Bioactive Botanics against Pathogenic and Mycotoxigenic Fungi Isolated from Rice

[EN] Cereal fungal contamination causes both economic and human health problems. In the present study, the chemical composition of commercial bay leaf, cinnamon, clove and oregano essential oils, and antifungal activity against three pathogenic fungi isolated from rice, were investigated. Essential oils presented a high percentage of oxygenated components: 78.8% in bay leaf (eucalyptol 52%); 90.5% in clove (eugenol 90%); 92% in cinnamon (eugenol 60% and eugenyl acetate 18.5%); 72% in oregano (carvacrol 50% and thymol 20%). Monoterpenes and sesquiterpenes were: 18% in bay leaf, 9% in clove, 5% in cinnamon, 25% in oregano. Cinnamon, and clove essential oils reduced fungal growth by 70%, 80% and almost 90%. Moreover, there was total inhibition using oregano until the seventeenth day. Oregano, clove and cinnamon oils could provide an alternative for controlling Bipolaris spicifer, Fusarium culmorum and Fusarium sambucinum in stored grains and seeds, so extending their shelf life.Giménez, S.; Rosello Caselles, J.; Santamarina Siurana, MP. (2019). Bioactive Botanics against Pathogenic and Mycotoxigenic Fungi Isolated from Rice. Agricultural Research & Technology. 23(3):00326-00331. http://hdl.handle.net/10251/153363S003260033123

Chemical composition of essential oils of three Mentha species and their antifungal activity against selected phytopathogenic and post-harvest fungi

This is an Accepted Manuscript of an article published by Taylor & Francis in All Life (Online) on 05 Jan 2022, available online: http://www.tandfonline.com/10.1080/26895293.2021.2022007[EN] The postharvest life of most fruit, vegetables and cereals is limited by fungal proliferation. The chemical composition of Mentha piperita, M. spicata and M. suaveolens essential oils (EO), and the antifungal activity against four pathogenic and post-harvest fungi isolated from food, were herein investigated to evaluate their potential as natural food preservatives. The EO were obtained by hydrodistillation of aerial parts leaves, stems and inflorescences (except for peppermint oil, which was purchased in a specialized store) and submitted to GC-MS and GC-FID analysis. Regarding the EO composition, carvone (41.1%) and limonene (14.1%) were the major compounds in M. spicata, menthol (47.0%) and menthone (23.1%), as well as other menthol derivatives (neomenthol -3.6%- and menthofurane -3.7%-) in M. piperita, and piperitone oxide (40.2%) and piperitenone oxide (31.4%) in M. suaveolens. Botryotinia fuckeliana was the most sensitive fungus. The three studied EO inhibited growth by 92¿100%. The highest dose of M. suaveolens EO, 400 ¿g/mL, produced 100% MGI in all the studied fungi, except Fusarium oxysporum with 94.21%. The M. suaveolens EO can be considered to develop a low-risk enviro-friendly botanical biofungicide.The authors also thank the Spanish Type Culture Collection (CECT) for providing the molecular strain identification equipment. This study has been financed by MINECO, Ministerio de Economía y competitividad `Materiales biodegradables multicapa de alta barrera para el envasado activo de alimentos¿ (AGL2016-76699-R).Santamarina Siurana, MP.; Llorens Molina, JA.; Sempere Ferre, F.; Santamarina Siurana, MC.; Rosello Caselles, J.; Giménez Santamarina, S. (2022). Chemical composition of essential oils of three Mentha species and their antifungal activity against selected phytopathogenic and post-harvest fungi. All Life (Online). 15(1):64-73. https://doi.org/10.1080/26895293.2021.2022007647315

Effect of water activity and temperature on growth and sporulation of Aspergillus niger P. E. L. van Tieghem

El efecto de la actividad de agua (0,85, 0,90, 0,95, 0,98, 0,995) sobre el crecimiento y la esporulación de Aspergillus niger fue estudiado a distintas temperaturas (15 y 25 ºC). La máxima ratio de crecimiento de la especie se registró a una actividad de agua de 0,98, y se observó desarrollo en todo el intervalo ensayado salvo a 0,85 y 15 ºC. Además, se investigaron sus características morfológicas y culturales en las distintas condiciones de experimentación. A. niger esporuló en casi todas las actividades de agua y temperatura. Ambos factores y su interacción tuvieron un efecto significativo sobre el desarrollo de la especie.The effect of water activity (0.85, 0.90, 0.95, 0.98, 0.995) on the growth and sporulation of Aspergillus niger was studied at different temperatures (15 and 25 ºC). Maximum fungal growth rate occurred at water activity of 0.98, with growth being observed throughout the tested interval except at 0.85 and 15 ºC. In addition, morphological and cultural features were studied in the different experimental conditions. This species sporulated at almost all conditions. Both factors and their interaction had a significant effect on the development of the species.Ciencias Experimentale

