Kim, Da-Woon;Kim, Gi-Yong;Kim, Hee-Kyoung;Kim, Jueun;Jeon, Sun Jeong;Lee, Chul Won;Lee, Hyang Burm;Yun, Sung-Hwan
The Plant Pathology Journal
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v.32
no.3
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pp.182-189
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2016
Together with the Fusarium graminearum species complex, F. culmorum is a major member of the causal agents of Fusarium head blight on cereals such as wheat, barley and corn. It causes significant yield and quality losses and results in the contamination of grain with mycotoxins that are harmful to humans and animals. In Korea, F. culmorum is listed as a quarantine fungal species since it has yet to be found in the country. In this paper, we report that two isolates (J1 and J2) of F. culmorum were collected from the air at a rice paddy field in Korea. Species identification was confirmed by phylogenetic analysis using multilocus sequence data derived from five genes encoding translation elongation factor, histone H3, phosphate permease, a reductase, and an ammonia ligase and by morphological comparison with reference strains. Both diagnostic PCR and chemical analysis confirmed that these F. culmorum isolates had the capacity to produce nivalenol, the trichothecene mycotoxin, in rice substrate. In addition, both isolates were pathogenic on wheat heads and corn stalks. This is the first report on the occurrence of F. culmorum in Korea.
Barley worldwide is affected seriously by Fusarium seedling blight (FSB) and Fusarium head blight (FHB) diseases caused by the Fusarium species. The objective of this study was to facilitate the resistance of hulled and hull-less barley at different growth stages to F. culmorum according to direct parameters: disease rating (DR), fresh weight of leaves and roots, kernel weight per spike, kernel number per spike, plump kernels, and indirect parameters - chlorophyll a fluorescence (CF). Plate assay, greenhouse and field tests were performed on 30 spring barley doubled haploid (DH) lines and their parents infected with Fusarium culmorum. Direct parameters proved that hulled genotypes show less symptoms. Most studied chlorophyll a fluorescence (CF) parameters (apart from DIo/CS - amount of energy dissipated from PSII for laboratory test, TRo/CS - amount of excitation energy trapped in PSII reaction centers, ETo/CS - amount of energy used for electron transport and RC/CS - number of active reaction centres in the state of fully reduced PSII reaction center in field experiment) were significantly affected by F. culmorum infection. In all experiments, hulled genotypes had higher values of CF parameters compared to hull-less ones. Significant correlations were detected between direct and indirect parameters and also between various environments. It was revealed that ABS/CS, TRo/CS, and RC/CS have significant positive correlation in greenhouse test and field experiment. Significant correlations suggest the possibility of applying the CF parameters in selection of barley DH lines resistant to F. culmorum infection.
Fusarium head blight is one of the most important and most common diseases of winter wheat. In order to better understanding this disease and to assess the correlations between different factors, 30 cultivars of this cereal were evaluated in a two-year period. Fusarium head blight resistance was evaluated and the concentration of trichothecene mycotoxins was analysed. Grain samples originated from plants inoculated with Fusarium culmorum and naturally infected with Fusarium species. The genetic distance between the tested cultivars was determined and data were analysed using multivariate data analysis methods. Genetic dissimilarity of wheat cultivars ranged between 0.06 and 0.78. They were grouped into three distinct groups after cluster analysis of genetic distance. Wheat cultivars differed in resistance to spike and kernel infection and in resistance to spread of Fusarium within a spike (type II). Only B trichothecenes (deoxynivalenol, 3-acetyldeoxynivalenol and nivalenol) produced by F. culmorum in grain samples from inoculated plots were present. In control samples trichothecenes of groups A (H-2 toxin, T-2 toxin, T-2 tetraol, T-2 triol, scirpentriol, diacetoxyscirpenol) and B were detected. On the basis of Fusarium head blight assessment and analysis of trichothecene concentration in the grain relationships between morphological characters, Fusarium head blight resistance and mycotoxins in grain of wheat cultivars were examined. The results were used to create of matrices of distance between cultivars - for trichothecene concentration in inoculated and naturally infected grain as well as for FHB resistance Correlations between genetic distance versus resistance/mycotoxin profiles were calculated using the Mantel test. A highly significant correlation between genetic distance and mycotoxin distance was found for the samples inoculated with Fusarium culmorum. Significant but weak relationships were found between genetic distance matrix and FHB resistance or trichothecene concentration in naturally infected grain matrices.
