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Plants (Basel, Switzerland) May 2021Grapevine bunch rot, caused by and , causes important economic losses every year in grape production. In the present study, we examined the plant protective activity of...
Grapevine bunch rot, caused by and , causes important economic losses every year in grape production. In the present study, we examined the plant protective activity of the biological control agents, K165, sp. FP12 and sp. FP15 against and on grapes. The in vitro experiments showed that strain K165 significantly reduced the growth of both fungi, while FP15 restricted the growth of and FP12 was ineffective. Following the in vitro experiments, we conducted in planta experiments on grape berries. It was shown that K165, FP12 and FP15 reduced rot severity by 81%, 57% and 37%, respectively, compared to the control, whereas, in the case of , the only protective treatment was that with K165, which reduced rot by 75%. The transcriptomic analysis of the genes encoding the pathogenesis-related proteins PR2, PR3, PR4 and PR5 indicates the activation of multiple defense responses involved in the biocontrol activity of the examined biocontrol agents.
PubMed: 34068090
DOI: 10.3390/plants10050970 -
Journal of Bacteriology Feb 2013Paenibacillus alvei CCM 2051(T) cells are decorated with a two-dimensional (2D) crystalline array comprised of the glycosylated S-layer protein SpaA. At its N terminus,...
Paenibacillus alvei CCM 2051(T) cells are decorated with a two-dimensional (2D) crystalline array comprised of the glycosylated S-layer protein SpaA. At its N terminus, SpaA possesses three consecutive surface layer (S-layer) homology (SLH) domains containing the amino acid motif TRAE, known to play a key role in cell wall binding, as well as the TVEE and TRAQ variations thereof. SpaA is predicted to be anchored to the cell wall by interaction of the SLH domains with a peptidoglycan (PG)-associated, nonclassical, pyruvylated secondary cell wall polymer (SCWP). In this study, we have analyzed the role of the three predicted binding motifs within the SLH domains by mutating them into TAAA motifs, either individually, pairwise, or all of them. Effects were visualized in vivo by homologous expression of chimeras made of the mutated S-layer proteins and enhanced green fluorescent protein and in an in vitro binding assay using His-tagged SpaA variants and native PG-containing cell wall sacculi that either contained SCWP or were deprived of it. Experimental data indicated that (i) the TRAE, TVEE, and TRAQ motifs are critical for the binding function of SLH domains, (ii) two functional motifs are sufficient for cell wall binding, regardless of the domain location, (iii) SLH domains have a dual-recognition function for the SCWP and the PG, and (iv) cell wall anchoring is not necessary for SpaA glycosylation. Additionally, we showed that the SLH domains of SpaA are sufficient for in vivo cell surface display of foreign proteins at the cell surface of P. alvei.
Topics: Amino Acid Motifs; Amino Acid Sequence; Bacterial Proteins; Cell Membrane; Cell Wall; Gene Expression Regulation, Bacterial; Glycoproteins; Glycosylation; Membrane Glycoproteins; Paenibacillus; Plasmids; Protein Structure, Tertiary
PubMed: 23204458
DOI: 10.1128/JB.01487-12 -
Glycobiology Jun 2010Glycosylation is a frequent and heterogeneous posttranslational protein modification occurring in all domains of life. While protein N-glycosylation at asparagine and...
