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The tomato leafminer (Tuta absoluta) is one of the main pests of the tomato cultivation. The insect feeds on the leaves and fruits, thus much of its production is lost. The aim of the present work was the molecular and morphological identification of the tomato leafminer species and to determine whether the insecticides used to combat T. absoluta in greenhouses harm and to what extent the populations of natural enemies Nesidiocoris tenuis and Macrolophus pygmaeus. Extensive searches of Greek and international bibliography. Adult insects were collected from the areas of Dionysios Halkidiki and Trilofos in Thessaloniki, Greece, followed by their morphological identification through identification key. Their DNA was then isolated and amplified by the polymerase chain reaction (PCR) method of the mitochondrial genome of the cytochrome oxidase (COI) subunit I, which was selected for identification. The PCR products were sequenced and analyzed in GenBank database. The database and morphological identification indicated that the insects were of T. absoluta (Lepidoptera: Gelechiidae) species. For the second part of the experiment, insects of the natural enemies N. tenuis and M. pygmaeus from a mass-breeding unit were used. The most widely used synthetic and natural insecticides were used and their insect toxicity was tested and after daily observations, the toxicity rates for each insecticide were found.