Climate change challenges, plant science solutions.

Climate change is a defining challenge of the 21st century, and this decade is a critical time for action to mitigate the worst effects on human populations and ecosystems. Plant science can play an important role in developing crops with enhanced resilience to harsh conditions (e.g. heat, drought, salt stress, flooding, disease outbreaks) and engineering efficient carbon-capturing and carbon-sequestering plants. Here, we present examples of research being conducted in these areas and discuss challenges and open questions as a call to action for the plant science community.

Time of day and genotype sensitivity adjust molecular responses to temperature stress in sorghum.

Sorghum is one of the four major C4 crops that are considered to be tolerant to environmental extremes. Sorghum shows distinct growth responses to temperature stress depending on the sensitivity of the genetic background. About half of the transcripts in sorghum exhibit diurnal rhythmic expressions emphasizing significant coordination with the environment. However, an understanding of how molecular dynamics contribute to genotype-specific stress responses in the context of the time of day is not known. We examined whether temperature stress and the time of day impact the gene expression dynamics in thermo-sensitive and thermo-tolerant sorghum genotypes. We found that time of day is highly influencing the temperature stress responses, which can be explained by the rhythmic expression of most thermo-responsive genes. This effect is more pronounced in thermo-tolerant genotypes, suggesting a stronger regulation of gene expression by the time of day and/or by the circadian clock. Genotypic differences were mostly observed on average gene expression levels, which may be responsible for contrasting sensitivities to temperature stress in tolerant versus susceptible sorghum varieties. We also identified groups of genes altered by temperature stress in a time-of-day and genotype-specific manner. These include transcriptional regulators and several members of the Ca2+ -binding EF-hand protein family. We hypothesize that expression variation of these genes between genotypes along with time-of-day independent regulation may contribute to genotype-specific fine-tuning of thermo-responsive pathways. These findings offer a new opportunity to selectively target specific genes in efforts to develop climate-resilient crops based on their time-of-day and genotype variation responses to temperature stress.

Warm nights disrupt transcriptome rhythms in field-grown rice panicles.

In rice, a small increase in nighttime temperature reduces grain yield and quality. How warm nighttime temperatures (WNT) produce these detrimental effects is not well understood, especially in field conditions where the typical day-to-night temperature fluctuation exceeds the mild increase in nighttime temperature. We observed genome-wide disruption of gene expression timing during the reproductive phase in field-grown rice panicles acclimated to 2 to 3 °C WNT. Transcripts previously identified as rhythmically expressed with a 24-h period and circadian-regulated transcripts were more sensitive to WNT than were nonrhythmic transcripts. The system-wide perturbations in transcript levels suggest that WNT disrupt the tight temporal coordination between internal molecular events and the environment, resulting in reduced productivity. We identified transcriptional regulators whose predicted targets are enriched for sensitivity to WNT. The affected transcripts and candidate regulators identified through our network analysis explain molecular mechanisms driving sensitivity to WNT and identify candidates that can be targeted to enhance tolerance to WNT.

Carbon dioxide responsiveness mitigates rice yield loss under high night temperature.

Increasing night-time temperatures are a major threat to sustaining global rice (Oryza sativa L.) production. A simultaneous increase in [CO2] will lead to an inevitable interaction between elevated [CO2] (e[CO2]) and high night temperature (HNT) under current and future climates. Here, we conducted field experiments to identify [CO2] responsiveness from a diverse indica panel comprising 194 genotypes under different planting geometries in 2016. Twenty-three different genotypes were tested under different planting geometries and e[CO2] using a free-air [CO2] enrichment facility in 2017. The most promising genotypes and positive and negative controls were tested under HNT and e[CO2] + HNT in 2018. [CO2] responsiveness, measured as a composite response index on different yield components, grain yield, and photosynthesis, revealed a strong relationship (R2 = 0.71) between low planting density and e[CO2]. The most promising genotypes revealed significantly lower (P < 0.001) impact of HNT in high [CO2] responsive (HCR) genotypes compared to the least [CO2] responsive genotype. [CO2] responsiveness was the major driver determining grain yield and related components in HCR genotypes with a negligible yield loss under HNT. A systematic investigation highlighted that active selection and breeding for [CO2] responsiveness can lead to maintained carbon balance and compensate for HNT-induced yield losses in rice and potentially other C3 crops under current and future warmer climates.

