Evidence for partial functional overlap of KEA and MSL transport proteins in the chloroplast inner envelope of Arabidopsis thaliana.

Arabidopsis thaliana possesses two different ion-export mechanisms in the plastid inner envelope membrane. Due to a genome duplication, the transport proteins are encoded by partly redundant loci: K+-efflux antiporter1 (KEA1) and KEA2 and mechanosensitive channel of small conductance-like2 (MSL2) and MSL3. Thus far, a functional link between these two mechanisms has not been established. Here, we show that kea1msl2 loss-of-function mutants exhibit phenotypes such as slow growth, reduced photosynthesis and changes in chloroplast morphology, several of which are distinct from either single mutants and do not resemble kea1kea2 or msl2msl3 double mutants. Our data suggest that KEA1 and MSL2 function in concert to maintain plastid ion homeostasis and osmoregulation. Their interplay is critical for proper chloroplast development, organelle function, and plant performance.

Probing the physiological role of the plastid outer-envelope membrane using the oemiR plasmid collection.

Plastids are the site of complex biochemical pathways, most prominently photosynthesis. The organelle evolved through endosymbiosis with a cyanobacterium, which is exemplified by the outer envelope membrane that harbors more than 40 proteins in Arabidopsis. Their evolutionary conservation indicates high significance for plant cell function. While a few proteins are well-studied as part of the protein translocon complex the majority of outer envelope protein functions is unclear. Gaining a deeper functional understanding has been complicated by the lack of observable loss-of-function mutant phenotypes, which is often rooted in functional genetic redundancy. Therefore, we designed outer envelope-specific artificial micro RNAs (oemiRs) capable of downregulating transcripts from several loci simultaneously. We successfully tested oemiR function by performing a proof-of-concept screen for pale and cold-sensitive mutants. An in-depth analysis of pale mutant alleles deficient in the translocon component TOC75 using proteomics provided new insights into putative compensatory import pathways. The cold stress screen not only recapitulated 3 previously known phenotypes of cold-sensitive mutants but also identified 4 mutants of additional oemiR outer envelope loci. Altogether our study revealed a role of the outer envelope to tolerate cold conditions and showcasts the power of the oemiR collection to research the significance of outer envelope proteins.

Biophysical and proteomic analyses of Pseudomonas syringae pv. tomato DC3000 extracellular vesicles suggest adaptive functions during plant infection.

Vesiculation is a process employed by Gram-negative bacteria to release extracellular vesicles (EVs) into the environment. EVs from pathogenic bacteria play functions in host immune modulation, elimination of host defenses, and acquisition of nutrients from the host. Here, we observed EV production of the bacterial speck disease causal agent, Pseudomonas syringae pv. tomato (Pto) DC3000, as outer membrane vesicle release. Mass spectrometry identified 369 proteins enriched in Pto DC3000 EVs. The EV samples contained known immunomodulatory proteins and could induce plant immune responses mediated by bacterial flagellin. Having identified two biomarkers for EV detection, we provide evidence for Pto DC3000 releasing EVs during plant infection. Bioinformatic analysis of the EV-enriched proteins suggests a role for EVs in antibiotic defense and iron acquisition. Thus, our data provide insights into the strategies this pathogen may use to develop in a plant environment. IMPORTANCE The release of extracellular vesicles (EVs) into the environment is ubiquitous among bacteria. Vesiculation has been recognized as an important mechanism of bacterial pathogenesis and human disease but is poorly understood in phytopathogenic bacteria. Our research addresses the role of bacterial EVs in plant infection. In this work, we show that the causal agent of bacterial speck disease, Pseudomonas syringae pv. tomato, produces EVs during plant infection. Our data suggest that EVs may help the bacteria to adapt to environments, e.g., when iron could be limiting such as the plant apoplast, laying the foundation for studying the factors that phytopathogenic bacteria use to thrive in the plant environment.

