A new species of the Drosophila (Drosophila) melanica species group (Diptera: Drosophilidae), Drosophila denruoi Suwito & Watabe, sp. nov., is described from northern Vietnam. It is proved by crossing experiments that reproductive isolation is complete between this species and other morphologically similar species. Larval ganglion cells of D. denruoi possess 2n = 12 chromosomes, comprised of four pairs of acrocentric, one pair of metacentric and one pair of micro-chromosomes. This is the largest number of acrocentric chromosomes in the group and implies that the species has the most primitive chromosomal configuration in the melanica group. Based on the results of crossing experiments and close morphological examination, Drosophila pengi Okada & Kurokawa, 1957 from central Japan, once regarded as a synonym of Drosophila tsigana Burla & Gloor, 1952, is shown to be distinct from D. tsigana. Drosophila pengi is thus resurrected as a member of the melanica group. We synonymize Drosophila bisetata Toda, 1988, described from central Myanmar, with D. pengi. In addition, a population from Guizhou, southwestern China, once regarded as D. tsigana, is also identified as D. pengi. These populations from central Myanmar and southwestern China share the diagnostic characters of D. pengi. At the same time, it is reconfirmed that the Hokkaido population is conspecific with European D. tsigana. Furthermore, another species, Drosophila longiserrata Toda, 1988 described from central Myanmar, is synonymized with Drosophila afer Tan, Hsu & Sheng, 1949 from China, based on their having identical karyotypes and external morphology.