Fusarium culmorum is one of the most important causal agents of root rot of wheat. In this study, 10 F. culmorum isolates were collected from farms located in five agro-ecological regions of Morocco. These were used to challenge 20 durum wheat genotypes via artificial inoculation of plant roots under controlled conditions. The isolate virulence was determined by three traits (roots browning index, stem browning index, and severity of root rot). An alpha-lattice design with three replicates was used, and the resulting ANOVA revealed a significant (P < 0.01) effect of isolate (I), genotype (G), and G × I interaction. A total of four response types were observed (R, MR, MS, and S) revealing that different genes in both the pathogen and the host were activated in 53% of interactions. Most genotypes were susceptible to eight or more isolates, while the Moroccan cultivar Marouan was reported resistant to three isolates and moderately resistant to three others. Similarly, the Australian breeding line SSD1479-117 was reported resistant to two isolates and moderately resistant to four others. The ICARDA elites Icaverve, Berghisyr, Berghisyr2, Amina, and Icaverve2 were identified as moderately resistant. Principal component analysis based on the genotypes responses defined two major clusters and two sub-clusters for the 10 F. culmorum isolates. Isolate Fc9 collected in Khemis Zemamra was the most virulent while isolate Fc3 collected in Haj-Kaddour was the least virulent. This work provides initial results for the discovery of differential reactions between the durum lines and isolates and the identification of novel sources of resistance.
Cepacidine A was previously isolated as a novel antifungal antibiotic from the culture broth of Pseudomonas cepacia AF2001. It exhibits a potent in vitro antifungal activity against various plant pathogenic fungi, such as Plasmopora veticola on grapes, Septoria nodorum and Fusarium culmorum on wheat, as well as Colletotrichum lagenarium on cucumbers. Accordingly, this study was conducted to evaluate the potential crop protection activity of strain P. cepacia AF2001. The strain was tested in semi-greenhouse biocontrol assays, and showed an excellent biological activity against Pythium ultimum in cotton and cucumbers; however, only a minor activity against Rhizoctonia aolani in cotton was observed. Furthermore, the nematodes Haemonchus contortus and Trichostrongylus only exhibited a moderated activity in the in vitro larval development assay with no activity in the in vivo animal model.
Seventy four microorganisms, which have antagonistic activity against to Fusarium culmorum, were tested for their inhibitory effect on colony development of obligate biotroph Erysiphe graminis f. sp. hordei Marchal, the causal agent of powdery mildew on barley plants. Of these, 13 actinomycetes isolates were shown to reduce the colony development of mildew completely by application of their 10% cell-free culture filtrates on barley leaves. An Isolate, A252, was the most powerful antagonist and its antifungal activity was further assessed. The colony development of mildew was significantly reduced by application of the 1% cell-free culture filtrate of isolate A252. In comparison to the control, the protective and curative application of 10% cell-free culture filtrate from A252 showed 88.5% and 96.1% reduction of colony numbers respectively. By the protective application, 68.3% of the inhibition was observed after 9 days of treatment, thus showed prolonged inhibitory effect. In vitro test, complete inhibition of the mycelial growth of Microdochium nivale was achieved by the treatment of 1% A252 culture filtrate and 80.2% of inhibition was observed by the 0.1% treatment.
Rye was used here to dissect molecular mechanisms of resistance to Fusarium head blight (FHB) and to go deeper with our understanding of that process in cereals. F. culmorum-damaged kernels of two lines different in their potential of resistance to FHB were analyzed using two-dimensional gel electrophoresis and mass spectrometry to identify resistance markers. The proteome profiling was accompanied by measurements of ${\alpha}-$ and ${\beta}-amylase$ activities and mycotoxin content. The proteomic studies indicated a total of 18 spots with clear differences in protein abundance between the more resistant and more susceptible rye lines after infection. Eight proteins were involved in carbohydrate metabolism of which six proteins showed a significantly higher abundance in the resistant line. The other proteins recognized here were involved in stress response and redox homeostasis. Three remaining proteins were associated with protease inhibition/resistance and lignin biosynthesis, revealing higher accumulation levels in the susceptible rye line. After inoculation, the activities of ${\alpha}-$ and ${\beta}-amylases$, higher in the susceptible line, were probably responsible for a higher level of starch decomposition after infection and a higher susceptibility to FHB. The presented results could be a good reference for further research to improve crop resistance to FHB.
