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1. Terpenes modulate bacterial and fungal growth and sorghum rhizobiome communities

   https://doi.org/10.1128/spectrum.01332-23  

   Chou, Ming-Yi; Andersen, Trine B.; Mechan Llontop, Marco E.; Beculheimer, Nick; Sow, Alassane; Moreno, Nick; Shade, Ashley; Hamberger, Bjoern; Bonito, Gregory; Calatrava, Victoria (October 2023, Microbiology Spectrum)

   Hom, Erik F. (Ed.)

   ABSTRACT Terpenes are among the oldest and largest class of plant-specialized bioproducts that are known to affect plant development, adaptation, and biological interactions. While their biosynthesis, evolution, and function in aboveground interactions with insects and individual microbial species are well studied, how different terpenes impact plant microbiomes belowground is much less understood. Here we designed an experiment to assess how belowground exogenous applications of monoterpenes (1,8-cineole and linalool) and a sesquiterpene (nerolidol) delivered through an artificial root system impacted its belowground bacterial and fungal microbiome. We found that the terpene applications had significant and variable impacts on bacterial and fungal communities, depending on terpene class and concentration; however, these impacts were localized to the artificial root system and the fungal rhizosphere. We complemented this experiment with pure culture bioassays on responsive bacteria and fungi isolated from the sorghum rhizobiome. Overall, higher concentrations (200 µM) of nerolidol were inhibitory toFerrovibriumand tested Firmicutes. While fungal isolates ofPenicilliumandPericoniawere also more inhibited by higher concentrations (200 µM) of nerolidol,Clonostachyswas enhanced at this higher level and together withHumicolawas inhibited by the lower concentration tested (100 µM). On the other hand, 1,8-cineole had an inhibitory effect onOrbiliaat both tested concentrations but had a promotive effect at 100 µM onPenicilliumandPericonia. Similarly, linalool at 100 µM had significant growth promotion inMortierella, but an inhibitory effect forOrbilia. Together, these results highlight the variable direct effects of terpenes on single microbial isolates and demonstrate the complexity of microbe-terpene interactions in the rhizobiome. ImportanceTerpenes represent one of the largest and oldest classes of plant-specialized metabolism, but their role in the belowground microbiome is poorly understood. Here, we used a “rhizobox” mesocosm experimental set-up to supply different concentrations and classes of terpenes into the soil compartment with growing sorghum for 1 month to assess how these terpenes affect sorghum bacterial and fungal rhizobiome communities. Changes in bacterial and fungal communities between treatments belowground were characterized, followed by bioassays screening on bacterial and fungal isolates from the sorghum rhizosphere against terpenes to validate direct microbial responses. We found that microbial growth stimulatory and inhibitory effects were localized, terpene specific, dose dependent, and transient in time. This work paves the way for engineering terpene metabolisms in plant microbiomes for improved sustainable agriculture and bioenergy crop production. 

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2. DiI staining of sensory neurons in the entomopathogenic nematode Steinernema hermaphroditum

   https://doi.org/10.17912/micropub.biology.000516  

   Garg P, Tan CH (February 2022, microPublication biology)

   Steinernema hermaphroditum entomopathogenic nematodes (EPN) and their Xenorhabdus griffiniae symbiotic bacteria have recently been shown to be a genetically tractable system for the study of both parasitic and mutualistic symbiosis. In their infective juvenile (IJ) stage, EPNs search for insect hosts to invade and quickly kill them with the help of the symbiotic bacteria they contain. The mechanisms behind these behaviors have not been well characterized, including how the nematodes sense their insect hosts. In the well-studied free‑living soil nematode Caenorhabditis elegans, ciliated amphid neurons enable the worms to sense their environment, including chemosensation. Some of these neurons have also been shown to control the decision to develop as a stress-resistant dauer larva, analogous to the infective juveniles of EPNs, or to exit from dauer and resume larval development. In C. elegans and other nematodes, dye-filling with DiI is an easy and efficient method to label these neurons. We developed a protocol for DiI staining of S. hermaphroditum sensory neurons. Using this method, we could identify neurons positionally analogous to the C. elegans amphid neurons ASI, ADL, ASK, ASJ, as well as inner labial neurons IL1 and IL2. Similar to findings in other EPNs, we also found that the IJs of S. hermaphroditum are dye-filling resistant. 

