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1.
Heliyon ; 10(6): e27881, 2024 Mar 30.
Article in English | MEDLINE | ID: mdl-38545156

ABSTRACT

Isolates of Rhizoctonia solani (AG-3 PT, AG-4 HG-I, AG-4 HG-II) and one binucleate Rhizoctonia sp. (BNR) belonging to AG-Bb were investigated for pathogenicity on tomato cultivar Mobil. The BNR isolate revealed the lowest virulence and it was used as biocontrol agent against R. solani AG-4 HG-II, which showed the highest virulence on tomato. Inoculation of tomato plants with the hypovirulent BNR isolate reduced the disease symptoms of R. solani and induced resistance. Resistance induction was observed not only on the plants simultaneously inoculated with BNR and R. solani, but also when the plants were inoculated by the BNR and R. solani with time intervals. The peroxidase (POX), superoxide dismutase (SOD) and catalase (CAT) activities and expression levels of the corresponding genes in tomato plants increased after R. solani or BNR inoculation. The highest level of antioxidant activities and expression of their genes, lignin and callose formation were observed in the plants inoculated with the BNR and R. solani, simultaneously. The BNR inoculation reduced H2O2 accumulation. The highest level of priming was observed for the POX among other antioxidants tested via application of the BNR. Treatment with potassium cyanide (as a POX inhibitor) reduced basal resistance and BNR-induced resistance (BNR-IR) via reduction of lignification and callose deposition in tomato plants. These findings demonstrated the role of antioxidant enzymes, mainly the POX, in both basal resistance and BNR-IR. Therefore, redox state and antioxidants are involved in cell wall strengthening via lignin and callose formation, as important defense components which decrease the pathogen progress in plant tissues.

2.
J Appl Microbiol ; 135(2)2024 Feb 01.
Article in English | MEDLINE | ID: mdl-38346851

ABSTRACT

AIM: To investigate antifungal activity of the extract and major metabolite of the endophytic fungus Acrophialophora jodhpurensis (belonging to Chaetomiaceae) against crown and root rot caused by Rhizoctonia solani (teleomorph: Thanatephorus cucumeris), as an important pathogen of tomato. METHODS AND RESULTS: The endophytic fungus A. jodhpurensis, has high inhibitory effect against R. solani AG4-HG II in vitro and in vivo. The media conditions were optimized for production of the endophyte's metabolites. The highest amounts of secondary metabolites were produced at pH 7, 30°C temperature, and in the presence of 0.5% glucose, 0.033% sodium nitrate, and 1 gl-1 asparagine as the best carbon, nitrogen, and amino acid sources, respectively. The mycelia were extracted by methanol and the obtained extract was submitted to various chromatography techniques. Phytochemical analysis via thin-layer chromatography (TLC) and nuclear magnetic resonance (NMR) spectroscopy showed that ergosterol peroxide was the major component in the extract of this endophyte. Antifungal activities of the methanolic extract and ergosterol peroxide in the culture media were studied against R. solani. Minimum inhibitory concentrations of the extract and ergosterol peroxide against the pathogen were 600 and 150 µg ml-1, respectively. Ergosterol peroxide revealed destructive effects on the pathogen structures in microscopic analyses and induced sclerotia production. Histochemical analyses revealed that it induced apoptosis in the mycelia of R. solani via superoxide production and cell death. Application of ergosterol peroxide in the leaf disc assay reduced the disease severity in tomato leaves. CONCLUSIONS: Antifungal metabolites produced by A. jodhpurensis, such as ergosterol peroxide, are capable of controlling destructive Rhizoctonia diseases on tomato.


Subject(s)
Antifungal Agents , Ergosterol/analogs & derivatives , Rhizoctonia , Sordariales , Antifungal Agents/pharmacology , Antifungal Agents/metabolism , Plant Extracts/pharmacology , Plant Diseases/prevention & control , Plant Diseases/microbiology
3.
Mol Biol Rep ; 51(1): 306, 2024 Feb 16.
Article in English | MEDLINE | ID: mdl-38363387

