One new Brazilian species of Drosophila belonging to the nesiota subgroup of the flavopilosa group, which is recorded for the first time from the State of São Paulo, is illustrated and described as D. mariaehelenae, sp. nov. It has been found breeding on flowers of Cestrum intermedium Sendtn., together with D. cestri Brncic, D. cordeiroi Brncic, D. cordeiroi Brncic, D. flavopilosa Frey and D. incompta Wheeler & Takada, Zygothrica vittimaculosa Burla and one undetermined species of the Drosophila tripunctata species group. Photographs are provided of the male genitalia of the first four species.