Detection of the fumonisin-producing Fusarium fujikuroi species complex (FFSC) associated with wild grasses in Iran (original) (raw)
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Twenty one Fusarium isolates namely, F. verticillioides (15 isolates), F. proliferatum (3isolates), F. anthophilum (3 isolates), isolated from different types of animal feeds, collected from different animal farms in Cairo, Egypt, were subjected to PCR analysis. Depending on the data obtained from the random amplified polymorphic DNA (RAPD) analysis it was possible to discriminate between the three Fusarium species. The data indicated that 8 RAPD markers distinguished the F. verticillioides from the other species. Similarly 7 and 2 markers were found to be specific to F. anthophilum and F. proliferatum respectively. These markers can be considered as useful markers for proper identification of the three Fusarium species. In the present study a PCR-based detection kit for identifying fumonisin-producing Fusarium species was developed. One set of primers designed from the internal transcription spacer region (ITS) of the genus Fusarium was used. The data indicated that, all of the 21 isolates showed clear band corresponding to the molecular size of the ITS region (431bp), where it was absent in the control sample (Aspergillus flavus). The other set of primers specific to fumonisin producing fum1 gene region of Fusarium was used to differentiate the fumonisin producing Fusarium species from non-fumonisin producers. The data indicated that all of the Fusarium isolates amplified fragments with the molecular size of 183bp corresponding to the correct size of fum1 gene, while this band could not be detected in the control. The results of this study indicated that PCR-based technique could be used not only to differentiate the Fusarium species from other genera of fungi but also to identify fumonisin-producing F. verticillioides, F. proliferatum and F. anthophilum.
Brazilian Journal of Microbiology, 2009
The study reports the occurrence of fumonisin producing Fusarium verticillioides in 90 samples of stored paddy (Oryza sativa L.) collected from different geographical regions of Karnataka, India. Fumonisin producing F. verticillioides was identified based on micromorphological characteristics and PCR using two sets of primers. One set of primers was F. verticillioides species specific, which selectively amplified the intergenic space region of rDNA. The other set of primers was specific to fumonisin producing F. verticillioides. Eight paddy samples were positive for F. verticillioides. Eleven isolates obtained from these samples were capable of producing fumonisin.
Twenty one Fusarium isolates namely, F. verticillioides (15 isolates), F. proliferatum (3isolates), F. anthophilum (3 isolates), isolated from different types of horse feeds, collected from different horse farms in Cairo, Egypt, were subjected to PCR analysis. Depend on the data obtained from the random amplified polymorphic DNA (RAPD) analysis it is possible to discriminate between the three Fusarium species. The data indicate that 8 RAPD markers distinguish the F. verticillioides from the other species. Similarly 7 and 2 markers were found to be specific to F. anthophilum and F. proliferatum respectively. These markers can be considered as useful markers for proper identification of the three Fusarium species. In the present study a PCR-based detection kit for identifying fumonisin-producing Fusarium species was developed. One set of primers designed from the internal transcription spacer region (ITS) of the genus Fusarium was used. The data indicated that, all of the 21 isolates showed a clear bands corresponding to the molecular size of the ITS region (431bp), where it was absent in the control sample (Aspergillus flavus). The other set of primers specific to fumonisin producing fum1 gene region of Fusarium was used to differentiate the fumonisin producing Fusarium species from non-fumonisin producers. The data indicate that all of the Fusarium isolates amplified fragments with the molecular size of 183bp corresponding to the correct size of fum1 gene while this band could not be detected in the control. The results of this study indicated that PCR-based technique could be used not only to 2 differentiate the Fusarium species from other genera of fungi but also to identify fumonisin-producing F. verticillioides, F. proliferatum and F. anthophilum.
