculated plants by 30% and 50%, respectively, and induced phosphate (P) limitation compared to the well-watered control. The P nutritional status was significantly increased above the deficiency threshold by three single-strain inoculants and eight consortia. This was associated with the presence of the arbuscular mycorrhizal fungus (AMF) inoculant Rhizophagus irregularis MUCL41833 (five cases) and stimulation of root growth (five cases). Additionally, Bacillus amyloliquefaciens FZB42 and AMF + Pseudomonas brassicacearum 3Re2-7 significantly reduced irreversible drought-induced leaf damage after recovery to well-watered conditions. However, the microbial inoculants did not mitigate drought-induced reductions in tuber biomass, neither in greenhouse nor in field experiments. By contrast, NH4+-dominated fertilization significantly increased tuber biomass under drought stress (534%), which was further increased by additional AMF inoculation (951%). This coincided with (i) improved enzymatic detoxification of drought-induced reactive oxygen species (ROS), (ii) improved osmotic adjustment in the shoot tissue (glycine betaine accumulation), (iii) increased shoot concentrations of ABA, jasmonic acid, and indole acetic acid, involved in drought stress signaling and tuberization, and (iv) reduced irreversible drought-induced leaf damage. Additional application of bacterial inoculants further improved ROS detoxification by increasing the production of antioxidants but stimulated biomass allocation towards shoot growth at the expense of tuber development. The results demonstrated that microbial consortia could increase the probability of drought protection effects influenced by the form of N supply. However, protective effects on vegetative growth do not necessarily translate into yield benefits, which can be achieved by adequate combination of inoculants and fertilizers.

Characterization of three new plant growth-promoting microbes and effects of the interkingdom interactions on plant growth and disease prevention.

The novel interkingdom PGPM consortia enhanced the ability of plant growth promotion and disease resistance, which would be beneficial to improve plant growth in sustainable agriculture through engineering microbiome. Plant growth-promoting microbes (PGPMs) play important roles in promoting plant growth and bio-controlling of pathogens. Much information reveals that the plant growth-promoting ability of individual PGPM affects plant growth. However, the effects of the PGPM consortia properties on plant growth remain largely unexplored. Here, we characterized three new PGPM strains including Rhodotorula graminis JJ10.1 (termed as J), Pseudomonas psychrotolerans YY7 (termed as Y) and P. chlororaphis T8 (termed as T), and assessed their effects in combination with Bacillus amyloliquefaciens FZB42 (termed as F) on plant growth promotion and disease prevention in Arabidopsis thaliana and tomato (Solanum lycopersicum) plants by investigating morphological changes, whole-genome sequencing and plant growth promoting (PGP) characterization. Results revealed that the three new strains R. graminis JJ10.1, P. psychrotolerans YY7 and P. chlororaphis T8 had the potential for being combined with B. amyloliquefaciens FZB42 to form interkingdom PGPM consortia. The combinations of R. graminis JJ10.1, B. amyloliquefaciens FZB42, and P. psychrotolerans YY7, i. e. JF and JYF, exhibited the strongest ability of synergetic biofilm production. Furthermore, the growth-promotion abilities of the consortia were significantly enhanced compared with those of individual strains under both inoculation and volatile organic compounds (VOCs) treatment. Importantly, the consortia showed stronger abilities of in planta disease prevention than individual strains. Findings of our study may provide future guidance for engineering the minimal microbiome communities to improve plant growth and/or disease resistance in sustainable agriculture.

