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1.
Molecules ; 28(2)2023 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-36677848

RESUMO

Tobacco target spot disease is caused by Rhizoctonia solani AG-3 TB, which causes serious harm to the quality and yield of tobacco. In this study, thin layer chromatography (TLC), high performance liquid chromatography (HPLC), infrared absorption spectroscopy (IR), and nuclear magnetic resonance spectroscopy (NMR) were used to purify and identify the potential phytotoxin produced by R. solani AG-3 TB. The result indicated that the purified toxin compound was 3-methoxyphenylacetic acid (3-MOPAA) (molecular formula: C9H10O3). The exogenous purified compound 3-MOPAA was tested, and the results revealed that 3-MOPAA can cause necrosis in tobacco leaves. 3-MOPAA is a derivative of phenylacetic acid (PAA), which should be produced by specific enzymes, such as hydroxylase or methylase, in the presence of PAA. These results enrich the research on the pathogenic phytotoxins of R. solani and provide valuable insights into the pathogenic mechanism of AG-3 TB.


Assuntos
Tabaco , Toxinas Biológicas , Pirrolidinonas , Rhizoctonia
2.
Lett Appl Microbiol ; 76(1)2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36688753

RESUMO

In genome analyses of Rhizoctonia solani AG1-IA causing sheath blight (ShB) of rice, many genes were identified to have a hypothetical role in pathogenesis. To understand their roles in pathogenesis, their expressions during fungal infection were studied. An aggressive R. solani strain, RIRS-K, was first identified among six isolates, RIRS-K, RIRS-17, RIRS-S, RIRS-T, RIRS-MU and RIRS-FD, for inducing a maximum relative lesion height (RLH) of 32.7% on a ShB susceptible cultivar, Pusa Basmati-1. Hypothetical pathogenicity genes (52 nos) identified by in silico analyses of the publicly available genomic database of the pathogen strain were evaluated in Pathogen-Host Interaction (PHI) blast and RIRS-K. Though PHI blast identified 26 genes as potential ones, only 8 were constitutively expressive in RIRS-K cultured in a minimal broth. Among them, only expressions of AG1IA_06195, AG02692, AG04508, and AG05730 were induced in the rice plant inoculated with RIRS-K and, hence, were identified as the candidate ones. The candidate genes were highly expressed in the aggressive strain (RIRS-K) in comparison to the less aggressive one (RIRS-17). In further testing of their expressions in the highly aggressive fungal strain, RIRS-K infecting PB-1 pre-colonized by a potent biocontrol consortium comprising of Bacillus subtilis (S17TH), Pseudomonas putida (TEPF-Sungal-1), and Trichoderma harzianum (S17TH), the disease scoring and gene expression studies indicated that the candidate genes were downregulated. The studies, therefore, speculated that these genes might play a role in pathogen aggressiveness and ShB development.


Assuntos
Oryza , Oryza/microbiologia , Doenças das Plantas/microbiologia , Genoma Fúngico , Rhizoctonia/genética
3.
J Biotechnol ; 362: 1-11, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36513313

RESUMO

qRT-PCR is a globally accepted technique for assaying gene expression in relative terms which compares the difference between critical threshold (CT) values of a gene calculated form two independently isolated RNA samples. Independent RNA isolations, however, include error due to batch effect which must be normalized for error-free calculation of relative gene expression. Hence, CT values of internal control (IC) genes are used for normalization during the calculation of expression fold-change in gene expression analysis. The expression of ICs genes expected to be stable in all the experimental conditions. However, it is almost impossible to find such a gene which do not depict expression fluctuation in response to the changes in experimental conditions. Hence, it is necessary to identify suitable IC gene(s) for any given experimental condition before conducting any particular gene expression study. Here, we examined the suitability of eight candidate IC genes, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), eukaryotic elongation factor-1 (eEF-1α), 25 S rRNA (25 S), 18 S rRNA (18 S), ubiquitin C E2 ligase (UBC), Actin (Act), ubiquitin 5 (UBQ5) and ubiquitin 10 (UBQ10), for assaying gene expression in rice during sheath blight infection. Our analysis suggest that GAPDH might be the IC of choice when expression studies include contrasting genotypes differing in their tolerance to sheath blight pathogen as well as progressive infection time. While if expression analysis have to be performed only in one genotype but under progressive sheath blight infection, UBQ5 might be chosen as IC because of its high expression stability under the proposed experimental setup.