Molecular Detection of Fumonisin-producing Fusarium Species in Animal Feeds Using Polymerase Chain Reaction (PCR) 1Department of Microbiology, Faculty of Veterinary Medicine, Cairo University- Giza-Egypt. 2Department of Mycology and Mycotoxins, Animal Health Research Institute, Giza, Egypt). 3Department of Genetics, Faculty of Agriculture, Cairo University, Giza-Egypt. ABSTRACT Twenty one Fusarium isolates namely, F. verticillioides (15 isolates), F. proliferatum (3isolates), F. anthophilum (3 isolates), isolated from different types of animal feeds, collected from different animal farms in Cairo, Egypt, were subjected to PCR analysis. Depending on the data obtained from the random amplified polymorphic DNA (RAPD) analysis it was possible to discriminate between the three Fusarium species. The data indicated that 8 RAPD markers distinguished the F. verticillioides from the other species. Similarly 7 and 2 markers were found to be specific to F. anthophilum and F. proliferatum respectively. These markers can be considered as useful markers for proper identification of the three Fusarium species. In the present study a PCR-based detection kit for identifying fumonisin-producing Fusarium species was developed. One set of primers designed from the internal transcription spacer region (ITS) of the genus Fusarium was used. The data indicated that, all of the 21 isolates showed clear band corresponding to the molecular size of the ITS region (431bp), where it was absent in the control sample (Aspergillus flavus). The other set of primers specific to fumonisin producing fum1 gene region of Fusarium was used to differentiate the fumonisin producing Fusarium species from non-fumonisin producers. The data indicated that all of the Fusarium isolates amplified fragments with the molecular size of 183bp corresponding to the correct size of fum1 gene, while this band could not be detected in the control. The results of this study indicated that PCR-based technique could be used not only to differentiate the Fusarium species from other genera of fungi but also to identify fumonisin-producing F. verticillioides, F. proliferatum and F. anthophilum. Key words: Fusarium species, Animal feeds, Fumonisin-pro
… of agricultural and …, 2006
A total of 52 corn samples collected in 2000 from four main corn production provinces of Iran (Fars, Kermanshah, Khuzestan, and Mazandaran) were analyzed for contamination with Fusarium verticillioides and fumonisins (FB 1 , FB 2 , FB 3 , and 3-epi-FB 3 ). The mean incidence of F. verticillioides (percent of kernels infected) for these four areas was 26.7, 21.4, 24.9, and 59.0%, respectively. The incidence in Mazandaran was significantly (p < 0.05) above that of the other areas. All samples from Mazandaran were contaminated with fumonisins with a mean level of total fumonisins of 10674 µg/ kg. In contrast, the incidence of fumonisin contamination above 10 µg/kg was 53 (8/15), 42 (5/12), and 57% (8/14) in the samples from Fars, Kermanshah, and Khuzestan, respectively, and the corresponding mean total fumonisin levels were 215, 71, and 174 µg/kg, respectively. No statistical differences (p > 0.05) were observed in the fumonisin levels of the corn samples from these three provinces, which were significantly (p < 0.05) lower than the fumonisin contamination in samples from Mazandaran.
Fusarium verticillioides produces Fumonisins are a group of mycotoxins that contaminate food and feed products posses maximum threat to human and animal health. In this work Eighty two strains of Fusarium species collected from infected rice samples were subjected to PCR assay to discriminate fumonisin producing and nonproducing strains with Inter Generic Spacer region (IGS) of rDNA coding units specific primer named as VERTF-1/VERTF-2 were used. 21 isolates of F. verticillioides scored positive for VERTF-1/ VERTF-2 pair of primers proves to be potential fumonisin production and 25 isolates were scored negative. Specific primers for polyketide synthase (PKS) gene FUM1-(previously FUM5) were used to all 83 strains resulted in positive signals observed in 21 strains of F. verticillioides. This present study proves the efficiency of IGS and gene specific primer also represents well for Fusarium strains isolated from rice also. For both primers PCR detection was consistent even at 100 pg/μl concentration of genomic DNA. This quite rapid and specific method helps in accurate discrimination of Fumonisin producing strains.
