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At least 199 records · Page 11

Generating Ectropis obliqua axenic larvae in support of functional microbiome studies

Abstract Ectropis obliqua Prout (Lepidoptera: Geometridae) is a serious leaf‐eating pest in tea gardens throughout China, with rich bacterial communities in the larval gut. Few studies have investigated whether these bacteria play an important role in growth, development, and adaptability of E. obliqua . Axenic larval models are a useful tool for the study of interactions between gut bacteria and host insects. At present, artificial diet and antibiotics are typically used to remove larval gut bacteria for study; however, this method may change the natural food of insects and the habitat provided by their host plants, and cannot completely eliminate insect gut bacteria. Therefore, in order to establish an axenic insect feeding model, we first investigated the effects of egg age, different disinfectant concentration, and treatment time on the eggs of E. obliqua through orthogonal experiments. Next, we fed E. obliqua larvae tissue‐cultured tea seedlings under aseptic conditions. Thus, we successfully generated a feeding model of gut axenic larvae in the laboratory. Based on it, we then compared the mortality of axenic and non‐axenic E. obliqua larvae infected with the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill. These results demonstrated the feasibility of this axenic larval model and that B. bassiana is a broad‐spectrum entomogenous fungus with a toxic effect on E. obliqua .

Su, Qingqing↗

The role of height–driven constraints and compensations on tree vulnerability to drought

Frequent observations of higher mortality in larger trees than in smaller ones during droughts have sparked an increasing interest in size–dependent drought–induced mortality. However, the underlying physiological mechanisms are not well understood, with height–associated hydraulic constraints often being implied as the potential mechanism driving increased drought vulnerability. Here we performed a quantitative synthesis on how key traits that drive plant water and carbon economy change with tree height within species and assessed the implications that the different constraints and compensations may have on the interacting mechanisms (hydraulic failure, carbon starvation and/or biotic–agent attacks) affecting tree vulnerability to drought. While xylem tension increases with tree height, taller trees present a range of structural and functional adjustments, including more efficient water use and transport and greater water uptake and storage capacity, that mitigate the path–length–associated drop in water potential. These adaptations allow taller trees to withstand episodic water stress. Conclusive evidence for height–dependent increased vulnerability to hydraulic failure and carbon starvation, and their coupling to defence mechanisms and pest and pathogen dynamics, is still lacking. Further research is needed, particularly at the intraspecific level, to ascertain the specific conditions and thresholds above which height hinders tree survival under drought.

54 ENVIRONMENTAL SCIENCES↗

Engineering the production of conjugated fatty acids in Arabidopsis thaliana leaves

Summary The seeds of many nondomesticated plant species synthesize oils containing high amounts of a single unusual fatty acid, many of which have potential usage in industry. Despite the identification of enzymes for unusual oxidized fatty acid synthesis, the production of these fatty acids in engineered seeds remains low and is often hampered by their inefficient exclusion from phospholipids. Recent studies have established the feasibility of increasing triacylglycerol content in plant leaves, which provides a novel approach for increasing energy density of biomass crops. Here, we determined whether the fatty acid composition of leaf oil could be engineered to accumulate unusual fatty acids. Eleostearic acid ( ESA ) is a conjugated fatty acid produced in seeds of the tung tree ( Vernicia fordii ) and has both industrial and nutritional end‐uses. Arabidopsis thaliana lines with elevated leaf oil were first generated by transforming wild‐type, cgi‐58 or pxa1 mutants (the latter two of which contain mutations disrupting fatty acid breakdown) with the diacylglycerol acyltransferases ( DGAT 1 or DGAT 2 ) and/or oleosin genes from tung. High‐leaf‐oil plant lines were then transformed with tung FADX , which encodes the fatty acid desaturase/conjugase responsible for ESA synthesis. Analysis of lipids in leaves revealed that ESA was efficiently excluded from phospholipids, and co‐expression of tung FADX and DGAT 2 promoted a synergistic increase in leaf oil content and ESA accumulation. Taken together, these results provide a new approach for increasing leaf oil content that is coupled with accumulation of unusual fatty acids. Implications for production of biofuels, bioproducts, and plant–pest interactions are discussed.

