Freshwater ecosystems support biological communities with high species richness and conservation interest. However, these ecosystems are highly altered by human intervention and threatened worldwide, making them a priority in conservation planning and biodiversity monitoring. Bryophytes, including several conservation-interest taxa, are recognized indicators of ecological status in freshwaters. We aimed to develop a framework for designing monitoring networks to detect trends in aquatic and semi-aquatic bryophyte communities, prioritizing high-conservation interest communities in different contexts of human pressure (specifically, resulting from the intersection of two criteria: (i) protection status and (ii) presence of a potential impact area).The framework consists of three steps: (1) Spatial modelling of biodiversity; (2) Spatial conservation prioritization; and (3) Model-assisted monitoring network design. Community-level modelling was used to model the distribution of the main bryophyte assemblages in the study area. A conservation prioritization software was utilized to identify areas with high conservation value. The monitoring network was designed using stratified random sampling and unequal-probability sampling techniques to target high conservation value sites distributed across different contexts of human pressure.We have identified four distinct community types, each characterized both by a small group of common and dominant species, and by small group of rarer, conservation-interest species. This typification of four species assemblages occurring in the study area, also highlighted those with potentially higher conservation-interest. The most valuable areas for the conservation of aquatic and semi-aquatic bryophyte communities coincide with specific environmental zones: mountainous areas in Lusitania, large watercourses in the Mediterranean North and some locations in the Mediterranean Mountains. Finally, we obtained a potential monitoring network consisting of 64 monitoring points, unequally distributed across different contexts of human pressure, privileging locations with higher conservation value.The framework presented here illustrates the potential of combining biodiversity modelling, spatial conservation prioritization and monitoring design in the development of monitoring networks. Namely, this framework allowed us to counter data deficiencies, to identify high priority areas to monitor and to design a monitoring network considering different scenarios of human pressure at a regional scale.This framework can also be valuable for conservation efforts as an approach to monitoring conservation-interest biodiversity features in anthropogenically modified riverscapes, which present different degrees of human pressure and the cumulative effects of these different impact elements. Moreover, this approach allows for the comprehensive monitoring of biodiversity values important for management at the national and regional levels. In addition, this framework is one of the first efforts in the development of monitoring networks that target aquatic and semi-aquatic bryophyte communities, a long-neglected plant group of high ecological and conservation importance in freshwater ecosystems.