Glycosylation is a frequent and heterogeneous posttranslational protein modification occurring in all domains of life. While protein N-glycosylation at asparagine and O-glycosylation at serine, threonine or hydroxyproline residues have been studied in great detail, only few data are available on O-glycosidic attachment of glycans to the amino acid tyrosine. In this study, we describe the identification and characterization of a bacterial protein tyrosine O-glycosylation system. In the Gram-positive, mesophilic bacterium Paenibacillus alvei CCM 2051(T), a polysaccharide consisting of [-->3)-beta-d-Galp-(1[alpha-d-Glcp-(1-->6)] -->4)-beta-d-ManpNAc-(1-->] repeating units is O-glycosidically linked via an adaptor with the structure -[GroA-2-->OPO(2)-->4-beta-d-ManpNAc-(1-->4)] -->3)-alpha-l-Rhap-(1-->3)-alpha-l-Rhap-(1-->3)-alpha-l-Rhap-(1-->3)-beta-d-Galp-(1--> to specific tyrosine residues of the S-layer protein SpaA. A +AH4-24.3-kb S-layer glycosylation (slg) gene cluster encodes the information necessary for the biosynthesis of this glycan chain within 18 open reading frames (ORF). The corresponding translation products are involved in the biosynthesis of nucleotide-activated monosaccharides, assembly and export as well as in the transfer of the completed polysaccharide chain to the S-layer target protein. All ORFs of the cluster, except those encoding the nucleotide sugar biosynthesis enzymes and the ATP binding cassette (ABC) transporter integral transmembrane proteins, were disrupted by the insertion of the mobile group II intron Ll.LtrB, and S-layer glycoproteins produced in mutant backgrounds were analyzed by mass spectrometry. There is evidence that the glycan chain is synthesized in a process comparable to the ABC-transporter-dependent pathway of the lipopolysaccharide O-polysaccharide biosynthesis. Furthermore, with the protein WsfB, we have identified an O-oligosaccharyl:protein transferase required for the formation of the covalent beta-d-Gal-->Tyr linkage between the glycan chain and the S-layer protein.
Topics: Bacillus; Bacterial Proteins; Glycosylation; Mutation; Polysaccharides; Reverse Transcriptase Polymerase Chain Reaction; Tyrosine
PubMed: 20200052
DOI: 10.1093/glycob/cwq035 -
MicrobiologyOpen Mar 2019European foulbrood is a globally distributed brood disease affecting honey bees. It may lead to lethal infections of larvae and, in severe cases, even to colony...
European foulbrood is a globally distributed brood disease affecting honey bees. It may lead to lethal infections of larvae and, in severe cases, even to colony collapse. Lately, a profound genetic and phenotypic diversity was documented for the causative agent Melissococcus plutonius. However, experimental work on the impact of diverse M. plutonius strains on hosts with different genetic background is completely lacking and the role of secondary invaders is poorly understood. Here, we address these issues and elucidate the impact and interaction of both host and pathogen on one another. Moreover, we try to unravel the role of secondary bacterial invasions in foulbrood-diseased larvae. We employed in vitro infections with honey bee larvae from queens with different genetic background and three different M. plutonius strains. Larvae infection experiments showed host-dependent survival dynamics although M. plutonius strain 49.3 consistently had the highest virulence. This pattern was also reflected in significantly reduced weights of 49.3 strain-infected larvae compared to the other treatments. No difference was found in groups additionally inoculated with a secondary invader (Enterococcus faecalis or Paenibacillus alvei) neither in terms of larval survival nor weight. These results suggest that host background contributes markedly to the course of the disease but virulence is mainly dependent on pathogen genotype. Secondary invaders following a M. plutonius infection do not increase disease lethality and therefore may just be a colonization of weakened and immunodeficient, or dead larvae.
Topics: Animals; Bees; Enterococcaceae; Gram-Positive Bacterial Infections; Host-Pathogen Interactions; Larva; Paenibacillus; Survival Analysis
PubMed: 29799173
DOI: 10.1002/mbo3.649 -
Frontiers in Plant Science 2021The sustainable development of agriculture can be stimulated by the great market availability of bio-inputs, including phosphate-solubilizing microbial strains. However,...
The sustainable development of agriculture can be stimulated by the great market availability of bio-inputs, including phosphate-solubilizing microbial strains. However, these strains are currently selected using imprecise and questionable solubilization methodologies in solid or liquid media. We hypothesized that the hydroponic system could be a more efficient methodology for selecting phosphate-solubilizing strains as plant growth promoters. This methodology was tested using the plant as a model. The growth-promoting potential of the strains was compared with that of the Biomaphos® commercial microbial mixture. The obtained calcium phosphate (CaHPO) solubilization results using the hydroponic system were inconsistent with those observed in solid and liquid media. However, the tests in liquid medium demonstrated poor performances of sp. (328EF) and (33EF) in reducing pH and solubilizing CaHPO, which corroborates with the effects of biotic stress observed in plants inoculated with these strains. Nevertheless, the hydroponic system allowed the characterization of (PA12), which is also efficient in solubilization in a liquid medium. The bacterium (PA26) was the most effective in CaHPO solubilization owing to the higher phosphorus (P) absorption, growth promotion, and physiological performance observed in plants inoculated with this bacterium. The hydroponic method proved to be superior in selecting solubilizing strains, allowing the assessment of multiple patterns, such as nutritional level, growth, photosynthetic performance, and anatomical variation in plants, and even the detection of biotic stress responses to inoculation, obtaining strains with higher growth promotion potential than Biomaphos®. This study proposed a new approach to confirm the solubilizing activity of microorganisms previously selected and potentially intended for the bio-input market that are useful in P availability for important crops, such as soybeans.
