Species of the genus Fusarium are well-researched in many fields, and a commonly problem by researchers interested in Fusarium species is the probable taxonomic system and the identification method of this genus. The traditional taxonomic system for fungi has been proposed based on the mainly morphological species concept, including the genus Fusarium. Recently, many researchers have applied molecular markers to examine the taxonomy and identification of Fusarium species. This review shows some recent findings from our studies about molecular phylogeny and identification of Fusarium species based on analyses with nucleotide sequences. First, the genetic markers were evaluated for identifying Fusarium isolates by calculation of the homologies with pairwise comparison of all tested strains, and of the ratio of nucleotide substitution rate. It was suggested that aminoadipate reductase gene (lys2) is notionally the most appropriate genetic marker for identifying isolates among the six genes examined. Second, actual identification of food-borne isolates of the genus Fusarium based on the nucleotide sequence homology was performed, and the results were evaluated. In terms of accuracy and ease, b-tubulin gene, not lys2, is the most useful genetic maker among the six genes examined. Finally, the genetic markers were evaluated for the phylogenetic analysis of Fusarium species. It was suggested the lys 2 have a singular evolutionary history than other genes. To obtain a reliable phylogeny for Fusarium species, the lys2 sequences were excluded from the dataset, and the species tree was inferred. The reliable Fusarium species tree was reconstructed, and some interesting relationships were newly described.
{"title":"遺伝子塩基配列を用いての Fusarium 属菌の同定と分子系統学的位置付けに関する研究","authors":"渡辺 麻衣子","doi":"10.2520/MYCO.63.133","DOIUrl":"https://doi.org/10.2520/MYCO.63.133","url":null,"abstract":"Species of the genus Fusarium are well-researched in many fields, and a commonly problem by researchers interested in Fusarium species is the probable taxonomic system and the identification method of this genus. The traditional taxonomic system for fungi has been proposed based on the mainly morphological species concept, including the genus Fusarium. Recently, many researchers have applied molecular markers to examine the taxonomy and identification of Fusarium species. This review shows some recent findings from our studies about molecular phylogeny and identification of Fusarium species based on analyses with nucleotide sequences. First, the genetic markers were evaluated for identifying Fusarium isolates by calculation of the homologies with pairwise comparison of all tested strains, and of the ratio of nucleotide substitution rate. It was suggested that aminoadipate reductase gene (lys2) is notionally the most appropriate genetic marker for identifying isolates among the six genes examined. Second, actual identification of food-borne isolates of the genus Fusarium based on the nucleotide sequence homology was performed, and the results were evaluated. In terms of accuracy and ease, b-tubulin gene, not lys2, is the most useful genetic maker among the six genes examined. Finally, the genetic markers were evaluated for the phylogenetic analysis of Fusarium species. It was suggested the lys 2 have a singular evolutionary history than other genes. To obtain a reliable phylogeny for Fusarium species, the lys2 sequences were excluded from the dataset, and the species tree was inferred. The reliable Fusarium species tree was reconstructed, and some interesting relationships were newly described.","PeriodicalId":19069,"journal":{"name":"Mycotoxins","volume":"11 1","pages":"133-142"},"PeriodicalIF":0.0,"publicationDate":"2013-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84191527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
C. G. Afolabi, E. J. A. Ekpo, Ranajit Bandyopadhyay
{"title":"ナイジェリアにおけるゼアラレノンと T-2 トキシンによるとうもろこし汚染とヒトへの暴露","authors":"C. G. Afolabi, E. J. A. Ekpo, Ranajit Bandyopadhyay","doi":"10.2520/MYCO.63.143","DOIUrl":"https://doi.org/10.2520/MYCO.63.143","url":null,"abstract":"","PeriodicalId":19069,"journal":{"name":"Mycotoxins","volume":"83 1","pages":"143-149"},"PeriodicalIF":0.0,"publicationDate":"2013-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74182206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Y. Nakajima, Hinayo Ichikawa, Kazuyuki Maeda, T. Nishiuchi, Tetsuo Kobayashi, M. Kimura
Marker genes are required for the genetic manipulation of living organisms. For selecting transformants after the process of transformation, two types of marker genes are employed: one for the positive selection of the transformants carrying the positive selection marker gene and another for the negative selection of the transformants lacking the negative selection marker gene. Here we have summarized the current advancements in genome engineering of Fusarium species. We have focused on introducing arbitrary mutations by applying both positive and negative selection approaches that result in the exclusion of the marker genes from the target genome. We have discussed the advantages and necessity of using this technique in studies on the regulation of biosynthetic gene expression in mycotoxin production.
