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ABSTRACT Advanced genome editing technologies have enabled rapid and flexible rewriting of theEscherichia coligenome, benefiting fundamental biology and biomanufacturing. Unfortunately, some of the most useful technologies to advance genome editing inE. colihave not yet been ported into other bacterial species. For instance, the addition of bacterial retrons to the genome editing toolbox has increased the efficiency of recombineering inE. coliby enabling sustained, abundant production of ssDNA recombineering donors by reverse transcription that install flexible, precise edits in the prokaryotic chromosome. To extend the utility of this technology beyondE. coli, we surveyed the portability and versatility of retron-mediated recombineering across three different bacterial phyla (Proteobacteria, BacillotaandActinomycetota) and a total of 15 different species. We found that retron recombineering is functional in all species tested, reaching editing efficiencies above 20% in six of them, above 40% in three of them, and above 90% in two of them. We also tested the extension of the recombitron architecture optimizations and strain backgrounds in a subset of hosts to additionally increase editing rates. The broad recombitron survey carried out in this study forms the basis for widespread use of retron-derived technologies through the whole Bacteria domain. 

Abstract We analyze 330 ks of Chandra X-ray imaging and spectra of the nearby, edge-on starburst and Seyfert type 2 galaxy NGC 4945 to measure the hot gas properties along the galactic outflows. We extract and model spectra from 15 regions extending from −0.55 to +0.85 kpc above and below the galactic disk to determine the best-fit parameters and metal abundances. We find that the hot gas temperatures and number densities peak in the central regions and decrease along the outflows. These profiles are inconsistent with a spherical, adiabatically expanding wind model, suggesting the need to include mass loading and/or a nonspherical outflow geometry. We estimate the mass outflow rate of the hot wind to be 1.6M_⊙yr⁻¹. Emission from charge exchange is detected in the northern outflow, and we estimate it contributes 12% to the emitted, broadband (0.5–7 keV) X-ray flux. 

Abstract We present a detailed analysis of nearly two decades of optical/UV and X-ray data to study the multi-wavelength pre-explosion properties and post-explosion X-ray properties of nearby SN2023ixf located in M101. We find no evidence of precursor activity in the optical to UV down to a luminosity of$$\lesssim$$$$1.0\times10^{5}\, \textrm{L}_{\odot}$$, while X-ray observations covering nearly 18 yr prior to explosion show no evidence of luminous precursor X-ray emission down to an absorbed 0.3–10.0 keV X-ray luminosity of$$\sim$$$$6\times10^{36}$$erg s$$^{-1}$$. ExtensiveSwiftobservations taken post-explosion did not detect soft X-ray emission from SN2023ixf within the first$$\sim$$3.3 days after first light, which suggests a mass-loss rate for the progenitor of$$\lesssim$$$$5\times10^{-4}\,\textrm{M}_{\odot}$$yr$$^{-1}$$or a radius of$$\lesssim$$$$4\times10^{15}$$cm for the circumstellar material. Our analysis also suggests that if the progenitor underwent a mass-loss episode, this had to occur$$>$$0.5–1.5 yr prior to explosion, consistent with previous estimates.Swiftdetected soft X-rays from SN2023ixf$$\sim$$$$4.25$$days after first light, and it rose to a peak luminosity of$$\sim10^{39}$$erg s$$^{-1}$$after 10 days and has maintained this luminosity for nearly 50 days post first light. This peak luminosity is lower than expected, given the evidence that SN2023ixf is interacting with dense material. However, this might be a natural consequence of an asymmetric circumstellar medium. X-ray spectra derived from merging allSwiftobservations over the first 50 days are best described by a two-component bremsstrahlung model consisting of a heavily absorbed and hotter component similar to that found usingNuSTAR, and a less-absorbed, cooler component. We suggest that this soft component arises from cooling of the forward shock similar to that found in Type IIn SN2010jl. 

Abstract We analyze image and spectral data from ≈365 ks of observations from the Chandra X-ray Observatory of the nearby, edge-on starburst galaxy NGC 253 to constrain properties of the hot phase of the outflow. We focus our analysis on the −1.1 to +0.63 kpc region of the outflow and define several regions for spectral extraction where we determine best-fit temperatures and metal abundances. We find that the temperatures and electron densities peak in the central ∼250 pc region of the outflow and decrease with distance. These temperature and density profiles are in disagreement with an adiabatic spherically expanding starburst wind model and suggest the presence of additional physics such as mass loading and nonspherical outflow geometry. Our derived temperatures and densities yield cooling times in the nuclear region of a few million years, which may imply that the hot gas can undergo bulk radiative cooling as it escapes along the minor axis. Our metal abundances of O, Ne, Mg, Si, S, and Fe all peak in the central region and decrease with distance along the outflow, with the exception of Ne, which maintains a flat distribution. The metal abundances indicate significant dilution outside of the starburst region. We also find estimates of the mass outflow rates, which are 2.8M_⊙yr⁻¹in the northern outflow and 3.2M_⊙yr⁻¹in the southern outflow. Additionally, we detect emission from charge exchange and find it makes a significant contribution (20%–42%) to the total broadband (0.5–7 keV) X-ray emission in the central and southern regions of the outflow. 