ABSTRACT

BACKGROUND: Identification and characterization of the endophytic microorganism, is gaining their underestimated significance in influencing health, performance, and other biological attributions of plants in general and forest tree species in particular. Because of the scarcity of information on the endophytic microbiome of the Hyrcanian forests species, including hornbeam (Carpinus betulus L.) trees, as a major constituent thereof, the present study aimed at the identification and partial characterization of the endophytic Bacillus species of Carpinus betulus as the first step in this context. METHODS AND RESULTS: Shoot samples were collected from the Hyrcanian forest locations of Mazandaran and Golestan provinces in Iran. Bacterial strains were isolated from the surface-disinfected shoot segments and subjected to phenotypic characterization. Following assessment of the genetic diversity of the isolates by BOX-PCR fingerprinting, the representative isolates of each of the 15 groups were used for further characterization. Analysis of the nucleotide sequences of the 16S rDNA and HSP60 gene of the isolates led to the identification of 10 species. The predominant species was B. cereus followed by B. subtilis. The other species encountered were B. thuringiensis, Priestia filamentosa, B. velezensis, B. mojavensis, B. amyloliquefaciens, B. safensis, P. aryabhattai, and Gottfriedia acidiceleris. Most isolates possessed characteristics which could contribute to the biocontrol potential of the isolates, including formation of biofilm, production of hydrogen cyanide, tolerant to relatively high concentration of sodium chloride, and antibacterial activity. CONCLUSIONS: Ten Bacillus species were identified as the prevailing endophytic species of C. betulus in the Hyrcanian forest of northern Iran, most turned up to possess biological activities involved in biocontrol capability of the isolates against some plant pathogens. These potentially capable bacteria could be implemented in the promotion of plant growth as well as in the biological control of pathogens. This is the first report on the characterization and elucidation of the diversity of the potentially beneficial endophytic species of Bacillus and the closely related genera living in the internal tissues of hornbeam trees.


Subject(s)
Bacillus , Iran , Phylogeny , Bacillus/genetics , Bacteria/genetics , Forests , Trees , Plants , Betulaceae
4.
Microb Drug Resist ; 28(11): 1003-1018, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36219761

ABSTRACT

Pseudomonas aeruginosa is an opportunistic pathogen that causes several serious health problems and numerous forms of virulence. During the treatment of P. aeruginosa infections, the development of multidrug-resistant isolates creates significant clinical problems. Using antivirulence compounds to disrupt pathogenicity rather than killing the bacterium may be an interesting strategy to overcome this problem, because less harsh conditions will exist for the development of resistance. To reduce pathogenicity and biofilm formation, newly synthesized analogs of imidazolyl (8n) and previously synthesized analogs (8a-8m) with a similar backbone [the 5-(imidazolyl-methyl) thiazolidinediones] were tested against pyoverdine and pyocyanin production, protease activity, and biofilm formation. Compared to the positive control group, the best compounds reduced the production of pyoverdine (8n) by 89.57% and pyocyanin (8i) by 22.68%, and protease activity (8n) by 2.80% for PAO1 strain, at a concentration of 10 µM. Moreover, the biofilm formation assay showed a reduction of 87.94% (8i) for PAO1, as well as 30.53% (8d) and 44.65% (8m) for 1074 and 1707 strains, respectively. The compounds used in this study did not show any toxicity in the human dermal fibroblasts and 4T1 cells (viability higher than 90%). The in silico study of these compounds revealed that their antivirulence activity could be due to their interaction with the PqsR, PqsE, and LasR receptors.


Subject(s)
Pseudomonas aeruginosa , Thiazolidinediones , Humans , Pyocyanine/pharmacology , Quorum Sensing , Biofilms , Anti-Bacterial Agents/pharmacology , Virulence Factors , Thiazolidinediones/pharmacology , Peptide Hydrolases/pharmacology , Bacterial Proteins/pharmacology
5.
Front Plant Sci ; 13: 984583, 2022.
Article in English | MEDLINE | ID: mdl-36212286

