Tolypella intricata is one of the rarest charophytes from the genus Tolypella that can be found both in Poland and in Europe. Rarely found in the last 200 years, it was recently rediscovered in two localities in eastern Poland in the Lublin region. In this article, we present two new locations of T. intricata in Poland that make is a significant contribution to a closer understanding of the distribution of the biology and ecology of this species. The article provides basic information on the autecology of T. intricata together with the associated macrophyte vegetation. The morphology and ecology data are compared with the literature, along with a critical review of such information found in the literature. The results of molecular tests confirming the new position of T. intricata are also presented.