Novel DNA Barcoding and Multiplex PCR Strategy for the Molecular Identification and Mycotoxin Gene Detection of Fusarium spp. in Maize from Bulgaria.

IF 2 Q3 BIOCHEMICAL RESEARCH METHODS
Daniela Stoeva, Deyana Gencheva, Georgi Radoslavov, Peter Hristov, Rozalina Yordanova, Georgi Beev
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引用次数: 0

Abstract

Fusarium spp. represent a critical threat to maize production and food safety due to their mycotoxin production. This study introduces a refined molecular identification protocol integrating four genomic regions-ITS1, IGS, TEF-1α, and β-TUB-for robust species differentiation of Fusarium spp. isolates from post-harvest maize in Bulgaria. The protocol enhances species resolution, especially for closely related taxa within the Fusarium fujikuroi species complex (FFSC). A newly optimized multiplex PCR strategy was developed using three primer sets, each designed to co-amplify a specific pair of toxigenic genes: fum6/fum8, tri5/tri6, and tri5/zea2. Although all five genes were analyzed, they were detected through separate two-target reactions, not in a single multiplex tube. Among 17 identified isolates, F. proliferatum (52.9%) dominated, followed by F. verticillioides, F. oxysporum, F. fujikuroi, and F. subglutinans. All isolates harbored at least one toxin biosynthesis gene, with 18% co-harboring genes for both fumonisins and zearalenone. This dual-protocol approach enhances diagnostic precision and supports targeted mycotoxin risk management strategies.

Abstract Image

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Abstract Image

保加利亚玉米镰刀菌分子鉴定和霉菌毒素基因检测的新型DNA条形码和多重PCR策略。
镰刀菌因其产生的霉菌毒素对玉米生产和食品安全构成严重威胁。本研究引入了一种包含四个基因组区域(its1、IGS、TEF-1α和β- tub)的精细分子鉴定方案,用于保加利亚收获后玉米镰刀菌分离株的强大物种分化。该方案提高了物种分辨力,特别是对密切相关的Fusarium fujikuroi物种复合体(FFSC)中的分类群。采用三组引物,设计了一种新的优化多重PCR策略,每组引物可共同扩增一对特定的产毒基因:fum6/fum8、tri5/tri6和tri5/zea2。虽然分析了所有五个基因,但它们是通过单独的双靶反应检测到的,而不是在一个单一的多重管中检测到的。鉴定的17株分离株中,以增殖F.(52.9%)居多,其次为黄萎病F.、尖孢F.、富士F.和亚粘滞F.。所有分离株至少含有一种毒素生物合成基因,其中18%同时含有伏马菌素和玉米赤霉烯酮基因。这种双重方案方法提高了诊断精度,并支持有针对性的霉菌毒素风险管理战略。
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来源期刊
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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