ABSTRACT

In this study, efficiency of the endophytic fungal isolate Msh5 was evaluated on promoting tomato plant growth and controlling Alternaria alternata, the causal agent of early blight in tomatoes. Morphological and molecular (ITS and tub2 sequences) analyses revealed that the fungal isolate, Msh5, was Acrophialophora jodhpurensis (Chaetomium jodhpurense Lodha). This beneficial fungus was capable of producing indole-3-acetic acid (IAA), urease, siderophore, extracellular enzymes, and solubilized phosphate. Under laboratory conditions, the Msh5 isolate of A. jodhpurensis inhibited A. alternata growth in dual culture, volatile and non-volatile metabolites assays. The supernatant of this endophytic fungus was capable of reducing spore germination and altering the hyphal structure of A. alternata and the spores produced germ tubes showed vacuolization and abnormal structure compared to the control. Also, the effect of A. jodhpurensis on plant growth parameters (such as shoot and root weight and length) and suppressing A. alternata was investigated in vivo via seed inoculation with spores of A. jodhpurensis using 1% sugar, 0.5% carboxymethyl cellulose (CMC) or 0.5% molasses solution as stickers. Colonization of tomato roots by the endophytic fungus resulted in significant increasing plant growth parameters and reduction in the progress of the diseases caused by A. alternata compared to the controls. Among the different coating materials used as stickers, sugar was found to be the most effective for enhancing plant growth parameters and decreasing the disease progress. Therefore, A. jodhpurensis isolate Msh5 can be suggested as a potential biofertilizer and biocontrol agent for protecting tomato plants against A. alternata.

6.
Biology (Basel) ; 11(1)2022 Jan 14.
Article in English | MEDLINE | ID: mdl-35053136

ABSTRACT

P. aeruginosa strain FG106 was isolated from the rhizosphere of tomato plants and identified through morphological analysis, 16S rRNA gene sequencing, and whole-genome sequencing. In vitro and in vivo experiments demonstrated that this strain could control several pathogens on tomato, potato, taro, and strawberry. Volatile and non-volatile metabolites produced by the strain are known to adversely affect the tested pathogens. FG106 showed clear antagonism against Alternaria alternata, Botrytis cinerea, Clavibacter michiganensis subsp. michiganensis, Phytophthora colocasiae, P. infestans, Rhizoctonia solani, and Xanthomonas euvesicatoria pv. perforans. FG106 produced proteases and lipases while also inducing high phosphate solubilization, producing siderophores, ammonia, indole acetic acid (IAA), and hydrogen cyanide (HCN) and forming biofilms that promote plant growth and facilitate biocontrol. Genome mining approaches showed that this strain harbors genes related to biocontrol and growth promotion. These results suggest that this bacterial strain provides good protection against pathogens of several agriculturally important plants via direct and indirect modes of action and could thus be a valuable bio-control agent.

7.
J Appl Microbiol ; 132(2): 1319-1329, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34480830

ABSTRACT

AIMS: This study aimed to identify virulence-associated genes and functions that affect disease development on pear caused by Erwinia amylovora EaUMG3 isolated from Iran. METHODS AND RESULTS: A mini-Tn5 transposon library was generated in EaUMG3. An E. amylovora mutant that had lost its ability to cause lesions on immature pear fruits, was selected for further analysis. This mutant was shown to have a transposon insertion in yqhC, a gene belongs to the AraC family of transcriptional regulators. A mutant of the wild-type EaUMG3 carrying an unmarked deletion of the yqhC gene was created using pDMS197. The Ea∆yqhC mutant showed reduced disease progression on immature pear fruits and pear plants, reduced motility and significantly lower levels of the virulence factors siderophore and amylovoran. Complementation with yqhC cloned in pBBR1MCS restored disease progression and the level of virulence factors to near wild type. CONCLUSION: YqhC transcriptional regulator is necessary for full virulence of E. amylovora. In addition, this regulator affects virulence factors such as siderophore production, amylovoran production, and motility. SIGNIFICANCE AND IMPACT OF STUDY: The identification of a novel transcriptional regulator with strong impact in the pathogenesis of E. amylovora, an organism causing significant economic losses in fruit production.


Subject(s)
Bacterial Proteins , Erwinia amylovora , Plant Diseases/microbiology , Pyrus , Bacterial Proteins/genetics , Erwinia amylovora/genetics , Erwinia amylovora/pathogenicity , Genes, Bacterial , Pyrus/microbiology , Virulence/genetics
8.
Syst Appl Microbiol ; 42(4): 468-480, 2019 Jul.
Article in English | MEDLINE | ID: mdl-31122691