{"title":"Genome engineering of Fusarium species by using positive and negative selection approaches for studying regulation of mycotoxin production","authors":"Y. Nakajima, Hinayo Ichikawa, Kazuyuki Maeda, T. Nishiuchi, Tetsuo Kobayashi, M. Kimura","doi":"10.2520/MYCO.63.85","DOIUrl":"https://doi.org/10.2520/MYCO.63.85","url":null,"abstract":"Marker genes are required for the genetic manipulation of living organisms. For selecting transformants after the process of transformation, two types of marker genes are employed: one for the positive selection of the transformants carrying the positive selection marker gene and another for the negative selection of the transformants lacking the negative selection marker gene. Here we have summarized the current advancements in genome engineering of Fusarium species. We have focused on introducing arbitrary mutations by applying both positive and negative selection approaches that result in the exclusion of the marker genes from the target genome. We have discussed the advantages and necessity of using this technique in studies on the regulation of biosynthetic gene expression in mycotoxin production.","PeriodicalId":19069,"journal":{"name":"Mycotoxins","volume":"145 1","pages":"85-92"},"PeriodicalIF":0.0,"publicationDate":"2013-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88865282","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
C. Ezekiel, C. Nwangburuka, G. Chioma, M. Sulyok, B. Warth, C. Afolabi, Adenike A. Osiberu, Michael O. Oladimeji, O. A. Denton, G. Tayo, R. Krska
Seeds of two okra accessions and 17 samples of sesame seeds were examined for contamination by Fusarium species. Altogether, 37 Fusarium isolates were obtained from the two crops: 6 isolates from two okra accessions and 31 isolates from 10 sesame samples. The isolates belonged to three species: F. oxysporum, F. semitectum and F. verticillioides. All isolates from okra were identified as F. semitectum while the three species of Fusarium occurred in sesame. Six randomly selected isolates from the two crops were screened for their ability to produce mycotoxins in ofada rice culture and the crude extracts of the mycotoxins were tested on week-old Clarias gariepinus fingerlings. Six metabolites were produced by the isolates in ofada rice: equisetin (EQUS), fumonisin B1 (FB1), FB2, methyl-equisetin (M-EQUS), moniliformin (MON) and zearalenone (ZEA). All isolates produced EQUS at concentrations ranging 454-29,983 μg/kg. All isolates except F. semitectum BUFC 041 and F. oxysporum BUFC 024 produced MON and ZEA, while three F. semitectum isolates produced M-EQUS. Only F. verticillioides isolates produced fumonisins. There was evidence of variation within species obtained from both crops based on toxin profile and level of toxin produced. The culture extracts of all isolates induced 100% lethality to C. gariepinus fingerlings except for extracts of F. semitectum BUFC 041 which killed only 62.2% of the fingerlings, possibly due to the absence of MON. Our data suggest that okra and sesame may be potential sources of toxigenic Fusarium.