Synergistic combination of arbuscular mycorrhizal fungi and plant growth-promoting rhizobacteria modulates morpho-physiological characteristics and soil structure in Nitraria tangutorum bobr. Under saline soil conditions

Nitraria tangutorum Bobr., a typical xero-halophyte, can be used for vegetation restoration and reconstruction in arid and semiarid regions affected by salinity. However, global climate change and unreasonable human activity have exacerbated salinization in arid and semi-arid regions, which in turn has led to the growth inhibition of halophytes, including N. tangutorum. Arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) have the potential to improve the salt tolerance of plants and their adaptation to saline soil environments. In this study, the effects of single and combined inoculations of AMF (Glomus mosseae) and PGPR (Bacillus amyloliquefaciens FZB42) on N. tangutorum were evaluated in severe saline soil conditions. The results indicate that AMF and PGPR alone may not adapt well to the real soil environment, and cannot ensure the effect of either growth promotion or salt-tolerance induction on N. tangutorum seedlings. However, the combination of AMF and PGPR significantly promoted mycorrhizal colonization, increased biomass accumulation, improved morphological development, enhanced photosynthetic performance, stomatal adjustment ability, and the exchange of water and gas. Co-inoculation also significantly counteracted the adverse effect of salinity on the soil structure of N. tangutorum seedlings. It is concluded that the effectiveness of microbial inoculation on the salt tolerance of N. tangutorum seedlings depends on the functional compatibility between plants and microorganisms as well as the specific combinations of AMF and PGPR.

Suppression of Tomato Bacterial Wilt Incited by Ralstonia pseudosolanacearum Using Polyketide Antibiotic-Producing Bacillus spp. Isolated from Rhizospheric Soil

Bacillus spp. has the potential to control bacterial and fungal diseases of crops. In vitro study, Bacillus amyloliquefaciens DSBA-11 showed best to inhibit the growth of Ralstonia pseudosolanacearum as compared to Bacillus cereus JHTBS-7, B. pumilus MTCC-7092, B. subtilis DTBS-5 and B. licheniformis DTBL-6.Three primers sets from nucleotide sequences of polyketide antibiotic synthase genes viz., macrolactin, difficidin and bacillaene of B. amyloliquefaciens FZB42 were designed and standardized protocol for simultaneous detection of polyketide antibiotics-producing strains of Bacillus spp. by multiplex—PCR with products size of 792 bp, 705 bp and 616 bp respectively. All the strains of B. amyloliquefaciens contained three polyketide antibiotic synthase genes, and B. subtilis possessed difficidin and macrolactin, whereas B. cereus JHTBS-7, B. pumilus MTCC-7092 and B. licheniformis DTBL-6 did not contain any polyketide antibiotic genes. By using this technique, polyketide-producing strains of Bacillus spp. were screened within a short period with high accuracy. These polyketide synthase genes were cloned by using a T&A vector to study the role of these genes in producing antibiotics that suppressed the growth of R. pseudosolanacearum under both in vitro and in vivo conditions. Bio-efficacy of cloned products of these genes macrolactin, bacillaene, and difficidin along with parent strain B. amyloliquefaciens DSBA-11 inhibited the growth of R. pseudosolanacearum and formed 1.9 cm2, 1.9 cm2, 1.7 cm2 and 3.3 cm2 inhibition area under in vitro conditions respectively. Minimum bacterial wilt disease intensity (29.3%) with the highest biocontrol efficacy (57.72%) was found in tomato cv. Pusa Ruby (susceptible to wilt disease) was treated with B. amyloliquefaciens DSBA-11 followed by cloned products of difficidin and macrolactin under glasshouse conditions. Hence, the developed multiplex protocol might be helpful for screening polyketide antibiotics producing potential strains of Bacillus spp. from soil which can apply for managing the wilt disease of tomatoes. The polyketide antibiotics produced by bacteria might have a significant role suppression of R. pseudosolanacearum due to the disintegration of cells.

Genome Sequence Resource of Bacillus velezensis Strain HC-8, a Native Bacterial Endophyte with Biocontrol Potential Against the Honeysuckle Powdery Mildew Causative Pathogen Erysiphe lonicerae var. lonicerae.

Genome Sequence Resource of Bacillus velezensis Strain HC-8, a Native Bacterial Endophyte with Biocontrol Potential Against the Honeysuckle Powdery Mildew Causative Pathogen Erysiphe lonicerae var. lonicerae.

