Root-knot nematode disease， a global plant root disease caused by parasitic root-knot nematodes （Meloidogyne spp.）， severely impacts tobacco production. Recently， population shifts have occurred resulting in the prevalence of M. arenaria in Yunnan province， which has now become an urgent issue for tobacco production. However， studies on root-knot nematodes has historically focused on M. incognita， with relatively limited studies on M. arenaria. Based on our previous research， we developed a STARP molecular marker tightly linked to a resistant QTL. By deployment of this marker to screen 387 germplasm resources， we identified 18 positive tobacco germplasms which were subsequently evaluated for resistance to M. arenaria under three different environments. Phenotypic assessments revealed five tobacco varieties or lines with stable resistance to M. arenaria： Coker 316， TI 706， SA1224， SA1215， and Y3. To our knowledge， it is the first report identifying TI 706 and Coker 316 as resistance to M. arenaria. Furthermore， Coker 316， TI 706， and SA1224 exhibit stable and reliable resistance and thus serve as valuable sources for resistance breeding.