Sustaining productivity gains in the face of climate change: A research agenda for US wheat.

Wheat is a globally important crop and one of the "big three" US field crops. But unlike the other two (maize and soybean), in the United States its development is commercially unattractive, and so its breeding takes place primarily in public universities. Troublingly, the incentive structures within these universities may be hindering genetic improvement just as climate change is complicating breeding efforts. "Business as usual" in the US public wheat-breeding infrastructure may not sustain productivity increases. To address this concern, we held a multidisciplinary conference in which researchers from 12 US (public) universities and one European university shared the current state of knowledge in their disciplines, aired concerns, and proposed initiatives that could facilitate maintaining genetic improvement of wheat in the face of climate change. We discovered that climate-change-oriented breeding efforts are currently considered too risky and/or costly for most university wheat breeders to undertake, leading to a relative lack of breeding efforts that focus on abiotic stressors such as drought and heat. We hypothesize that this risk/cost burden can be reduced through the development of appropriate germplasm, relevant screening mechanisms, consistent germplasm characterization, and innovative models predicting the performance of germplasm under projected future climate conditions. However, doing so will require coordinated, longer-term, inter-regional efforts to generate phenotype data, and the modification of incentive structures to consistently reward such efforts.

Genetic control of source-sink relationships in grain sorghum.

MAIN CONCLUSION: QTL hotspots identified for selected source-sink-related traits provide the opportunity for pyramiding favorable alleles for improving sorghum productivity under diverse environments. A sorghum bi-parental mapping population was evaluated under six different environments at Hays and Manhattan, Kansas, USA, in 2016 and 2017, to identify genomic regions controlling source-sink relationships. The population consisted of 210 recombinant inbred lines developed from US elite post-flowering drought susceptible (RTx430) and a known post-flowering drought tolerant cultivar (SC35). Selected physiological traits related to source (effective quantum yield of photosystem II and chlorophyll index), sink (grain yield per panicle) and panicle neck diameter were recorded during grain filling. The results showed strong phenotypic and genotypic association between panicle neck diameter and grain yield per panicle during mid-grain filling and at maturity. Multiple QTL model revealed 5-12 including 2-5 major QTL for each trait. Among them 3, 7 and 8 QTL for quantum yield, panicle neck diameter and chlorophyll index, respectively, have not been identified previously in sorghum. Phenotypic variation explained by QTL identified across target traits ranged between 5.5 and 25.4%. Panicle neck diameter and grain yield per panicle were positively associated, indicating the possibility of targeting common co-localized QTL to improve both traits simultaneously through marker-assisted selection. Three major QTL hotspots, controlling multiple traits were identified on chromosome 1 (52.23-61.18 Mb), 2 (2.52-11.43 Mb) and 3 (1.32-3.95 Mb). The identified genomic regions and underlying candidate genes can be utilized in pyramiding favorable alleles for improving source-sink relationships in sorghum under diverse environments.

Classical phenotyping and deep learning concur on genetic control of stomatal density and area in sorghum.

Stomatal density (SD) and stomatal complex area (SCA) are important traits that regulate gas exchange and abiotic stress response in plants. Despite sorghum (Sorghum bicolor) adaptation to arid conditions, the genetic potential of stomata-related traits remains unexplored due to challenges in available phenotyping methods. Hence, identifying loci that control stomatal traits is fundamental to designing strategies to breed sorghum with optimized stomatal regulation. We implemented both classical and deep learning methods to characterize genetic diversity in 311 grain sorghum accessions for stomatal traits at two different field environments. Nearly 12,000 images collected from abaxial (Ab) and adaxial (Ad) leaf surfaces revealed substantial variation in stomatal traits. Our study demonstrated significant accuracy between manual and deep learning methods in predicting SD and SCA. In sorghum, SD was 32%-39% greater on the Ab versus the Ad surface, while SCA on the Ab surface was 2%-5% smaller than on the Ad surface. Genome-Wide Association Study identified 71 genetic loci (38 were environment-specific) with significant genotype to phenotype associations for stomatal traits. Putative causal genes underlying the phenotypic variation were identified. Accessions with similar SCA but carrying contrasting haplotypes for SD were tested for stomatal conductance and carbon assimilation under field conditions. Our findings provide a foundation for further studies on the genetic and molecular mechanisms controlling stomata patterning and regulation in sorghum. An integrated physiological, deep learning, and genomic approach allowed us to unravel the genetic control of natural variation in stomata traits in sorghum, which can be applied to other plants.