59 BASIC BIOLOGICAL SCIENCES↗

Towards engineering hybrid incompatibility in plants

The potential for gene flow between genetically modified organisms (GMOs) and non-GMO relatives poses a significant challenge to the development and regulatory approval of GMO crops (Wedger et al., 2024), for example, the spread of herbicide resistance transgenes from crops such as rice or sorghum to cross-pollinating weedy species. Addressing this concern, we developed Engineered Genetic Incompatibility (EGI) (Maselko et al., 2017), a system that establishes species-like barriers to gene flow between otherwise sexually compatible populations. EGI employs Programmable Transcriptional Activators (PTAs) to drive lethal over- and/or ectopic expression of tightly regulated genes following undesired hybridization events (Figure 1a,b). A benign mutation of the target promoter in the EGI organism protects it from ill effects of the PTA, which acts as a sentinel for the wild-type (WT) promoter sequence. Given numerous potential PTA targets, multiple mutually incompatible subpopulations are feasible (Maselko et al., 2020). EGI has been demonstrated in yeast as a proof-of-concept (Maselko et al., 2017) and in insects as a strategy for genetic biocontrol of pests (Maselko et al., 2020; Upadhyay et al., 2022). EGI in plants would provide a strategy to halt gene flow between engineered crops and their domestic and wild relatives without altering normal cultivation or propagation practices. Here, we present promising results towards the demonstration of EGI in plants and highlight technical challenges that still need to be overcome.

CRISPRa↗

Similar local, but different systemic, metabolomics responses of closely related pine subspecies to folivory by caterpillars of the processionary moth

Plants respond locally and systemically to herbivore attack. Most of the research conducted on plant-herbivore relationships at elemental and molecular levels have focused on nutrients or/and certain molecular compounds or specific families of defensive metabolites showing that herbivores tend to select plant individuals or species with higher nutrient concentrations and to avoid those with higher levels of phenolics and terpenes. Unfortunately, the defensive role of phenolics in conifers is still unclear. We performed stoichiometric and metabolomics, local and systemic, analyses in two subspecies of Pinus sylvestris under the herbivorous attack by the caterpillars of the pine processionary moth, an important pest in the Mediterranean Basin. Herbivorous attack was not associated with any of the elements analyzed. Both pine subspecies responded locally to folivory mainly by increasing the concentrations of various terpenes and phenolics. Systemic responses differed between subspecies and most of the metabolites presented intermediate concentrations between those of the affected parts and unattacked trees. Contrary as usually thought, foliar nutrient concentrations did not show to be a main factor of an alleged plant selection by adult female processionary moths for oviposition. Local increases in phenolics were more associated with antioxidant function for protection against oxidative damage produced by folivory. On the other hand, terpenes were directly related to defense against herbivores. Herbivory attack produced a general systemic shift in pines, including both primary and secondary metabolisms, that was, however, less intense and chemically different from the local responses. Subspecies responded similarly locally but differently to folivory at systemic level.

Environmental Molecular Sciences Laboratory↗

Bacillus thuringiensisCry1Da_7 and Cry1B.868 Protein Interactions with Novel Receptors Allow Control of Resistant Fall Armyworms,Spodoptera frugiperda(J.E. Smith)

Two new modified Bacillus thuringiensis (Bt) proteins, Cry1Da_7 and Cry1B.868, with activity against fall armyworms (FAW), Spodoptera frugiperda (J.E. Smith), were evaluated for their potential to bind new insect receptors compared to proteins currently deployed as plant-incorporated protectants (PIPs) in row crops. Results from resistant insect bioassays, disabled insecticidal protein (DIP) bioassays, and cell-based assays using insect cells expressing individual receptors demonstrate that receptor utilizations of the newly modified Cry1Da_7 and Cry1B.868 proteins are distinct from each other and from those of commercially available Bt proteins such as Cry1F, Cry1A.105, Cry2Ab, and Vip3A. Accordingly, these two proteins target different insect proteins in FAW midgut cells and when pyramided together should provide durability in the field against this economically important pest.