Bacillus amyloliquefaciens Rescues Glycyrrhizic Acid Loss Under Drought Stress in Glycyrrhiza uralensis by Activating the Jasmonic Acid Pathway.

Drought is a major factor limiting the production of the perennial medicinal plant Glycyrrhiza uralensis Fisch. (Fabaceae) in Northwest China. In this study, 1-year-old potted plants were inoculated with the strain Bacillus amyloliquefaciens FZB42, using a gradient of concentrations (CFU), to test for microbe-induced host tolerance to drought condition treatments in a greenhouse experiment. At the concentration of 108 CFU ml–1, FZB42 had significant growth-promoting effect on G. uralensis: the root biomass was 1.52, 0.84, 0.94, and 0.38 times that under normal watering and mild, moderate, and severe drought stress conditions, respectively. Under moderate drought, the positive impact of FZB42 on G. uralensis growth was most pronounced, with both developing axial and lateral roots strongly associated with indoleacetic acid (IAA) accumulation. An untargeted metabolomic analysis and physiological measurements of mature roots revealed that FZB42 improved the antioxidant system of G. uralensis through the accumulation of proline and sucrose, two osmotic adjustment solutes, and by promoting catalase (CAT) activity under moderate drought stress. Furthermore, significantly higher levels of total flavonoids, liquiritin, and glycyrrhizic acid (GA), the pharmacologically active substances of G. uralensis, were found in the roots of inoculated plants after FZB42 inoculation under all imposed drought conditions. The jasmonic acid (JA) content, which is closely related to plant defense responses and secondary metabolites’ production, was greatly increased in roots after the bacterial inoculations, indicating that FZB42 activated the JA pathway. Taken together, our results demonstrate that inoculation with FZB42 alleviates the losses in production and pharmacological metabolites of G. uralensis caused by drought via the JA pathway’s activation. These results provide a developed prospect of a microbial agent to improve the yield and quality of medical plants in arid and semi-arid regions.

Impact of Soil Inoculation with Bacillus amyloliquefaciens FZB42 on the Phytoaccumulation of Germanium, Rare Earth Elements, and Potentially Toxic Elements.

Bioaugmentation promises benefits for agricultural production as well as for remediation and phytomining approaches. Thus, this study investigated the effect of soil inoculation with the commercially available product RhizoVital®42, which contains Bacillus amyloliquefaciens FZB42, on nutrient uptake and plant biomass production as well as on the phytoaccumulation of potentially toxic elements, germanium, and rare earth elements (REEs). Zea mays and Fagopyrum esculentum were selected as model plants, and after harvest, the element uptake was compared between plants grown on inoculated versus reference soil. The results indicate an enrichment of B. amyloliquefaciens in inoculated soils as well as no significant impact on the inherent bacterial community composition. For F. esculentum, inoculation increased the accumulation of most nutrients and As, Cu, Pb, Co, and REEs (significant for Ca, Cu, and Co with 40%, 2042%, and 383%, respectively), while it slightly decreased the uptake of Ge, Cr, and Fe. For Z. mays, soil inoculation decreased the accumulation of Cr, Pb, Co, Ge, and REEs (significant for Co with 57%) but showed an insignificant increased uptake of Cu, As, and nutrient elements. Summarily, the results suggest that bioaugmentation with B. amyloliquefaciens is safe and has the potential to enhance/reduce the phytoaccumulation of some elements and the effects of inoculation are plant specific.

An Endophytic Strain of Bacillus amyloliquefaciens Suppresses Fusarium oxysporum Infection of Chinese Wolfberry by Altering Its Rhizosphere Bacterial Community.