PubMed: 34777440
DOI: 10.3389/fpls.2021.759463 -
Metabolites Sep 2019Priming is a natural phenomenon that pre-conditions plants for enhanced defence against a wide range of pathogens. It represents a complementary strategy, or sustainable...
Priming is a natural phenomenon that pre-conditions plants for enhanced defence against a wide range of pathogens. It represents a complementary strategy, or sustainable alternative that can provide protection against disease. However, a comprehensive functional and mechanistic understanding of the various layers of priming events is still limited. A non-targeted metabolomics approach was used to investigate metabolic changes in plant growth-promoting rhizobacteria (PGPR)-primed seedlings infected with the anthracnose-causing fungal pathogen, , with a focus on the post-challenge primed state phase. At the 4-leaf growth stage, the plants were treated with a strain of at 10 cfu mL. Following a 24 h PGPR application, the plants were inoculated with a spore suspension (10 spores mL), and the infection monitored over time: 1, 3, 5, 7 and 9 days post-inoculation. Non-infected plants served as negative controls. Intracellular metabolites from both inoculated and non-inoculated plants were extracted with 80% methanol-water. The extracts were chromatographically and spectrometrically analysed on an ultra-high performance liquid chromatography (UHPLC) system coupled to high-definition mass spectrometry. The acquired multidimensional data were processed to create data matrices for chemometric modelling. The computed models indicated time-related metabolic perturbations that reflect primed responses to the fungal infection. Evaluation of orthogonal projection to latent structure-discriminant analysis (OPLS-DA) loading shared and unique structures (SUS)-plots uncovered the differential stronger defence responses against the fungal infection observed in primed plants. These involved enhanced levels of amino acids (tyrosine, tryptophan), phytohormones (jasmonic acid and salicylic acid conjugates, and zeatin), and defence-related components of the lipidome. Furthermore, other defence responses in both naïve and primed plants were characterised by a complex mobilisation of phenolic compounds and biosynthesis of the flavones, apigenin and luteolin and the 3-deoxyanthocyanidin phytoalexins, apigeninidin and luteolinidin, as well as some related conjugates.
PubMed: 31547091
DOI: 10.3390/metabo9100194 -
Applied and Environmental Microbiology Jul 2019Recent papers have reported dipeptides containing d-amino acids to have novel effects that cannot be observed with ll-dipeptides, and such dipeptides are expected to be...
Recent papers have reported dipeptides containing d-amino acids to have novel effects that cannot be observed with ll-dipeptides, and such dipeptides are expected to be novel functional compounds for pharmaceuticals and food additives. Although the functions of d-amino acid-containing dipeptides are gaining more attention, there are few reports on the synthetic enzymes that can accept d-amino acids as substrates, and synthetic methods for d-amino acid-containing dipeptides have not yet been constructed. Previously, we developed a chemoenzymatic system for amide synthesis that comprised enzymatic activation and a subsequent nucleophilic substitution reaction. In this study, we demonstrated the application of the system for d-amino acid-containing-dipeptide synthesis. We chose six adenylation domains as targets according to our newly constructed hypothesis, i.e., an adenylation domain located upstream from the epimerization domain may activate d-amino acid as well as l-amino acid. We successfully synthesized over 40 kinds of d-amino acid-containing dipeptides, including ld-, dl-, and dd-dipeptides, using only two adenylation domains, TycA-A from tyrocidine synthetase and BacB2-A from bacitracin synthetase. Furthermore, this study offered the possibility that the epimerization domain could be a clue to the activity of the adenylation domains toward d-amino acid. This paper provides additional information regarding d-amino acid-containing-dipeptide synthesis through the combination of enzymatic adenylation and chemical nucleophilic reaction, and this system will be a useful tool for dipeptide synthesis. Because almost all amino acids in nature are l-amino acids, the functioning of d-amino acids has received little attention. Thus, there is little information available on the activity of enzymes toward d-amino acids or synthetic methods for d-amino acid-containing dipeptides. Recently, d-amino acids and d-amino acid-containing peptides have attracted attention as novel functional compounds, and d-amino acid-activating enzymes and synthetic methods are required for the development of the d-amino acid-containing-peptide industry. This study provides additional knowledge regarding d-amino acid-activating enzymes and proposes a unique synthetic method for d-amino acid-containing peptides, including ld-, dl-, and dd-dipeptides.
