Effect of maximum tolerable stresses with time on the histostructure of the liver in monkeys
Optimal tolerable stress-time effects of acceleration on histology of monkey liver
Kotovskiy, Ye. F.↗
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Using supervised machine-learning approaches to understand abiotic stress tolerance and design resilient crops
Abiotic stresses such as drought, heat, cold, salinity and flooding significantly impact plant growth, development and productivity. As the planet has warmed, these abiotic stresses have increased in frequency and intensity, affecting the global food supply and making it imperative to develop stress-resilient crops. In the past 20 years, the development of omics technologies has contributed to the growth of datasets for plants grown under a wide range of abiotic environments. Integration of these rapidly growing data using machine-learning (ML) approaches can complement existing breeding efforts by providing insights into the mechanisms underlying plant responses to stressful conditions, which can be used to guide the design of resilient crops. In this review, we introduce ML approaches and provide examples of how researchers use these approaches to predict molecular activities, gene functions and genotype responses under stressful conditions. Finally, we consider the potential and challenges of using such approaches to enable the design of crops that are better suited to a changing environment. This article is part of the theme issue ‘Crops under stress: can we mitigate the impacts of climate change on agriculture and launch the ‘Resilience Revolution’?’.
abiotic stress↗
Increased Cuticle Waxes by Overexpression of WSD1 Improves Osmotic Stress Tolerance in Arabidopsis thaliana and Camelina sativa
To ensure global food security under the changing climate, there is a strong need for developing ‘climate resilient crops’ that can thrive and produce better yields under extreme environmental conditions such as drought, salinity, and high temperature. To enhance plant productivity under the adverse conditions, we constitutively overexpressed a bifunctional wax synthase/acyl-CoA:diacylglycerol acyltransferase (WSD1) gene, which plays a critical role in wax ester synthesis in Arabidopsis stem and leaf tissues. The qRT-PCR analysis showed a strong upregulation of WSD1 transcripts by mannitol, NaCl, and abscisic acid (ABA) treatments, particularly in Arabidopsis thaliana shoots. Gas chromatography and electron microscopy analyses of Arabidopsis seedlings overexpressing WSD1 showed higher deposition of epicuticular wax crystals and increased leaf and stem wax loading in WSD1 transgenics compared to wildtype (WT) plants. WSD1 transgenics exhibited enhanced tolerance to ABA, mannitol, drought and salinity, which suggested new physiological roles for WSD1 in stress response aside from its wax synthase activity. Transgenic plants were able to recover from drought and salinity better than the WT plants. Furthermore, transgenics showed reduced cuticular transpirational rates and cuticle permeability, as well as less chlorophyll leaching than the WT. The knowledge from Arabidopsis was translated to the oilseed crop Camelina sativa (L.) Crantz. Similar to Arabidopsis, transgenic Camelina lines overexpressing WSD1 also showed enhanced tolerance to drought stress. Our results clearly show that the manipulation of cuticular waxes will be advantageous for enhancing plant productivity under a changing climate.
59 BASIC BIOLOGICAL SCIENCES↗
Species‐Specific Epigenetic Signature Associates With Heat Stress Tolerance in the Perennial Tree Species Populus
Epigenetic regulation in annual plants is recognized as a key component of recurring stress acclimation and adaptation, but reports on perennial tree species are limited. In this study, two contrasting tree species, Populus trichocarpa and Populus deltoides, and an F1 hybrid cross between them showed species-specific epigenetic and physiological responses to heat stress (42°C) following priming (35°C). By analyzing whole-genome methylation, transcriptomics, proteomics, metabolomics, and photosynthesis parameters, we found that P. deltoides expresses specific epigenetic signatures in response to heat, resulting in improved photosynthetic efficiency compared to P. trichocarpa. Conversely, P. trichocarpa displayed stress signaling and defense mechanisms that could not sustain a net assimilation rate despite maintaining higher gas exchange. Heat stress following priming in hybrid plants increased transcript levels of thermotolerance-related transcription factors, such as SPL12. Selected regions in the promoter of SPL12 showed differential methylation between direct heat stress and priming followed by heat stress. As a result, upregulation of downstream genes and associated increases in protein and metabolite abundance for stress adaptation were exhibited. Consequently, hybrid plants showed enhanced photosynthesis and gas exchange rates, a trait lacking in P. trichocarpa. These results imply that priming may not be universally effective in enhancing plant performance under stress, particularly in perennial tree species. However, priming can acclimate the perennial tree species P. deltoides to withstand elevated temperature stress better. Our study has demonstrated that priming-based stress adaptation is species-specific but can be attained through crossbreeding, indicating its potential use in breeding programs.
