Summary: Plant can acquire tolerance to environmental stresses via transcriptome reprogramming at transcriptional and alternative splicing (AS) levels. However, how AS coordinates with transcriptional regulation to contribute to abiotic stresses responses is still ambiguous. In this study, we performed genome‐wide analyses of AS responses to drought stress (DS), heat stress (HS) and their combination (HD) in wheat seedlings, and further compared them with transcriptional responses. In total, we found 200, 3576 and 4056 genes exhibiting significant AS pattern changes in response to DS, HS and HD, respectively, and combined drought and heat stress can induce specific AS compared with individual one. In addition, wheat homeologous genes exhibited differential AS responses under stress conditions that more AS events occurred on B subgenome than on A and D genomes. Comparison of genes regulated at AS and transcriptional levels showed that only 12% of DS‐induced AS genes were subjected to transcriptional regulation, whereas the proportion increased to ~40% under HS and HD. Functional enrichment analysis revealed that abiotic stress‐responsive pathways tended to be highly overrepresented among these overlapped genes under HS and HD. Thus, we proposed that transcriptional regulation may play a major role in response to DS, which coordinates with AS regulation to contribute to HS and HD tolerance in wheat.

Harnessing genetic diversity in major staple crops through the development of new breeding capabilities is essential to ensure food security¹. Here we examined the genetic and phenotypic diversity of the A. E. Watkins landrace collection² of bread wheat (Triticum aestivum), a major global cereal, by whole-genome re-sequencing of 827 Watkins landraces and 208 modern cultivars and in-depth field evaluation spanning a decade. We found that modern cultivars are derived from two of the seven ancestral groups of wheat and maintain very long-range haplotype integrity. The remaining five groups represent untapped genetic sources, providing access to landrace-specific alleles and haplotypes for breeding. Linkage disequilibrium-based haplotypes and association genetics analyses link Watkins genomes to the thousands of identified high-resolution quantitative trait loci and significant marker–trait associations. Using these structured germplasm, genotyping and informatics resources, we revealed many Watkins-unique beneficial haplotypes that can confer superior traits in modern wheat. Furthermore, we assessed the phenotypic effects of 44,338 Watkins-unique haplotypes, introgressed from 143 prioritized quantitative trait loci in the context of modern cultivars, bridging the gap between landrace diversity and current breeding. This study establishes a framework for systematically utilizing genetic diversity in crop improvement to achieve sustainable food security.

Wheat (Triticum aestivum) is particularly vulnerable to heat stress during the grain filling stage, and this can adversely affect the final yield. However, the underlying physiological and molecular mechanisms are largely unknown. In this study, the effects of heat stress on grain filling were investigated using wheat varieties with different levels of thermotolerance. Decreased grain weights and filling durations, increased protein contents, and stable filling rates across diverse varieties under different heat regimes suggested a general mechanism for heat adaptation. Proteomic analysis identified 309 heat-responsive proteins (HRPs), and revealed a general decrease in protein synthesis components and metabolic proteins, but a significant increase in stress-response proteins and storage proteins. Metabolomic analysis identified 98 metabolites specifically changed by heat stress, and suggested a global decrease in the content of carbohydrate metabolites, an increased content of amino acids, and stable levels of starch synthesis precursors. The energy-consuming HRPs suggested that less energy was channelled into metabolism and protein synthesis, whereas more energy was allocated to the stress response under elevated heat conditions. Collectively, the data demonstrated a widely distributed mechanism for heat adaptation of metabolism, in which the assimilation and energy required for metabolism and protein synthesis are reallocated to heat protection and deposition of reserves, resulting in increased storage protein accumulation and a stable filling rate.