Root rot disease is a serious infection leading to production loss of Chinese wolfberry (Lycium barbarum). This study tested the potential for two bacterial biological control agents, Bacillus amyloliquefaciens HSB1 and FZB42, against five fungal pathogens that frequently cause root rot in Chinese wolfberry. Both HSB1 and FZB42 were found to inhibit fungal mycelial growth, in vitro and in planta, as well as to promote the growth of wolfberry seedlings. In fact, a biocontrol experiment showed efficiency of 100% with at least one treatment involving each biocontrol strain against Fusarium oxysporum. Metagenomic sequencing was used to assess bacterial community shifts in the wolfberry rhizosphere upon introduction of each biocontrol strain. Results showed that HSB1 and FZB42 differentially altered the abundances of different taxa present and positively influenced various functions of inherent wolfberry rhizosphere bacteria. This study highlights the application of biocontrol method in the suppression of fungal pathogens that cause root rot disease in wolfberry, which is useful for agricultural extension agents and commercial growers.

Novel Genetic Dysregulations and Oxidative Damage in Fusarium graminearum Induced by Plant Defense Eliciting Psychrophilic Bacillus atrophaeus TS1.

This study elaborates inter-kingdom signaling mechanisms, presenting a sustainable and eco-friendly approach to combat biotic as well as abiotic stress in wheat. Fusarium graminearum is a devastating pathogen causing head and seedling blight in wheat, leading to huge yield and economic losses. Psychrophilic Bacillus atrophaeus strain TS1 was found as a potential biocontrol agent for suppression of F. graminearum under low temperature by carrying out extensive biochemical and molecular studies in comparison with a temperate biocontrol model strain Bacillus amyloliquefaciens FZB42 at 15 and 25 °C. TS1 was able to produce hydrolytic extracellular enzymes as well as antimicrobial lipopeptides, i.e., surfactin, bacillomycin, and fengycin, efficiently at low temperatures. The Bacillus strain-induced oxidative cellular damage, ultrastructural deformities, and novel genetic dysregulations in the fungal pathogen as the bacterial treatment at low temperature were able to downregulate the expression of newly predicted novel fungal genes potentially belonging to necrosis inducing protein families (fgHCE and fgNPP1). The wheat pot experiments conducted at 15 and 25 °C revealed the potential of TS1 to elicit sudden induction of plant defense, namely, H2O2 and callose enhanced activity of plant defense-related enzymes and induced over-expression of defense-related genes which accumulatively lead to the suppression of F. graminearum and decreased diseased leaf area.

Chemical interplay and complementary adaptative strategies toggle bacterial antagonism and co-existence.

SummaryBacterial communities are in a continuous adaptive and evolutionary race for survival. In this work we expand our knowledge on the chemical interplay and specific mutations that modulate the transition from antagonism to co-existence between two plant-beneficial bacteria, Pseudomonas chlororaphis PCL1606 and Bacillus amyloliquefaciens FZB42. We reveal that the bacteriostatic activity of bacillaene produced by Bacillus relies on an interaction with the protein elongation factor FusA of P. chlororaphis and how mutations in this protein lead to tolerance to bacillaene and other protein translation inhibitors. Additionally, we describe how the unspecific tolerance of B. amyloliquefaciens to antimicrobials associated with mutations in the glycerol kinase GlpK is provoked by a decrease of Bacillus cell membrane permeability, among other pleiotropic responses. We conclude that nutrient specialization and mutations in basic biological functions are bacterial adaptive dynamics that lead to the coexistence of two primary competitive bacterial species rather than their mutual eradication.

Isolation, heterologous expression, and purification of a novel antifungal protein from Bacillus subtilis strain Z-14

