Abstract
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.
Highlights
Agriculture will face significant challenges in the 21st century, largely due to the need to increase global food supply under the declining availability of soil, land and water resources, the environmental impacts of chemical inputs and the declining resources of mineral phosphate fertilizers
In this study we rigorously evaluated the possibility to identifying Arbuscular mycorrhizal fungi (AMF) spores of nineteen isolates belonging to fourteen species, seven genera and five families by MALDI-TOF-MS biotyping
The partial 18S-5.8S-partial 28 S region of nuclear rRNA or the partial 28 S region of all isolates analysed by MALDI was used for maximum likelihood phylogenetic analyses in MEGA and PhyML (Fig. 1)
Summary
Agriculture will face significant challenges in the 21st century, largely due to the need to increase global food supply under the declining availability of soil, land and water resources, the environmental impacts of chemical inputs and the declining resources of mineral phosphate fertilizers. As a means towards developing sustainable agriculture, soil organisms that are considered plant bio-fertilizer have been used for biotechnological applications to agronomy and the environment. Fungi of the phylum Glomeromycota are ubiquitous hypogeous microorganisms that live in symbiosis with 80% of the world’s vascular land plants[3,4]. There are three possible reasons why unknown taxa have not been described to date They may represent isolates that are difficult to grow[10]. Large intra-isolate variation of nuclear ribosomal genes[15,16] complicates assignment of a single marker gene sequence to a fungal strain and assessment of the diversity of AMF in molecular field studies. A quick, accurate taxonomic identification of AMF isolates is necessary for culture collections (IBG, INVAM, SAF, CICG, and GINCO) and research and for industrial certification
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