Plant-parasitic nematodes represent one of the most significant threats to global agriculture, causing substantial losses every year. This is particularly true for sugarcane cultivation, which holds vital socio-economic importance, especially for the sugarcane-ethanol industry. The presence of plant-parasitic nematodes, particularly from the genus Meloidogyne, can result in productivity losses of up to 40% in this crop. Given this problem, it is crucial to identify and understand the genetic variability of species that strongly impact this cultivation. Such knowledge is essential to guide effective nematode management planning. For this group, the application of robust genomic methodologies, in addition to the analysis of a larger set of isolates, can significantly contribute to understanding these organisms' intrinsic variability. A promising technique is Genotyping-by-Sequencing (GBS), based on next-generation sequencing (NGS) associated with restriction enzymes. This technique has demonstrated great potential for studies on the genetic diversity of Meloidogyne species. Currently, there are sugarcane varieties resistant to root-knot nematodes, but these are only resistant to one species, complicating control in the field, where several species simultaneously affect the crop. Thus, the use of genetically modified plants, conferring resistance to multiple species, is an attractive alternative. This makes the search for effective genes and the production of genetically modified plants an urgent necessity. In this context, the aim of this work is to identify the Meloidogyne species present in sugarcane fields across the country, using advanced molecular techniques. In addition, it is proposed to study the genetic variability of these pathogens using the GBS technique. The aim is also to identify essential or parasitism-associated genes that, once silenced, may interrupt these plant-parasitic nematodes' life cycle. Ultimately, the aim is to generate genetically modified plants resistant to these plant pathogens.