DNA methylation↗
Development of Genetic Countermeasures for Enhancing Cellular Stress Tolerance on a Lunar Surface Mission
The Lunar Explorer Instrument for space biology Applications (LEIA) LEIA investigates the response to partial gravity and ionizing radiation of: Different DNA damage and stress response pathways and Bioproduction of antioxidants LEIA utilizes: Various strains of the yeast Saccharomyces cerevisiae, which will be desiccated in fluidic cards and rehydrated on the lunar surface LEIA develops: Genetic countermeasures to improve tolerance to the desiccation process and the constraints associated with long duration missions beyond low Earth orbit (LEO)
Neha Lingam↗
Discovering Innovations in Stress Tolerance through Comparative Gene Regulatory Network Analysis and Cell-Type Specific Expression Maps (Final Technical Report with Cover Page)
Through this grant, we developed a comparative framework to elucidate the mechanisms behind variations in environmental stress responses among a diverse group of species within the Brassicaceae family. Our focus was on the differences in physiological and transcriptomic responses to abscisic acid (ABA), a hormone associated with water stress. We examined the differential growth responses of four Brassicaceae species, finding that most exhibited reduced root growth correlated with smaller meristem size. In contrast, Schrenkiella parvula showed accelerated growth due to increased root cell elongation. We employed RNA sequencing to analyze the transcriptional responses to ABA across these species, and innovative bioinformatics techniques were used to pinpoint biological pathways with significant divergence. Additionally, we utilized DAP-seq to map the gene regulatory networks associated with ABAresponsive transcription factors, revealing that variations in the regulation of growth hormone biosynthesis play a critical role in the distinct ABA effects on root growth among the species. This research sets a new standard for comparative physiology by integrating comparative genomics and transcriptomics to uncover pathway divergences.
59 BASIC BIOLOGICAL SCIENCES↗
Decreases in polyunsaturated fatty acid content improve heat stress tolerance during flowering and silicle development in pennycress (Thlaspi arvense L.)
Introduction: Pennycress (Thlaspi arvense L.) is an emerging intermediate oilseed crop grown in the offseason between primary summer crops to produce three cash crops in two years. Previous efforts to improve seed oil quality produced Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) genome-edited lines with decreased polyunsaturated fatty acids (PUFAs) levels, through loss of function of the FATTY ACID DESATURASE 2 (FAD2), REDUCED OLEATE DESATURATION1 (ROD1), and FATTY ACID ELONGASE1 (FAE1) genes. While seed oil compositions were previously characterized, it remains unknown how vegetative and reproductive tissue compositions might differ and affect tolerance to high temperature (HT) conditions.Methods: In four growth chamber experiments, we explored HT tolerance during flowering and silicle development. Plants were subjected to a 34 °C day/28 °C night regime and compared to control plants maintained at 20 °C. Pollen grain viability at a range of temperatures, lipid peroxidation and proline content in leaves and silicles following HT, and seed yield were measured.Results: Both fad2 and rod1 mutant lines had relatively higher pollen viability (71% and 54% respectively) under moderately elevated temperature (28 °C) compared to wild-type controls (37%). They also showed smaller decreases in seed yield (0% and 40% for fad2 and rod1 respectively, compared to 61% for wild type), following HT exposure during late flowering and early silicle development. Silicles of fad2 plants experienced 65% less lipid peroxidation under HT and 55% less buildup of proline, signifying less stress.Discussion: The differential results of fad2 and rod1 are likely due to the role of FAD2 in membranes in all tissues, whereas ROD1 predominantly affects triacylglycerol (TAG) composition in oil-accumulating tissues including pollen. Our results indicate that decreasing PUFAs, through gene editing, can increase heat tolerance in reproductive tissues as an auxiliary benefit accompanying improved seed oil quality.