Assuntos
Oryza , Oryza/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Genes de Plantas , Perfilação da Expressão Gênica/métodos , Gliceraldeído-3-Fosfato Desidrogenases , Ubiquitina/genética , Expressão Gênica , RNA , Rhizoctonia , Doenças das Plantas/genética
4.
Planta ; 257(1): 13, 2022 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-36522558

RESUMO

MAIN CONCLUSION: Screening for resistance in 40 potato genotypes to Rhizoctonia solani AG-3PT-stem-canker, antioxidant enzymes activity as well as total phenol compounds were documented. Rhizoctonia solani AG-3PT-stem-canker is one of the most devastating diseases that leads to severe economic losses in potatoes, Solanum tuberosum globally. Crop management and eugenic practices, especially the use of resistance can be effective in reducing the disease incidence. However, the information about potato-R. Solani interaction is still limited. This study explored screening for resistance in forty potato genotypes to R. solani, analyzing biomass growth parameters (BGPs), as well as antioxidant enzymes activity of which peroxidase/peroxide-reductases (POXs), superoxide dismutase (SOD), polyphenol oxidase (PPO), catalase (CAT), phenylalanine ammonia-lyase (PAL), ß-1,3-glucanase (GLU) and total phenol compounds (TPCs) were taken into account. In addition, we analyzed up-regulation of two gene markers (PR-1 and Osmotin), using reverse transcription quantitative PCR (RT-qPCR). For which, the resistant 'Savalan', partially resistant 'Agria', partially susceptible 'Sagita' and susceptible 'Pashandi' were selected to explore the trails in their roots and leaves over the time courses of 1, 2 and 3-weeks post inoculation (wpi) following inoculation. Cluster analysis divided potatoes into four distinct groups, based on disease severity scales (0-100%) significance. The BGPs, shoot and root length, fresh and dry weight, and root volume were also significantly higher in infected potatoes compared to non-inoculated controls. Antioxidant enzymes activity also indicated the highest increased levels for POX (fourfold at 3wpi), CAT (1.5-fold at 3wpi), SOD (6.8-fold at 1wpi), and PAL (2.7-fold at 3wpi) in the resistant genotype, 'Savalan', whereas the highest activity was recorded in TPC (twofold at 1 wpi), PPO (threefold at 3wpi), and GLU (2.3-fold at 1wpi) in partially resistant genotypes. Although the defense-related enzymatic activities were sharply elevated in the resistant and partially resistant genotypes following inoculation, no significant correlations were between the activity trends of the related enzymes. The two related gene markers also showed comprehensive transcriptional responses up to 3.4-fold, predominantly in resistant genotypes. Surprisingly, the PR-1 gene marker, basically resistant to Wilting agent Verticillium dahlia was overexpressed in resistant 'Savalan' and 'Agria' against R. solani AG3-PT. Similar results were obtained on Osmotin gene marker resistant to late-blight P. infestans, and early-blight Alternaria solani that similarly modulates immunity against R. solani. Furthermore, there was a significant correlation between resistance, enzyme activity, and gene expression in the aforesaid cultivars. Studying the physiological metabolic pathways of antioxidant enzymes activity appears to be an important direction in research to elucidate resistance to R. solani in potatoes.


Assuntos
Solanum tuberosum , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Resistência à Doença/genética , Antioxidantes , Doenças das Plantas , Rhizoctonia/fisiologia , Fenilalanina Amônia-Liase/genética , Fenilalanina Amônia-Liase/metabolismo , Catecol Oxidase/metabolismo , Superóxido Dismutase , Fenóis , Mecanismos de Defesa
5.
Molecules ; 27(21)2022 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-36364004

RESUMO

This study aimed to synthesize, characterize, and explore the eco-friendly and antifungal potential of precocenes and their derivatives. The organic synthesis of the mono-O-alkyl-2,2-dimethyl 2H-1-chromene series, including the natural product precocene I, and the di-O-alkyl 2,2-dimethyl-2H-1-chromene series, including the natural 2H-1-chromenes precocenes II and III, was achieved. The synthetic compounds were subjected to spectroscopic analysis, 1HNMR,13CNMR, and mass characterization. The antifungal activity of synthesized precocenes I, II, and III, as well as their synthetic intermediates, was evaluated by the poison food technique. Precocene II (EC50 106.8 µg × mL-1 and 4.94 µg mL-1), and its regioisomers 7a (EC50 97.18 µg × mL-1 and 35.30 µg × mL-1) and 7d (EC50 170.58 × µg mL-1), exhibited strong fungitoxic activity against Aspergillus niger and Rhizoctonia solani. Some of the novel chromenes, 11a and 11b, which had never been evaluated before, yielded stronger fungitoxic effects. Finally, docking simulations for compounds with promising fungitoxic activity were subjected to structure-activity relationship analyses against the polygalactouronases and voltage-dependent anion channels. Conclusively, precocenes and their regioisomers demonstrated promising fungitoxic activity; such compounds can be subjected to minor structural modifications to yield promising and novel fungicides.


