Freshwater biodiversity is declining at an unprecedented rate. Freshwater conservationists and environmental managers have enough evidence to demonstrate that action must not be delayed but have insufficient evidence to identify those actions that will be most effective in reversing the current trend. Here, the focus is on identifying essential research topics that, if addressed, will contribute directly to restoring freshwater biodiversity through supporting ‘bending the curve’ actions (i.e. those actions leading to the recovery of freshwater biodiversity, not simply deceleration of the current downward trend). The global freshwater research and management community was asked to identify unanswered research questions that could address knowledge gaps and barriers associated with ‘bending the curve’ actions. The resulting list was refined into six themes and 25 questions. Although context‐dependent and potentially limited in global reach, six overarching themes were identified: (i) learning from successes and failures; (ii) improving current practices; (iii) balancing resource needs; (iv) rethinking built environments; (v) reforming policy and investments; and (vi) enabling transformative change. Bold, efficient, science‐based actions are necessary to reverse biodiversity loss. We believe that conservation actions will be most effective when supported by sound evidence, and that research and action must complement one another. These questions are intended to guide global freshwater researchers and conservation practitioners, identify key projects and signal research needs to funders and governments. Our questions can act as springboards for multidisciplinary and multisectoral collaborations that will improve the management and restoration of freshwater biodiversity.

Environmental unpredictability is known to result in the evolution of bet‐hedging traits. Variable dormancy enhances survival through harsh conditions, and is widely cited as a diversification bet‐hedging trait. The floating aquatic plant, Spirodela polyrhiza (Greater Duckweed), provides an opportunity to study diversification because although partially reliable seasonal cues exist, its growing season is subject to an unpredictable and literally “hard” termination when the surface water freezes, and overwinter survival depends on a switch from production of normal daughter fronds to production of dense, sinking “turions” prior to freeze‐over. The problem for S. polyrhiza is that diversified dormancy behavior must be generated among clonally produced, genetically identical offspring. Variation in phenology has been observed in the field, but its sources are unknown. Here, we investigate sources of phenological variation in turion production, and test the hypothesis that diversification in turion phenology is generated within genetic lineages through effects of parental birth order. As expected, phenotypic plasticity to temperature is expressed along a thermal gradient; more interestingly, parental birth order was found to have a significant and strong effect on turion phenology: Turions are produced earlier by late birth‐order parents. These results hold regardless of whether turion phenology is measured as first turion birth order, time to first turion, or turion frequency. This study addresses a question of current interest on potential mechanisms generating diversification, and suggests that consistent phenotypic differences across birth orders generate life history variation.

Sediment DNA (sedDNA) analyses are rapidly emerging as powerful tools for the reconstruction of environmental and evolutionary change. While there are an increasing number of studies using molecular genetic approaches to track changes over time, few studies have compared the coherence between quantitative polymerase chain reaction (PCR) methods and metabarcoding techniques. Primer specificity, bioinformatic analyses, and PCR inhibitors in sediments could affect the quantitative data obtained from these approaches. We compared the performance of droplet digital polymerase chain reaction (ddPCR) and high-throughput sequencing (HTS) for the quantification of target genes of cyanobacteria in lake sediments and tested whether the two techniques similarly reveal expected patterns through time. Absolute concentrations of cyanobacterial 16S rRNA genes were compared between ddPCR and HTS using dated sediment cores collected from two experimental (Lake 227, fertilized since 1969 and Lake 223, acidified from 1976 to 1983) and two reference lakes (Lakes 224 and 442) in the Experimental Lakes Area (ELA), Canada. Relative abundances of Microcystis 16S rRNA (MICR) genes were also compared between the two methods. Moderate to strong positive correlations were found between the molecular approaches among all four cores but results from ddPCR were more consistent with the known history of lake manipulations. A 100-fold increase in ddPCR estimates of cyanobacterial gene abundance beginning in ~1968 occurred in Lake 227, in keeping with experimental addition of nutrients and increase in planktonic cyanobacteria. In contrast, no significant rise in cyanobacterial abundance associated with lake fertilization was observed with HTS. Relative abundances of Microcystis between the two techniques showed moderate to strong levels of coherence in top intervals of the sediment cores. Both ddPCR and HTS approaches are suitable for sedDNA analysis, but studies aiming to quantify absolute abundances from complex environments should consider using ddPCR due to its high tolerance to PCR inhibitors.

Analyses of sedimentary DNA (sedDNA) have increased exponentially over the last decade and hold great potential to study the effects of anthropogenic stressors on lake biota over time. Herein, we synthesise the literature that has applied a sedDNA approach to track historical changes in lake biodiversity in response to anthropogenic impacts, with an emphasis on the past c. 200 years. We identified the following research themes that are of particular relevance: (1) eutrophication and climate change as key drivers of limnetic communities; (2) increasing homogenisation of limnetic communities across large spatial scales; and (3) the dynamics and effects of invasive species as traced in lake sediment archives. Altogether, this review highlights the potential of sedDNA to draw a more comprehensive picture of the response of lake biota to anthropogenic stressors, opening up new avenues in the field of paleoecology by unrevealing a hidden historical biodiversity, building new paleo‐indicators, and reflecting either taxonomic or functional attributes. Broadly, sedDNA analyses provide new perspectives that can inform ecosystem management, conservation, and restoration by offering an approach to measure ecological integrity and vulnerability, as well as ecosystem functioning.