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Quantitative descriptions of foodweb structure based on isotope niche space require knowledge of producers’ isotopic signatures. In freshwater ecosystems charophytes are one of the main components of submerged vegetation and the feeding base for many herbivorous consumers, but knowledge about their organic carbon isotopic signatures is sparse. In this study, the δ13C organic values (and organic %C and %N) of the four species of submerged macrophytes (three charophytes - Chara hispida, Nitella hyalina and Tolypella glomerata - and one angiosperm, Myriophyllum spicatum) growing in a newly created shallow pond were measured monthly over a period of one year, to discern if i) all charophyte species susceptible to being food for consumers and growing in the same waterbody have the same C isotopic composition; ii) the δ13C values of a charophyte species change on a seasonal and spatial scale; iii) the different parts (apical nodes, internodes, rhizoids, reproductive organs, oospores) of a charophyte species have the same isotopic composition. The δ13C, %C and %N values of organic matter in the sediments where the plants were rooted were also measured as well as several limnological variables. The δ13C values for the angiosperm (-13.7±0.7‰) indicated 13C-enrichment, whereas the N. hyalina δ13C values were the most negative (-22.4±0.7‰). The mean δ13C value for C. hispida was -19.0±1.0‰ and -20.7±0.8‰ for T. glomerata. C. hispida δ13C values had a significant seasonal variation with 13C-poor values in the cold season, and slight spatial differences. Statistically significant differences were found between charophyte rhizoids (13C-enriched) and the other parts of the thalli. The δ13C values in the sediments varied throughout time (-13‰ to -26‰). The C content was lower in the charophytes than in the angiosperm and there were no large differences among the charophytes. Charophyte fructifications were enriched in organic C compared to the thalli parts. The study provides an isotopic baseline for further studies for the elucidation of higher trophic-level relationships which are particularly complex in shallow water bodies where interactions between the pelagic and the benthic zones are intricate.