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Bacillus pumilus improved drought tolerance in Glycyrrhiza uralensis G5 seedlings through enhancing primary and secondary metabolisms
Abstract It has been reported that drought stress adversely affects the growth and yield of Glycyrrhiza uralensis, Chinese liquorice, in agricultural production. Bacillus pumilus , an important plant growth‐promoting bacterium, play a significant role in improving plant tolerance to abiotic stress. However, the role of Bacillus pumilus G5 in resisting drought stress is largely unknown. In the present study, we found that drought stress significantly inhibited the growth and reduced the biomass of G. uralensis seedlings by restraining C‐ and N‐metabolism, while this could be effectively reversed by B. pumilus G5 inoculation. Specifically, B. pumilus G5 significantly increased the content of primary metabolites such as soluble sugar, soluble protein, and free amino acids by regulating the C and N metabolic processes in G. uralensis seedlings. Moreover, B. pumilus G5 increased the content of glycyrrhizic acid, one of the important secondary metabolites, likely mediated through the increased content of primary metabolites and by recovering the expression of three key enzymes, HMGR, SQS, and β‐AS, in the biosynthesis of glycyrrhizic acid. Interestingly, the regulating effect of B. pumilus G5 inoculation on promoting the accumulation of glycyrrhizic acid and increasing the expression of synthesis‐related genes is spatially selective. In summary, our findings suggest that B. pumilus G5 could alleviate adverse effects induced by drought stress on the growth of G. uralensis seedlings by regulating C‐ and N‐metabolisms that further triggered the accumulation of secondary metabolites, and this finally improved the drought tolerance of cultivated G. uralensis seedlings.
Symbiosis Under Stress: How Arbuscular Mycorrhizal Fungi and Sorghum Metabolism Shift Under Drought
As drought becomes more prevalent across the globe, causing billions of dollars in agricultural loss, the need to maintain crop health and productivity grows increasingly important. Out of the most important cereal crops, sorghum shows the greatest drought tolerance, and plant-microbiome interactions at the root region play a crucial role in this. A key microbial player is arbuscular mycorrhizal fungi (AMF), which deliver water and nutrients to plants in exchange for nutrients they cannot produce. Using sorghum as a model plant, we combine mass-spectrometry based proteomics and metabolomics to examine metabolic interactions between sorghum roots and AMF under drought stress. With AMF, sorghum downregulated lipid-related biological processes involving fatty acid biosynthesis and metabolism. Similarly, amino acid biosynthesis pathways were also suppressed; however, metabolite analysis revealed increased amino acid abundance related to the phenylpropanoid pathway. AMF also enhanced the upregulation of drought-protective osmolytes, such as mannitol and sorbitol, suggesting their key role in mediating sorghum’s response to drought stress. AMF hyphal biomass also had an increased abundance of key osmoprotectant amino acids, indicating similar mechanisms of drought tolerance between sorghum and AMF. Metabolomic evidence also suggested that carbohydrate exchange between sorghum and AMF shifted under stress, indicating an altered exudation pattern likely driven by drought response. Our results demonstrate the molecular mechanisms through which AMF modulate sorghum metabolism under drought conditions, highlighting their promising role in improving crop resilience. By identifying the molecular targets that can improve drought tolerance, we can begin engineering drought resistant agricultural biosystems.
