A novel approach to identify plant-parasitic nematodes using maldi-tof mass spectrometry
Perera, M.R., Vanstone, V.A. and Jones, M.G.K. (2005) A novel approach to identify plant-parasitic nematodes using maldi-tof mass spectrometry. In: 44th Annual Meeting of the Society of Nematologists, 9 - 13 July 2005, Fort Lauderdale, Florida.
The ability to accurately identify nematodes at the species level requires specific skills in nematode taxonomy. The most common approach used is microscopy, which takes time and requires specific training: molecular approaches to plant nematode identification can also be used. Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) is being used as a new approach to identify microorganisms. Identification is based on the characteristic protein profile of the organism generated by MALDI-TOF MS. In MALDI-TOF MS, pulses from a UV laser are absorbed by a crystalline matrix to which macromolecules have been adsorbed, and this results in relatively gentle desorption and ionization of the macromolecules, which are then accelerated using a high voltage and travel along the flight tube. The time taken to reach the detector is directly related to the mass/charge ratio of the molecule. MALDI-TOF is a promising approach for rapid identification of organisms because of the simple sample preparation and the rapidity of the technique. The aim of this work was to develop a rapid, simple method to identify plant parasitic nematodes, based on analysis of protein profiles of nematodes generated by MALDI-TOF MS. Two methods have been used: grinding and direct analysis of intact nematodes. Both methods were standardized using the wheat seed-gall nematode Anguina tritici as a model. The standardized methods were applied to analyze the seed-gall nematodes A. tritici and A. funesta and the root-knot nematode, Meloidogyne javanica, which infects many horticultural crops. Characteristic protein profiles and diagnostic peaks were identified for individual nematode species and for mixtures of these species. The results provide proof-of-concept that these nematode species can be identified by protein profiling using MALDI-TOF MS. This approach could be extended to identify other plant and non-plant parasitic nematodes by generating unique species-specific protein profiles.
|Publication Type:||Conference Item|
|Murdoch Affiliation:||Western Australian State Agricultural Biotechnology Centre
School of Biological Sciences and Biotechnology
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