59 BASIC BIOLOGICAL SCIENCES↗
Use of Tardigrade Proteins for Enhancing Cellular Stress Tolerance on the Moon
With sights set on returning a human presence to the Moon and plans to explore even farther beyond, it is imperative we understand the impact of deep space radiation, partial gravity, and the lunar surface on biology. The need for countermeasures to protect future astronauts from the unique conditions of space is ever-pressing and an assessment of bioproduction capabilities in space is a crucial step for sustaining long-term missions. The Lunar Explorer Instrument for space biology Applications (LEIA) aims to investigate the cell’s response to the lunar environment and test strategies for enhancing cellular tolerance on the Moon, utilizing the budding yeast Saccharomyces cerevisiae as a biosensor. S. cerevisiae serves as an excellent model organism due to its widespread use as an analog for human cells, well-characterized set of genetic tools, and ability to survive desiccation. However, these cells must remain desiccated for up to one year prior to launch and the radiation-sensitive strains designed for LEIA will likely be more vulnerable to desiccation effects. In this work, we introduce tardigrade proteins known for their protective mechanisms into potential LEIA strains to test genetic strategies for improving desiccation and radiation tolerance. Tardigrades are extremotolerant animals well-known for their ability to survive in harsh environments. Notably, they can survive extreme desiccation and radiation due to the upregulation of intrinsically disordered proteins (IDPs). It has been shown that integration of these tardigrade IDPs, such as cytosolic abundant heat-soluble (CAHS) proteins, into yeast can confer increased desiccation tolerance in synergy with trehalose. Our ongoing testing and assessment of bioengineered radiation and desiccation tolerance will help us gain insight into optimizing strain design and selection for protecting LEIA’s biological payload as well as genetic engineering strategies for enhancing bioproduction potential for future missions beyond low Earth orbit.
Genetics↗
Use of Tardigrade Proteins for Enhancing Cellular Stress Tolerance on the Moon
With sights set on returning a human presence to the Moon and plans to explore even farther beyond, it is imperative we understand the impact of deep space radiation, partial gravity, and the lunar surface on biology. The need for countermeasures to protect future astronauts from the unique conditions of space is ever-pressing and an assessment of bioproduction capabilities in space is a crucial step for sustaining long-term missions. The Lunar Explorer Instrument for space biology Applications (LEIA) aims to investigate the cell’s response to the lunar environment and test strategies for enhancing cellular tolerance on the Moon, utilizing the budding yeast Saccharomyces cerevisiae as a biosensor. S. cerevisiae serves as an excellent model organism due to its widespread use as an analog for human cells, well-characterized set of genetic tools, and ability to survive desiccation. However, these cells must remain desiccated for up to one year prior to launch and the radiation-sensitive strains designed for LEIA will likely be more vulnerable to desiccation effects. In this work, we introduce tardigrade proteins known for their protective mechanisms into potential LEIA strains to test genetic strategies for improving desiccation and radiation tolerance. Tardigrades are extremotolerant animals well-known for their ability to survive in harsh environments. Notably, they can survive extreme desiccation and radiation due to the upregulation of intrinsically disordered proteins (IDPs). It has been shown that integration of these tardigrade IDPs, such as cytosolic abundant heat-soluble (CAHS) proteins, into yeast can confer increased desiccation tolerance in synergy with trehalose. Our ongoing testing and assessment of bioengineered radiation and desiccation tolerance will help us gain insight into optimizing strain design and selection for protecting LEIA’s biological payload as well as genetic engineering strategies for enhancing bioproduction potential for future missions beyond low Earth orbit.
Genetics↗
Genome of Paspalum vaginatum and the role of trehalose mediated autophagy in increasing maize biomass
Abstract A number of crop wild relatives can tolerate extreme stress to a degree outside the range observed in their domesticated relatives. However, it is unclear whether or how the molecular mechanisms employed by these species can be translated to domesticated crops. Paspalum ( Paspalum vaginatum ) is a self-incompatible and multiply stress-tolerant wild relative of maize and sorghum. Here, we describe the sequencing and pseudomolecule level assembly of a vegetatively propagated accession of P. vaginatum . Phylogenetic analysis based on 6,151 single-copy syntenic orthologues conserved in 6 related grass species places paspalum as an outgroup of the maize-sorghum clade. In parallel metabolic experiments, paspalum, but neither maize nor sorghum, exhibits a significant increase in trehalose when grown under nutrient-deficit conditions. Inducing trehalose accumulation in maize, imitating the metabolic phenotype of paspalum, results in autophagy dependent increases in biomass accumulation.
59 BASIC BIOLOGICAL SCIENCES↗
Combined environmental stresses
Tolerance levels, physiological effects, and performance degradation during simultaneous or sequential exposures to two environmental stresses, and also three or more simultaneous stresses are described. Environmental stress combinations are characterized by four descriptors: order of occurrence, duration of exposure, severity of exposure, and type of interaction. Combined stress data and facilities for combined stress study are briefly mentioned.