59 BASIC BIOLOGICAL SCIENCES↗

Full Genome of Phialocephala scopiformis DAOMC 229536, a Fungal Endophyte of Spruce Producing the Potent Anti-Insectan Compound Rugulosin

ABSTRACT We present the full genome of Phialocephala scopiformis DAOMC 229536 (Helotiales, Ascomycota), a foliar endophyte of white spruce from eastern Quebec. DAOMC 229536 produces the anti-insectan compound rugulosin, which inhibits a devastating forestry pest, the spruce budworm. This genome will enable fungal genotyping and host-endophyte evolutionary genomics in inoculated trees.

59 BASIC BIOLOGICAL SCIENCES↗

iTOUGH2 v7.1

iTOUGH2 (inverse TOUGH2) provides inverse modeling capabilities for TOUGH2, a simulator for multi-dimensional, multi-phase, multi-component, non-isothermal flow and transport in fractured porous media. iTOUGH2 performs sensitivity analyses, data-worth analyses, parameter estimation, and uncertainty propagation analyses in geosciences and reservoir engineering and other application areas. iTOUGH2 supports a number of different combinations of fluids and components (equation-of-state (EOS) modules). In addition, the optimization routines implemented in iTOUGH2 can also be used for sensitivity analysis, automatic model calibration, and uncertainty quantification of any external code that uses text-based input and output files using the PEST protocol. iTOUGH2 solves the inverse problem by minimizing a non-linear objective function of the weighted differences between model output and the corresponding observations. Multiple minimization algorithms (derivative-free, gradient-based, and second-order; local and global) are available. iTOUGH2 also performs Latin Hypercube Monte Carlo simulations for uncertainty propagation analyses. A detailed residual and error analysis is provided. This upgrade includes (a) global sensitivity analysis methods, (b) dynamic memory allocation (c) additional input features and output analyses, (d) increased forward simulation capabilities, (e) parallel execution on multicore PCs and Linux clusters, and (f) bug fixes.

Finsterle, Stefan↗

LLNL Automized Surface Titration Model

The LLNL Automized Surface Titration Model (L-ASTM) is a community data-driven surface complexation modeling workflow for simulating potentiometric titration of mineral surfaces. The model accepts raw experimental potentiometric titration data formatted in a findable, accessible, interoperable, and reusable (FAIR) structure. The workflow was coded in Python and coupled to PHREEQC for surface complexation modeling and PEST for data fitting and parameter estimation.

Solchan, Han↗

Survival, healing, and swim performance of juvenile migratory sea lamprey ( Petromyzon marinus ) implanted with a new acoustic microtransmitter designed for small eel-like fishes

