Wetlands are important sites for biological conservation because they support rich biodiversity and present high productivity. Species-area relationship is an important tool in conservation planning and it has been extensively used for wetland management. This study had as aims: (1) to analyse macrophyte and macroinvertebrate diversity in the fragmented wetlands of the Sinos River basin; and (2) to investigate whether wetland areas could work as a tool for selecting the important habitats for biodiversity conservation. Throughout the study, 56 species of macrophytes and 57taxa of macroinvertebrates were identified. Macrophyte richness was related to the wetland area, but macroinvertebrate richness, however, was not related to the wetland area. The macrophyte and macroinvertebrate composition were not related to the wetland area. Species composition varied between the regions of the basin and the difference in the species composition occurred mainly by the distribution of macroinvertebrates. With regard to the biodiversity conservation of the Sinos River basin, the wetland area must not be a priority criterion for choosing the important wetlands for conservation. The environmental policies for biodiversity conservation must include management actions focused also on the protection of small wetlands. Moreover, other criteria should be analysed in further research such as habitat diversity, hydroperiod, geographic distribution and connectivity.