{"title":"Occurrence, mycotoxins and toxicity of Fusarium species from Abelmoschus esculentus and Sesamum indicum seeds","authors":"C. Ezekiel, C. Nwangburuka, G. Chioma, M. Sulyok, B. Warth, C. Afolabi, Adenike A. Osiberu, Michael O. Oladimeji, O. A. Denton, G. Tayo, R. Krska","doi":"10.2520/MYCO.63.27","DOIUrl":"https://doi.org/10.2520/MYCO.63.27","url":null,"abstract":"Seeds of two okra accessions and 17 samples of sesame seeds were examined for contamination by Fusarium species. Altogether, 37 Fusarium isolates were obtained from the two crops: 6 isolates from two okra accessions and 31 isolates from 10 sesame samples. The isolates belonged to three species: F. oxysporum, F. semitectum and F. verticillioides. All isolates from okra were identified as F. semitectum while the three species of Fusarium occurred in sesame. Six randomly selected isolates from the two crops were screened for their ability to produce mycotoxins in ofada rice culture and the crude extracts of the mycotoxins were tested on week-old Clarias gariepinus fingerlings. Six metabolites were produced by the isolates in ofada rice: equisetin (EQUS), fumonisin B1 (FB1), FB2, methyl-equisetin (M-EQUS), moniliformin (MON) and zearalenone (ZEA). All isolates produced EQUS at concentrations ranging 454-29,983 μg/kg. All isolates except F. semitectum BUFC 041 and F. oxysporum BUFC 024 produced MON and ZEA, while three F. semitectum isolates produced M-EQUS. Only F. verticillioides isolates produced fumonisins. There was evidence of variation within species obtained from both crops based on toxin profile and level of toxin produced. The culture extracts of all isolates induced 100% lethality to C. gariepinus fingerlings except for extracts of F. semitectum BUFC 041 which killed only 62.2% of the fingerlings, possibly due to the absence of MON. Our data suggest that okra and sesame may be potential sources of toxigenic Fusarium.","PeriodicalId":19069,"journal":{"name":"Mycotoxins","volume":"38 1","pages":"27-38"},"PeriodicalIF":0.0,"publicationDate":"2013-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90587271","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. T. Amatulli, F. Fanelli, A. Moretti, G. Mulè, A. Logrieco
Mycotoxins are secondary metabolites produced by several fungal species and represent a great concern for the economical and healthy implications on food and feed chain. Cereals are the primary source of human diet, wheat being the third most produced grain worldwide. Although Fusarium still represents the main source of mycotoxin contamination of wheat, in recent years, due also to evident climate changes that influence agricultural environment, other mycotoxingenic fungi have been pointed out as important wheat contaminants. Among these a disease called "Black Point", caused by Alternaria spp., is increasing it importance as re-emerging risk. Diseases and mycotoxins (alternariol, altenuene, alternariol methyl-ether and tenuazolic acid) associated with Alternaria infection have been reported in several countries suggesting to deepen the knowledge about this genus. This paper summarizes the recent findings on wheat contamination by Alternaria spp and their related toxins.
{"title":"Alternaria species and mycotoxins associated to black point of cereals","authors":"M. T. Amatulli, F. Fanelli, A. Moretti, G. Mulè, A. Logrieco","doi":"10.2520/MYCO.63.39","DOIUrl":"https://doi.org/10.2520/MYCO.63.39","url":null,"abstract":"Mycotoxins are secondary metabolites produced by several fungal species and represent a great concern for the economical and healthy implications on food and feed chain. Cereals are the primary source of human diet, wheat being the third most produced grain worldwide. Although Fusarium still represents the main source of mycotoxin contamination of wheat, in recent years, due also to evident climate changes that influence agricultural environment, other mycotoxingenic fungi have been pointed out as important wheat contaminants. Among these a disease called \"Black Point\", caused