ABSTRACT

Bacteria of the Pseudomonas putida group are studied for a large panel of properties ranging from plant growth promotion and bioremediation to pathogenicity. To date, most of the classification of individual pseudomonads from this group relies on 16S RNA gene analysis, which is insufficient for accurate taxonomic characterization within bacterial species complexes of the Pseudomonas putida group. Here, a collection of 20 of these bacteria, isolated from various soils, was assessed via multi-locus sequence analysis of rpoD, gyrB and rrs genes. The 20 strains clustered in 7 different clades of the P. putida group. One strain per cluster was sequenced and results were compared to complete genome sequences of type strains of the P. putida group. Phylogenetic analyses, average nucleotide identity data and digital DNA hybridizations, combined to phenotypic characteristics, resulted in the proposition and description of four new species i.e. Pseudomonas alloputida Kh7 T (= LMG 29756 T = CFBP 8484 T) sp. nov., Pseudomonas inefficax JV551A3 T (= DSM108619 T = CFBP 8493 T) sp. nov., Pseudomonas persica RUB6 T (= LMG 29757 T = CFBP 8486 T) sp. nov. and Pseudomonas shirazica VM14 T (= LMG 29953 T = CFBP 8487 T) sp. nov.


Subject(s)
Genome, Bacterial/genetics , Phylogeny , Pseudomonas putida/classification , DNA, Bacterial/genetics , Genes, Bacterial/genetics , Genes, Essential/genetics , Nucleic Acid Hybridization , Phenotype , Pseudomonas putida/genetics , Sequence Analysis, DNA , Soil Microbiology , Species Specificity
9.
Mycologia ; 111(3): 359-370, 2019.
Article in English | MEDLINE | ID: mdl-31021706

ABSTRACT

Farnesol induces morphological changes characteristic of apoptosis in filamentous fungi. Growth-inhibitory effect and induced features of apoptosis on Rhizoctonia solani AG1-IA were observed in our study by addition of exogenous farnesol to the culture. The obtained results implied that farnesol triggered apoptosis-like features, such as production of reactive oxygen species (ROS), in R. solani AG1-IA and that there was increased superoxide dismutase (SOD) activity in the presence of farnesol, as well as decreased fungal biomass. Light microscopic analysis showed that farnesol disrupted the cytoplasm and deformed the hyphae of R. solani AG1-IA. The diameter of the hyphal cross-section in the fungus treated with farnesol decreased compared with control. Transmission electron microscopy (TEM) showed marked alternations in the cell wall, cell membrane, parenthesome, septum, and septal pore of the fungal cells. The findings of this work suggest that farnesol is deleterious to R. solani and has potential for use as an antifungal compound against this destructive phytopathogenic fungus.


Subject(s)
Antifungal Agents/pharmacology , Farnesol/pharmacology , Respiratory Burst/drug effects , Rhizoctonia/drug effects , Rhizoctonia/physiology , Hyphae/drug effects , Microbial Sensitivity Tests , Plant Diseases/microbiology , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolism
10.
Res Microbiol ; 168(8): 760-772, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28851671

ABSTRACT

Fluorescent pseudomonads from bean root and rhizosphere in Iran were investigated for biocontrol of the fungal pathogen Rhizoctonia solani. Phylogenetic analysis of concatenated 16S rRNA, gyrB and rpoD sequences for 33 Pseudomonas isolates showed that 15 belonged to four clusters within the 'P. fluorescens' group, i.e. one corresponding to P. thivervalensis, two others including P. moraviensis or P. baetica, and the last one without closely-related established species. The 18 other isolates belonged to five clusters within the 'P. putida' group, one including P. mosselii and P. entomophila, another including strains currently described as P. putida, and three without closely-related species described. Ten isolates were selected based on in vitro inhibition of R. solani. Cellulase activity was identified in three pseudomonads, chitinase activity in two pseudomonads, extracellular protease activity in nine pseudomonads and hydrogen cyanide production in two pseudomonads. Genes coding for production of phenazine, pyoluteorin, pyrrolnitrin and 2,4-diacetylphloroglucinol were not found, whereas the 1-aminocyclopropane-1-carboxylate deamination gene acdS was present in three pseudomonads. The antagonistic acdS+ strain VKh13 from the 'P. putida' group effectively protected soil-grown bean from R. solani AG 4-HGI. Results show that pseudomonads from uncharacterized taxa were readily obtained from Iranian soils and displayed biocontrol potential against R. solani.