A new biocontrol agent of Drechslera oryzae

El hongo patógeno del arroz Drechslera oryzae se inoculó en un mismo sustrato junto a Trichoderma harzianum en distintas condiciones medioambientales. Los mecanismos ejercidos por T. harzianum para antagonizar a D. oryzae y que en algunos casos actuaron sinérgicamente fueron: competencia por el espacio y los nutrientes, micoparasitismo y posible antibiosis. La capacidad antagonista de T. harzianum aumentó con los valores de temperatura y actividad del agua.The rice pathogen Drechslera oryzae and the antagonistic fungus Trichoderma harzianum were inoculated in the same substratum at different environmental conditions. The mechanisms exerted by T. harzianum as antagonist over D. oryzae and that on some occasions acted sinergically were: competitiveness for space and nutrients, mycoparasitism and a possible antibiosis. The antagonistic capacity of T. harzianum was higher as the values of water activity and temperature were increasing.Ciencias Experimentale

Effect of water activity and temperature on growth and sporulation of Aspergillus niger P. E. L. van Tieghem

[ES] El efecto de la actividad de agua (0,85, 0,90, 0,95, 0,98, 0,995) sobre el crecimiento y la esporulación de Aspergillus niger fue estudiado a distintas temperaturas (15 y 25 ºC). La máxima ratio de crecimiento de la especie se registró a una actividad de agua de 0,98, y se observó desarrollo en todo el intervalo ensayado salvo a 0,85 y 15 ºC. Además, se investigaron sus características morfológicas y culturales en las distintas condiciones de experimentación. A. niger esporuló en casi todas las actividades de agua y temperatura. Ambos factores y su interacción tuvieron un efecto significativo sobre el desarrollo de la especie[EN] The effect of water activity (0.85, 0.90, 0.95, 0.98, 0.995) on the growth and sporulation of Aspergillus niger was studied at different temperatures (15 and 25 ºC). Maximum fungal growth rate occurred at water activity of 0.98, with growth being observed throughout the tested interval except at 0.85 and 15 ºC. In addition, morphological and cultural features were studied in the different experimental conditions. This species sporulated at almost all conditions. Both factors and their interaction had a significant effect on the development of the speciesSempere Ferre, F.; Santamarina Siurana, MP. (2021). Efecto de la actividad de agua y la temperatura sobre el crecimiento y la esporulación de Aspergillus niger P.E.L. van Tieghem. Nereis. Revista Iberoamericana Interdisciplinar de Métodos, Modelización y Simulación. 13:127-133. https://doi.org/10.46583/nereis_2021.13.8171271331

In vitro antagonistic activity of Trichoderma harzianum against Fusarium sudanense causing seedling Blight and seed rot on wheat