BackgroundWheat sheath blight, a soil borne fungal disease caused by Rhizoctonia cerealis, is considered as one of the most serious threats to wheat worldwide. Bacillus subtilis Z-14 was isolated from soil sampled from a wheat rhizosphere and was confirmed to have strong antifungal activity against R. cerealis.ResultsAn antifungal protein, termed F2, was isolated from the culture supernatant of Z-14 strain using precipitation with ammonium sulfate, anion exchange chromatography, and reverse phase chromatography. Purified F2 had a molecular mass of approximately 8 kDa, as assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Edman degradation was used to determine the amino acid sequence of the N-terminus, which was NH2ASGGTVGIYGANMRS. This sequence is identical to a hypothetical protein RBAM_004680 (YP_001420098.1) synthesized by B. amyloliquefaciens FZB42. The recombinant F2 protein (rF2) was heterologously expressed in the yeast host Pichia pastoris, purified using a Niaffinity column, and demonstrated significant antifungal activity against R. cerealis. The purified rF2 demonstrated broad spectrum antifungal activity against different varieties of fungi such as Fusarium oxysporum, Verticillium dahliae, Bipolaris papendorfii, and Fusarium proliferatum. rF2 was thermostable, retaining 91.5% of its activity when incubated for 30 min at 100 °C. Meanwhile, rF2 maintained its activity under treatment by proteinase K and trypsin and over a wide pH range from 5 to 10.ConclusionsA novel antifungal protein, F2, was purified from biocontrol Bacillus subtilis Z-14 strain fermentation supernatant and heterologously expressed in Pichia pastoris to verify its antifungal activity against R. cerealis and the validity of the gene encoding F2. Considering its significant antifungal activity and stable characteristics, protein F2 presents an alternative compound to resist fungal infections caused by R. cerealis.

Induction of Salt Tolerance in Arabidopsis thaliana by Volatiles From Bacillus amyloliquefaciens FZB42 via the Jasmonic Acid Signaling Pathway.

Previously, we showed that Bacillus amyloliquefaciens FZB42 can confer salt tolerance in plants by root inoculation under salt stress condition, and the FZB42 volatile organic compounds (VOCs) promoted plant growth and development under non-salt stress condition. In the present study, we investigated the mechanism that allows FZB42 VOCs to confer salt tolerance in Arabidopsis without colonization of plant roots. We found that FZB42 VOCs significantly increased the biomass of Arabidopsis and also maintained the leaf chlorophyll content under salt stress condition. Physiological tests showed that the plant anti-oxidation system was activated by FZB42 VOCs, where higher peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) activities were detected in plants exposed to FZB42 VOCs compared with non-exposed plants. In addition, FZB42 VOCs increased the leaf total soluble sugars (TSS) content but decreased the proline content compared with the non-exposed plants. Moreover, FZB42 VOCs significantly decreased the Na+ contents of the whole plants and induced the expression of genes (NHX1; Na+/H+ exchanger 1 and HKT1; high-affinity K+ transporter 1) that function to alleviate Na+ toxicity. Furthermore, analysis of mutants with defects in specific hormone pathways showed that FZB42 VOCs induced salt tolerance in plants by modulating jasmonic acid (JA) signaling, which was confirmed by the up-regulation of JA synthesis, defense-related genes, and JA biosynthesis inhibitor tests. The results of this study provide new insights into the molecular mechanism related to the interactions between plant growth-promoting rhizobacteria and plants under salt stress condition.

Screening for Candidate Genes Associated with Biocontrol Mechanisms of Bacillus pumilus DX01 Using Tn5 Transposon Mutagenesis and a 2-DE-Based Comparative Proteomic Analysis.

A total of 1467 mutants of the biocontrol bacterium Bacillus pumilus DX01 were obtained by Tn5 insertional mutagenesis and subjected to the determination of antagonistic capabilities. Compared with the wild-type strain DX01, the mutant M25 was identified to have the most significant reduction in antagonistic capability against the phytopathogen Bipolaris maydis and extracellular proteinase activity. The integration site of the exogenous T-DNA in the genome of mutant M25 was revealed in the coding region of malony CoA-ACP transacylase (MCAT) gene (mcat), which belongs to a polyketide synthase (PKS) gene cluster, DX01pks of B. pumilus DX01. Furthermore, the whole DX01pks gene cluster was cloned using Illumina Solexa sequencing technology, and it has a modular framework different from the other two gene clusters involved in polyketide synthesis in B. amyloliquefaciens FZB42 (pks1) and B. subtilis 168 (pksX). Finally, in order to gain more insights into the molecular mechanisms of biocontrol of B. pumilus DX01, the changes in the relative level of expression of total proteins between the original strain DX01 and the mutant M25 were detected by 2-DE-based proteomic analysis. A total of twenty differentially expressed proteins were identified upon the mcat gene transposition mutagenesis. Of these proteins, seven proteins were up-regulated in expression level and the other proteins were down-regulated. Taken together, the results in this study showed that Tn5 transposon mutagenesis of B. pumilus DX01 can lead to a significant change of antiphytopathogen ability, and the DX01pks gene cluster possibly play a potential role in the biocontrol processes of this bacterium.

