Ancient lakes are among the most interesting models for evolution studies, because their biodiversity is the result of a complex combination of migration and speciation. Here, we investigate the origin of single celled planktonic eukaryotes from the oldest lake in the world – Lake Baikal. By using 18S rDNA metabarcoding we recovered 1,427 Operational Taxonomic Units (OTUs) belonging to protists populating surface waters (1-50 meters) and belonging to pico- and nano-plankton size fractions. The recovered communities resembled other lacustrine freshwater assemblages found elsewhere, especially the unclassified protists. However, our results suggest that a fraction of Baikal protists originated evolutionary recently from marine/brackish ancestors. Moreover, our results suggest that rapid radiation may have occurred among some protist taxa, partially mirroring what was already shown for multicellular organisms in Lake Baikal. We found potential species flocks in Stramenopiles, Alveolata, Opisthokonta, Archaeplastida, Rhizaria and Hacrobia. Putative flocks predominated in Chrysophytes, which are highly diverse in Lake Baikal. Some of the species, especially those from these flocks, may be endemic from Lake Baikal, because their 18S rDNA differed > 10% from known DNA. Overall, our study points to novel diversity of planktonic protists in Lake Baikal, some of which may have emerged in situ after evolutionary diversification.