Assuntos
Antifúngicos , Fungicidas Industriais , Antifúngicos/química , Rhizoctonia , Fungicidas Industriais/farmacologia , Relação Estrutura-Atividade , Benzopiranos/farmacologia
6.
Plant Physiol Biochem ; 193: 78-89, 2022 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-36343463

RESUMO

Banded leaf and sheath blight (BLSB) is a devasting disease caused by the necrotrophic fungus Rhizoctonia solani that affects maize (Zea mays L.) fields worldwide, especially in China and Southeast Asia. Understanding how maize plants respond to R. solani infection is a key step towards controlling the spread of this fungal pathogen. In this study, we determined the transcriptome of maize plants infected by a low-virulence strain (LVS) and a high-virulence strain (HVS) of R. solani for 3 and 5 days by transcriptome deep-sequencing (RNA-seq). We identified 3,015 (for LVS infection) and 1,628 (for HVS infection) differentially expressed genes (DEGs). We confirmed the expression profiles of 10 randomly selected DEGs by quantitative reverse transcription PCR. We also performed a Gene Ontology (GO) enrichment analysis to establish which biological processes are associated with these DEGs, which revealed the enrichment of defense-related GO terms in LVS- and HVS-regulated genes. We selected 388 DEGs upregulated upon fungal infection as possible candidate genes. Among them, the overexpression of ZmNAC41 (encoding NAC transcription factor 41) or ZmBAK1 (encoding BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1) in rice enhanced resistance to R. solani. In addition, overexpressing ZmBAK1 in rice also increased plant height, plant weight, thousand-grain weight, and grain length. The identification of 388 potential key maize genes related to resistance to R. solani provides significant insights into improving BLSB resistance.


Assuntos
Oryza , Zea mays , Zea mays/genética , Zea mays/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Rhizoctonia/genética , Oryza/genética , Perfilação da Expressão Gênica , Transcriptoma/genética
7.
Int J Mol Sci ; 23(21)2022 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-36361711

RESUMO

Rhizoctonia solani is one of the important pathogenic fungi causing several serious crop diseases, such as maize and rice sheath blight. Current methods used to control the disease mainly depend on spraying fungicides because there is no immunity or high resistance available in crops. Spraying double-strand RNA (dsRNA) for induced-gene silencing (SIGS) is a new potentially sustainable and environmentally friendly tool to control plant diseases. Here, we found that fluorescein-labelled EGFP-dsRNA could be absorbed by R. solani in co-incubation. Furthermore, three dsRNAs, each targeting one of pathogenicity-related genes, RsPG1, RsCATA, and RsCRZ1, significantly downregulated the transcript levels of the target genes after co-incubation, leading to a significant reduction in the pathogenicity of the fungus. Only the spray of RsCRZ1 dsRNA, but not RsPG1 or RsCATA dsRNA, affected fungal sclerotium formation. dsRNA stability on leaf surfaces and its efficiency in entering leaf cells were significantly improved when dsRNAs were loaded on layered double hydroxide (LDH) nanosheets. Notably, the RsCRZ1-dsRNA-LDH approach showed stronger and more lasting effects than using RsCRZ1-dsRNA alone in controlling pathogen development. Together, this study provides a new potential method to control crop diseases caused by R. solani.