[EN] Fusarium sudanense is a novel fungus recently isolated from asymptomatic samples of wheat grains in Argentina. The fungus caused symptoms of seedling blight and seed rot on wheat after artificial inoculations. It is known that the production of mycotoxins by pathogens belonging to the Fusarium genus is harmful to human and animal health. Moreover, the warm and humid conditions that are favorable for growth and mycotoxin production of these species put the Argentinian wheat production area at a high risk of mycotoxin contamination with this novel pathogen. The aim of this work was to evaluate the antagonistic effect of Trichoderma harzianum against F. sudanense under in vitro tests at different environmental conditions. Fungi were screened in dual culture at different water activities (alpha(w)) (0.995, 0.98, 0.95, and 0.90) and temperatures (25 and 15 degrees C). The growth rate of the fungi, interaction types, and dominance index were evaluated. Also, the interaction between T. harzianum and F. sudanense was examined by light and cryo-scanning microscopy. T. harzianum suppressed the growth of F. sudanense at 0.995, 0.98, and 0.95 alpha(w) at 25 degrees C and 0.995 and 0.98 alpha(w) at 15 degrees C. Macroscopic study revealed different interaction types between F. sudanense and T. harzianum on dual culture. Dominance on contact where the colonies of T. harzianum overgrew the pathogen was the most common interaction type determined. The competitive capacity of T. harzianum was diminished by decreasing the temperature and alpha(w). At 0.95 alpha(w) and 15 degrees C, both fungi grew slowly, and interaction type "A" was assigned. Microscopic analysis from the interaction zone of dual cultures revealed an attachment of T. harzianum to the F. sudanense hyphae, penetration with or without formation of appressorium-like structures, coiling, plasmolysis, and a veil formation. According to our results, T. harzianum demonstrated capability to antagonize F. sudanense and could be a promising biocontrol agent.This work was supported by the Escuela Tecnica Superior de Ingenieria Agronomica y del Medio Natural (ETSIANM), UPV, Spain, and by Facultad de Ciencias Agrarias y Forestales of the Universidad Nacional de La Plata (grant no. 11A 296), Argentina.Larran, S.; Santamarina Siurana, MP.; Rosello Caselles, J.; Simón, MR.; Perelló, A. (2020). In vitro antagonistic activity of Trichoderma harzianum against Fusarium sudanense causing seedling Blight and seed rot on wheat. ACS Omega. 5(36):23276-23283. https://doi.org/10.1021/acsomega.0c03090S2327623283536Arnold, A. E. (2007). Understanding the diversity of foliar endophytic fungi: progress, challenges, and frontiers. Fungal Biology Reviews, 21(2-3), 51-66. doi:10.1016/j.fbr.2007.05.003Porras-Alfaro, A., & Bayman, P. (2011). Hidden Fungi, Emergent Properties: Endophytes and Microbiomes. Annual Review of Phytopathology, 49(1), 291-315. doi:10.1146/annurev-phyto-080508-081831Keswani, C., Singh, H. B., Hermosa, R., García-Estrada, C., Caradus, J., He, Y.-W., … Sansinenea, E. (2019). Antimicrobial secondary metabolites from agriculturally important fungi as next biocontrol agents. Applied Microbiology and Biotechnology, 103(23-24), 9287-9303. doi:10.1007/s00253-019-10209-2Mesa Vanegas, A. M., Calle Osorno, J., & Marín Pavas, D. A. (2020). Metabolitos secundarios en Trichoderma spp. y sus aplicaciones biotecnológicas agrícolas. Actualidades Biológicas, 41(111). doi:10.17533/udea.acbi.v41n111a02Moussa, T. A. A., Al-Zahrani, H. S., Kadasa, N. M. S., Ahmed, S. A., de Hoog, G. S., & Al-Hatmi, A. M. S. (2017). Two new species of the Fusarium fujikuroi species complex isolated from the natural environment. Antonie van Leeuwenhoek, 110(6), 819-832. doi:10.1007/s10482-017-0855-1Larran, S., Santamarina Siurana, M. P., Roselló Caselles, J., Simón, M. R., & Perelló, A. (2020). Fusarium sudanense, endophytic fungus causing typical symptoms of seedling blight and seed rot on wheat. Journal of King Saud University - Science, 32(1), 468-474. doi:10.1016/j.jksus.2018.07.005Larran, S. Estudio de la micobiota endofítica asociada al trigo y soja y su significancia en la interacción con patógenos fúngicos; Facultad de Ciencias Agrarias y Forestales, Universidad Nacional de La Plata. 2016 http://sedici.unlp.edu.ar/handle/10915/54499.Shi, W., Tan, Y., Wang, S., Gardiner, D., De Saeger, S., Liao, Y., … Wu, A. (2016). Mycotoxigenic Potentials of Fusarium Species in Various Culture Matrices Revealed by Mycotoxin Profiling. Toxins, 9(1), 6. doi:10.3390/toxins9010006Bockus, W. W., Bowden, R. L., Hunger, R. M., Morrill, W. L., Murray, T. D., & Smiley, R. W. (Eds.). (2010). Compendium of Wheat Diseases and Pests, Third Edition. doi:10.1094/9780890546604Leslie, J. F., & Summerell, B. A. (Eds.). (2006). The Fusarium Laboratory Manual. doi:10.1002/9780470278376Perincherry, L., Lalak-Kańczugowska, J., & Stępień, Ł. (2019). Fusarium-Produced Mycotoxins in Plant-Pathogen Interactions. Toxins, 11(11), 664. doi:10.3390/toxins11110664Edwards, J., Auer, D., de Alwis, S.