Morphological and molecular identification and PCR amplification to determine the toxigenic potential of Fusarium spp. isolated from maize ears in southern Poland

Lenart, Anna; Klimek-Kopyra, Agnieszka; Boroń, P

doi:10.1007/s12600-012-0284-7

Cited by 7 publications

(4 citation statements)

References 29 publications

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“…Some Aspergillus strains ( A. flavus NRRL 3518, A. flavus SS2, A. carbonarius NRRL 368, and A. ochraceus NRRL 35018) did not show amplification in any target genes ( Figure 2 A,C). The probability of a particular toxin being produced can be predicted according to the presence or absence of an amplification product, but the effective biosynthesis of the toxin remains to be confirmed by analytical chemistry analysis [ 32 , 33 ].…”

Section: Resultsmentioning

confidence: 99%

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

Sudharsan

Britzi²,

Zakin

et al. 2017

Toxins

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This study aimed to assess the occurrence of toxigenic fungi and mycotoxin contamination in stored wheat grains by using advanced molecular and analytical techniques. A multiplex polymerase chain reaction (PCR) strategy was established for rapid identification of mycotoxigenic fungi, and an improved analytical method was developed for simultaneous multi-mycotoxin determination in wheat grains by liquid chromatography-tandem mass spectrometry (LC/MS/MS) without the need for any clean-up. The optimized multiplex PCR method was highly specific in detecting fungal species containing species-specific and mycotoxin metabolic pathway genes. The method was applied for evaluation of 34 wheat grain samples collected from storage warehouses for the presence of mycotoxin-producing fungi, and a few samples were found positive for Fusarium and Aspergillus species. Further chemical analysis revealed that 17 samples contained mycotoxins above the level of detection, but only six samples were found to be contaminated over the EU regulatory limits with at least one mycotoxin. Aflatoxin B1, fumonisins, and deoxynivalenol were the most common toxins found in these samples. The results showed a strong correlation between the presence of mycotoxin biosynthesis genes as analyzed by multiplex PCR and mycotoxin detection by LC/MS/MS. The present findings indicate that a combined approach might provide rapid, accurate, and sensitive detection of mycotoxigenic species and mycotoxins in wheat grains.

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Section: Resultsmentioning

confidence: 99%

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

Sudharsan

Britzi²,

Zakin

et al. 2017

Toxins

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show abstract

“…PCR amplification was carried out in the Peltier Thermal Cycler, PTC-100 ® (MJ Research, Inc. USA) according to temperature profiles described by Lenc et al (2008) and Lenart et al (2013). To visualize the PCR products, 1X TBE electrophoresis in ethidium-bromide-stained and 1% agarose gel were used.…”

Section: Methodsmentioning

confidence: 99%

Detection of nivalenol and deoxynivalenol chemotypes produced by Fusarium graminearum species complex isolated from barley in Iran using specific PCR assays

Chehri¹,

Godini²

2017

Journal of Plant Protection Research

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In order to identify trichothecenes chemotypes produced by Fusarium graminearum species complex (FGSC) isolated from barley, 68 barley samples were collected from markets in Kermanshah and Hamedan provinces, Iran. Thirty-one Fusarium isolates were obtained from grains and morphologically classified into three species FGSC (14), F. equiseti (9), and F. proliferatum (8). The identification of the members of FGSC was confirmed molecularly using Fg16F/Fg16R primers. Fusarium asiaticum isolates (4) were distinguished from other FGSC using Fg6CTPSf177/Fg16R primers. Polymerase chain reaction-based (PCRbased) detection of mycotoxin-synthesis-pathway gene was also used to determine the potential of the analysed strains to produce deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-AcDON), 3-acetyldeoxynivalenol (3-AcDON), and nivalenol (NIV). Of 14 tested isolates, 10 and 4 isolates belonged to DON and NIV chemotype, respectively. Also, the results of DON chemotype survey using specific primers MinusTri7F/R and Tri315F/R showed 1 and 9 isolates produced 3-AcDON and 15-AcDON, respectively. These results show that DON was the most common chemotype in western Iran. To our knowledge, this is the first report on 15-AcDON, 3-AcDON, and NIV isolated from barley in Iran.