Subject(s)
Antibiosis , Fabaceae/microbiology , Phylogeny , Plant Diseases/microbiology , Pseudomonas/isolation & purification , Pseudomonas/physiology , Rhizoctonia/physiology , Iran , Plant Roots/microbiology , Pseudomonas/classification , Pseudomonas/genetics , Rhizosphere , Soil Microbiology
11.
Molecules ; 22(4)2017 Apr 14.
Article in English | MEDLINE | ID: mdl-28420101

ABSTRACT

Citrus bacterial canker (CBC) caused by Xanthomonas citri subsp. citri (Xcc), is the most devastating of the citrus diseases worldwide. During our study, we found that Essential oils (EOs) of some citrus cultivars are effective on Xcc. Therefore, it prompted us to determine the plant metabolites responsible for the antibacterial properties. We obtained EOs from some locally cultivated citrus by using a Clevenger apparatus and their major constituents were identified by gas chromatography/mass spectrometry (GC-MS). The effect of Citrus aurantium, C. aurantifolia, Fortunella sp. EOs and their major constituents were evaluated against Xcc-KVXCC1 using a disk diffusion assay. Minimal inhibitory and bactericidal concentration of the EOs and their constituents were determined using the broth microdilution method. C. aurantium, C. aurantifolia Eos, and their major constituents including citral, linalool, citronellal, geraniol, α-terpineol, and linalyl acetate indicated antibacterial effects against Xcc. The C. aurantifolia EO and citral showed the highest antibacterial activity among the tested EOs and constituents with inhibition zones of 15 ± 0.33 mm and 16.67 ± 0.88 mm, respectively. Synergistic effects of the constituents were observed between α-terpineol-citral, citral-citronellal, citral-geraniol, and citronellal-geraniol by using a microdilution checkerboard assay. Transmission electron microscopy revealed that exposure of Xcc cells to citral caused cell wall damage and altered cytoplasmic density. We introduced C. aurantifolia and C. aurantium EOs, and their constituents citral, α-terpineol, citronellal, geraniol, and linalool as possible control agents for CBC.


Subject(s)
Citrus/chemistry , Oils, Volatile/pharmacology , Plant Extracts/pharmacology , Xanthomonas/drug effects , Xanthomonas/growth & development , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Drug Synergism , Gas Chromatography-Mass Spectrometry , Microbial Sensitivity Tests , Oils, Volatile/chemistry , Phytochemicals/chemistry , Plant Extracts/chemistry , Xanthomonas/ultrastructure
12.
Genome Announc ; 5(13)2017 Mar 30.
Article in English | MEDLINE | ID: mdl-28360179

ABSTRACT

Here, we report the first draft whole-genome sequence of Pseudomonas fluorescens strain EK007-RG4, which was isolated from the phylloplane of a pear tree. P. fluorescens EK007-RG4 displays strong antagonism against Erwinia amylovora, the causal agent for fire blight disease, in addition to several other pathogenic and non-pathogenic bacteria.

13.
J Plant Physiol ; 199: 29-38, 2016 Jul 20.
Article in English | MEDLINE | ID: mdl-27302004

ABSTRACT

Nitric oxide (NO) is one of the main signal molecules, which is involved in plant growth and development and can change regular physiological activity in biotic and abiotic stresses. In this study, the role of NO in induced resistance with Pseudomonas fluorescent (CHA0) and basal resistance against Rhizoctonia solani in bean plant was investigated. Our results revealed that P. fluorescent and R. solani can increase NO production at 6h post inoculation (hpi). Also, using the NO donor S-nitroso-N-acetyl D-penicillamine (SNAP) led to increase NO and bean plant resistance against R. solani. Utilizing the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethy-limidazoline-1-oxyl-3-oxide (cPTIO), not only decreased basal resistance but also reduced induced resistance. In continue, the activity of antioxidant enzymes was studied in the former treatments. SNAP, CHA0 and R. solani increased the activity of peroxidase (POX), catalase (CAT) and ascorbate peroxidase (APX) at 6, 12 and 24h post inoculation (hpi). In contrast, using cPTIO and R. solani simultaneously (cPTIO+R) showed reduction in activity of POX and APX at 6 hpi. The cPTIO+R treatment increased POX, APX and CAT activity at 12 and 24 hpi. Hydrogen peroxide (H2O2) monitoring in the leaf discs clarified that SNAP can increase H2O2 production like CHA0 and R. solani. On the other hand, SNAP increased the resistance level of leaf discs against R. solani. Treating the leaf discs with cPTIO led to decrease resistance against the pathogen. These leaf discs showed reduction in H2O2 production at 6 hpi and suddenly enhanced H2O2 generation was observed at 24hpi. This study showed that CHA0 can increase NO level in bean plants. NO induced H2O2 generation and regulated redox state of the host plant. This interaction resulted in significant defense against the pathogen.