Assuntos
Oryza , Rhizoctonia , Rhizoctonia/genética , Doenças das Plantas/genética , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia , RNA de Cadeia Dupla/genética , Oryza/genética
8.
J Agric Food Chem ; 70(43): 13839-13848, 2022 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-36270026

RESUMO

A series of pyrazol-5-yl-benzamide derivatives containing the oxazole group were designed and synthesized as potential SDH inhibitors. According to the results of the bioassays, most target compounds displayed moderate-to-excellent in vitro antifungal activities against Valsa mali, Sclerotinia scleotiorum, Alternaria alternata, and Botrytis cinerea. Among them, compounds C13, C14, and C16 exhibited more excellently inhibitory activities against S. sclerotiorum than boscalid (EC50 = 0.96 mg/L), with EC50 values of 0.69, 0.26, and 0.95 mg/L, respectively. In vivo experiments on rape leaves and cucumber leaves showed that compounds C13 and C14 exhibited considerable protective effects against S. sclerotiorum than boscalid. SEM analysis indicated that compounds C13 and C14 significantly destroyed the typical structure and morphology of S. scleotiorum hyphae. In the respiratory inhibition effect assays, compounds C13 (28.0%) and C14 (33.9%) exhibited a strong inhibitory effect on the respiration rate of S. sclerotiorum mycelia, which was close to boscalid (30.6%). The results of molecular docking indicated that compounds C13 and C14 could form strong interactions with the key residues TRP O:173, ARG P:43, TYR Q:58, and MET P:43 of the SDH. Furthermore, the antifungal mechanism of these derivatives was demonstrated by the SDH enzymatic inhibition assay. These results demonstrate that compounds C13 and C14 can be developed into novel SDH inhibitors for crop protection.


Assuntos
Rhizoctonia , Succinato Desidrogenase , Succinato Desidrogenase/metabolismo , Antifúngicos/farmacologia , Simulação de Acoplamento Molecular , Relação Estrutura-Atividade , Oxazóis/farmacologia , Benzamidas/farmacologia
9.
J Agric Food Chem ; 70(42): 13464-13472, 2022 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-36250688

RESUMO

Succinate dehydrogenase (SDH) is known as an ideal target for the development of novel fungicides. Over the years, a series of novel pyrazole carboxamides containing a diarylamine scaffold have been reported as potent SDH inhibitors (SDHIs) in our laboratory. Among them, compound SCU3038 (EC50 = 0.016 mg/L) against in vitro Rhizoctonia solani was better than fluxapyroxad (EC50 = 0.033 mg/L). However, its mechanism of action is still unclear. In this paper, in pot tests, bioactivity evaluation indicated that in vivo antifungal activity of compound SCU3038 (EC50 = 0.95 mg/L) against R. solani was better than that of fluxapyroxad (EC50 = 2.29 mg/L) and thifluzamide (EC50 = 1.88 mg/L). In field trials, control efficacy of compound SCU3038 (74.10%) at 200 g ai/ha against rice sheath blight was better than that of thifluzamide (71.40%). Furthermore, target evaluation showed that compound SCU3038 could inhibit the fungal SDH from R. solani and fix in the binding site of SDH by molecular docking, thereby it could dissolve and reduce mitochondria of R. solani as observed by electron microscopy. In addition, transcriptome results showed that compound SCU3038 affected the TCA cycle pathway in mitochondria, and this was manifested in the downregulation of eight genes and upregulation of one gene. The most important phenomenon was the repressed expression of SDH2 confirmed by qRT-PCR. It was observed that compound SCU3038 was a potent SDHI, and these results afforded further research on pyrazole carboxamides.


Assuntos
Fungicidas Industriais , Succinato Desidrogenase , Antifúngicos/farmacologia , Antifúngicos/química , Fungicidas Industriais/farmacologia , Fungicidas Industriais/química , Simulação de Acoplamento Molecular , Rhizoctonia/metabolismo , Pirazóis/farmacologia , Pirazóis/química , Relação Estrutura-Atividade , Doenças das Plantas
10.
J Basic Microbiol ; 62(11): 1346-1359, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36122185

RESUMO

Phytotoxins produced by Rhizoctonia solani AG1-1A (Anastomosis Group 1 Subgroup 1A) play a significant role in developing sheath blight disease in rice. A phytotoxin in the partially purified ethyl acetate fraction from the culture filtrate of a highly aggressive R. solani (RIRS-K) isolate, with Indian Type Culture Collection (ITCC) number 7479, infecting rice that could incite necrotic symptoms characteristic of the fungus was identified. The role of the crude toxin in the pathogenicity and virulence of the fungal pathogen on rice was first established by artificial inoculation assay under controlled conditions. The crude ethyl acetate extract obtained from the culture filtrate of RIRS-K was first fractionated by column chromatography. Further purification of the bioactive fraction was carried out by using bioassay-guided fractionation, and a toxic fraction was obtained. The most bioactive fraction was analyzed by GC-MS analysis, and 3-butylpyridine (3-BP) was identified as a major compound in the active fraction by comparing its mass spectrum with NIST library and its standard. The purified bioactive fraction and standard (3-BP) toxicity was further validated and compared at 1000 ppm. The result showed that both the bioactive fraction and the 3-BP have caused necrosis, similar to the one incited by R. solani. This study showed that 3-BP is one of the major compounds responsible for the necrosis development in the rice plant during ShB disease and is a hitherto unexplored toxin of R. solani in rice.