Genetic and molecular mechanisms underlying root architecture and function under heat stress-A hidden story.

Heat stress events are resulting in a significant negative impact on global food production. The dynamics of cellular, molecular and physiological homoeostasis in aboveground parts under heat stress are extensively deciphered. However, root responses to higher soil/air temperature or stress signalling from shoot to root are limited. Therefore, this review presents a holistic view of root physio-morphological and molecular responses to adapt under hotter environments. Heat stress reprogrammes root cellular machinery, including crosstalk between genes, phytohormones, reactive oxygen species (ROS) and antioxidants. Spatio-temporal regulation and long-distance transport of phytohormones, such as auxin, cytokinin and abscisic acid (ABA) determine the root growth and development under heat stress. ABA cardinally integrates a signalling pathway involving heat shock factors, heat shock proteins and ROS to govern heat stress responses. Additionally, epigenetic modifications by transposable elements, DNA methylation and acetylation also regulate root growth under heat stress. Exogenous application of chemical compounds or biological agents such as ascorbic acid, metal ion chelators, fungi and bacteria can alleviate heat stress-induced reduction in root biomass. Future research should focus on the systemic effect of heat stress from shoot to root with more detailed investigations to decipher the molecular cues underlying the roots architecture and function.

Metabolomic and transcriptomic analyses reveal that sucrose synthase regulates maize pollen viability under heat and drought stress.

Maize pollen is highly sensitive to heat and drought, but few studies have investigated the combined effects of heat and drought on pollen viability. In this study, pollen's structural and physiological characteristics were determined after heat, drought, and combined stressors. Furthermore, integrated metabolomic and transcriptomic analyses of maize pollen were conducted to identify potential mechanisms of stress responses. Tassel growth and spikelet development were considerably suppressed, pollen viability was negatively impacted, and pollen starch granules were depleted during anthesis under stress. The inhibitory effects were more significant due to combined stresses than to heat or drought individually. The metabolic analysis identified 71 important metabolites in the combined stress compared to the other treatments, including sugars and their derivatives related to pollen viability. Transcriptomics also revealed that carbohydrate metabolism was significantly altered under stress. Moreover, a comprehensive metabolome-transcriptome analysis identified a central mechanism in the biosynthesis of UDP-glucose involved in reducing the activity of sucrose synthase SH-1 (shrunken 1) and sus1 (sucrose synthase 1) that suppressed sucrose transfer to UDP-glucose, leading to pollen viability exhaustion under stress. In conclusion, the lower pollen viability after heat and drought stress was associated with poor sucrose synthase activity due to the stress treatments.

Analysis of differential gene expression and alternative splicing is significantly influenced by choice of reference genome.

RNA-seq analysis has enabled the evaluation of transcriptional changes in many species including nonmodel organisms. However, in most species only a single reference genome is available and RNA-seq reads from highly divergent varieties are typically aligned to this reference. Here, we quantify the impacts of the choice of mapping genome in rice where three high-quality reference genomes are available. We aligned RNA-seq data from a popular productive rice variety to three different reference genomes and found that the identification of differentially expressed genes differed depending on which reference genome was used for mapping. Furthermore, the ability to detect differentially used transcript isoforms was profoundly affected by the choice of reference genome: Only 30% of the differentially used splicing features were detected when reads were mapped to the more commonly used, but more distantly related reference genome. This demonstrated that gene expression and splicing analysis varies considerably depending on the mapping reference genome, and that analysis of individuals that are distantly related to an available reference genome may be improved by acquisition of new genomic reference material. We observed that these differences in transcriptome analysis are, in part, due to the presence of single nucleotide polymorphisms between the sequenced individual and each respective reference genome, as well as annotation differences between the reference genomes that exist even between syntenic orthologs. We conclude that even between two closely related genomes of similar quality, using the reference genome that is most closely related to the species being sampled significantly improves transcriptome analysis.