Cell Wall Compositions of Sorghum bicolor Leaves and Roots Remain Relatively Constant Under Drought Conditions
Renewable fuels are needed to replace fossil fuels in the immediate future. Lignocellulosic bioenergy crops provide a renewable alternative that sequesters atmospheric carbon. To prevent displacement of food crops, it would be advantageous to grow biofuel crops on marginal lands. These lands will likely face more frequent and extreme drought conditions than conventional agricultural land, so it is crucial to see how proposed bioenergy crops fare under these conditions and how that may affect lignocellulosic biomass composition and saccharification properties. We found that while drought impacts the plant cell wall of Sorghum bicolor differently according to tissue and timing of drought induction, drought-induced cell wall compositional modifications are relatively minor and produce no negative effect on biomass conversion. This contrasts with the cell wall-related transcriptome, which had a varied range of highly variable genes (HVGs) within four cell wall-related GO categories, depending on the tissues surveyed and time of drought induction. Further, many HVGs had expression changes in which putative impacts were not seen in the physical cell wall or which were in opposition to their putative impacts. Interestingly, most pre-flowering drought-induced cell wall changes occurred in the leaf, with matrix and lignin compositional changes that did not persist after recovery from drought. Most measurable physical post-flowering cell wall changes occurred in the root, affecting mainly polysaccharide composition and cross-linking. This study couples transcriptomics to cell wall chemical analyses of a C4 grass experiencing progressive and differing drought stresses in the field. As such, we can analyze the cell wall-specific response to agriculturally relevant drought stresses on the transcriptomic level and see whether those changes translate to compositional or biomass conversion differences. Our results bolster the conclusion that drought stress does not substantially affect the cell wall composition of specific aerial and subterranean biomass nor impede enzymatic hydrolysis of leaf biomass, a positive result for biorefinery processes. Coupled with previously reported results on the root microbiome and rhizosphere and whole transcriptome analyses of this study, we can formulate and test hypotheses on individual gene candidates’ function in mediating drought stress in the grass cell wall, as demonstrated in sorghum.
Divergences in hydraulic conductance and anatomical traits of stems and leaves in three temperate tree species coping with drought, N addition and their interactions
Drought and N addition have been shown to affect tree hydraulic traits, but few studies have been made on their interactions across species with different wood types or leaf forms. We examined the responses of hydraulic conductance and xylem anatomical traits of Quercus mongolica (ring-porous with simple-leaves), Fraxinus mandshurica (ring-porous with compound-leaves), and Tilia amurensis (diffuse-porous with simple-leaves) to drought, N addition and their interactions. Drought stress decreased current-year xylem-specific conductivity in stems (Ksx) and leaf hydraulic conductance (Kleaf), but N addition affected Ksx and Kleaf differently among species and watering regimes. These divergent effects were associated with different responses of anatomical traits and leaf forms. Higher mean vessel diameter in stems (Dstem) and lower vessel density in leaves (VDvein) were observed with N addition. The three-way interactive effects of drought, N addition and tree species were significant for most values of anatomical traits. These results were also reflected in large differences in vessel diameter and density among species with different wood types or leaf forms. The two-way interactive effects of drought and N addition were significant on Kleaf and predawn water potential (?pd), but not Ksx, indicating that leaves were more sensitive than stems to a combination of drought stress and N addition. Our results provide mechanistic insight into the variable responses of xylem water transport to the interactions of drought and N availability.
Developmentally-specific physiological and metabolic responses support drought resilience in switchgrass and constrains biofuel yield
Switchgrass (Panicum virgatum) is a promising bioenergy crop due in part to its resilience to drought stress. However, the significance of drought timing remains poorly understood, both from a plant biology perspective and its impact on downstream biofuel production. This study determines the developmental stage-specific physiological and metabolic responses of switchgrass to drought stress and its implications for biofuel production using a custom-built programmable irrigation system. Vegetative, flowering, and senescence-stage drought significantly reduced carbon dioxide assimilation, and stomatal conductance without affecting biomass yield. Metabolic profiling revealed significant accumulation of glucose, fructose, quinic acid, shikimate and GABA during vegetative-stage drought, while flowering and senescence stages exhibited limited metabolic changes. Similarly, specialized metabolites also displayed distinct developmental patterns, with vegetative-stage drought driving the most pronounced metabolic alterations. Thermochemically-treated and hydrolyzed switchgrass biomass from vegetative-stage drought showed elevated lignocellulose-derived compounds and saponins with the latter most positively correlating with fermentation lag times. Conversely, senescence-stage drought enhanced ethanol yields while lowering saponin levels in the hydrolysates. While vegetative-stage drought enhanced physiological resilience, it compromises downstream biofuel production by introducing fermentation inhibitors, particularly saponins.
