Volatile-mediated inhibitory activity of Rhizobacteria as a result of multiple factors interaction: the case of Lysobacter capsici AZ78

Plant beneficial rhizobacteria may antagonize soilborne plant pathogens by producing a vast array of volatile organic compounds (VOCs). The production of these compounds depends on the medium composition used for bacterial cell growth. Accordingly, Lysobacter capsici AZ78 (AZ78) grown on a protein-rich medium was previously found to emit volatile pyrazines with toxic activity against soilborne phypathogenic fungi and oomycetes. However, the discrepancy between the quantity of pyrazines in the gaseous phase and the minimum quantity needed to achieve inhibition of plant pathogens observed, lead us to further investigate the volatile-mediated inhibitory activity of AZ78. Here, we show that, besides VOCs, AZ78 cells produce ammonia in increased amounts when a protein-rich medium is used for bacterial growth. The production of this volatile compound caused the alkalinization of the physically separated culture medium where Rhizoctonia solani was inoculated subsequently. Results achieved in this work clearly demonstrate that VOC, ammonia and the growth medium alkalinization contribute to the overall inhibitory activity of AZ78 against R. solani. Thus, our findings suggest that the volatile-mediated inhibitory activity of rhizobacteria in protein-rich substrates can be regarded as a result of multiple factors interaction, rather than exclusively VOCs productio

Ecological role of volatile organic compounds emitted by Pantoea agglomerans as interspecies and interkingdom signals

Volatile organic compounds (VOCs) play an essential role in microbe–microbe and plant–microbe interactions. We investigated the interaction between two plant growth-promoting rhizobacteria, and their interaction with tomato plants. VOCs produced by Pantoea agglomerans MVC 21 modulates the release of siderophores, the solubilisation of phosphate and potassium by Pseudomonas (Ps.) putida MVC 17. Moreover, VOCs produced by P. agglomerans MVC 21 increased lateral root density (LRD), root and shoot dry weight of tomato seedlings. Among the VOCs released by P. agglomerans MVC 21, only dimethyl disulfide (DMDS) showed effects similar to P. agglomerans MVC 21 VOCs. Because of the effects on plants and bacterial cells, we investigated how P. agglomerans MVC 21 VOCs might influence bacteria–plant interaction. Noteworthy, VOCs produced by P. agglomerans MVC 21 boosted the ability of Ps. putida MVC 17 to increase LRD and root dry weight of tomato seedlings. These results could be explained by the positive effect of DMDS and P. agglomerans MVC 21 VOCs on acid 3-indoleacetic production in Ps. putida MVC 17. Overall, our results clearly indicated that P. agglomerans MVC 21 is able to establish a beneficial interaction with Ps. putida MVC 17 and tomato plants through the emission of DMD

Trichoderma spp. volatile organic compounds protect grapevine plants by activating defence-related processes against downy mildew

7openInternationalInternational coauthor/editorVolatile organic compounds (VOCs) are produced by soil-borne microorganisms and play crucial roles in fungal interactions with plants and phytopathogens. Although VOCs have been characterized in Trichoderma spp., the mechanisms against phytopathogens strongly differ according to the strain and pathosystem. This study aimed at characterizing VOCs produced by three Trichoderma strains used as biofungicides and to investigate their effects against grapevine downy mildew (caused by Plasmopara viticola). A VOC-mediated reduction of downy mildew severity was found in leaf disks treated with Trichoderma asperellum T34 (T34), T. harzianum T39 (T39) and T. atroviride SC1 (SC1) and 31 compounds were detected by head space-solid phase microextraction gas chromatography-mass spectrometry. Among the Trichoderma VOCs annotated, -farnesene, cadinene, 1,3-octadiene, 2-pentylfuran and 6-pentyl-2H-pyran-2-one reduced downy mildew severity on grapevine leaf disks. In particular, 6-pentyl-2H-pyran-2-one and 2-pentylfuran increased the accumulation of callose and enhanced the modulation of defence-related genes after P. viticola inoculation, indicating an induction of grapevine defence mechanisms. Moreover, 6-pentyl-2H-pyran-2-one activated the hypersensitive response after P. viticola inoculation, possibly to reinforce the grapevine defence reaction. These results indicate that Trichoderma VOCs can induce grapevine resistance, and these molecules could be further applied to control grapevine downy mildew.openValentina Lazazzara, Bianca Vicelli, Christoph Bueschl, Alexandra Parich, Ilaria Pertot, Rainer Schuhmacher, Michele PerazzolliLazazzara, V.; Vicelli, B.; Bueschl, C.; Parich, A.; Pertot, I.; Schuhmacher, R.; Perazzolli, M