Plant heat stress: Concepts directing future research.

Predicted increases in future global temperatures require us to better understand the dimensions of heat stress experienced by plants. Here we highlight four key areas for improving our approach towards understanding plant heat stress responses. First, although the term 'heat stress' is broadly used, that term encompasses heat shock, heat wave and warming experiments, which vary in the duration and magnitude of temperature increase imposed. A greater integration of results and tools across these approaches is needed to better understand how heat stress associated with global warming will affect plants. Secondly, there is a growing need to associate plant responses to tissue temperatures. We review how plant energy budgets determine tissue temperature and discuss the implications of using leaf versus air temperature for heat stress studies. Third, we need to better understand how heat stress affects reproduction, particularly understudied stages such as floral meristem initiation and development. Fourth, we emphasise the need to integrate heat stress recovery into breeding programs to complement recent progress in improving plant heat stress tolerance. Taken together, we provide insights into key research gaps in plant heat stress and provide suggestions on addressing these gaps to enhance heat stress resilience in plants.

The neglected other half - role of the pistil in plant heat stress responses.

Heat stress coinciding with reproductive stage leads to a significant loss in reproductive organs viability, resulting in lower seed-set and crop productivity. Successful fertilization and seed formation are determined by the viability of male and female reproductive organs. The impact of heat stress on the male reproductive organ (pollen) is studied more often compared to the female reproductive organ (pistil). This is attributed to easier accessibility of the pollen coupled with the notion that the pistil's role in fertilization and seed-set under heat stress is negligible. However, depending on species and developmental stages, recent studies reveal varying degrees of sensitivity of the pistil to heat stress. Remarkably, in some cases, the vulnerability of the pistil is even greater than the pollen. This article summarizes the current knowledge of the impact of heat stress on three critical stages of pistil for successful seed-set, that is, female reproductive organ development (gametogenesis), pollen-pistil interactions including pollen capture on stigma and pollen tube growth in style, as well as fertilization and early embryogenesis. Further, future research directions are suggested to unravel molecular basis of heat stress tolerance in pistil, which is critical for sustaining crop yields under predicted warming scenarios.

Physiological and molecular attributes contribute to high night temperature tolerance in cereals.

Asymmetric warming resulting in a faster increase in night compared to day temperatures affects crop yields negatively. Physiological characterization and agronomic findings have been complemented more recently by molecular biology approaches including transcriptomic, proteomic, metabolomic and lipidomic investigations in crops exposed to high night temperature (HNT) conditions. Nevertheless, the understanding of the underlying mechanisms causing yield decline under HNT is still limited. The discovery of significant differences between HNT-tolerant and HNT-sensitive cultivars is one of the main research directions to secure continuous food supply under the challenge of increasing climate change. With this review, we provide a summary of current knowledge on the physiological and molecular basis of contrasting HNT tolerance in rice and wheat cultivars. Requirements for HNT tolerance and the special adaptation strategies of the HNT-tolerant rice cultivar Nagina-22 (N22) are discussed. Putative metabolite markers for HNT tolerance useful for marker-assisted breeding are suggested, together with future research directions aimed at improving food security under HNT conditions.

High night temperature effects on wheat and rice: Current status and way forward.