Subject(s)
Disease Resistance , Nitric Oxide/metabolism , Phaseolus/immunology , Plant Diseases/immunology , Pseudomonas fluorescens/physiology , Rhizoctonia/physiology , Antioxidants/metabolism , Ascorbate Peroxidases/metabolism , Benzoates/pharmacology , Biological Control Agents , Catalase/metabolism , Hydrogen Peroxide/metabolism , Imidazoles/pharmacology , Nitric Oxide Donors/pharmacology , Peroxidases/metabolism , Phaseolus/cytology , Phaseolus/physiology , Plant Diseases/microbiology , Plant Leaves/cytology , Plant Leaves/immunology , Plant Leaves/physiology , S-Nitroso-N-Acetylpenicillamine/pharmacology
14.
World J Microbiol Biotechnol ; 31(10): 1619-27, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26250545

ABSTRACT

To identify novel virulence associated genes in Xanthomonas oryzae pv. oryzae (Xoo), a Xoo isolate (XooIR42), obtained from north of Iran, was selected to generate a mini-Tn5 transposon mutation library. One mutant (XooM176) that indicated reduced virulence on rice plants, while grew similar to wild type was selected. This mutant had an insertion in a coding region with 96% amino acid identity to a response regulator of Xoo KACC10331, citB (Xoo_RS12710). Genome analysis of Xoo KACC10331 indicated several genes including a flagelin protein (FlgL) and a chemotaxis protein (Xoo_RS12720) which were identified as virulence genes 4297 and 1403 nucleotides from the citB, respectively. The swarming motility, resistance to hydrogen peroxide, induced a hypersensitive response, in planta growth and pathogenicity were reduced in XooM176 mutant compared to that of wild-type. A plasmid containing the full citB gene of Xoo KACC10331was sufficient to complement the XooM176 mutant for lesion formation and resistance to hydrogen peroxide. We therefore propose that Xoo requires CitB for full pathogenicity in rice plants and also for protection against oxidative stress.


Subject(s)
Plant Diseases/microbiology , Virulence Factors/genetics , Virulence Factors/metabolism , Xanthomonas/pathogenicity , DNA Transposable Elements , Drug Tolerance , Gene Deletion , Genetic Complementation Test , Hydrogen Peroxide/toxicity , Iran , Mutagenesis, Insertional , Oryza/microbiology , Oxidative Stress , Stress, Physiological , Virulence , Xanthomonas/drug effects , Xanthomonas/genetics , Xanthomonas/growth & development
15.
J Microbiol ; 53(2): 176-80, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25564444

ABSTRACT

Ferula (Ferula asafoetida L.) and Dorema (Dorema aucheri Bioss.) both from Apiaceae family were tested for their anti-quorum sensing (QS) activity against Pseudomonas aeruginosa. Both essential oils exhibited anti-QS activity at 25 µg/ml of concenteration. At this concenteration Ferula fully abolished and Dorema reduced the violacein production by C. violaceum. Pyocyanin, pyoverdine, elastase and biofilm production were decreased in Ferula oil treatments. Dorema oil reduced pyoverdine and elastase production, while pyocyanin and biofilm production were not affacted. Expresion analysis of QS-dependent genes confirmed our phenotypic data. Our data introduced native Dorema and Ferula plants as novel QS and virulence inhibitors.