Murray, R. H.↗
Tomato deploys defence and growth simultaneously to resist bacterial wilt disease
Abstract Plant disease limits crop production, and host genetic resistance is a major means of control. Plant pathogenic Ralstonia causes bacterial wilt disease and is best controlled with resistant varieties. Tomato wilt resistance is multigenic, yet the mechanisms of resistance remain largely unknown. We combined metaRNAseq analysis and functional experiments to identify core Ralstonia ‐responsive genes and the corresponding biological mechanisms in wilt‐resistant and wilt‐susceptible tomatoes. While trade‐offs between growth and defence are common in plants, wilt‐resistant plants activated both defence responses and growth processes. Measurements of innate immunity and growth, including reactive oxygen species production and root system growth, respectively, validated that resistant plants executed defence‐related processes at the same time they increased root growth. In contrast, in wilt‐susceptible plants roots senesced and root surface area declined following Ralstonia inoculation. Wilt‐resistant plants repressed genes predicted to negatively regulate water stress tolerance, while susceptible plants repressed genes predicted to promote water stress tolerance. Our results suggest that wilt‐resistant plants can simultaneously promote growth and defence by investing in resources that act in both processes. Infected susceptible plants activate defences, but fail to grow and so succumb to Ralstonia , likely because they cannot tolerate the water stress induced by vascular wilt.
Meline, Valerian↗
Trade-Offs Between Growth Rate and Other Fungal Traits
If we better understand how fungal responses to global change are governed by their traits, we can improve predictions of fungal community composition and ecosystem function. Specifically, we can examine trade-offs among traits, in which the allocation of finite resources toward one trait reduces the investment in others. We hypothesized that trade-offs among fungal traits relating to rapid growth, resource capture, and stress tolerance sort fungal species into discrete life history strategies. We used the Biolog Filamentous Fungi database to calculate maximum growth rates of 37 fungal species and then compared them to their functional traits from the fun fun database. In partial support of our hypothesis, maximum growth rate displayed a negative relationship with traits related to resource capture. Moreover, maximum growth rate displayed a positive relationship with amino acid permease, forming a putative Fast Growth life history strategy. A second putative life history strategy is characterized by a positive relationship between extracellular enzymes, including cellobiohydrolase 6, cellobiohydrolase 7, crystalline cellulase AA9, and lignin peroxidase. These extracellular enzymes were negatively related to chitosanase 8, an enzyme that can break down a derivative of chitin. Chitosanase 8 displayed a positive relationship with many traits that were hypothesized to cluster separately, forming a putative Blended life history strategy characterized by certain resource capture, fast growth, and stress tolerance traits. These trait relationships complement previously explored microbial trait frameworks, such as the Competitor-Stress Tolerator-Ruderal and the Yield-Resource Acquisition-Stress Tolerance schemes.
54 ENVIRONMENTAL SCIENCES↗
Exploring Trait Trade-Offs for Fungal Decomposers in a Southern California Grassland
Fungi are important decomposers in terrestrial ecosystems, so their responses to climate change might influence carbon (C) and nitrogen (N) dynamics. We investigated whether growth and activity of fungi under drought conditions were structured by trade-offs among traits in 15 fungal isolates from a Mediterranean Southern California grassland. We inoculated fungi onto sterilized litter that was incubated at three moisture levels (4, 27, and 50% water holding capacity, WHC). For each isolate, we characterized traits that described three potential lifestyles within the newly proposed “YAS” framework: growth yield, resource acquisition, and stress tolerance. Specifically, we measured fungal hyphal length per unit litter decomposition for growth yield; the potential activities of the extracellular enzymes cellobiohydrolase (CBH), β -glucosidase (BG), β -xylosidase (BX), and N-acetyl- β - D -glucosaminidase (NAG) for resource acquisition; and ability to grow in drought vs. higher moisture levels for drought stress tolerance. Although, we had hypothesized that evolutionary and physiological trade-offs would elicit negative relationships among traits, we found no supporting evidence for this hypothesis. Across isolates, growth yield, drought stress tolerance, and extracellular enzyme activities were not significantly related to each other. Thus, it is possible that drought-induced shifts in fungal community composition may not necessarily lead to changes in fungal biomass or decomposer ability in this arid grassland.
59 BASIC BIOLOGICAL SCIENCES↗
Investigation of a Novel Salt Stress-Responsive Pathway Mediated by Arabidopsis DEAD-Box RNA Helicase Gene AtRH17 Using RNA-Seq Analysis
Previously, we reported that overexpression of AtRH17, an Arabidopsis DEAD-box RNA helicase gene, confers salt stress-tolerance via a pathway other than the well-known salt stress-responsive pathways. To decipher the salt stress-responsive pathway in AtRH17-overexpressing transgenic plants (OXs), we performed RNA-Sequencing and identified 397 differentially expressed genes between wild type (WT) and AtRH17 OXs. Among them, 286 genes were upregulated and 111 genes were downregulated in AtRH17 OXs relative to WT. Gene ontology annotation enrichment and KEGG pathway analysis showed that the 397 upregulated and downregulated genes are involved in various biological functions including secretion, signaling, detoxification, metabolic pathways, catabolic pathways, and biosynthesis of secondary metabolites as well as in stress responses. Genevestigator analysis of the upregulated genes showed that nine genes, namely, LEA4-5, GSTF6, DIN2/BGLU30, TSPO, GSTF7, LEA18, HAI1, ABR, and LTI30, were upregulated in Arabidopsis under salt, osmotic, and drought stress conditions. In particular, the expression levels of LEA4-5, TSPO, and ABR were higher in AtRH17 OXs than in WT under salt stress condition. Taken together, our results suggest that a high AtRH17 expression confers salt stress-tolerance through a novel salt stress-responsive pathway involving nine genes, other than the well-known ABA-dependent and ABA-independent pathways.