Rapid increases in minimum night temperature than in maximum day temperature is predicted to continue, posing significant challenges to crop productivity. Rice and wheat are two major staples that are sensitive to high night-temperature (HNT) stress. This review aims to (i) systematically compare the grain yield responses of rice and wheat exposed to HNT stress across scales, and (ii) understand the physiological and biochemical responses that affect grain yield and quality. To achieve this, we combined a synthesis of current literature on HNT effects on rice and wheat with information from a series of independent experiments we conducted across scales, using a common set of genetic materials to avoid confounding our findings with differences in genetic background. In addition, we explored HNT-induced alterations in physiological mechanisms including carbon balance, source-sink metabolite changes and reactive oxygen species. Impacts of HNT on grain developmental dynamics focused on grain-filling duration, post-flowering senescence, changes in grain starch and protein composition, starch metabolism enzymes and chalk formation in rice grains are summarized. Finally, we highlight the need for high-throughput field-based phenotyping facilities for improved assessment of large-diversity panels and mapping populations to aid breeding for increased resilience to HNT in crops.

Bottlenecks and opportunities in field-based high-throughput phenotyping for heat and drought stress.

Flowering and grain-filling stages are highly sensitive to heat and drought stress exposure, leading to significant loss in crop yields. Therefore, phenotyping to enhance resilience to these abiotic stresses is critical for sustaining genetic gains in crop improvement programs. However, traditional methods for screening traits related to these stresses are slow, laborious, and often expensive. Remote sensing provides opportunities to introduce low-cost, less biased, high-throughput phenotyping methods to capture large genetic diversity to facilitate enhancement of stress resilience in crops. This review focuses on four key physiological traits and processes that are critical in understanding crop responses to drought and heat stress during reproductive and grain-filling periods. Specifically, these traits include: (i) time of day of flowering, to escape these stresses during flowering; (ii) optimizing photosynthetic efficiency; (iii) storage and translocation of water-soluble carbohydrates; and (iv) yield and yield components to provide in-season yield estimates. Moreover, we provide an overview of current advances in remote sensing in capturing these traits, and discuss the limitations with existing technology as well as future direction of research to develop high-throughput phenotyping approaches. In the future, phenotyping these complex traits will require sensor advancement, high-quality imagery combined with machine learning methods, and efforts in transdisciplinary science to foster integration across disciplines.

Heat stress during flowering in cereals - effects and adaptation strategies.

Heat stress during flowering has differential impact on male and female reproductive organ viability leading to yield losses in field crops. Unlike flooded rice, dryland cereals such as sorghum, pearl millet and wheat have optimised their flower opening during cooler early morning or late evening hours to lower heat stress damage during flowering. Although previous studies have concluded that pollen viability determines seed set under heat stress, recent findings have revealed pearl millet and sorghum pistils to be equally sensitive to heat stress. Integrating flower opening time during cooler hours with increased pollen and pistil viability will overcome heat stress-induced damage during flowering under current and future hotter climatic conditions.

Deterioration of ovary plays a key role in heat stress-induced spikelet sterility in sorghum.

In sorghum (Sorghum bicolor [L.] Moench), the impact of heat stress during flowering on seed set is known, but mechanisms that lead to tolerance are not known. A diverse set of sorghum genotypes was tested under controlled environment and field conditions to ascertain the impact of heat stress on time-of-day of flowering, pollen viability, and ovarian tissue. A highly conserved early morning flowering was observed, wherein >90% of spikelets completed flowering within 30 min after dawn, both in inbreds and hybrids. A strong quantitative impact of heat stress was recorded before pollination (reduced pollen viability) and post pollination (reduced pollen tube growth and linear decline in fertility). Although viable pollen tube did reach the micropylar region, 100% spikelet sterility was recorded under 40/22°C (day/night temperatures), even in the tolerant genotype Macia. Heat stress induced significant damage to the ovarian tissue near the micropylar region, leading to highly condensed cytoplasmic contents and disintegrated nucleolus and nucleus in the susceptible genotype RTx430. Whereas, relatively less damages to ovarian cell organelles were observed in the tolerant genotype Macia under heat stress. Integrating higher tolerance in female reproductive organ will help in effective utilization of the early morning flowering mechanism to enhance sorghum productivity under current and future hotter climate.

High night temperature induced changes in grain starch metabolism alters starch, protein, and lipid accumulation in winter wheat.

