Melatonin (MEL) is defined as a plant master regulator. It regulates plantgrowth and development in various stress environments by increasing in amount. Effects of external MEL application on the content of the secretion products in micro-morphological structures of leaf trichomesof Melissa officinalis L. (lemon balm) without the presence of external stress factors were characterized. The relationships between differentMEL concentrations and changes in type, density and size of trichomes, total chlorophyll and proline amounts, and secretion products were defined. Furthermore, monoterpenoids, which are the main products of trichomes that determine the quality of essential oils in lemon balm, varied under the effects of MEL. Especially 100 mM MEL concentration had the most dramatic impact. The highest number of glandular trichomes (GT) were observed at this concentration. 150 mM concentration of MEL had the opposite effect. At this concentration, significant decreases in secondary metabolite content and glandular structures were observed. Thus, it was concluded that MEL had a positive effect on these parameters up to a certain concentration. Changes in the size and number of GT due to MEL application is greatly reflected by the quality and content of secondary metabolites. Total chlorophyll, proline and monoterpene numbers were highest at 100 mM MEL concentration. The effects of MEL application without the presence of any stress conditions were supported by total chlorophyll and proline content. Important information was obtained about the relationship of these effects with the density of peltate trichomes and the chemical compounds and quality of essential oils.