Transcriptional Regulation of Drought Response in Arabidopsis and Woody Plants
Within the context of global warming, long-living plants such as perennial woody species endure adverse conditions. Among all of the abiotic stresses, drought stress is one of the most detrimental stresses that inhibit plant growth and productivity. Plants have evolved multiple mechanisms to respond to drought stress, among which transcriptional regulation is one of the key mechanisms. In this review, we summarize recent progress on the regulation of drought response by transcription factor (TF) families, which include abscisic acid (ABA)-dependent ABA-responsive element/ABRE-binding factors (ABRE/ABF), WRKY, and Nuclear Factor Y families, as well as ABA-independent AP2/ERF and NAC families, in the model plant Arabidopsis. We also review what is known in woody species, particularly Populus, due to its importance and relevance in economic and ecological processes. We discuss opportunities for a deeper understanding of drought response in woody plants with the development of high-throughput omics analyses and advanced genome editing techniques.
Comparative genomics analysis of drought response between obligate CAM and C 3 photosynthesis plants
Crassulacean acid metabolism (CAM) plants exhibit elevated drought and heat tolerance compared to C 3 and C 4 plants through an inverted pattern of day/night stomatal closure and opening for CO 2 assimilation. However, the molecular responses to water-deficit conditions remain unclear in obligate CAM species. In this study, we presented genome-wide transcription sequencing analysis using leaf samples of an obligate CAM species Kalanchoë fedtschenkoi under moderate and severe drought treatments at two-time points of dawn (2-h before the start of light period) and dusk (2-h before the dark period). Differentially expressed genes were identified in response to environmental drought stress and a whole genome wide co-expression network was created as well. We found that the expression of CAM-related genes was not regulated by drought stimuli in K. fedtschenkoi. Our comparative analysis revealed that CAM species (K. fedtschenkoi) and C 3 species (Arabidopsis thaliana, Populus deltoides ‘WV94’) share some common transcriptional changes in genes involved in multiple biological processes in response to drought stress, including ABA signaling and biosynthesis of secondary metabolites.
Co-occurrence networks reveal more complexity than community composition in resistance and resilience of microbial communities
Plant response to drought stress involves fungi and bacteria that live on and in plants and in the rhizosphere, yet the stability of these myco- and micro-biomes remains poorly understood. We investigate the resistance and resilience of fungi and bacteria to drought in an agricultural system using both community composition and microbial associations. Here we show that tests of the fundamental hypotheses that fungi, as compared to bacteria, are (i) more resistant to drought stress but (ii) less resilient when rewetting relieves the stress, found robust support at the level of community composition. Results were more complex using all-correlations and co-occurrence networks. In general, drought disrupts microbial networks based on significant positive correlations among bacteria, among fungi, and between bacteria and fungi. Surprisingly, co-occurrence networks among functional guilds of rhizosphere fungi and leaf bacteria were strengthened by drought, and the same was seen for networks involving arbuscular mycorrhizal fungi in the rhizosphere. We also found support for the stress gradient hypothesis because drought increased the relative frequency of positive correlations.
Genome-resolved metagenomics reveals role of iron metabolism in drought-induced rhizosphere microbiome dynamics
Recent studies have demonstrated that drought leads to dramatic, highly conserved shifts in the root microbiome. At present, the molecular mechanisms underlying these responses remain largely uncharacterized. Here we employ genome-resolved metagenomics and comparative genomics to demonstrate that carbohydrate and secondary metabolite transport functionalities are overrepresented within drought-enriched taxa. These data also reveal that bacterial iron transport and metabolism functionality is highly correlated with drought enrichment. Using time-series root RNA-Seq data, we demonstrate that iron homeostasis within the root is impacted by drought stress, and that loss of a plant phytosiderophore iron transporter impacts microbial community composition, leading to significant increases in the drought-enriched lineage, Actinobacteria. Finally, we show that exogenous application of iron disrupts the drought-induced enrichment of Actinobacteria, as well as their improvement in host phenotype during drought stress. Collectively, our findings implicate iron metabolism in the root microbiome’s response to drought and may inform efforts to improve plant drought tolerance to increase food security.
