Rapid identification of Fusarium species causing head blight in Canada using MALDI-TOF mass spectrometry.

IF 1.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Shimosh Kurera, Bhaktiben Patel, Tehreem Ashfaq, Maria Eckhardt, Janice Bamforth, Niradha Withana Gamage, Gurcharn Singh Brar, Linda J Harris, Maria Antonia Henriquez, Rajagopal Subramaniam, Xiben Wang, Zamir Punja, Curtis Pozniak, Matthew G Bakker, Sean Walkowiak
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引用次数: 0

Abstract

Fusarium head blight (FHB) is a limitation to grain production and can be caused by several different Fusarium species. We evaluated the ability of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) to perform species identifications. The method generates a unique peptide mass fingerprint (PMF) for each sample that can be matched to a reference library. We first created a reference library of PMF profiles for Fusarium species from across Canada. Then, we tested the library to perform identifications using two validation panels. The first panel consisted of 820 fungal isolates from wheat (2021-2023 harvest years) and the second was 74 fungal isolates from oat and barley (2022 harvest year). The species identity of samples from the validation panels was confirmed with high-throughput quantitative PCR (HT-qPCR) using species-specific DNA markers. The first validation panel was mostly F. graminearum and there was 95% overlap between the MALDI-TOF MS and DNA-based identifications. The second panel was mostly F. poae and the identifications from the two methods had 86% overlap. Our findings indicate that MALDI-TOF MS biotyping is sensitive enough to identify Fusarium strains to their species complexes and certain Fusarium strains to the species level.

用MALDI-TOF质谱法快速鉴定加拿大引起头疫病的镰刀菌。
赤霉病(Fusarium head blight, FHB)是一种限制粮食生产的病害,可由几种不同的赤霉病种引起。我们评估了基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)进行物种鉴定的能力。该方法为每个样品生成独特的肽质量指纹图谱(PMF),可以与参考库相匹配。我们首先创建了一个来自加拿大各地镰刀菌的PMF档案参考库。然后,我们测试了库,使用两个验证面板执行识别。第一个小组包括820个小麦(2021-2023收获年)的真菌分离株,第二个小组包括74个燕麦和大麦(2022收获年)的真菌分离株。利用物种特异性DNA标记,用高通量定量PCR (HT-qPCR)确认验证板样品的物种身份。第一个验证小组主要是F. graminearum, MALDI-TOF MS和dna鉴定之间有95%的重叠。第二组主要为F. poae,两种方法的鉴定有86%的重叠。我们的研究结果表明,MALDI-TOF质谱生物分型对镰刀菌菌种复合体和某些镰刀菌菌种水平具有足够的敏感性。
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来源期刊
CiteScore
4.80
自引率
0.00%
发文量
71
审稿时长
2.5 months
期刊介绍: Published since 1954, the Canadian Journal of Microbiology is a monthly journal that contains new research in the field of microbiology, including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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