Assuntos
Oryza , Oryza/microbiologia , Doenças das Plantas/microbiologia , Rhizoctonia , Necrose
11.
Int J Mol Sci ; 23(18)2022 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-36142601

RESUMO

Rhizoctonia cerealis is the causal agent of sharp eyespot, a devastating disease of cereal crops including wheat. Several metalloproteases have been implicated in pathogenic virulence, but little is known about whole-genome metalloproteases in R. cerealis. In this study, a total of 116 metalloproteases-encoding genes were identified and characterized from the R. cerealis Rc207 genome. The gene expression profiles and phylogenetic relationship of 11 MEP36/fungalysin metalloproteases were examined during the fungal infection to wheat, and function of an upregulated secretory MEP36 named RcFL1 was validated. Of 11 MEP36 family metalloproteases, ten, except RcFL5, were predicted to be secreted proteins and nine encoding genes, but not RcFL5 and RcFL2, were expressed during the R. cerealis infection process. Phylogenetic analysis suggested that MEP36 metalloproteases in R. cerealis were closely related to those of Rhizoctonia solani but were remote to those of Bipolaris sorokiniana, Fusarium graminearum, F. pseudograminearum, and Pyricularia oryzae. Expression of RcFL1 was significantly upregulated during the infection process and induced plant cell death in wheat to promote the virulence of the pathogen. The MEP36 domain was necessary for the activities of RcFL1. Furthermore, RcFL1 could repress the expression of wheat genes coding for the chitin elicitor receptor kinase TaCERK1 and chitinases. These results suggest that this MEP36 metalloprotease RcFL1 may function as a virulence factor of R. cerealis through inhibiting host chitin-triggered immunity and chitinases. This study provides insights on pathogenic mechanisms of R. cerealis. RcFL1 likely is an important gene resource for improving resistance of wheat to R. cerealis through host-induced gene silencing strategy.


Assuntos
Quitinases , Triticum , Basidiomycota , Quitina/metabolismo , Quitinases/metabolismo , Metaloproteases/genética , Metaloproteases/metabolismo , Filogenia , Doenças das Plantas/microbiologia , Rhizoctonia/fisiologia , Triticum/metabolismo , Virulência/genética , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
12.
Molecules ; 27(17)2022 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-36080312

RESUMO

Root rot is one of the most significant soil and seed-borne fungal diseases, limiting the cultivation of fenugreek plants. Endophytic bacteria and their natural bioproducts have emerged as growth promoters and disease suppressors in the current era. Despite limited research, seeds are a good funder of endophytic microbiomes, which are transmitted from them to other seedling parts, thereby providing a shield against biotic and abiotic anxiety and promoting the growth at early germination and later stages. The current study evaluated the hypothesis that seed endophytic bacteria and their lytic enzymes, growth promotors, and antifungal molecules can induce growth, and inhibit root rot disease development at the same time. The isolation trial from fenugreek seeds revealed a lytic Achromobacter sp., which produces indole acetic acid, has antifungal compounds (e.g., 2-Butanol, 3,3'-oxybis-), and reduces the growth of Rhizoctonia solani by 43.75%. Under the greenhouse and natural field conditions, bacterial cells and/or supernatant improved the growth, physiology, and yield performance of fenugreek plants, and effectively suppressed the progress of root rot disease; this is the first extensive study that uses a new seed-borne endophytic bacterium as a plant-growth-promoting, and biocontrol tool against the sclerotia-forming; R. solani; the causative of fenugreek root rot.