Cell wall ester modifications and volatile emission signatures of plant response to abiotic stress
Abstract Growth suppression and defence signalling are simultaneous strategies that plants invoke to respond to abiotic stress. Here, we show that the drought stress response of poplar trees ( Populus trichocarpa ) is initiated by a suppression in cell wall derived methanol (MeOH) emissions and activation of acetic acid (AA) fermentation defences. Temperature sensitive emissions dominated by MeOH (AA/MeOH <30%) were observed from physiologically active leaves, branches, detached stems, leaf cell wall isolations and whole ecosystems. In contrast, drought treatment resulted in a suppression of MeOH emissions and strong enhancement in AA emissions together with volatiles acetaldehyde, ethanol, and acetone. These drought‐induced changes coincided with a reduction in stomatal conductance, photosynthesis, transpiration, and leaf water potential. The strong enhancement in AA/MeOH emission ratios during drought (400%–3500%) was associated with an increase in acetate content of whole leaf cell walls, which became significantly 13 C 2 ‐labelled following the delivery of 13 C 2 ‐acetate via the transpiration stream. The results are consistent with both enzymatic and nonenzymatic MeOH and AA production at high temperature in hydrated tissues associated with accelerated primary cell wall growth processes, which are downregulated during drought. While the metabolic source(s) require further investigation, the observations are consistent with drought‐induced activation of aerobic fermentation driving high rates of foliar AA emissions and enhancements in leaf cell wall O ‐acetylation. We suggest that atmospheric AA/MeOH emission ratios could be useful as a highly sensitive signal in studies investigating environmental and biological factors influencing growth‐defence trade‐offs in plants and ecosystems.
Characterizing proteases in sorghum drought response using activity-based protein profiling (ABPP)
Sorghum (Sorghum bicolor) is a C4 grass species that is a major food and bioenergy crop grown worldwide. Understanding the response of sorghum to climate change is therefore critical to future sustainable food and bioenergy production. Major advancements in genome sequencing have provided a wealth of genomic data for DOE-relevant plant species, but the functions of many of these genes remain unknown. Genotype to phenotype characterization of sorghum and other bioenergy crops is needed to identify key genes responsible for plant resilience to drought and other environmental stressors. Proteases, which modulate protein degradation and cellular signaling processes by catalyzing the hydrolysis of proteins into smaller proteins, peptides, or amino acids, are highly implicated in plant drought stress responses. We developed a method for using activity-based protein profiling (ABPP) of selected protease activities (serine and cysteine proteases) to characterize drought stress responses in sorghum roots and leaves. Analysis of these ABPP data identified several known serine and cysteine proteases as well as uncharacterized sorghum proteins which may have protease activities that can be explored in future validation studies.
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.
Springtime Drought Shifts Carbon Partitioning of Recent Photosynthates in 10-Year Old Picea mariana Trees, Causing Restricted Canopy Development
Springtime bud-break and shoot development induces substantial carbon (C) costs in trees. Drought stress during shoot development can impede C uptake and translocation. This is therefore a channel through which water shortage can lead to restricted shoot expansion and physiological capacity, which in turn may impact annual canopy C uptake. We studied effects of drought and re-hydration on early season shoot development, C uptake and partitioning in five individual 10-year old Picea mariana [black spruce] trees to identify and quantify dynamics of key morphological/physiological processes. Trees were subjected to one of two treatments: (i) well-watered control or (ii) drought and rehydration. We monitored changes in morphological [shoot volume, leaf mass area (LMA)], biochemical [osmolality, non-structural carbohydrates (NSC)] and physiological [rates of respiration (R d ) and light-saturated photosynthesis ( A sat )] processes during shoot development. Further, to study functional compartmentalization and use of new assimilates, we 13 C-pulse labeled shoots at multiple development stages, and measured isotopic signatures of leaf respiration, NSC pools and structural biomass. Shoot water potential dropped to a minimum of −2.5 MPa in shoots on the droughted trees. Development of the photosynthetic apparatus was delayed, as shoots on well-watered trees broke-even 14 days prior to shoots from trees exposed to water deficit. R d decreased with shoot maturation as growth respiration declined, and was lower in shoots exposed to drought. We found that shoot development was delayed by drought, and while rehydration resulted in recovery of A sat to similar levels as shoots on the well-watered trees, shoot volume remained lower. Water deficit during shoot expansion resulted in longer, yet more compact (i.e., with greater LMA) shoots with greater needle osmolality. The 12 C: 13 C isotopic patterns indicated that internal C partitioning and use was dependent on foliar developmental and hydration status. Shoots on drought-stressed trees prioritized allocating newly fixed C to respiration over structural components. In conclusion, temporary water deficit delayed new shoot development and resulted in greater LMA in black spruce. Since evergreen species such as black spruce retain active foliage for multiple years, impacts of early season drought on net primary productivity could be carried forward into subsequent years.
