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Nematodes are highly abundant soil organisms, and their presence can have profound effects on soil health and plant growth. Among them,
Rotylenchus
species are known for their economic importance as root ectoparasites or semi-endoparasites, inflicting damage on a wide variety of economically important plants. Their impact on agricultural crops, ornamentals, and fruit and forest trees makes them significant subjects for study. In this paper, we present an updated species list of
Rotylenchus
spp., a genus of spiral plant-parasitic nematodes belonging to the family Hoplolaimidae. As of the current research, 107 species within the
Rotylenchus
genus have been recognized. To facilitate the identification of
Rotylenchus
species, we introduce a novel browser-based interactive key for the identification of such huge number of species. This web-assisted tool utilizes a list of 48 diagnostic character-states belonging to 11 characters for identifying 107
Rotylenchus
species, providing an easy and accurate method for the identification of these plant-parasitic nematodes. This paper contributes to the understanding of
Rotylenchus
species’ diversity and their taxonomy while offering a valuable tool to aid researchers, agricultural professionals, and plant pathologists in accurate species identification and subsequent management strategies.