Acquisition of rock phosphate by combined application of ammonium fertilizers and Bacillus amyloliquefaciens FZB42 in maize as affected by soil pH.

The use of plant growth-promoting micro-organisms (PGPMs) to improve plant-nutrient acquisition has a long history but reproducibility remains a challenge. Recent findings suggest an important role of suitable inoculant-fertilizer combinations for the expression of PGPM-effects, particularly with respect to nitrogen (N) supply. In face of the well-documented N form effects on rhizosphere pH, this study addressed the impact of ammonium-assisted PGPM-interactions on the acquisition of sparingly soluble calcium-phosphates as affected by soil pH. The effects of stabilized ammonium fertilization combined with the PGPM inoculant Bacillus amyloliquefaciens FZB42 on the acquisition of rock phosphate in maize were examined on two soils (moderately acidic-pH 5·6 and alkaline-pH 7·8). On the two contrasting soils, FZB42 improved the P status and promoted plant growth by different mechanisms. On the acidic soil, a combination of ammonium-fertilization with FZB42 increased P-acquisition by Rock P solubilization via rhizosphere acidification but P-supply in the noninoculated control was already sufficient to meet the plant demands. By contrast, on the alkaline soil, plant growth-promotion was associated with FZB42-induced root growth stimulation. The results suggest a significant impact of soil pH on performance and the mode of action of PGPM inoculants, to be considered for practical applications. The study advanced existing knowledge on PGPM-assisted P solubilization as affected by different soil properties. The results suggest perspectives for management options to be considered for efficient use of PGPMs in terms of selecting application strategies with compatible PGPM-fertilizer combinations, depending on soil pH conditions.

Biosynthesis of poly‐γ‐glutamic acid in Escherichia coli by heterologous expression of pgsBCAE operon from Bacillus

Poly-γ-glutamic acid (γ-PGA) is an excellent water-soluble biosynthesis material. To confirm the rate-limiting steps of γ-PGA biosynthesis pathway, we introduced a heterologous Bacillus strain pathway and employed an enzyme-modulated dismemberment strategy in Escherichia coli. In this study, we heterologously introduced the γ-PGA biosynthesis pathway of two laboratory-preserved strains-Bacillus amyloliquefaciens FZB42 and Bacillus subtilis 168 into E. coli, and compared their γ-PGA production levels. Next,by changing the plasmid copy numbers and supplying sodium glutamate, we explored the effects of gene expression levels and concentrations of the substrate l-glutamic acid on γ-PGA production. We finally employed a two-plasmid induction system using an enzyme-modulated dismemberment of pgsBCAE operon to confirm the rate-limiting genes of the γ-PGA biosynthesis pathway. Through heterologously over-expressing the genes of the γ-PGA biosynthesis pathway and exploring gene expression levels, we produced 0·77gl-1 γ-PGA in strain RSF-EBCAE(BS). We also confirmed that the rate-limiting genes of the γ-PGA biosynthesis pathway were pgsB and pgsC. This work is beneficial to increase γ-PGA production and study the mechanism of γ-PGA biosynthesis enzymes.

Functional Analysis of a Gene Cluster from Chitinophaga pinensis Involved in Biosynthesis of the Pyrrolidine Azasugar DAB-1.