Background: Little is known about the transformer stage of the parasitic lampreys, a brief but critical period that encompasses juvenile out-migration from rivers to lakes or oceans to begin parasitic feeding. Information about this life stage could have significant conservation implications for both imperiled and invasive lampreys. We investigated tag retention, survival, wound healing, and swim performance of newly transformed sea lamprey (Petromyzon marinus) implanted with a new micro-acoustic transmitter, the eel–lamprey acoustic transmitter (ELAT), in a controlled laboratory environment. Results: The 61-day survival of our tagged subjects was 71%, within the range reported in similar studies of juvenile lampreys. However, survival was significantly lower in the tagged animals (vs control), with no effect statistically attributable to measures of animal length, mass, condition, or population of origin (Great Lakes vs. Atlantic drainage). Mortality in tagged fish was concentrated in the first four days post-surgery, suggesting injury from the surgical process. An unusually long recovery time from anesthesia may have contributed to the increased mortality. In a simple burst swim assay, tagged animals swam significantly slower (– 22.5%) than untagged animals, but were not significantly different in endurance swim tests. A composite wound healing score at day four was a significant predictor of maximum burst swim speed at day 20, and wound condition was related to animal mass, but not length, at the time of tagging. Conclusions: Impairments to survival and swim performance of juvenile sea lamprey implanted with the ELAT transmitter were within currently reported ranges for telemetry studies with small, difficult to observe fishes. Our results could be improved with more refined anesthesia and surgical techniques. The ability to track migratory movements of imperiled and pest populations of parasitic lampreys will improve our ability to estimate vital rates that underlie recruitment to the adult population (growth, survival) and to investigate the environmental factors that regulate the timing and rates of movement, in wild populations.

59 BASIC BIOLOGICAL SCIENCES↗

Molecular and Serological Survey of Selected Viruses in Free-Ranging Wild Ruminants in Iran

A molecular and serological survey of selected viruses in free-ranging wild ruminants was conducted in 13 different districts in Iran. Samples were collected from 64 small wild ruminants belonging to four different species including 25 Mouflon (Ovis orientalis), 22 wild goat (Capra aegagrus), nine Indian gazelle (Gazella bennettii) and eight Goitered gazelle (Gazella subgutturosa) during the national survey for wildlife diseases in Iran. Serum samples were evaluated using serologic antibody tests for Peste de petits ruminants virus (PPRV), Pestiviruses [Border Disease virus (BVD) and Bovine Viral Diarrhoea virus (BVDV)], Bluetongue virus (BTV), Bovine herpesvirus type 1 (BHV-1), and Parainfluenza type 3 (PI3). Sera were also ELISA tested for Pestivirus antigen. Tissue samples including spleen, liver, lung, tonsils, mesenteric and mediastinal lymph nodes and white blood cells (WBCs) were tested using polymerase chain reaction (PCR) for PPRV, Foot and Mouth Disease virus (FMDV), Pestivirus, BTV, Ovine herpesvirus type 2 (OvHV-2) and BHV-1. Serologic tests were positive for antibodies against PPRV (17%), Pestiviruses (2%) and BTV (2%). No antibodies were detected for BHV-1 or PI3, and no Pestivirus antigen was detected. PCR results were positive for PPRV (7.8%), FMDV (11%), BTV (3%), OvHV-2 (31%) and BHV-1 (1.5%). Finally, none of the samples were positive for Pestiviruses.

59 BASIC BIOLOGICAL SCIENCES↗

Varroa destructor mites vector and transmit pathogenic honey bee viruses acquired from an artificial diet

The ectoparasitic mite Varroa destructor is one of the most destructive pests of the honey bee (Apis mellifera) and the primary biotic cause of colony collapse in many regions of the world. These mites inflict physical injury on their honey bee hosts from feeding on host hemolymph and fat body cells/cellular components, and serve as the vector for deadly honey bee viruses, including Deformed wing virus (DWV) and the related Varroa destructor virus-1 (VDV-1) (i.e., DWV-like viruses). Studies focused on elucidating the dynamics of Varroa-mediated vectoring and transmission of DWV-like viruses may be confounded by viruses present in ingested host tissues or the mites themselves. Here we describe a system that includes an artificial diet free of insect tissue-derived components for maintaining Varroa mites for in vitro experimentation. Using this system, together with the novel engineered cDNA clone-derived genetically tagged VDV-1 and wild-type DWV, we demonstrated for the first time that Varroa mites provided an artificial diet supplemented with engineered viruses for 36 hours could acquire and transmit sufficient numbers of virus particles to establish an infection in virus-naïve hosts. While the in vitro system described herein provides for only up to five days of mite survival, precluding study of the long-term impacts of viruses on mite health, the system allows for extensive insights into the dynamics of Varroa-mediated vectoring and transmission of honey bee viruses.