Assuntos
Achromobacter , Trigonella , Antifúngicos/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Rhizoctonia , Sementes
13.
Mol Plant Microbe Interact ; 35(9): 803-813, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36102883

RESUMO

Rhizoctonia cerealis is a soilborne fungus that can cause sharp eyespot in wheat, resulting in massive yield losses found in many countries. Due to the lack of resistant cultivars, fungicides have been widely used to control this pathogen. However, chemical control is not environmentally friendly and is costly. Meanwhile, the lack of genetic transformation tools has hindered the functional characterization of virulence genes. In this study, we attempted to characterize the function of virulence genes by two transient methods, host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS), which use RNA interference to suppress the pathogenic development. We identified ten secretory orphan genes from the genome. After silencing these ten genes, only the RcOSP1 knocked-down plant significantly inhibited the growth of R. cerealis. We then described RcOSP1 as an effector that could impair wheat biological processes and suppress pathogen-associated molecular pattern-triggered immunity in the infection process. These findings confirm that HIGS and SIGS can be practical tools for researching R. cerealis virulence genes. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Assuntos
Fungicidas Industriais , Triticum , Basidiomycota , Inativação Gênica , Padrões Moleculares Associados a Patógenos , Doenças das Plantas/microbiologia , Rhizoctonia/genética , Triticum/microbiologia
14.
Arch Microbiol ; 204(10): 629, 2022 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-36115881

RESUMO

This work aims at exploring an antagonistic actinobacterial strain isolated from the roots of Ziziphus lotus in bioformulation processes and the biocontrol of Rhizoctonia solani damping-off of tomato seedlings. The strain Streptomyces caeruleatus ZL-2 was investigated for the principal in vitro biocontrol mechanisms and then formulated in three different biofungicides: wettable talcum powder (WTP), sodium alginate propagules (SAP) and clay sodium alginate propagules (CAP). Compared to a marketed control products (Serenade® and Acil 060FS®), the formulated biofungicides were investigated against the R. solani damping-off of tomato cv. Aïcha seedlings. The strain ZL-2 produced chitinases, cellulases, ß-1,3-glucanases, cyanhydric acid and siderophores. It also showed strong antagonistic effect on the mycelial growth of R. solani. Bioautographic and HPLC analysis revealed the production of a single or several co-migrating antifungal compounds. The biofungicide WTP presented an attractive biocontrol effect by significantly reducing the disease severity index (DSI) compared to untreated seeds. No significant differences were obtained compared to the chemical treatment with Acil 060FS®. The viability of spores and biocontrol efficacy of the WTP were confirmed after 1-year storage. Strain ZL-2 has never been reported in the bioformulation of active biofungicides against Rhizoctonia solani damping-off and this work opens up very attractive prospects in the fields of biocontrol and crop improvement.


Assuntos
Celulases , Quitinases , Alginatos , Antifúngicos/farmacologia , Argila , Nitrazepam , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Rhizoctonia , Plântula/microbiologia , Sideróforos , Esporos Fúngicos , Streptomyces , Talco
15.
J Environ Sci Health B ; 57(10): 821-834, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36127826

RESUMO

Rhizoctonia solani AG1-IA is a necrotrophic fungus that causes rice sheath blight and results in severe yield and quality reductions in rice worldwide. Differences of genetic structure and fungicide sensitivity of the pathogen have significant effects on the severity and control effect of this disease in the field. To determine correlations among population genetic structure, geographic origin, growth rate, and fungicide resistance of the pathogen, 293 strains of R. solani were isolated from diseased rice collected from 13 cities of Jiangsu Province and five regions of China. Simple sequence repeat (SSR) molecular marker technology was used to analyze the genetic diversity of these strains, and a total of 74 bands were amplified by nine pairs of primers. Population genetic structure analysis showed that strains from Central China and northern Jiangsu had the highest Nei's gene diversity index and Shannon diversity index. The vast majority of strains grew fast with colony diameters of more than 60.0 mm cultured at 28 °C for 36 h. The half-maximal effective concentration (EC50) of them to tebuconazole, thifluzamide, and propiconazole varied ∼16.2-, 3.8-, and 7.5-fold. However, the genetic diversity of R. solani had no significant correlation with their geographic origin, growth rate or fungicide sensitivity.