Subject(s)
Anti-Bacterial Agents/pharmacology , Apiaceae , Ferula , Oils, Volatile/pharmacology , Pseudomonas aeruginosa/drug effects , Quorum Sensing/drug effects , Quorum Sensing/genetics , Bacterial Proteins/metabolism , Biofilms/drug effects , Chromobacterium/drug effects , Chromobacterium/metabolism , Indoles/metabolism , Iron Chelating Agents , Metalloendopeptidases/metabolism , Oils, Volatile/isolation & purification , Oligopeptides/metabolism , Phenotype , Plant Leaves/chemistry , Pseudomonas aeruginosa/physiology , Pyocyanine/metabolism , Virulence/drug effects
16.
Plant Dis ; 96(3): 398-408, 2012 Mar.
Article in English | MEDLINE | ID: mdl-30727144

ABSTRACT

Rhizoctonia spp. are the main causal agents of root and crown rot on sugar beet. In this study, isolates of Rhizoctonia spp. were obtained from diseased sugar beet in Iran over 2 years. Of 68 isolates, 61 were R. solani and 7 were R. cerealis. The anastomosis group (AG) of all isolates was determined on glass slides against the testers. Characterization of intraspecific groups (ISGs) of R. solani isolates revealed that, of 61 isolates, 43 were AG2-2 IIIB and 18 were AG2-2 IV. Amplified fragment length polymorphism (AFLP) analyses were used to investigate genetic structure of Rhizoctonia populations. Principal coordinate plots and cluster analysis differentiated R. solani from R. cerealis isolates and separated the R. solani isolates belonging to different ISGs. AFLP data indicated that the R. solani and R. cerealis populations are not clonal. Analysis of molecular variance in AG2-2 IIIB isolates showed that geographic region was the main factor determining genetic structure of the populations. Sampling year had no significant effect on the genotypes. Pathogenicity tests on Beta vulgaris 'FD0432' revealed that R. solani AG2-2 IIIB and AG2-2 IV isolates were more virulent than R. cerealis.

17.
Arch Virol ; 157(2): 343-7, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22075917

ABSTRACT

Peach latent mosaic viroid isolates from peach and plum in Iran have been compared with an Australian isolate from nectarine. Thirteen sequence variants 336-338 nt in size were obtained. All variants clustered phylogenetically with variants reported from several hosts and countries. A total nucleic acid extract, a slightly longer than full-length RT-PCR amplicon, and a recombinant plasmid clone from the Australian isolate were all infectious to, and symptomatic in, mechanically inoculated peach seedlings. The infectious clone generated two progeny viroid molecules, which each showed 10 different mutations compared with the parent clone inoculated 30 days previously.


Subject(s)
Evolution, Molecular , Genetic Variation , Plant Diseases/virology , Prunus/virology , Viroids/genetics , Viroids/isolation & purification , Australia , Base Sequence , Iran , Molecular Sequence Data , Nucleic Acid Conformation , Phylogeny , Viroids/chemistry , Viroids/classification
18.
PLoS One ; 6(12): e29276, 2011.
Article in English | MEDLINE | ID: mdl-22195037

ABSTRACT

BACKGROUND: Small colony variants (SCVs) are slow-growing bacteria, which often show increased resistance to antibiotics and cause latent or recurrent infections. It is therefore important to understand the mechanisms at the basis of this phenotypic switch. METHODOLOGY/PRINCIPAL FINDINGS: One SCV (termed PAO-SCV) was isolated, showing high resistance to gentamicin and to the cephalosporine cefotaxime. PAO-SCV was prone to reversion as evidenced by emergence of large colonies with a frequency of 10(-5) on media without antibiotics while it was stably maintained in presence of gentamicin. PAO-SCV showed a delayed growth, defective motility, and strongly reduced levels of the quorum sensing Pseudomonas quinolone signal (PQS). Whole genome expression analysis further suggested a multi-layered antibiotic resistance mechanism, including simultaneous over-expression of two drug efflux pumps (MexAB-OprM, MexXY-OprM), the LPS modification operon arnBCADTEF, and the PhoP-PhoQ two-component system. Conversely, the genes for the synthesis of PQS were strongly down-regulated in PAO-SCV. Finally, genomic analysis revealed the presence of mutations in phoP and phoQ genes as well as in the mexZ gene encoding a repressor of the mexXY and mexAB-oprM genes. Only one mutation occurred only in REV, at nucleotide 1020 of the tufA gene, a paralog of tufB, both encoding the elongation factor Tu, causing a change of the rarely used aspartic acid codon GAU to the more common GAC, possibly causing an increase of tufA mRNA translation. High expression of phoP and phoQ was confirmed for the SCV variant while the revertant showed expression levels reduced to wild-type levels. CONCLUSIONS: By combining data coming from phenotypic, gene expression and proteome analysis, we could demonstrate that resistance to aminoglycosides in one SCV mutant is multifactorial including overexpression of efflux mechanisms, LPS modification and is accompanied by a drastic down-regulation of the Pseudomonas quinolone signal quorum sensing system.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial/genetics , Genome, Bacterial/genetics , Pseudomonas aeruginosa/growth & development , Pseudomonas aeruginosa/genetics , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Colony Count, Microbial , Down-Regulation/drug effects , Down-Regulation/genetics , Drug Resistance, Bacterial/drug effects , Gene Expression Profiling , Gene Expression Regulation, Bacterial/drug effects , Genes, Bacterial/genetics , Genetic Variation/drug effects , Gentamicins/pharmacology , Microbial Sensitivity Tests , Oligonucleotide Array Sequence Analysis , Phenotype , Proteome/metabolism , Pseudomonas aeruginosa/drug effects , Quinolones/metabolism , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Transcription, Genetic/drug effects
19.
J Plant Physiol ; 168(10): 1114-22, 2011 Jul 01.
Article in English | MEDLINE | ID: mdl-21269732