Star formation in galaxies is regulated by turbulence, outflows, gas heating and cloud dispersal -- processes which depend sensitively on the properties of the interstellar medium (ISM) into which supernovae (SNe) explode. Unfortunately, direct measurements of ISM environments around SNe remain scarce, as SNe are rare and often distant. Here we demonstrate a new approach: mapping the ISM around the massive stars that are soon to explode. This provides a much larger census of explosion sites than possible with only SNe, and allows comparison with sensitive, high-resolution maps of the atomic and molecular gas from the Jansky VLA and ALMA. In the well-resolved Local Group spiral M33, we specifically observe the environments of red supergiants (RSGs, progenitors of Type II SNe), Wolf-Rayet stars (WRs, tracing stars >30 M⊙, and possibly future stripped-envelope SNe), and supernova remnants (SNRs, locations where SNe have exploded). We find that massive stars evolve not only in dense, molecular-dominated gas (with younger stars in denser gas), but also a substantial fraction (∼45\% of WRs; higher for RSGs) evolve in lower-density, atomic-gas-dominated, inter-cloud media. We show that these measurements are consistent with expectations from different stellar-age tracer maps, and can be useful for validating SN feedback models in numerical simulations of galaxies. Along with the discovery of a 20-pc diameter molecular gas cavity around a WR, these findings re-emphasize the importance of pre-SN/correlated-SN feedback evacuating the dense gas around massive stars before explosion, and the need for high-resolution (down to pc-scale) surveys of the multi-phase ISM in nearby galaxies. 

Abstract We present the ALMA detection of molecular outflowing gas in the central regions of NGC 4945, one of the nearest starbursts and also one of the nearest hosts of an active galactic nucleus (AGN). We detect four outflow plumes in CO J = 3 − 2 at ∼0.″3 resolution that appear to correspond to molecular gas located near the edges of the known ionized outflow cone and its (unobserved) counterpart behind the disk. The fastest and brightest of these plumes has emission reaching observed line-of-sight projected velocities of over 450 km s −1 beyond systemic, equivalent to an estimated physical outflow velocity v ≳ 600 km s −1 for the fastest emission. Most of these plumes have corresponding emission in HCN or HCO + J = 4 − 3. We discuss a kinematic model for the outflow emission where the molecular gas has the geometry of the ionized gas cone and shares the rotation velocity of the galaxy when ejected. We use this model to explain the velocities we observe, constrain the physical speed of the ejected material, and account for the fraction of outflowing gas that is not detected due to confusion with the galaxy disk. We estimate a total molecular mass outflow rate M ̇ mol ∼ 20 M ⊙ yr −1 flowing through a surface within 100 pc of the disk midplane, likely driven by a combination of the central starburst and AGN. 

Abstract During photosynthesis, electrons travel from light-excited chlorophyll molecules along the electron transport chain to the final electron acceptor nicotinamide adenine dinucleotide phosphate (NADP) to form NADPH, which fuels the Calvin–Benson–Bassham cycle (CBBC). To allow photosynthetic reactions to occur flawlessly, a constant resupply of the acceptor NADP is mandatory. Several known stromal mechanisms aid in balancing the redox poise, but none of them utilizes the structurally highly similar coenzyme NAD(H). Using Arabidopsis (Arabidopsis thaliana) as a C3-model, we describe a pathway that employs the stromal enzyme PHOSPHOGLYCERATE DEHYDROGENASE 3 (PGDH3). We showed that PGDH3 exerts high NAD(H)-specificity and is active in photosynthesizing chloroplasts. PGDH3 withdrew its substrate 3-PGA directly from the CBBC. As a result, electrons become diverted from NADPH via the CBBC into the separate NADH redox pool. pgdh3 loss-of-function mutants revealed an overreduced NADP(H) redox pool but a more oxidized plastid NAD(H) pool compared to wild-type plants. As a result, photosystem I acceptor side limitation increased in pgdh3. Furthermore, pgdh3 plants displayed delayed CBBC activation, changes in nonphotochemical quenching, and altered proton motive force partitioning. Our fluctuating light-stress phenotyping data showed progressing photosystem II damage in pgdh3 mutants, emphasizing the significance of PGDH3 for plant performance under natural light environments. In summary, this study reveals an NAD(H)-specific mechanism in the stroma that aids in balancing the chloroplast redox poise. Consequently, the stromal NAD(H) pool may provide a promising target to manipulate plant photosynthesis.