Volatile-Mediated Inhibitory Activity of Rhizobacteria as a Result of Multiple Factors Interaction: The Case of Lysobacter capsici AZ78

Plant beneficial rhizobacteria may antagonize soilborne plant pathogens by producing a vast array of volatile organic compounds (VOCs). The production of these compounds depends on the medium composition used for bacterial cell growth. Accordingly, Lysobacter capsici AZ78 (AZ78) grown on a protein-rich medium was previously found to emit volatile pyrazines with toxic activity against soilborne phypathogenic fungi and oomycetes. However, the discrepancy between the quantity of pyrazines in the gaseous phase and the minimum quantity needed to achieve inhibition of plant pathogens observed, lead us to further investigate the volatile-mediated inhibitory activity of AZ78. Here, we show that, besides VOCs, AZ78 cells produce ammonia in increased amounts when a protein-rich medium is used for bacterial growth. The production of this volatile compound caused the alkalinization of the physically separated culture medium where Rhizoctonia solani was inoculated subsequently. Results achieved in this work clearly demonstrate that VOC, ammonia and the growth medium alkalinization contribute to the overall inhibitory activity of AZ78 against R. solani. Thus, our findings suggest that the volatile-mediated inhibitory activity of rhizobacteria in protein-rich substrates can be regarded as a result of multiple factors interaction, rather than exclusively VOCs production.</jats:p

Towards a broader view of the metabolome: untargeted profiling of soluble and bound polyphenols in plants

AbstractPhenylalanine (Phe) is a central precursor for numerous secondary plant metabolites with a multitude of biological functions. Recent studies on the fungal disease Fusarium head blight in wheat showed numerous Phe-derived defence metabolites to be induced in the presence of the pathogen. These studies also suggest a partial incorporation of Phe-derived secondary metabolites into the cell wall. To broaden the view of the metabolome to bound Phe derivatives, an existing approach using 13C-labelled Phe as tracer was extended. The developed workflow consists of three successive extractions with an acidified acetonitrile-methanol-water mixture to remove the soluble plant metabolites, followed by cell wall hydrolysis with 4M aqueous NaOH, acidification with aqueous HCl, and liquid-liquid extraction of the hydrolysate with ethyl acetate. The untargeted screening of Phe-derived metabolites revealed 156 soluble compounds and 90 compounds in the hydrolysed samples including known cell wall constituents like ferulic acid, coumaric acid, and tricin. Forty-nine metabolites were found exclusively in the hydrolysate. The average cumulative extraction yield of the soluble metabolites was 99.6%, with a range of 91.8 to 100%. Repeatability coefficients of variation of the protocol ranged from 10.5 to 25.9%, with a median of 16.3%. To demonstrate the suitability of the proposed method for a typical metabolomics application, mock-treated and Fusarium graminearum-treated wheat samples were compared. The study revealed differences between the hydrolysates of the two sample types, confirming the differential incorporation of Phe-derived metabolites into the cell wall under infection conditions.</jats:p

Evaluating the Performance of Lateral Flow Devices for Total Aflatoxins with Special Emphasis on Their Robustness under Sub-Saharan Conditions