by Alternaria spp., is increasing it importance as re-emerging risk. Diseases and mycotoxins (alternariol, altenuene, alternariol methyl-ether and tenuazolic acid) associated with Alternaria infection have been reported in several countries suggesting to deepen the knowledge about this genus. This paper summarizes the recent findings on wheat contamination by Alternaria spp and their related toxins.","PeriodicalId":19069,"journal":{"name":"Mycotoxins","volume":"66 1","pages":"39-46"},"PeriodicalIF":0.0,"publicationDate":"2013-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85718497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Penicillium expansum, one of the patulin producing fungi that causes decay on apple, is recognized as the main source of patulin contamination on apple and apple products. The widely used method for patulin analysis in apple juice is liquid-liquid extraction with ethyl acetate followed by HPLC-UV or LC-MS detection. Previous studies have shown cyclodextrin polymers to exhibit favorable adsorption properties for several classes of small organic molecules, including patulin in apple juice. In this study, an insoluble polymer composed of cyclodextrin crosslinked with 4 ,4 ʼ -methylenebis(phenyl isocyanate) was synthesized for use in the solid phase extraction of patulin from apple juice. Conditions investigated for this method were solvent for column conditioning, sample volume to load patulin on the column, solvent for washing, and solvent and volume for patulin elution and optimized recovery of patulin from the column. At the optimized conditions, the recovery and relative standard deviation (RSD) of patulin from apple juice spiked at 10, 20, 50, 80 and 100 ng mL were 78 and 20 %, 71 and 13 %, 78 and 17 %, 71 and 7.1 %, 67 and 2.9 %, respectively. Limit of quantitation (LOQ) of patulin in apple juice by this method was 10 ng mL.
膨胀青霉是一种产生棒曲霉素的真菌，能引起苹果的腐烂，被认为是苹果和苹果产品棒曲霉素污染的主要来源。常用的苹果汁中棒曲霉素分析方法是乙酸乙酯液液萃取- HPLC-UV或LC-MS检测。先前的研究表明，环糊精聚合物对几类小有机分子具有良好的吸附性能，包括苹果汁中的展霉素。以环糊精与4,4′-亚甲基双(苯基异氰酸酯)交联为原料合成了一种不溶性聚合物，用于固相萃取苹果汁中的展霉素。考察了色谱柱的溶剂条件、柱上装展青霉素的进样量、洗涤溶剂条件、展青霉素洗脱的溶剂和体积条件，并优化了展青霉素的回收率。在优化条件下，在10、20、50、80和100 ng mL加标条件下，苹果汁中展青霉素的回收率和相对标准偏差分别为78%和20%、71%和13%、78%和17%、71%和7.1%、67%和2.9%。该方法在苹果汁中棒曲霉素的定量限为10 ng mL。
{"title":"Use of cyclodextrin-based polymer for patulin analysis in apple juice","authors":"Takashi Shirasawa, M. Ueda, M. Appell, T. Goto","doi":"10.2520/MYCO.63.1","DOIUrl":"https://doi.org/10.2520/MYCO.63.1","url":null,"abstract":"Penicillium expansum, one of the patulin producing fungi that causes decay on apple, is recognized as the main source of patulin contamination on apple and apple products. The widely used method for patulin analysis in apple juice is liquid-liquid extraction with ethyl acetate followed by HPLC-UV or LC-MS detection. Previous studies have shown cyclodextrin polymers to exhibit favorable adsorption properties for several classes of small organic molecules, including patulin in apple juice. In this study, an insoluble polymer composed of cyclodextrin crosslinked with 4 ,4 ʼ -methylenebis(phenyl isocyanate) was synthesized for use in the solid phase extraction of patulin from apple juice. Conditions investigated for this method were solvent for column conditioning, sample volume to load patulin on the column, solvent for washing, and solvent and volume for patulin elution and optimized recovery of patulin from the column. At the optimized conditions, the recovery and relative standard deviation (RSD) of patulin from apple juice spiked at 10, 20, 50, 80 and 100 ng mL were 78 and 20 %, 71 and 13 %, 78 and 17 %, 71 and 7.1 %, 67 and 2.9 %, respectively. Limit of quantitation (LOQ) of patulin in apple juice by this method was 10 ng mL.","PeriodicalId":19069,"journal":{"name":"Mycotoxins","volume":"36 1","pages":"1-8"},"PeriodicalIF":0.0,"publicationDate":"2013-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77963139","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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