A KBase Case Study on Genome-wide Transcriptomics and Plant Primary Metabolism in Sorghum
A better understanding of the genetic and metabolic mechanisms that confer stress resistance and tolerance in plants is key to engineering new crops through advanced breeding technologies. This requires a systems biology approach that builds on a genome-wide understanding of the regulation of gene expression, plant metabolism, physiology and growth. In this study, we examine the response to drought stress in Sorghum, as we leverage the tools for transcriptomics and plant metabolic modeling we have implemented at the U.S. Department of Energy Systems Biology Knowledgebase (KBase). KBase enables researchers worldwide to collaborate and advance research by allowing them to upload private or public data into the KBase Narrative Interface, empowering them to analyze this data using a rich, extensible array of computational and data-analytics tools, and allows them to securely share scientific workflows and conclusions. We demonstrate how to use the current RNA-seq tools in KBase, applicable to both plants and microbes, to assemble and quantify long transcripts and identify differentially expressed genes effectively. More specifically, we demonstrate the utility of the platform by identifying key genes that are differentially expressed during drought-stress in Sorghum bicolor, which is an important sustainable production crop plant. We then show how to use KBase tools to predict the membership of genes in metabolic pathways and examine expression data in the context of metabolic subsystems. We demonstrate the power of the platform by making the data analysis and interpretation available to the biologists in the reproducible, re-usable, point-and-click format of a KBase Narrative thus promoting FAIR (Findable, Accessible, Interoperable and Reusable) guiding principles for scientific data management and stewardship.
Data from: "Warming and the dependence of limber pine (Pinus flexilis) establishment on summer soil moisture within and above its current elevation range"
This data package contains data that were used for analysis in “Warming and the dependence of limber pine (Pinus flexilis) establishment on summer soil moisture within and above its current elevation range”, by Moyes et al. 2013. All data collection and field research were completed on Niwot Ridge, Colorado, USA.This data package contains nine comma-separated-values (.csv) files, one text (.txt) file, and two zipped seedling folders that were used for leaf area analysis. One zipped folder contains 468 .jpg photographs of seedlings, and the second contains 468 corresponding Image J-processed .jpg images that include silhouette leaf area values. .csv and .txt files can be opened using any compatible simple text-editor software such as TextEdit (Mac) and Notepad (Windows); .csv’s can also be opened using R and Microsoft Excel. Image files can be opened using Preview (Mac) and Photos (Windows). In addition, there are a total of 31 Microsoft Excel files: three .xlsx files, and 28 raw Li-Cor output .xls files. This data user’s guide is available in .pdf format, and can be opened using Adobe Acrobat Reader, or any other compatible file viewing software. Geospatial data showing field site locations are also included in the archive for use and reference. There are two geospatial formats in this archive: ESRI shapefiles (.shp) and keyhole markup-language (.kml) files. Both file types contain bounding box information, with the former being polygons, and the latter containing corner coordinates for each site. ESRI shapefiles can be opened using any geospatial software compatible with the file type (such as ESRI’s ArcGIS suite and QGIS), and .kml files are compatible with Google Earth and Google Maps.--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Continued changes in climate are projected to alter the geographic distributions of plant species, in part by affecting where individuals can establish from seed. We tested the hypothesis that warming promotes uphill redistribution of subalpine tree populations by reducing cold limitation at high elevation and enhancing drought stress at low elevation. We seeded limber pine (Pinus flexilis) into plots with combinations of infrared heating and water addition treatments, at sites positioned in lower subalpine forest, the treeline ecotone, and alpine tundra. In 2010, first-year seedlings were assessed for physiological performance and survival over the snow-free growing