ABSTRACT

We examined basal defense responses and cytomolecular aspects of riboflavin-induced resistance (IR) in sugar beet-Rhizoctonia solani pathsystem by investigating H(2)O(2) burst, phenolics accumulation and analyzing the expression of phenylalanine ammonia-lyase (PAL) and peroxidase (cprx1) genes. Riboflavin was capable of priming plant defense responses via timely induction of H(2)O(2) production and phenolics accumulation. A correlation was found between induction of resistance by riboflavin and upregulation of PAL and cprx1 which are involved in phenylpropanoid signaling and phenolics metabolism. Application of peroxidase and PAL inhibitors suppressed not only basal resistance, but also riboflavin-IR of sugar beet to the pathogen. Treatment of the leaves with each inhibitor alone or together with riboflavin reduced phenolics accumulation which was correlated with higher level of disease progress. Together, these results demonstrate the indispensability of rapid H(2)O(2) accumulation, phenylpropanoid pathway and phenolics metabolism in basal defense and riboflavin-IR of sugar beet against R. solani.


Subject(s)
Beta vulgaris/drug effects , Photosensitizing Agents/pharmacology , Plant Immunity/drug effects , Rhizoctonia/immunology , Riboflavin/pharmacology , Beta vulgaris/immunology , Beta vulgaris/microbiology , Gene Expression Regulation, Enzymologic/drug effects , Gene Expression Regulation, Plant/drug effects , Host-Pathogen Interactions , Hydrogen Peroxide/analysis , Hydrogen Peroxide/metabolism , Peroxidases/antagonists & inhibitors , Peroxidases/genetics , Phenols/metabolism , Phenylalanine Ammonia-Lyase/antagonists & inhibitors , Phenylalanine Ammonia-Lyase/genetics , Plant Diseases/immunology , Plant Diseases/microbiology , Plant Immunity/physiology , Plant Leaves/drug effects , Plant Leaves/microbiology , Plant Leaves/physiology , Respiratory Burst/drug effects , Rhizoctonia/pathogenicity , Signal Transduction , Time Factors
20.
World J Microbiol Biotechnol ; 27(6): 1267-80, 2011 Jun.
Article in English | MEDLINE | ID: mdl-25187126

ABSTRACT

The communication or quorum-sensing signal molecules (QSSM) are specialized molecules used by numerous gram-negative bacterial pathogens of animals and plants to regulate or modulate bacterial virulence factor production. In plant-associated bacteria, genes encoding the production of these signal molecules, QSSMs, were discovered to be linked with the phenotype of bacterium, because mutation of these genes typically disrupts some behaviors of bacteria. There are other regulator genes which respond to the presence of signal molecule and regulate the production of signal molecule as well as some virulence factors. The synthesis and regulator genes (collectively called quorum-sensing genes hereafter) are repressed in low bacterial population but induced when bacteria reach to high cell density. Multiple regulatory components have been identified in the bacteria that are under control of quorum sensing. This review describes different communication signal molecules, and the various chemical, physical and genomic factors known to synthesize signals. Likewise, the role of some signal-degrading enzymes or compounds and the interaction of QSSMs with eukaryotic metabolism will be discussed here.

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