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“…e PCR amplification for determination of the toxigenic potential of the Fusarium spp. isolates was carried out in the Veriti ermal Cycler (Applied Biosystems, USA) according to the following program: 94 °C for 3 min, 94 °C for 45 sec, 58 °C for 45 sec, 72 °C for 1 min and 35 cycles, and final extension at 72 °C for 5 min as described by [32,33]. e PCR products were visualized by 1 × TBE electrophoresis in ethidiumbromide-stained 1% agarose gel.…”

Section: Pcr Assessment Of the Toxigenic Potential Of The Fusarium Sppmentioning

confidence: 99%

Molecular Determination of Toxigenic Potential of Fusarium spp. Isolated from Seeds of Wheat (Triticum aestivum) Genotypes and Evaluation of Levels of Fumonisins in the Grains at Harvest in Three Major Wheat Producing Counties in Kenya

Otieno

Imbahale

Wekesa

et al. 2022

International Journal of Agronomy

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Infestation of cereals such as wheat by pathogenic Fusarium species (Fusarium spp.) is associated with over 25% weight loss of the grain and accumulation of hazardous mycotoxins. Potential management strategies against the pathogens include the development of more resistant wheat genotypes and the development of rapid and accurate Fusarium spp. identification methods, among other control measures. This study evaluated the toxigenic potential in populations of Fusarium spp. isolated from seeds of locally developed wheat genotypes and levels of fumonisins in the grains at harvest in three counties in Kenya. The sampling of the wheat grains took place between September 2016 and October 2017. Determination of toxigenic potential was PCR based using Tri13F/Tri13DONR and FUM1F/FUM1R specific primer pairs while detection of fumonisin levels was done using Total Fumonisin Assay 0.25/5.0 ELISA kit. Data were analyzed using ANOVA and the Tukey HSD test. FUM1 gene was detected in 60% of the Fusarium spp. analyzed. The distribution of the gene within the isolates across the three regions was as follows: Narok: 54%, Uasin Gishu: 25%, and Nakuru: 21%. Tri13DON gene was not detected in the assessed potential deoxynivalenol (DON) producers. Fumonisin levels in the wheat samples from the three counties were significantly different ( p < 0.001 ). The highest fumonisin levels, 9.6 ppm, occurred in 30.7% of the grains of the studied wheat genotypes. Relatively high fumonisin levels occurred in Njoro II, Eagle 10, Robin, and Korongo wheat genotypes with no significant difference at 0.05 confidence interval. In conclusion, the toxigenic potential amongst the Fusarium spp. studied was confirmed based on the occurrence of FUM1 gene and the detection of fumonisins in 76% of the sampled wheat grains. More research is recommended to ascertain the prevalence of the genes determining the production of DON and the other trichothecenes in Fusarium spp. prevalent in the developed wheat genotypes in the different agroecological regions in Kenya. In addition, the assessment of the occurrence and levels of the respectful mycotoxins also needs further research. These would provide additional information for the improvement of strategies put in place to manage the effects of the pathogenic Fusarium spp. in the crop and to ensure mycotoxin safety of the wheat food chain for both livestock and human consumption. It also shows the need for the development of more disease-resistant wheat varieties by wheat seed-producing companies.

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Morphological and molecular identification and PCR amplification to determine the toxigenic potential of Fusarium spp. isolated from maize ears in southern Poland

Cited by 7 publications

References 29 publications

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

Rapid Detection and Identification of Mycotoxigenic Fungi and Mycotoxins in Stored Wheat Grain

Detection of nivalenol and deoxynivalenol chemotypes produced by Fusarium graminearum species complex isolated from barley in Iran using specific PCR assays

Molecular Determination of Toxigenic Potential of Fusarium spp. Isolated from Seeds of Wheat (Triticum aestivum) Genotypes and Evaluation of Levels of Fumonisins in the Grains at Harvest in Three Major Wheat Producing Counties in Kenya

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