Azasugars, "nitrogen in the ring" analogues of monosaccharides, are known to be distributed in select plant, fungal. and bacterial species. We identify Chitinophaga pinensis DSM 2588 as the first bacterial source of the plant pyrrolidine azasugar 1,4-dideoxy-1,4-aminoarabinitol (DAB-1). Comparative sequence analyses identified C. pinensis as a putative azasugar producer, via observation of a three-gene cluster coding for putative aminotransferase, alcohol dehydrogenase, and sugar phosphatase enzymes, similar to the previously reported azasugar biosynthetic signature identified in Bacillus amyloliquefaciens FZB42. Multistep fractionation of C. pinensis culture media guided by a maltase inhibition assay yielded a component with a mass consistent with the structure of DAB-1. Heterologous expression of the three-gene cluster in E. coli, a non-azasugar producer, led to the isolation of nectrisine, a biosynthetic precursor to DAB-1, which displayed potent slow tight binding inhibition of maltase. Reduction of nectrisine with NaBH4 removed the slow tight binding inhibition kinetics, and MS analysis provided evidence for the production of a compound matching that of the isolated DAB-1 from C. pinensis. 1H NMR analysis of the nectrisine produced in E. coli after NaBD4 reduction produced a spectrum consistent with DAB-1 deuterated at C-1, primarily at the pro-S position. These results support the idea that the azasugar three-gene cluster represents a general biosynthetic path leading to several different compounds, which may prove useful for the identification of other azasugar-producing organisms.

Zn(II) suppresses biofilm formation in Bacillus amyloliquefaciens by inactivation of the Mn(II) uptake.

Biofilms are architecturally complex communities of microbial cells held together by a self-produced extracellular matrix. Considerable research has focused on the environmental signals that trigger or inhibit biofilm formation by affecting cellular signalling pathways; however, response to soil cues in plant-associated Bacillus has remained largely unaddressed. Therefore, we aimed to investigate the effect of Zn(II) ions in biofilm formation of Bacillus amyloliquefaciens FZB42. We demonstrated that the biofilm formation of B. amyloliquefaciens FZB42 was abolished by Zn(II) at non-deleterious concentrations. Moreover, Zn(II) blocked matrix exopolysaccharide and TasA accumulations. Furthermore, the presence of Zn(II) suppressed expression of the response regulator Spo0F but not of sensor histidine kinases KinA-D. Suppression of phosphorelay by excess Zn interferes with sinI induction under biofilm-inducing conditions, leading to repression of transcription of operons epsA-O and tapA-sigW-tasA. Addition of Zn(II) decreased the intracellular Mn(II) level by competing for binding to the solute-binding protein MntA during Mn(II) uptake. These results suggest that the metal ion Zn(II) has a negative effect on biofilm formation in the plant growth promoting and biocontrol bacterium B. amyloliquefaciens FZB42.

Suppression of Sclerotinia sclerotiorum by the Induction of Systemic Resistance and Regulation of Antioxidant Pathways in Tomato Using Fengycin Produced by Bacillus amyloliquefaciens FZB42.

Lipopeptides from Bacillus species exhibit promising biological control activity against plant pathogens. This study aimed to explore the potential of purified fengycin to induce systemic resistance in tomato against Sclerotinia sclerotiorum. Bacillus amyloliquefaciens FZB42, its mutant AK1S, and their corresponding metabolites showed in vitro inhibition of S. sclerotiorum mycelium. Fengycin derived from an AK1S mutant was purified and identified through HPLC and MALDI-TOF-MS, respectively. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed structural deformities in the fungal mycelium. Moreover, fengycin induced the accumulation of reactive oxygen species (ROS) in S. sclerotiorum mycelium and downregulated the expression of ROS-scavenging genes viz., superoxide dismutase (SsSOD1), peroxidase (SsPO), and catalase (SsCAT1) compared to the untreated control. Furthermore, the lesion size was dramatically reduced in fengycin-treated tomato plants compared to plants infected with S. sclerotiorum only in a greenhouse experiment. Additionally, the transcriptional regulation of defense-related genes GST, SOD, PAL, HMGR, and MPK3 showed the highest upsurge in expression at 48 h post-inoculation (hpi). However, their expression was subsequently decreased at 96 hpi in fengycin + S. sclerotiorum treatment compared to the plants treated with fengycin only. Conversely, the expression of PPO increased in a linear manner up to 96 hpi.