Unlike sporadic daytime heat spikes, a consistent increase in night-time temperatures can potentially derail the genetic gains being achieved. Ten winter wheat genotypes were exposed to six different night-time temperatures (15-27°C) during flowering and grain-filling stages in controlled environment chambers. We identified the night-time temperature of 23o C as the critical threshold beyond which a consistent decline in yields and quality was observed. Confocal laser scanning micrographs of central endosperm, bran, and germ tissue displayed differential accumulation of protein, lipid, and starch with increasing night-time temperatures. KS07077M-1 recorded a decrease in starch and an increase in protein and lipid in central endosperm with increasing night-time temperatures, whereas the same was significantly lower in the tolerant SY Monument. Expression analysis of genes encoding 21 enzymes (including isoforms) involved in grain-starch metabolism in developing grains revealed a high night-time temperature (HNT)-induced reduction in transcript levels of adenosine diphosphate glucose pyrophosphorylase small subunit involved in starch synthesis and a ≥2-fold increase in starch degrading enzymes isoamylase III, alpha-, and beta-amylase. The identified critical threshold, grain compositional changes, and the key enzymes in grain starch metabolism that lead to poor starch accumulation in grains establish the foundational knowledge for enhancing HNT tolerance in wheat.

Enhanced N-metabolites, ABA and IAA-conjugate in anthers instigate heat sensitivity in spring wheat.

Unraveling the metabolic and phytohormonal changes in anthers exposed to heat stress would help identify mechanisms regulating heat stress tolerance during the sensitive reproductive stage. Two spring wheat genotypes contrasting for heat tolerance were exposed to heat stress during heading in controlled environment chambers. Anthers were collected from main and primary spikes for metabolic and phytohormonal profiling. A significant reduction in seed set (38%), grain number (54%) and grain weight (52%) per plant was recorded in the sensitive (KSG1177) but not in the tolerant (KSG1214) genotype under heat stress compared to control. Anther metabolite accumulation did not vary quantitatively between main and primary spikes. Hierarchical clustering of the genotypes and treatments using metabolites and phytohormones revealed a distinct cluster for KSG1177 under heat stress from that of control and KSG1214. A significant increase in N-based amino acids, ABA, IAA-conjugate and a decrease in polyamines and organic acids were observed in wheat anthers exposed to heat stress. Unlike KSG1214, a significantly higher accumulation of amino acids, ABA and IAA-conjugate in anthers of the sensitive KSG1177 was recorded under heat stress. These findings provide the rationale and direction towards developing molecular markers for enhancing heat stress tolerance in wheat.

Disaggregating sorghum yield reductions under warming scenarios exposes narrow genetic diversity in US breeding programs.

Historical adaptation of sorghum production to arid and semiarid conditions has provided promise regarding its sustained productivity under future warming scenarios. Using Kansas field-trial sorghum data collected from 1985 to 2014 and spanning 408 hybrid cultivars, we show that sorghum productivity under increasing warming scenarios breaks down. Through extensive regression modeling, we identify a temperature threshold of 33 °C, beyond which yields start to decline. We show that this decline is robust across both field-trial and on-farm data. Moderate and higher warming scenarios of 2 °C and 4 °C resulted in roughly 17% and 44% yield reductions, respectively. The average reduction across warming scenarios from 1 to 5 °C is 10% per degree Celsius. Breeding efforts over the last few decades have developed high-yielding cultivars with considerable variability in heat resilience, but even the most tolerant cultivars did not offer much resilience to warming temperatures. This outcome points to two concerns regarding adaption to global warming, the first being that adaptation will not be as simple as producers' switching among currently available cultivars and the second being that there is currently narrow genetic diversity for heat resilience in US breeding programs. Using observed flowering dates and disaggregating heat-stress impacts, both pre- and postflowering stages were identified to be equally important for overall yields. These findings suggest the adaptation potential for sorghum under climate change would be greatly facilitated by introducing wider genetic diversity for heat resilience into ongoing breeding programs, and that there should be additional efforts to improve resilience during the preflowering phase.