season. Seedlings emerged in midsummer, about 5–8 weeks after snowmelt. Low temperature was not observed to limit seedling photosynthesis or respiration between emergence and October, and thus experimental warming did not appear to reduce cold limitation at high elevation. Instead, gas exchange and water potential from all sites indicated a prevailing effect of summer moisture stress on photosynthesis and carbon balance. Infrared heaters raised soil growing degree days (base 5 °C, p < 0.001) and August–September mean soil temperature (p < 0.001). Despite marked differences in vegetation cover and meteorological conditions across sites, volumetric soil moisture content (θ) at 5–10 cm below 0.16 and 0.08 m^3 m^(−3) consistently corresponded with moderate and severe indications of drought stress in midday stem water potential, stomatal conductance, photosynthesis, and respiration. Seedling survival was greater in watered plots than in heated plots (p = 0.01), and negatively related to soil growing degree days and duration of exposure to θ < 0.08 m^3 m^(−3) in a stepwise linear regression model (p < 0.0001). We concluded that seasonal moisture stress and high soil surface temperature imposed a strong limitation to limber pine seedling establishment across a broad elevation gradient, including at treeline, and that these limitations are likely to be enhanced by further climate warming.--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Maintenance log:Amended file name error in Data User's Guide May 19 2022
Potential for functional divergence in ectomycorrhizal fungal communities across a precipitation gradient
Abstract Functional traits influence the assembly of microbial communities, but identifying these traits in the environment has remained challenging. We studied ectomycorrhizal fungal (EMF) communities inhabiting Populus trichocarpa roots distributed across a precipitation gradient in the Pacific Northwest, USA. We profiled these communities using taxonomic (meta-barcoding) and functional (metagenomic) approaches. We hypothesized that genes involved in fungal drought-stress tolerance and fungal mediated plant water uptake would be most abundant in drier soils. We were unable to detect support for this hypothesis; instead, the abundance of genes involved in melanin synthesis, hydrophobins, aquaporins, trehalose-synthases, and other gene families exhibited no significant shifts across the gradient. Finally, we studied variation in sequence homology for certain genes, finding that fungal communities in dry soils are composed of distinct aquaporin and hydrophobin gene sequences. Altogether, our results suggest that while EMF communities exhibit significant compositional shifts across this gradient, coupled functional turnover, at least as inferred using community metagenomics is limited. Accordingly, the consequences of these distinct EMF communities on plant water uptake remain critically unknown, and future studies targeting the expression of genes involved in drought stress tolerance are required.
Storms Are an Important Driver of Change in Tropical Forests
Tropical forest dynamics and composition have changed over recent decades, but the proximate drivers of these changes remain unclear. Investigations into these trends have focused on increasing drought stress, CO 2 , temperature, and fires, whereas convective storms are generally overlooked. We argue that existing literature provides clear support for the importance of storms as drivers of forest change. We reanalyze the largest plot-based study of tropical forest carbon dynamics to show that lightning frequency—an indicator of storm activity—strongly predicts forest carbon storage and residence time, and its inclusion improves model fit and weakens evidence for the effects of high temperatures. Convective storm activity has increased 5%–25% per decade over the past half century. Extrapolating from historic trends, we estimate that storms likely contribute ca. 50% of the reported increases in biomass mortality across Amazonia, with all realistic combinations of assumptions indicating a possible range of 12%–118%. Spatial variation in storm activity shows weak relationships with drought, demonstrating that forests can experience high drought stress, high storm activity, or both. Accordingly, we hypothesise that convective storms are among the most important drivers of tropical forest change, and as such, they require significant research investment to avoid misguiding science, policy, and management.