59 BASIC BIOLOGICAL SCIENCES↗
SyPro Poplar: Improving Poplar Biomass Production under Abiotic Stress Conditions: an Integrated Omics, Bioinformatics, Synthetic Biology and Genetic Engineering Approach (Final Report)
SyPro Poplar: Improving Poplar Biomass Production under Abiotic Stress Conditions: an Integrated Omics, Bioinformatics, Synthetic Biology and Genetic Engineering Approach In the SyPro Poplar project, we aimed to integrate omics, bioinformatics, synthetic biology, and genetic engineering approaches to develop transgenic poplar trees with sustained photosynthetic activity and increased biomass production under individual and the simultaneous occurrence of water deficit, increased soil salinity, and elevated temperatures. Specifically, we intended to (i) study the functions of selected stress-responsive genes at tissue and cell type-specific levels, (ii) discover novel motifs and construct stress-responsive synthetic promoters, and (iii) use these promoters to drive the expression of genes shown to confer abiotic stress tolerance in a variety of crops and develop abiotic stress-tolerant poplars in a coordinated fashion.
09 BIOMASS FUELS↗
Ecological strategies of microbes: Thinking outside the triangle
Abstract I asked whether Grime's triangle of competitive, stress tolerance and ruderal ecological strategies—which was originally developed for plants—applies to microbes. I conducted a synthesis of empirical studies that tested relationships among microbial traits presumed to define the competitive, stress tolerance and ruderal, and other ecological strategies. There was broad support for Grime's triangle. However, the ecological strategies were inconsistently linked to shifts in microbial communities under environmental changes like nitrogen and phosphorus addition, warming, drought, etc. We may be missing important ecological strategies that more closely influence microbial community composition under shifting environmental conditions. We may need to start by documenting changes in microbial communities in response to environmental conditions at fine spatiotemporal scales relevant for microbes. We can then develop empirically based ecological strategies, rather than modifying those based on plant ecology. Synthesis . Microbes appear to sort into similar ecological strategies as plants. However, these microbial ecological strategies do not consistently predict how community composition will shift under environmental change. By starting ‘from the ground up’, we may be able to delineate ecological strategies more relevant for microbes.
54 ENVIRONMENTAL SCIENCES↗
Successional adaptive strategies revealed by correlating arbuscular mycorrhizal fungal abundance with host plant gene expression
The shifts in adaptive strategies revealed by ecological succession and the mechanisms that facilitate these shifts are fundamental to ecology. These adaptive strategies could be particularly important in communities of arbuscular mycorrhizal fungi (AMF) mutualistic with sorghum, where strong AMF succession replaces initially ruderal species with competitive ones and where the strongest plant response to drought is to manage these AMF. Although most studies of agriculturally important fungi focus on parasites, the mutualistic symbionts, AMF, constitute a research system of human-associated fungi whose relative simplicity and synchrony are conducive to experimental ecology. First, we hypothesize that, when irrigation is stopped to mimic drought, competitive AMF species should be replaced by AMF species tolerant to drought stress. We then, for the first time, correlate AMF abundance and host plant transcription to test two novel hypotheses about the mechanisms behind the shift from ruderal to competitive AMF. Surprisingly, despite imposing drought stress, we found no stress-tolerant AMF, probably due to our agricultural system having been irrigated for nearly six decades. Remarkably, we found strong and differential correlation between the successional shift from ruderal to competitive AMF and sorghum genes whose products (i) produce and release strigolactone signals, (ii) perceive mycorrhizal-lipochitinoligosaccharide (Myc-LCO) signals, (iii) provide plant lipid and sugar to AMF, and (iv) import minerals and water provided by AMF. These novel insights frame new hypotheses about AMF adaptive evolution and suggest a rationale for selecting AMF to reduce inputs and maximize yields in commercial agriculture.
59 BASIC BIOLOGICAL SCIENCES↗