59 BASIC BIOLOGICAL SCIENCES↗

Structural and functional insights into the first Bacillus thuringiensis vegetative insecticidal protein of the Vpb4 fold, active against western corn rootworm

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major maize pest in the United States causing significant economic loss. The emergence of field-evolved resistant WCR to Bacillus thuringiensis (Bt) traits has prompted the need to discover and deploy new insecticidal proteins in transgenic maize. In the current study we determined the crystal structure and mode of action (MOA) of the Vpb4Da2 protein (formerly known as Vip4Da2) from Bt, the first identified insecticidal Vpb4 protein with commercial level control against WCR. The Vpb4Da2 structure exhibits a six-domain architecture mainly comprised of antiparallel β-sheets organized into β-sandwich layers. The amino-terminal domains 1–3 of the protein share structural homology with the protective antigen (PA) PA14 domain and encompass a long β-pore forming loop as in the clostridial binary-toxB module. Domains 5 and 6 at the carboxyl-terminal half of Vpb4Da2 are unique as this extension is not observed in PA or any other structurally-related protein other than Vpb4 homologs. These unique Vpb4 domains adopt the topologies of carbohydrate-binding modules known to participate in receptor-recognition. Functional assessment of Vpb4Da2 suggests that domains 4–6 comprise the WCR receptor binding region and are key in conferring the observed insecticidal activity against WCR. The current structural analysis was complemented by in vitro and in vivo characterizations, including immuno-histochemistry, demonstrating that Vpb4Da2 follows a MOA that is consistent with well-characterized 3-domain Bt insecticidal proteins despite significant structural differences.

59 BASIC BIOLOGICAL SCIENCES↗

AmeriFlux US-CF1 CAF-LTAR Cook East

This is the AmeriFlux version of the carbon flux data for the site US-CF1 CAF-LTAR Cook East. Site Description - CF1 has operated since May 2017 at the R.J. Cook Agronomy Farm outside of Pullman, Washington, and monitors the no-till side of a paired-catchment study that is part of the Longterm Agroecosystem Research (LTAR) site common experiment. CF1 is located in “Cook East,” a field that has been in no-till management since 1998, and represents the Alternative treatment of the LTAR common experiment. It is paired with the CF2 tower location in “Cook West,” which has been in conventional tillage since prior to 1998 and represents the Prevalent treatments of the common experiment. Cook Agronomy Farm is in the high precipitation agroecological zone of the Columbia Plateau’s dryland cropping region. Crop rotations are wheat-based and include winter wheat, spring wheat, chickpea, spring canola, and winter peas. Wheat is the principal cash crop, with other crops grown in rotation for diversity, nutrient, and pest management. Soils are predominantly silt loam texture Mollisols in the Palouse, Thatuna, Naff soil series. The CF1 tower replaced US-RC1, which operated 2012-2016 in a neighboring field that was also in no-till management since 1998. CF1 and RC1 have distinct footprints, aspects, and soil series compositions.