Assuntos
Fungicidas Industriais , Oryza , Fungicidas Industriais/farmacologia , Estruturas Genéticas , Genética Populacional , Oryza/microbiologia , Doenças das Plantas/microbiologia , Rhizoctonia
16.
Plant Dis ; 106(10): 2730-2740, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36094426

RESUMO

As the excessive use of chemical fertilizers harms organisms and adversely affects the soil environment, the replacement of chemical fertilizers with biological fertilizers has attracted widespread attention as an environmental protection strategy. In this study, the effects of rhizosphere bacteria inoculation on growth of Pinus sylvestris var. mongolica seedlings, soil parameters, soil microbial community structure, and the biocontrol of damping-off were studied by pot experiments. The results showed that all three rhizosphere bacteria (Pseudomonas chlororaphis, Pseudomonas extremaustralis, and Acinetobacter lwoffii A07) tested exhibited growth-promoting properties, such as the production of indole-3-acetic acid, hydrolase, siderophores, and hydrogen cyanide; nitrogen fixation; and phosphorus solubilization. The application of the three bacteria increased plant biomass, root structure, and nutrient content and also increased soil nutrient content and enzyme activity. Bacterial inoculation promoted the growth of beneficial bacteria and antagonistic bacteria by adjusting the physicochemical properties of the soil, thereby improving the bacterial community structure. Among the soil features, available nitrogen, total nitrogen, available potassium, and urease activity were the main influencing factors. In addition, it was also found that bacterial inoculation significantly increased the activities of plant superoxide dismutase, catalase, peroxidase, and other defense enzymes; enhanced plant disease resistance; effectively inhibited damping-off; and promoted plant growth. In summary, the application of three rhizosphere bacteria systematically affected the interaction between plants, soil parameters, and soil microbial communities. These results provide a basis for understanding how rhizosphere bacteria promote the growth of P. sylvestris var. mongolica, thereby offering a promising sustainable alternative to chemical fertilizers.


Assuntos
Microbiota , Pinus sylvestris , Bactérias , Catalase , Fertilizantes , Cianeto de Hidrogênio , Nitrogênio/análise , Fósforo , Potássio , Rhizoctonia , Plântula/química , Sideróforos , Solo/química , Superóxido Dismutase , Urease
18.
Arch Microbiol ; 204(9): 587, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-36048258

RESUMO

Beauveria bassiana, a potential entomopathogenic biocontrol agent, has recently drawn attention worldwide for its other additional beneficial roles such as plant disease antagonist, beneficial rhizosphere colonizer, plant growth promoter and an endophyte. In the present study, endophytic colonizing behaviour of five (5) B. bassiana isolates viz., Bb4, Bb16, Bb25, Bb44 and Bb53 were studied in rice following three (3) artificial inoculation techniques viz., seed treatment, root inoculation and foliar spray and the endophytic colonizing ability were determined by culture-based assay. After B. bassiana inoculation, rice plants were challenged with Rhizoctonia solani and disease incidence and plant growth promotion were assessed. Per cent colonization of rice stems, leaves and roots were influenced by inoculation technique, post-inoculation time (7th, 14th, 21st and 28th dpi) and plant growth medium (sterile soil, non-sterile soil), recorded maximum on 14th-day post-inoculation (dpi) i.e., 96% in stems, 92% in leaves and 28% in roots, whereas, lower colonization was recorded on 7th, 21st and 28th dpi. Whereas, the foliar spray was found best as compared to seed and root inoculation techniques, and maximum fungal recovery was observed in stems and leaves and least in roots. Upon colonization, the physical presence of B. bassiana in rice was localized by light microscopy-based studies. Potential B. bassiana strains with endophytic ability were re-isolated and their identity was determined based on morphometric and PCR-based techniques. Further, the present study also identified several virulent genes viz., BbChit1, Cdep1, Bbhog1 and Bbjen1 and extracellular hydrolytic enzymes viz., α-amylase, cellulase, lipase, pectinase and xylanase secreted by endophytic B. bassiana strains as determinants responsible for establishing the endophytic association in rice. On the other hand, a significant reduction in disease incidence was observed in the endophytic B. bassiana Bb4-, Bb16- and Bb44-inoculated plants as compared to the non-endophytic B. bassiana Bb25- and Bb53-inoculated plants along with enhanced plant growth promotion. This is one of the few studies investigating the colonization of B. bassiana in rice and its promising role as a plant disease antagonist and plant growth promoter in rice.