Journal of The Korean Society of Grassland and Forage Science
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v.31
no.4
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pp.451-462
/
2011
The purpose of this study was to investigate fungi and mycotoxin contamination of the rice straw bale silage in Korean. It was tested the 33 samples of rice straw round bale silage with various condition which fed cattle in the farm. The level of fungal contamination was $2.1{\times}10^6\;cfu\;g^{-1}$ in the average and $9.2{\times}10^8\;cfu\;g^{-1}$ in the maximum. The fungal contamination was detected in the all of normal samples which good condition of rice straw bale silage. When the fungi was isolate and identify, it was found 28 species and mycotoxin producing fungi were 8 species as following as Aspergillus flavus, Aspergillus fumigatus, Fusarium culmorum, Fusarium verticillioides, Penicillium carneum, Penicillium paneum, Penicillium roqueforti, Penicillium viridicatum. Specially, Penicillium paneum was found 42% of samples and Aspergillus sp. (A. flavus, A. fumigatus) are 21% of samples. In case of mycotoxin contamination, the 42% of samples are detected more than one kind of mycotoxin. Some samples are contaminated three kinds of mycotoxin. This study was not found aflatoxin ($B_1$, $B_2$, $G_1$, $G_2$) and fumonisin ($B_1$, $B_2$), but were detected the contamination of ochratoxin A (1.0~5.8 ug/kg), deoxynivalenol (DON, 156.0~776.7 ug/kg) and zearalenone (ZON, 38.0~750.0 ug/kg). Therefore, the above results show that rice straw round bale silage expose on hazard factors as mycotoxigenic fungi and mycotoxin contamination, and than need more research about mycotoxin in animal feed to protect animal and human healthy.
A keratinolytic enzyme secreted by Aspergillus flavus K-03 cultured in feather meal basal medium (FMBM) containing 2% (w/v) chicken feather was purified and characterized. Keratinolytic enzyme secretion was the maximal at day 16 of the incubation period at pH 8 and $28^{\circ}C$. No relationship was detected between enzyme yield and increase of fungal biomass. The fraction obtained at 80% ammonium sulfate saturation showed 2.39-fold purification and was further purified by gel filtration in Sephadex G-100 followed by ion exchange chromatography on DEAE-Sephadex A-50, yielding an active protein peak showing 11.53-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymograms indicated that the purified keratinase is a monomeric enzyme with 31 kDa molecular weight. The extracellular keratinase of A. flavus was active in a board range of pH ($7{\sim}10$) and temperature ($30^{\circ}C{\sim}70^{\circ}C$) profiles with the optimal for keratinase activity at pH 8 and $45^{\circ}C$. The keratinase activity was totally inhibited by protease inhibitors such as phenylmethylsulfonyl fluoride (PMSF), iodoacetic acid, and ethylenediaminetetraacetate (EDTA) while no reduction of activity by the addition of dithiothreitol (DTT) was observed. N-terminal amino acid sequences were up to 80% homologous with the fungal subtilisins produced by Fusarium culmorum. Therefore, on the basis of these characteristics, the keratinase of A. flavus K-03 is determined to be subtilisins-like.
Journal of The Korean Society of Grassland and Forage Science
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v.30
no.1
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pp.77-88
/
2010
In order to ensure good animal health and performance, it is essential to produce forages with high feeding value and good hygienic quality. However, huge amounts of forages consumed by ruminants are contaminated with mold prior to harvest or during storage as hay, straw or silage. These mold can grow in forages only when nutrients are available, correct temperature exist, oxygen is present, and unbound water is available. Fungal 'species can be divided into two groups: field fungi and storage fungi. Field fungi invade the forages while the crop is still in the field, require high moisture conditions, and are such as species of Fusarium, Alternaria, Clodosporium, Diplodia, Gibberrella and Helminthosporium. Storage fungi invade forages during storage and need less moisture than field fungi. These such as species of Aspergillus and Penicillium usually do not occur any problem before harvest. Mold growth can spoil the nutritional aspects of the forages and also results in secondary metabolites that are highly toxic to animal, humans and plants. Moldy feeds are less palatable and may reduce dry matter intake. This, in turn, leads to a reduction of nutrition intake, reducing weight gains or milk production. Performance losses of 5 to 10 percent are typical with moldy feeds. Mycotoxins are toxic substances produced by fungi (molds) growing on crops in the field or storages. While greater than 400 mycotoxins have been chemically identified, the biological or veterinary medical impact of only several mycotoxins is known. Mycotoxins have attracted considerable attention as potential causes for poor performance and health disorders in domestic livestock. They can be carcinogenic, hepatotoxic, hematotoxic, immunosuppressive, estrogenic, or mutagenic. So, feeding moldy forages has adverse effects on animal health and milk consumers. Also, this author reported that rice straw hay was contaminated mycotoxigenic fungi such as Penicillium roqueforti and Fusarium culmorum in Korea. Therefore, it is an urgent need to develop an improved post harvest storage method to reduce nutrient loss and mycotoxin contamination of forages, which will have a positive impact on human health.
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