Phillips, Claire L.↗

AmeriFlux US-CF2 CAF-LTAR Cook West

This is the AmeriFlux version of the carbon flux data for the site US-CF2 CAF-LTAR Cook West. Site Description - CF2 has operated since May 2017 at the R.J. Cook Agronomy Farm outside of Pullman, Washington, and monitors the conventionally tilled side of a paired-catchment study that is part of the Longterm Agroecosystem Research (LTAR) site common experiment. CF2 is located in “Cook West,” a field that has been in conventional tillage since prior to 1998 and constitutes the Prevalent treatment of the common experiment. In this case, conventional tillage involves chisel plowing after the winter wheat phase of rotations (approximately every 3-4 years) and occasional harrowing. CF2 is paired with the CF1 tower location in “Cook East,” which has been in no-till since 1998 and represents the Alternative treatment of the common experiment. Both fields are planted in the same crops every year. Cook Agronomy Farm is in the high precipitation agroecological zone of the Columbia Plateau’s dryland cropping region. Crop rotations are wheat-based and include winter wheat, spring wheat, chickpea, spring canola, and winter peas. Wheat is the principal cash crop, with other crops grown in rotation for diversity, nutrient, and pest management. Soils are predominantly silt loam texture Mollisols in the Palouse, Thatuna, Naff soil series. The CF2 tower replaced US-RC2, which operated 2012-2016 in a neighboring field that was also conventionally tilled. CF2 and RC2 have distinct footprints, aspects, and soil series compositions.

Huggins, Dave↗

AmeriFlux FLUXNET-1F US-CF1 CAF-LTAR Cook East

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site US-CF1 CAF-LTAR Cook East. This is the FLUXNET version of the carbon flux data for the site US-CF1 CAF-LTAR Cook East produced by applying the standard ONEFlux (1F) software. Site Description - CF1 has operated since May 2017 at the R.J. Cook Agronomy Farm outside of Pullman, Washington, and monitors the no-till side of a paired-catchment study that is part of the Longterm Agroecosystem Research (LTAR) site common experiment. CF1 is located in “Cook East,” a field that has been in no-till management since 1998, and represents the Alternative treatment of the LTAR common experiment. It is paired with the CF2 tower location in “Cook West,” which has been in conventional tillage since prior to 1998 and represents the Prevalent treatments of the common experiment. Cook Agronomy Farm is in the high precipitation agroecological zone of the Columbia Plateau’s dryland cropping region. Crop rotations are wheat-based and include winter wheat, spring wheat, chickpea, spring canola, and winter peas. Wheat is the principal cash crop, with other crops grown in rotation for diversity, nutrient, and pest management. Soils are predominantly silt loam texture Mollisols in the Palouse, Thatuna, Naff soil series. The CF1 tower replaced US-RC1, which operated 2012-2016 in a neighboring field that was also in no-till management since 1998. CF1 and RC1 have distinct footprints, aspects, and soil series compositions.

Phillips, Claire L.↗

AmeriFlux FLUXNET-1F US-CF2 CAF-LTAR Cook West

This is the AmeriFlux Management Project (AMP) created FLUXNET-1F version of the carbon flux data for the site US-CF2 CAF-LTAR Cook West. This is the FLUXNET version of the carbon flux data for the site US-CF2 CAF-LTAR Cook West produced by applying the standard ONEFlux (1F) software. Site Description - CF2 has operated since May 2017 at the R.J. Cook Agronomy Farm outside of Pullman, Washington, and monitors the conventionally tilled side of a paired-catchment study that is part of the Longterm Agroecosystem Research (LTAR) site common experiment. CF2 is located in “Cook West,” a field that has been in conventional tillage since prior to 1998 and constitutes the Prevalent treatment of the common experiment. In this case, conventional tillage involves chisel plowing after the winter wheat phase of rotations (approximately every 3-4 years) and occasional harrowing. CF2 is paired with the CF1 tower location in “Cook East,” which has been in no-till since 1998 and represents the Alternative treatment of the common experiment. Both fields are planted in the same crops every year. Cook Agronomy Farm is in the high precipitation agroecological zone of the Columbia Plateau’s dryland cropping region. Crop rotations are wheat-based and include winter wheat, spring wheat, chickpea, spring canola, and winter peas. Wheat is the principal cash crop, with other crops grown in rotation for diversity, nutrient, and pest management. Soils are predominantly silt loam texture Mollisols in the Palouse, Thatuna, Naff soil series. The CF2 tower replaced US-RC2, which operated 2012-2016 in a neighboring field that was also conventionally tilled. CF2 and RC2 have distinct footprints, aspects, and soil series compositions.

Huggins, Dave↗