Assuntos
Beauveria , Oryza , Beauveria/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Plantas , Rhizoctonia , Solo
19.
BMC Genomics ; 23(1): 606, 2022 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-35986248

RESUMO

BACKGROUND: Rice sheath blight, which is caused by Rhizoctonia solani, is the most destructive disease affecting rice production, but the resistance mechanism to this pathogen has not been fully elucidated. RESULTS: In this study, we selected two rice cultivars based on their resistance to the pathogen and analyzed and compared the transcriptomic profiles of two cultivars, the moderately resistant variety Gangyuan8 and the highly susceptible variety Yanfeng47, at different time points after inoculation. The comparative transcriptome profiling showed that the expression of related genes gradually increased after pathogen inoculation. The number of differentially expressed genes (DEGs) in Yanfeng47 was higher than that in Gangyuan8, and this result revealed that Yanfeng47 was more susceptible to fungal attack. At the early stage (24 and 48 h), the accumulation of resistance genes and a resistance metabolism occurred earlier in Ganguan8 than in Yanfeng47, and the resistance enrichment entries were more abundant in Ganguan8 than in Yanfeng47. CONCLUSIONS: Based on the GO and KEGG enrichment analyses at five infection stages, we concluded that phenylalanine metabolism and the jasmonic acid pathway play a crucial role in the resistance of rice to sheath blight. Through a comparative transcriptome analysis, we preliminarily analyzed the molecular mechanism responsible for resistance to sheath blight in rice, and the results lay the foundation for the development of gene mining and functional research on rice resistance to sheath blight.


Assuntos
Oryza , Resistência à Doença/genética , Perfilação da Expressão Gênica , Oryza/genética , Oryza/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Rhizoctonia/genética , Transcriptoma
20.
J Appl Microbiol ; 133(5): 3215-3227, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35957552

RESUMO

AIM: To understand the mechanism of necrosis incited by a host-selective phytotoxin designated as Rhizoctonia solani toxin (RST) identified to be a potential pathogenic factor of R. solani AG1 IA, causing sheath blight (ShB) of rice. METHODS AND RESULTS: The metabolomic changes induced by the phytotoxic metabolite in a ShB susceptible rice cultivar were elucidated by gas chromatography-mass spectrometry analysis and compared with that of the pathogen to identify rice metabolites targeted by the phytotoxin. The profiles of about 29 metabolites with various physiological roles in rice plants have been identified worldwide. Unsupervised and supervised multivariate chemometrics (principal component analysis and partial least squares-discriminant analysis) and cluster (Heat maps) analyses were used to compare the metabolites obtained from chemical profiles of the treatments with sterile distilled water (SDW) control. The results indicated that the rice plant expressed more metabolites in response to the pathogen than the phytotoxin and was lowest in SDW control. The key metabolites expressed in rice in response to the treatments were investigated by the variable importance in projection (VIP) analysis using p < 0.05 VIP >15. The analysis identified 7 and 11 upregulating metabolites in the phytotoxin and the pathogen treatments, respectively, compared to the untreated control. Among the phytotoxin-treated and the pathogen inoculated samples, the phytotoxin-treated sample recorded upregulation of six metabolites, whereas nine metabolites were upregulated in the pathogen-inoculated samples. These upregulating metabolites are speculated for the necrotic symptoms characteristic to both the phytotoxin and pathogen. In this analysis, hexadecanoic acid and dotriacontane were highly expressed metabolites specific to the phytotoxin and pathogen-treated samples, respectively. Besides upregulation, the metabolites also have a VIP score of >1.5 and hence fulfilled the criteria of classifying them as reliable potential biomarkers. In the pathway analysis, hexadecanoic acid and dotriacontane were identified to be involved in several important biosynthetic pathways of rice, such as the biosynthesis of saturated fatty acid and unsaturated fatty acids cutin, suberin and wax. CONCLUSIONS: The study concludes that though certain metabolites induced by the phytotoxin in the susceptible variety during necrosis shares with that of the pathogen, the identification of metabolites specific to the phytotoxin in comparison to the pathogenic and SDW controls indicated that the phytotoxin modulates the host metabolism differently and hence can be a potential pathogenicity factor of the ShB fungus. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to lack of knowledge on the pathway genes of RST and in the absence of an ShB-resistant variety, understanding differentially expressed metabolic changes induced in the susceptible variety by the phytotoxin in comparison to that of the pathogenic and uninoculated controls enables us to identify the key metabolite changes during the ShB infection. Such metabolomic changes can further be used to infer gene functions for exploitation in ShB control.


Assuntos
Oryza , Oryza/microbiologia , Ácido Palmítico , Doenças das Plantas/microbiologia , Rhizoctonia/fisiologia , Fatores de Virulência , Água , Necrose
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