Impact of climate change on groundwater recharge in shallow young glacial aquifers in northern Poland.

We investigated the influence of climate change in the period 1951-2020 on shallow aquifers in the Brda and Wda outwash plains (Pomeranian Region, Northern Poland). There was a significant temperature rise (0.3 °C/10 years), which accelerated after 1980 (0.66 °C/10 years). Precipitation became increasingly irregular - extremely rainy years occurred right after or before extremely dry years, and intensive rainfall events became more frequent after 2000. The groundwater level decreased over the last 20 years, even though the average annual precipitation was higher than in the previous 50 years. We carried out numerical simulations of water flow in representative soil profiles for the years 1970-2020 using the HYDRUS-1D model, developed and calibrated during our earlier work at an experimental site in the Brda outwash plain (Gumula-Kawecka et al., 2022). We used a relationship between the water head and flux at the bottom of the soil profiles (the third-type boundary condition) to reproduce groundwater table fluctuations caused by recharge variability in time. The calculated daily recharge showed a decreasing linear trend for the last 20 years (0.05-0.06 mm d-1/10 years), and dropping trends in water table level and soil water content in the entire profile of vadose zone. Field tracer experiments were performed to estimate impact of extremely rain events on water flux in vadose zone. The results suggest that tracer travel times are strongly determined by water content in the unsaturated zone which is determined by precipitation amount in span of weeks, rather than extremely high precipitation events.

Towards a multi-basin SWAT model for the migration of nutrients and pesticides to Puck Bay (Southern Baltic Sea).

BACKGROUND: This paper analyzes the impact of changes in fertilization on crop yields and the runoff of nutrients from a small agricultural catchment (176 km2) to a shallow bay, using the SWAT model. Puck Bay is part of the Gulf of Gdansk and belongs to the Baltic Sea. The whole area of Puck Bay (364 km2) is protected (Natura 2000) yet despite this it suffers from eutrophication problems due to the relatively minimal depth and difficult water exchange. METHODS: The paper presents a comparison of the calculated yields and the runoff of nutrients and pesticides in the SWAT model, for a small agricultural coastal catchment. Calculations were made for 13 crop scenarios with weather data from 2011 to 2019. For each crop, an agriculture calendar was made. Two variants of fertilization were considered (autofertilization mode and according to the calendar). The nutrient runoff was calculated depending on the adopted scenario. In addition, the fate of selected pesticides was simulated. RESULTS: Depending on the crop, the annual load of NO3into the stream ranged from 0.74 to 3.65 kg ha-1. The annual load of organic phosphorous into the stream was between 0.686 and 3.64 kg ha-1. This is lower than in the majority of EU or Baltic countries. The surface runoff of dissolved Glyphosate was equal to 286 mg ha-1. The annual loads of nutrients from the catchment area are equivalent in both fertilization modes. Regardless of the selected fertilization mode, in addition to the dosage, the form of nutrients is important for the model.

Role of the Lakes in Groundwater Recharge and Discharge in the Young Glacial Area, Northern Poland.

The aim of this research was to delineate characteristic hydrogeological lake types in the Young Glacial Area (YGA). The YGA is in the central and east part of the Kashubian Lake District (KLD) in Northern Poland, an area covered by deposits of Quaternary glaciation. All the bigger lakes were investigated in the area of about 1500 km(2) (39 lakes). The role of lakes in groundwater recharge and discharge was determined from total dissolved solids (TDS) in lake waters and also from groundwater flow simulation. The general trend was that gaining lakes, as determined by flow modeling, had higher values of TDS than losing lakes. In addition to typical gaining lakes (with TDS > 250 mg/l), there were losing lakes perched on glacial till deposits with very low TDS (<100 mg/l). Two groups of losing lakes were delineated: ones with very low TDS and another group with slightly higher TDS (due to local contact with groundwater). Flow-through lakes with TDS of 170-200 mg/l were also delineated.