Topics: Amino Acids; Bacillus licheniformis; Bacterial Proteins; Biocatalysis; Dipeptides; Paenibacillus; Peptide Synthases; Protein Domains; Substrate Specificity
PubMed: 31003981
DOI: 10.1128/AEM.00120-19 -
Journal of Bacteriology Nov 2012Paenibacillus alvei is known as a secondary invader during European foulbrood of honeybees. Here, we announce the 6.83-Mb draft genome sequence of P. alvei type strain...
Paenibacillus alvei is known as a secondary invader during European foulbrood of honeybees. Here, we announce the 6.83-Mb draft genome sequence of P. alvei type strain DSM 29. Putative genes encoding an antimicrobial peptide, a binary toxin, a mosquitocidal toxin, alveolysin, and different polyketides and nonribosomal peptides were identified.
Topics: Animals; Bees; Disease Outbreaks; Europe; Genome, Bacterial; Molecular Sequence Data; Paenibacillus
PubMed: 23105091
DOI: 10.1128/JB.01698-12 -
Genomics Data Dec 2017Here we report the draft genome sequence of an endophytic strain isolated from the Universiti Kebangsaan Malaysia reserve forest, Malaysia. The genome size was...
Here we report the draft genome sequence of an endophytic strain isolated from the Universiti Kebangsaan Malaysia reserve forest, Malaysia. The genome size was approximately 8.04 Mb, and the assembly consisted of 107 scaffolds with 168 contigs, and had a G + C content of 53%. Phylogenetic analysis of strain SUK123 using the 16S rRNA gene revealed that it belonged to the family with the highest similarity to SD (99%). Whole genome comparison of SUK123 with related species using average nucleotide identity (ANI) analysis revealed a similarity of 98% to Mst1, 94% to B69, 91% to A2, 68% to SC2 and 69% to DMS29. The draft genome was deposited at the European Nucleotide Archive (PRJEB21373).
PubMed: 28856101
DOI: 10.1016/j.gdata.2017.08.005 -
Advanced Science (Weinheim,... Feb 2024Post-translational prenylations, found in eukaryotic primary metabolites and bacterial secondary metabolites, play crucial roles in biomolecular interactions. Employing...
Post-translational prenylations, found in eukaryotic primary metabolites and bacterial secondary metabolites, play crucial roles in biomolecular interactions. Employing genome mining methods combined with AlphaFold2-based predictions of protein interactions, PalQ , a prenyltransferase responsible for the tryptophan prenylation of RiPPs produced by Paenibacillus alvei, is identified. PalQ differs from cyanobactin prenyltransferases because of its evolutionary relationship to isoprene synthases, which enables PalQ to transfer extended prenyl chains to the indole C3 position. This prenylation introduces structural diversity to the tryptophan side chain and also leads to conformational dynamics in the peptide backbone, attributed to the cis/trans isomerization that arises from the formation of a pyrrolidine ring. Additionally, PalQ exhibited pronounced positional selectivity for the C-terminal tryptophan. Such enzymatic characteristics offer a toolkit for peptide therapeutic lipidation.
Topics: Dimethylallyltranstransferase; Tryptophan; Prenylation; Protein Processing, Post-Translational; Peptides
PubMed: 38059776
DOI: 10.1002/advs.202307372