Summary: Wheat (Triticum aestivumL.), a globally important crop, is challenged by increasing temperatures (heat stress,HS). However its polyploid nature, the incompleteness of its genome sequences and annotation, the lack of comprehensiveHS‐responsive transcriptomes and the unexplored heat sensing and signaling of wheat hinder our full understanding of its adaptations toHS. The recently released genome sequences of wheat, as well as emerging single‐molecular sequencing technologies, provide an opportunity to thoroughly investigate the molecular mechanisms of the wheat response toHS. We generated a high‐resolution spatio‐temporal transcriptome map of wheat flag leaves and filling grain underHSat 0 min, 5 min, 10 min, 30 min, 1 h and 4 h by combining full‐length single‐molecular sequencing and Illumina short reads sequencing. This hybrid sequencing newly discovered 4947 loci and 70 285 transcripts, generating the comprehensive and dynamic list ofHS‐responsive full‐length transcripts and complementing the recently released wheat reference genome. Large‐scale analysis revealed a global landscape of heat adaptations, uncovering unexpected rapid heat sensing and signaling, significant changes of more than half ofHS‐responsive genes within 30 min, heat shock factor‐dependent and ‐independent heat signaling, and metabolic alterations in earlyHS‐responses. Integrated analysis also demonstrated the differential responses and partitioned functions between organs and subgenomes, and suggested a differential pattern of transcriptional and alternative splicing regulation in theHSresponse. This study provided comprehensive data for dissecting molecular mechanisms of earlyHSresponses in wheat and highlighted the genomic plasticity and evolutionary divergence of polyploidy wheat.

Plants, as sessile organisms, evolved a complex and functionally diverse heat shock factor (HSF) gene family to cope with various environmental stresses. However, the limited evolution studies of the HSF gene family have hindered our understanding of environmental adaptations in plants. In this study, a comprehensive evolution analysis on the HSF gene family was performed in 51 representative plant species. Our results demonstrated that the HSFB group which lacks a typical AHA activation domain, was the most ancient, and is under stronger purifying selection pressure in the subsequent evolutionary processes. While, dramatic gene expansion and functional divergence occurred at evolution timescales corresponding to plant land inhabit, which contribute to the emergence and diversification of the HSFA and HSFC groups in land plants. During the plant evolution, the ancestral functions of HSFs were maintained by strong purifying pressure that acted on the DNA binding domain, while the variable oligomerization domain and motif organization of HSFs underwent functional divergence and generated novel subfamilies. At the same time, variations were further accumulated with plant evolution, and this resulted in remarkable functional diversification among higher plant lineages, including distinct HSF numbers and selection pressures of several HSF subfamilies between monocots and eudicots, highlighting the fundamental differences in different plant lineages in response to environmental stresses. Taken together, our study provides novel insights into the evolutionary origin, pattern and selection pressure of plant HSFs and delineates critical clues that aid our understanding of the adaptation processes of plants to terrestrial environments.

Hexaploid wheat (Triticum aestivum L.) is an important food crop but it is vulnerable to heat. The heat-responsive proteome of wheat remains to be fully elucidated because of previous technical and genomic limitations, and this has hindered our understanding of the mechanisms of wheat heat adaptation and advances in improving thermotolerance. Here, flag leaves of wheat during grain filling stage were subjected to high daytime temperature stress, and 258 heat-responsive proteins (HRPs) were identified with iTRAQ analysis. Enrichment analysis revealed that chlorophyll synthesis, carbon fixation, protein turnover, and redox regulation were the most remarkable heat-responsive processes. The HRPs involved in chlorophyll synthesis and carbon fixation were significantly decreased, together with severe membrane damage, demonstrating the specific effects of heat on photosynthesis of wheat leaves. In addition, the decrease in chlorophyll content may result from the decrease in HRPs involved in chlorophyll precursor synthesis. Further analysis showed that the accumulated effect of heat stress played a critical role in photosynthesis reduction, suggested that improvement in heat tolerance of photosynthesis, and extending heat tolerant period would be major research targets. The significantly accumulation of GSTs and Trxs in response to heat suggested their important roles in redox regulation, and they could be the promising candidates for improving wheat thermotolerance. In summary, our results provide new insight into wheat heat adaption and provide new perspectives on thermotolerance improvement.