-K., Summerell, B., Aoki, T., Proctor, R. H., … O’Donnell, K. (2016). Fusarium agapanthi sp. nov., a novel bikaverin and fusarubin-producing leaf and stem spot pathogen of Agapanthus praecox (African lily) from Australia and Italy. Mycologia, 108(5), 981-992. doi:10.3852/15-333Andrade, J. F., & Satorre, E. H. (2015). Single and double crop systems in the Argentine Pampas: Environmental determinants of annual grain yield. Field Crops Research, 177, 137-147. doi:10.1016/j.fcr.2015.03.008Ortega, L. M., Dinolfo, M. I., Astoreca, A. L., Alberione, E. J., Stenglein, S. A., & Alconada, T. M. (2015). Molecular and mycotoxin characterization of Fusarium graminearum isolates obtained from wheat at a single field in Argentina. Mycological Progress, 15(1). doi:10.1007/s11557-015-1147-7Perelló, A. E., & Dal Bello, G. M. (2011). Suppression of tan spot and plant growth promotion of wheat by synthetic and biological inducers under field conditions. Annals of Applied Biology, 158(3), 267-274. doi:10.1111/j.1744-7348.2011.00460.xHarman, G. E., Petzoldt, R., Comis, A., & Chen, J. (2004). Interactions Between Trichoderma harzianum Strain T22 and Maize Inbred Line Mo17 and Effects of These Interactions on Diseases Caused by Pythium ultimum and Colletotrichum graminicola. Phytopathology®, 94(2), 147-153. doi:10.1094/phyto.2004.94.2.147Perelló, A., Lampugnani, G., Abramoff, C., Slusarenko, A., & Bello, G. D. (2016). Suppression of seed-borneAlternaria arborescensand growth enhancement of wheat with biorational fungicides. International Journal of Pest Management, 63(2), 157-165. doi:10.1080/09670874.2016.1252478Sempere, F., & Santamarina, M. P. (2009). Antagonistic interactions between fungal rice pathogenFusarium Verticillioides (Sacc.) Nirenberg andTrichoderma harzianum Rifai. Annals of Microbiology, 59(2), 259-266. doi:10.1007/bf03178326SAMAPUNDO, S., DEVLIEHGERE, F., DE MEULENAER, B., & DEBEVERE, J. (2005). Effect of Water Activity and Temperature on Growth and the Relationship between Fumonisin Production and the Radial Growth of Fusarium verticillioides and Fusarium proliferatum on Corn. Journal of Food Protection, 68(5), 1054-1059. doi:10.4315/0362-028x-68.5.1054Sempere Ferre, F., & Santamarina, M. P. (2010). Efficacy of Trichoderma harzianum in suppression of Fusarium culmorum. Annals of Microbiology, 60(2), 335-340. doi:10.1007/s13213-010-0047-yLlorens, A., Mateo, R., Hinojo, M. J., Valle-Algarra, F. M., & Jiménez, M. (2004). Influence of environmental factors on the biosynthesis of type B trichothecenes by isolates of Fusarium spp. from Spanish crops. International Journal of Food Microbiology, 94(1), 43-54. doi:10.1016/j.ijfoodmicro.2003.12.017Magan, N., & Medina, A. (2016). Integrating gene expression, ecology and mycotoxin production by Fusarium and Aspergillus species in relation to interacting environmental factors. World Mycotoxin Journal, 9(5), 673-684. doi:10.3920/wmj2016.2076Pilar Santamarina, M., & Roselló, J. (2006). Influence of temperature and water activity on the antagonism of Trichoderma harzianum to Verticillium and Rhizoctonia. Crop Protection, 25(10), 1130-1134. doi:10.1016/j.cropro.2006.02.006Prasad, R. D., Rangeshwaran, R., Hegde, S. V., & Anuroop, C. P. (2002). Effect of soil and seed application of Trichoderma harzianum on pigeonpea wilt caused by Fusarium udum under field conditions. Crop Protection, 21(4), 293-297. doi:10.1016/s0261-2194(01)00100-4Perello, A. E., Monaco, C. I., Moreno, M. V., Cordo, C. A., & Simon, M. R. (2006). The effect ofTrichoderma harzianumandT. koningiion the control of tan spot(Pyrenophora tritici-repentis) and leaf blotch (Mycosphaerella graminicola) of wheat under field conditions in Argentina. Biocontrol Science and Technology, 16(8), 803-813. doi:10.1080/09583150600700099Larran, S., Simón, M. R., Moreno, M. V., Siurana, M. P. S., & Perelló, A. (2016). Endophytes from wheat as biocontrol agents against tan spot disease. Biological Control, 92, 17-23. doi:10.1016/j.biocontrol.2015.09.002Nakkeeran, S., Renukadevi, P., & Aiyanathan, K. E. A. (2016). Exploring the Potential of Trichoderma for the Management of Seed and Soil-Borne Diseases of Crops. Integrated Pest Management of Tropical Vegetable Crops, 77-130. doi:10.1007/978-94-024-0924-6_4Magan, N., & Lacey, J. (1984). Effect of water activity, temperature and substrate on interactions between field and storage fungi. Transactions of the British Mycological Society, 82(1), 83-93. doi:10.1016/s0007-1536(84)80214-4Sinclair, J. B. (1991). Latent Infection of Soybean Plants and Seeds by Fungi. Plant Disease, 75(3), 220. doi:10.1094/pd-75-0220Verhoeff, K. (1974). Latent Infections by Fungi. Annual Review of Phytopathology, 12(1), 99-110. doi:10.1146/annurev.py.12.090174.000531Sempere, F., & Santamarina, M. P. (2011). Cryo-scanning electron microscopy and light microscopy for the study of fungi interactions. Microscopy Research and Technique, 74(3), 207-211. doi:10.1002/jemt.2089

