Li, GuangjinChen, LeiPang, KeTang, QingWu, ChengxiYuan, XunlaiZhou, ChuanmingXiao, Shuhai2023-08-292023-08-292023-04399http://hdl.handle.net/10919/116159Macrofossils with unambiguous biogenic origin and predating the one-billion-year-old multicellular fossils Bangiomorpha and Proterocladus interpreted as crown-group eukaryotes are quite rare. Horodyskia is one of these few macrofossils, and it extends from the early Mesoproterozoic Era to the terminal Ediacaran Period. The biological interpretation of this enigmatic fossil, however, has been a matter of controversy since its discovery in 1982, largely because there was no evidence for the preservation of organic walls. Here we report new carbonaceous compressions of Horodyskia from the Tonian successions (similar to 950-720 Ma) in North China. The macrofossils herein with bona fide organic walls reinforce the biogenicity of Horodyskia. Aided by the new material, we reconstruct Horodyskia as a colonial organism composed of a chain of organic-walled vesicles that likely represent multinucleated (coenocytic) cells of early eukaryotes. Two species of Horodyskia are differentiated on the basis of vesicle sizes, and their co-existence in the Tonian assemblage provides a link between the Mesoproterozoic (H. moniliformis) and the Ediacaran (H. minor) species. Our study thus provides evidence that eukaryotes have acquired macroscopic size through the combination of coenocytism and colonial multicellularity at least similar to 1.48 Ga, and highlights an exceptionally long range and morphological stasis of this Proterozoic macrofossils.application/pdfenCreative Commons Attribution 4.0 Internationalbedding-plane markingsearly evolutiontestate amebasforaminiferasupergrouporiginsfossilscomplexsuccessionseukaryotesArticle - Refereedhttps://doi.org/10.1038/s42003-023-04740-261370460792399-3642