Demography and threats to population growth of Cirsium pitcheri, a threatened dune plant, in Wisconsin.

Multi-year and multi-site demographic data for rare plants allow researchers to observe threats and project population growth rates and thus long-term persistence of the species, generating knowledge, which allows for effective conservation planning. Demographic studies across more than a decade are extremely rare but allow for the effects of threats to be observed and assessed within the context of interannual environmental variation. We collected demographic data on the Threatened plant Cirsium pitcheri in two sites from 2011-2022. These sites were chosen because one exhibited the presence of non-native seed predators while the other did not, and we hypothesized that we would see declines and potentially extinction of the population threatened by predation. Over the course of our study, we observed additional threats, such as human trampling and high lake levels, which led to significant erosion, sand burial, and storm damage to plants. We find high interannual variation in vital rates and population growth rates for both populations, which mask the overall effects of predation. We observed dramatic declines in plant survivorship and population growth rates in both sites in the years with high lake levels. We conclude that high lake levels, which are expected to become more frequent with climate change, pose a significant threat to all near-shore populations of C. pitcheri.

Short interfering RNA strand selection is independent of dsRNA processing polarity during RNAi in Drosophila.

Short interfering RNAs (siRNAs) guide mRNA cleavage during RNA interference (RNAi). Only one siRNA strand assembles into the RNA-induced silencing complex (RISC), with preference given to the strand whose 5' terminus has lower base-pairing stability. In Drosophila, Dcr-2/R2D2 processes siRNAs from longer double-stranded RNAs (dsRNAs) and also nucleates RISC assembly, suggesting that nascent siRNAs could remain bound to Dcr-2/R2D2. In vitro, Dcr-2/R2D2 senses base-pairing asymmetry of synthetic siRNAs and dictates strand selection by asymmetric binding to the duplex ends. During dsRNA processing, Dicer (Dcr) liberates siRNAs from dsRNA ends in a manner dictated by asymmetric enzyme-substrate interactions. Because Dcr-2/R2D2 is unlikely to sense base-pairing asymmetry of an siRNA that is embedded within a precursor, it is not clear whether processed siRNAs strictly follow the thermodynamic asymmetry rules or whether processing polarity can affect strand selection. We use a Drosophila in vitro system in which defined siRNAs with known asymmetry can be generated from longer dsRNA precursors. These dsRNAs permit processing specifically from either the 5' or the 3' end of the thermodynamically favored strand of the incipient siRNA. Combined dsRNA-processing/mRNA-cleavage assays indicate that siRNA strand selection is independent of dsRNA processing polarity during Drosophila RISC assembly in vitro.