Chemodiversity of wild populations of aromatic plants as source of valuable essential oil profiles. A study on Thymus vulgaris L. from Valencia (Spain)

[EN] Chemodiversity of wild populations of aromatic plants is a valuable source of essential oils, whose composition may be suitable for specific purposes according their biological activity. Furthermore, knowing the intrapopulational variability based on individual analysis has allowed characterizing atypical profiles, which can reach high levels of active compounds. Obviously, it requires the treatment of a high number of individual samples. In this work, a methodology to characterize T. vulgaris profiles in an area of recognized biodiversity was proposed and applied. After Thin Layer Chromatography (TLC) screening data of 85 individual samples, 7 groups, and 13 individuals were classified. Then, 20 samples were subjected to GC/MS and GC/FID analysis, respectively. These data were subjected to Hierarchical Agglomerative, Discriminant Analysis and ANOVA, which finally highlighted five profiles: (1) based on the camphane skeleton (camphene, camphor and borneol), (2) rich in the oxygenated sesquiterpenic fraction, (3) rich in 1,8-cineole, with appreciable amounts of camphor and borneol (typical chemotype from Eastern Iberian Peninsula), (4) camphor and terpinen-4-ol as major compounds, and (5) linalool chemotype. It should be noted that the percentages of the main compounds in these groups were higher than some of those described in the literature for similar chemotypes. In summary, the preliminary screening by TLC, grouping individuals with similar profiles, allowed establishing a quick first approximation to the chemodiversity of T. vulgaris in the studied area. Furthermore, the analysis of unclassified and potentially atypical individuals has also provided valuable information to establish the final profiles.Llorens Molina, JA.; Vacas, S.; Burgals Royo, E.; Santamarina Siurana, MP.; Verdeguer Sancho, MM. (2020). Chemodiversity of wild populations of aromatic plants as source of valuable essential oil profiles. A study on Thymus vulgaris L. from Valencia (Spain). Natural Volatiles and Essential Oils. 7(3):29-50. https://doi.org/10.37929/nveo.722313S29507

In Vitro Antagonistic Activity of <i>Trichoderma harzianum</i> against <i>Fusarium sudanense</i> Causing Seedling Blight and Seed Rot on Wheat

Fusarium sudanense is a novel fungus recently isolated from asymptomatic samples of wheat grains in Argentina. The fungus caused symptoms of seedling blight and seed rot on wheat after artificial inoculations. It is known that the production of mycotoxins by pathogens belonging to the Fusarium genus is harmful to human and animal health. Moreover, the warm and humid conditions that are favorable for growth and mycotoxin production of these species put the Argentinian wheat production area at a high risk of mycotoxin contamination with this novel pathogen. The aim of this work was to evaluate the antagonistic effect of Trichoderma harzianum against F. sudanense under in vitro tests at different environmental conditions. Fungi were screened in dual culture at different water activities (αw) (0.995, 0.98, 0.95, and 0.90) and temperatures (25 and 15 °C). The growth rate of the fungi, interaction types, and dominance index were evaluated. Also, the interaction between T. harzianum and F. sudanense was examined by light and cryo-scanning microscopy. T. harzianum suppressed the growth of F. sudanense at 0.995, 0.98, and 0.95 αw at 25 °C and 0.995 and 0.98 αw at 15 °C. Macroscopic study revealed different interaction types between F. sudanense and T. harzianum on dual culture. Dominance on contact where the colonies of T. harzianum overgrew the pathogen was the most common interaction type determined. The competitive capacity of T. harzianum was diminished by decreasing the temperature and αw. At 0.95 αw and 15 °C, both fungi grew slowly, and interaction type "A" was assigned. Microscopic analysis from the interaction zone of dual cultures revealed an attachment of T. harzianum to the F. sudanense hyphae, penetration with or without formation of appressorium-like structures, coiling, plasmolysis, and a veil formation. According to our results, T. harzianum demonstrated capability to antagonize F. sudanense and could be a promising biocontrol agent.Centro de Investigación y Desarrollo en Fermentaciones Industriale