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New method for the identification of arbuscular mycorrhizal fungi by proteomic-based biotyping of spores using MALDI-TOF-MS

机译:基于蛋白质组学的孢子生物分型使用MALDI-TOF-MS鉴定丛枝菌根真菌的新方法

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摘要

Arbuscular mycorrhizal fungi (AMF, Glomeromycota) are mutualistic symbionts associated with majority of land plants. These fungi play an important role in plant growth, but their taxonomic identification remains a challenge for academic research, culture collections and inoculum producers who need to certify their products. Identification of these fungi was traditionally performed based on their spore morphology. DNA sequence data have successfully been used to study the evolutionary relationships of AMF, develop molecular identification tools and assess their diversity in the environment. However, these methods require considerable expertise and are not well-adapted for “routine” quality control of culture collections and inoculum production. Here, we show that Matrix-Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry proteomic-based biotyping is a highly efficient approach for AMF identification. Nineteen isolates belonging to fourteen species, seven genera and five families were clearly differentiated by MALDI biotyping at the species level, and intraspecific differentiation was achieved for the majority. AMF identification by MALDI biotyping could be highly useful, not only for research but also in agricultural and environmental applications. Fast, accurate and inexpensive molecular mass determination and the possibility of automation make MALDI-TOF-MS a real alternative to conventional morphological and molecular methods for AMF identification.
机译:丛枝菌根真菌(AMF,Glomeromycota)是与大多数陆地植物相关的共生共生体。这些真菌在植物生长中起着重要作用,但是它们的分类学鉴定仍然对需要对其产品进行认证的学术研究,培养物收集和接种物生产者构成挑战。这些真菌的鉴定传统上是根据它们的孢子形态进行的。 DNA序列数据已成功用于研究AMF的进化关系,开发分子鉴定工具并评估其在环境中的多样性。但是,这些方法需要大量的专业知识,因此不适用于培养物收集和接种物生产的“常规”质量控制。在这里,我们显示基于蛋白质组学的基质辅助激光解吸电离飞行时间质谱的生物型是一种用于AMF识别的高效方法。通过MALDI生物分型在物种水平上清楚地区分了14个物种,7个属和5个科的19个分离物,并且大多数实现了种内分化。通过MALDI生物分型鉴定AMF可能非常有用,不仅用于研究,而且在农业和环境应用中也是如此。快速,准确和廉价的分子量测定以及自动化的可能性使MALDI-TOF-MS成为用于AMF识别的常规形态学和分子方法的真正替代品。

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