As aflatoxins are a global risk for humans and animals, testing methods for rapid on-site screening are increasingly needed alongside the standard analytical laboratory tools. In the presented study, lateral flow devices (LFDs) for rapid total aflatoxin screening were thoroughly investigated with respect to their matrix effects, cross-reactivity, their performance under harsh conditions in Sub-Saharan Africa (SSA), and their stability, as well as when compared with liquid chromatography-tandem mass spectrometry (LC-MS/MS). To analyze the matrix effects, qualitative test kits offering a certain cutoff level were used to screen different nut samples. In addition, these tests were challenged on their cross-reactivity with 230 fungal toxins and metabolites. Furthermore, the resulting measurements performed under harsh tropical conditions (up to 38.4 °C and 91% relative humidity) in SSA, specifically Burkina Faso and Mozambique, were compared with the results from a well-established and validated LC-MS/MS-based reference method. The comparison of the on-site LFD results with the reference method showed a good agreement: 86.4% agreement, 11.8% non-agreement, and 1.8% invalid test results. To test the robustness of the cutoff tests, short- and long-term stability testing was carried out in Mozambique and Nigeria. For both experiments, no loss of test performance could be determined. Finally, a subset of African corn samples was shipped to Austria and analyzed under laboratory conditions using semiquantitative aflatoxin tests. A good correlation was found between the rapid strip tests and the LC-MS/MS reference method. Overall, the evaluated LFDs showed satisfying results regarding their cross-reactivity, matrix effects, stability, and robustness.</jats:p

Evaluating the Performance of Lateral Flow Devices for Total Aflatoxins with Special Emphasis on Their Robustness under Sub-Saharan Conditions

Open Access Article; Published online: 20 Oct 2021As aflatoxins are a global risk for humans and animals, testing methods for rapid on-site screening are increasingly needed alongside the standard analytical laboratory tools. In the presented study, lateral flow devices (LFDs) for rapid total aflatoxin screening were thoroughly investigated with respect to their matrix effects, cross-reactivity, their performance under harsh conditions in Sub-Saharan Africa (SSA), and their stability, as well as when compared with liquid chromatography-tandem mass spectrometry (LC-MS/MS). To analyze the matrix effects, qualitative test kits offering a certain cutoff level were used to screen different nut samples. In addition, these tests were challenged on their cross-reactivity with 230 fungal toxins and metabolites. Furthermore, the resulting measurements performed under harsh tropical conditions (up to 38.4 °C and 91% relative humidity) in SSA, specifically Burkina Faso and Mozambique, were compared with the results from a well-established and validated LC-MS/MS-based reference method. The comparison of the on-site LFD results with the reference method showed a good agreement: 86.4% agreement, 11.8% non-agreement, and 1.8% invalid test results. To test the robustness of the cutoff tests, short- and long-term stability testing was carried out in Mozambique and Nigeria. For both experiments, no loss of test performance could be determined. Finally, a subset of African corn samples was shipped to Austria and analyzed under laboratory conditions using semiquantitative aflatoxin tests. A good correlation was found between the rapid strip tests and the LC-MS/MS reference method. Overall, the evaluated LFDs showed satisfying results regarding their cross-reactivity, matrix effects, stability, and robustness

Quantitation of Mycotoxins in Food and Feed from Burkina Faso and Mozambique Using a Modern LC-MS/MS Multitoxin Method

In this study an LC-MS/MS multitoxin method covering a total of 247 fungal and bacterial metabolites was applied to the analysis of different foods and feedstuffs from Burkina Faso and Mozambique. Overall, 63 metabolites were determined in 122 samples of mainly maize and groundnuts and a few samples of sorghum, millet, rice, wheat, soy, dried fruits, other processed foods and animal feeds. Aflatoxin B1 was observed more frequently in maize (Burkina Faso, 50% incidence, median = 23.6 μg/kg; Mozambique, 46% incidence, median = 69.9 μg/kg) than in groundnuts (Burkina Faso, 22% incidence, median = 10.5 μg/kg; Mozambique, 14% incidence, median = 3.4 μg/kg). Fumonisin B1 concentrations in maize were higher in Mozambique (92% incidence, median = 869 μg/kg) than in Burkina Faso (81% incidence, median = 269 μg/kg). In addition, ochratoxin A, zearalenone, deoxynivalenol, nivalenol, and other less reported mycotoxins such as citrinin, alternariol, cyclopiazonic acid, sterigmatocystin, moniliformin, beauvericin, and enniatins were detected. Up to 28 toxic fungal metabolites were quantitated in a single sample, emphasizing the great variety of mycotoxin coexposure. Most mycotoxins have not been reported before in either country.Austrian Development Agencypeer Revie