The role of N form supply for PGPM‐host plant interactions in maize

AbstractThe form of nitrogen (N) supply has a significant impact on rhizosphere chemistry and root growth responses of higher plants. The respective effects are also employed as management options to improve nutrient acquisition and to minimize nutrient losses in cropping systems. However, surprisingly little is known concerning the interactions with rhizosphere biota. In this study, we investigated the effects of selected bacterial and fungal inoculants with proven plant growth‐promoting and phosphate (P)‐solubilizing potential (plant growth‐promoting microorganisms, PGPM) in maize with nitrate or stabilized ammonium supply, on soils with limited P availability and sparingly soluble rock phosphate (Rock‐P) applied as P fertilizer. The combination of the bacterial inoculants Pseudomonas sp. DSMZ 13134 (Proradix) and Bacillus amyloliquefaciens FZB42 with ammonium sulphate fertilization, stabilized with the nitrification inhibitor 3,4‐dimethylpyrazole‐phosphate (DMPP), resulted in a superior shoot biomass production (79–111%) and shoot P accumulation (109–235%) as compared with nitrate supply. This effect could be partially attributed to (1) ammonium‐induced rhizosphere acidification via increased root extrusion of protons, (2) promotion of root hair elongation, and (3) increased shoot concentrations of hormonal growth regulators (indole‐3‐acetic acid, zeatin, gibberellic acid). The effects, induced by the microbial inoculants were mainly related to increased root length development (43–44%), associated with a 60% increase in auxin production potential. No inoculant effects were detected on root hair elongation or on chemical modifications of the rhizosphere involved in P solubilisation, such as rhizosphere acidification, release of carboxylates or secretory phosphohydrolases. However, the ammonium‐induced stimulation of root hair elongation increased preferential sites for root colonization by the selected inoculants, which may explain the increase in rhizosphere abundance of PGPMs, exemplarily recorded for the fungal inoculant Trichoderma harzianum OMG16 (210%). The presented data suggest a network of positive interactions between stabilized ammonium fertilization and plant growth‐promoting functions of various bacterial and fungal PGPM inoculants. This offers perspectives to increase the efficiency and the reproducibility of PGPM‐assisted fertilization strategies.

Enhanced tomato plant growth in soil under reduced P supply through microbial inoculants and microbiome shifts.

Soil microbial communities interact with roots, affecting plant growth and nutrient acquisition. In the present study, we aimed to decipher the effects of the inoculants Trichoderma harzianum T-22, Pseudomonas sp. DSMZ 13134, Bacillus amyloliquefaciens FZB42 or Pseudomonas sp. RU47 on the rhizosphere microbial community and their beneficial effects on tomato plants grown in moderately low phosphorous soil under greenhouse conditions. We analyzed the plant mass, inoculant colony forming units and rhizosphere communities on 15, 22, 29 and 43 days after sowing. Selective plating showed that the bacterial inoculants had a good rhizocompetence and accelerated shoot and root growth and nutrient accumulation. 16S rRNA gene fingerprints indicated changes in the rhizosphere bacterial community composition. Amplicon sequencing revealed that rhizosphere bacterial communities from plants treated with bacterial inoculants were more similar to each other and distinct from those of the control and the Trichoderma inoculated plants at harvest time, and numerous dynamic taxa were identified. In conclusion, likely both, inoculants and the rhizosphere microbiome shifts, stimulated early plant growth mainly by improved spatial acquisition of available nutrients via root growth promotion. At harvest, all tomato plants were P-deficient, suggesting a limited contribution of inoculants and the microbiome shifts to the solubilization of sparingly soluble soil P.