Root-knot nematodes (Meloidogyne spp.) are among the biotic factors that limit tomato production worldwide. The objectives of this study were to assess the distribution and identify Meloidogyne spp. and associated problems from major tomato growing areas of Ethiopia. A total of 212-rhizosphere soil and 123 root samples were sampled from 40 localities during 2012/13 growing season. A total of 646 respondents participated in the questionnaire to assess knowledge and practice of farmers and factors associated with RKN damage on tomato. Out of the 212 soil samples collected, 47.2% were found infested by various Meloidogyne species 8 weeks after the start of the bioassay test. Out of the 123 root samples collected, 65% had root galls. The highest prevalence (100%) of RKN was found on samples collected from Adami Tullu, Babile, Erer Gota, Hurso, Jittu, Tikur Wuha, Tepo Choronke, Zeway and Koka. The highest incidence (100%) of RKN was found from Adami Tullu, Jara Weyo, Babile, Erer Gota, Hurso, Jittu, Tikur Wuha, Tony farm, Tepo Choronke, Zeway and Koka localities based on direct observation of galls on collected root samples. Out of the 646 respondents, 43.3% reported to have RKN damage symptoms when shown the symptoms while 56.7% of them did not report damage. The highest significant effect on the dependent variable RKN damage on tomato roots comes from the previous crop, soil texture, awareness about RKN and source of irrigation water used. The presence of Meloidogyne incognita, M. javanica, M. arenaria and M. hapla on tomato was confirmed using a combination of molecular and biochemical identification tools. Five isolates of Meloidogyne spp. were not properly identified and could be new species. The two tropical species, M. incognita (48.4%) and M. javanica (41.2%) were the most prevalent species. Both species were also co-infesting tomato plants. Meloidogyne hapla was detected for the first time in an open tomato production farmer's field at Zeway' locality with 1620m.a.s.l. elevation. The occurrence of these Meloidogyne species alone, or in mixed populations from samples collected, clearly shows that RKN are widespread in major tomato growing areas of Ethiopia. In the near future, the economic importance of M. arenaria and M. hapla on tomato production in Ethiopian agriculture should be investigated.