Al-Zaban Mayasar I, Alrokban Ahlam H, Mahmoud Mohamed A
Department of Biology, College of Science, Princess Nourah Bint Abdulrahman University, Riyadh, Saudi Arabia.
Central Laboratory of Biotechnology (CLB), Plant Pathology Research Institute, Agricultural Research Center, Giza, Egypt.
Mycology. 2023 May 24;14(3):227-238. doi: 10.1080/21501203.2023.2213704. eCollection 2023.
This study aimed to identify important mycotoxigenic fungi and accurate detection of mycotoxin in stored maize grains using molecular methods. The current study also optimised the real-time PCR (RT-PCR) assay. The melting curve was established to identify isolated fungal species of (4), (3), (3), and (one). A multiplex polymerase chain reaction (mPCR) technique was developed for the detection and characterisation of mycotoxin producing fungi, mycotoxin metabolic pathway genes, and the determination of eleven mycotoxins in stored maize grains using high-performance liquid chromatography (HPLC). The mPCR results indicated positive signals for potentially mycotoxigenic fungal species tested of , and . A protocol for multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was tested to distinguish between free and contaminated, stored maize with aflatoxin B1 (AFB1). The expression pattern of four aflatoxin biosynthetic pathway genes, AFB1 (, and ), was a good marker for contaminated, stored maize grains. HPLC analysis showed that maize grain samples were contaminated with mycotoxins, and the concentration was above the detection level. The results indicate that the polyphasic approach might provide a sensitive, rapid, and accurate method for detecting and identifying mycotoxigenic fungal species and mycotoxins in stored maize grains.
本研究旨在利用分子方法鉴定储存玉米籽粒中重要的产毒真菌并准确检测霉菌毒素。本研究还优化了实时荧光定量聚合酶链反应（RT-PCR）检测方法。建立了熔解曲线以鉴定分离出的（4种）、（3种）、（3种）和（1种）真菌物种。开发了一种多重聚合酶链反应（mPCR）技术，用于检测和鉴定产霉菌毒素的真菌、霉菌毒素代谢途径基因，并使用高效液相色谱（HPLC）测定储存玉米籽粒中的11种霉菌毒素。mPCR结果表明，所检测的潜在产毒真菌物种、和呈现阳性信号。测试了一种多重逆转录-聚合酶链反应（mRT-PCR）方法，以区分未受污染和受黄曲霉毒素B1（AFB1）污染的储存玉米。四种黄曲霉毒素生物合成途径基因（AFB1，和）的表达模式是受污染储存玉米籽粒的良好标志物。HPLC分析表明，玉米籽粒样品被霉菌毒素污染，且浓度高于检测水平。结果表明，多相方法可能为检测和鉴定储存玉米籽粒中产毒真菌物种和霉菌毒素提供一种灵敏、快速且准确的方法。