Food Biotechnology, 2008
One hundred and three Fusarium isolates from maize samples collected from different districts of Karnataka state, India, were analyzed with genus-specific, species-specific, and potential fumonisin specific oligonucleotide primers. One set of genus-specific primers ITS F and ITS R based on a highly conserved ITS region of the genus Fusarium were used to differentiate Fusarium species from closely related genera. All the Fusarium species tested scored positive with the ITS pair of primers. Detection and identification of Fusarium verticillioides species was done by using a newly designed reverse primer VERT-R (5′-CGA CTC ACG GCC AGG AAA CC −3′) based on an intergenic spacer sequence (IGS) combined with an already designed forward primer VERTF-1 (5′-GCG GGA ATT CAA AAG TGG CC-3′) published previously. Out of 103 Fusarium species tested, 83 isolates of F. verticillioides scored positive for VERTF-1/ VERT-R species-specific pair of primers. Further to discriminate potential fumonisinproducing and nonproducing strains of F. verticillioides, the VERTF-1/VERTF-2 set of primers [VERTF-1 (5′-GCG GGA ATT CAA AAG TGG CC-3′) and VERTF-2 (5′-GAG GGC GCG AAA CGG ATC GG-3′)] were used. 64 isolates of F. verticillioides scored positive for VERTF-1/ VERTF-2 pair of primers. In total, three primers, one forward primer VERTF-1 and two reverse primers VERT-R and VERTF-2, were used for the confirmation of F. verticillioides up to the species level
World Journal of Microbiology and Biotechnology, 2010
Fumonisins are a group of fungal toxins, occurring worldwide in maize infected mainly by Fusarium verticillioides. This paper describes the level of fumonisins in maize seed samples and the ability of F. verticillioides strains isolated from maize seeds grown in India to produce fumonisins. Forty-three seed samples intended to be used for consumption were collected from different regions of Karnataka and Andhra Pradesh. The samples were subjected to the agar plate method for the detection of F. verticillioides. Identification of F. verticillioides was done based on morphological characters and further confirmed by polymerase chain reaction. The majority of the samples were infected by F. verticillioides and infection percentage in the individual samples ranged from 5 to 51%. Twenty-three out of 35 (65%) strains were positive for fumonisin production in high performance liquid chromatography (HPLC) and competitive directenzyme linked immuno sorbent assay (CD-ELISA). Fumonisin level in seed samples ranged from 200 to 1,722 lg/g using CD-ELISA. HPLC could differentiate FB1 and FB2 toxins; out of 35 strains, 14 (40%) showed both FB1 and FB2 production. These findings indicate that there may be a risk of human exposure to fumonisins through the consumption of F. verticillioides infected cornbased foods in India.
European Journal of Biology
Objective: Fusarium spp. cause Fusarium head blight (FHB) and crown rot (CR) diseases. They also have harmful effects on animal and human health through their mycotoxins. Within the scope of this study, F. graminearum and F. culmorum isolates were purified from wheat ears and stalks contaminated with phytopathogens, which had been collected from various regions of Turkey, were identified and characterized by conventional and molecular methods. Materials and Methods: Sixty-eight Fusarium samples were isolated by single spore analysis and classified according to their macroconidia shape and size. Morphologically characterized samples were verified by amplification of SCAR markers. Their mating types (MAT) and chemotypes were also determined through polymerase chain reaction (PCR). Results: Thirty-eight F. graminearum and 30 F. culmorum isolates were identified via amplification of UBC85 and OPT18 SCAR markers, respectively. All isolates were determined as trichothecene producers by amplification of the tri5 gene. All F. graminarum isolates carry both MAT-1 and MAT-2 loci, whereas 7 of F. culmorum isolates were also determined as MAT-1 and 23 of them as MAT-2 mating types. Deoxynivalenol production capacity of all isolates was identified by tri13 amplification for chemotype determination. Conclusion: Routine monitoring of phytopathogens and their mycotoxin levels is a requirement since their annual levels may vary depending on environmental factors. This work provides knowledge about the distribution of Fusarium spp. leading to FHB and CR in different regions of Turkey between 2010 and 2020. Also, their chemotypes were demonstrated. Our studies will contribute to disease profiling and it is the first step in disease management.
Fumonisin B1-producing Fusarium species from agricultural crops in Malaysia
Many species of Fusarium are pathogenic as well toxigenic to a wide variety of plants. The present study was conducted to determine the ability of four members of Fusarium fujikuroi species complex and F. oxysporum from various crops to produce fumonisin B1 (FB 1). Isolates of Fusarium species from infected parts of asparagus, ginger, oil palm, mango, banana, maize, and rice were identified as F. verticillioides (11 isolates), F. proliferatum (50 isolates), F. fujikuroi (24 isolates), F. andiyazi (six isolates), and F. oxysporum (32 isolates). FUM1, a gene involved in fumonisin biosynthesis, was detected in 94 isolates of F. verticillioides (11 isolates), F. proliferatum (49 isolates), F. fujikuroi (24 isolates), and F. oxysporum (10 isolates) but only 61 were positive for FB 1 when tested using RIDA ® Quick Fumonisin test strip, indicating that the presence of FUM1 was not necessarily associated with FB 1 production. Based on ultra-high performance liquid chromatography, all the 61 isolates were detected to produce FB 1 , at variable levels, with concentrations ranging from 0.60 to 29.2 mg/g. Our results suggested that there is a potential risk of FB 1 contamination in agricultural crops in Malaysia.