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3. Microbial community structure in recovering forests of Mount St. Helens

   https://doi.org/10.3389/frmbi.2024.1399416  

   Maltz, Mia Rose; Allen, Michael F; Phillips, Michala L; Hernandez, Rebecca R; Shulman, Hannah B; Freund, Linton; Andrews, Lela V; Botthoff, Jon K; Aronson, Emma L (November 2024, Frontiers in Microbiomes)

   IntroductionThe 1980 eruption of Mount St. Helens had devastating effects above and belowground in forested montane ecosystems, including the burial and destruction of soil microbes. Soil microbial propagules and legacies in recovering ecosystems are important for determining post-disturbance successional trajectories. Soil microorganisms regulate nutrient cycling, interact with many other organisms, and therefore may support successional pathways and complementary ecosystem functions, even in harsh conditions. Historic forest management methods, such as old-growth and clearcut regimes, and locations of historic short-term gopher enclosures (Thomomys talpoides), to evaluate community response to forest management practices and to examine vectors for dispersing microbial consortia to the surface of the volcanic landscape. These biotic interactions may have primed ecological succession in the volcanic landscape, specifically Bear Meadow and the Pumice Plain, by creating microsite conditions conducive to primary succession and plant establishment. Methods and resultsUsing molecular techniques, we examined bacterial, fungal, and AMF communities to determine how these variables affected microbial communities and soil properties. We found that bacterial/archaeal 16S, fungal ITS2, and AMF SSU community composition varied among forestry practices and across sites with long-term lupine plots and gopher enclosures. The findings also related to detected differences in C and N concentrations and ratios in soil from our study sites. Fungal communities from previously clearcut locations were less diverse than in gopher plots within the Pumice Plain. Yet, clearcut meadows harbored fewer ancestral AM fungal taxa than were found within the old-growth forest. DiscussionBy investigating both forestry practices and mammals in microbial dispersal, we evaluated how these interactions may have promoted revegetation and ecological succession within the Pumice Plains of Mount St. Helens. In addition to providing evidence about how dispersal vectors and forest structure influence post-eruption soil microbiomes, this project also informs research and management communities about belowground processes and microbial functional traits in facilitating succession and ecosystem function. 

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4. Effects of drought and recovery on soil volatile organic compound fluxes in an experimental rainforest

   https://doi.org/10.1038/s41467-023-40661-8  

   Pugliese, Giovanni; Ingrisch, Johannes; Meredith, Laura K.; Pfannerstill, Eva Y.; Klüpfel, Thomas; Meeran, Kathiravan; Byron, Joseph; Purser, Gemma; Gil-Loaiza, Juliana; van Haren, Joost; et al (August 2023, Nature Communications)

   Abstract Drought can affect the capacity of soils to emit and consume biogenic volatile organic compounds (VOCs). Here we show the impact of prolonged drought followed by rewetting and recovery on soil VOC fluxes in an experimental rainforest. Under wet conditions the rainforest soil acts as a net VOC sink, in particular for isoprenoids, carbonyls and alcohols. The sink capacity progressively decreases during drought, and at soil moistures below ~19%, the soil becomes a source of several VOCs. Position specific¹³C-pyruvate labeling experiments reveal that soil microbes are responsible for the emissions and that the VOC production is higher during drought. Soil rewetting induces a rapid and short abiotic emission peak of carbonyl compounds, and a slow and long biotic emission peak of sulfur-containing compounds. Results show that, the extended drought periods predicted for tropical rainforest regions will strongly affect soil VOC fluxes thereby impacting atmospheric chemistry and climate. 

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5. Reversible kink instability drives ultrafast jumping in nematodes and soft robots

   https://doi.org/10.1101/2024.06.07.598012  

   Kumar, Sunny; Tiwari, Ishant; Ortega-Jimenez, Victor M; Dillman, Adler R; He, Dongjing; Hu, Yuhang; Bhamla, M Saad (June 2024, bioRxiv)

   Entomopathogenic nematodes (EPNs) exhibit a bending-elastic instability, or kink, before becoming airborne, a feature hypothesized but not proven to enhance jumping performance. Here, we provide the evidence that this kink is crucial for improving launch performance. We demonstrate that EPNs actively modulate their aspect ratio, forming a liquid-latched closed loop over a slow timescaleO(1 s), then rapidly open itO(10 µs), achieving heights of 20 body lengths (BL) and generating ∼ 10⁴W/Kg of power. Using jumping nematodes, a bio-inspired Soft Jumping Model (SoftJM), and computational simulations, we explore the mechanisms and implications of this kink. EPNs control their takeoff direction by adjusting their head position and center of mass, a mechanism verified through phase maps of jump directions in simulations and SoftJM experiments. Our findings reveal that the reversible kink instability at the point of highest curvature on the ventral side enhances energy storage using the nematode’s limited muscular force. We investigated the impact of aspect ratio on kink instability and jumping performance using SoftJM, and quantified EPN cuticle stiffness with AFM, comparing it withC. elegans. This led to a stiffness-modified SoftJM design with a carbon fiber backbone, achieving jumps of ∼25 BL. Our study reveals how harnessing kink instabilities, a typical failure mode, enables bidirectional jumps in soft robots on complex substrates like sand, offering a novel approach for designing limbless robots for controlled jumping, locomotion, and even planetary exploration. 

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