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Environmental change requires more crop production per water use to meet the rising global food demands. However, improving crop intrinsic water use efficiency (iWUE) usually comes at the expense of carbon assimilation. Sorghum is a key crop in many vulnerable agricultural systems with higher tolerance to water stress (WS) than most widely planted crops. To investigate physiological controls on iWUE and its inheritance in sorghum, we screened 89 genotypes selected based on inherited haplotypes from an elite line or five exotics lines, containing a mix of geographical origins and dry versus milder climates, which included different aquaporin (AQP) alleles. We found significant variation among key highly heritable gas exchange and hydraulic traits, with some being significantly affected by variation in haplotypes among parental lines. Plants with a higher proportion of the non-stomatal component of iWUE still maintained iWUE under WS by maintaining photosynthetic capacity, independently of reduction in leaf hydraulic conductance. Haplotypes associated with two AQPs (SbPIP1.1 and SbTIP3.2) influenced iWUE and related traits. These findings expand the range of traits that bridge the trade-off between iWUE and productivity in C4 crops, and provide possible genetic regions that can be targeted for breeding., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

In this study, we assessed 43 accessions of sorghum from 16 countries across three continents. Our objective was to identify stomatal and photosynthetic traits that could be exploited in breeding programmes to increase photosynthesis without increasing water use under dynamic light environments. Under field conditions, sorghum crops often have limited water availability and are exposed to rapidly fluctuating light intensities, which influences both photosynthesis and stomatal behaviour. Our results highlight a tight coupling between photosynthetic rate (A) and stomatal conductance (gs) even under dynamic light conditions that results in steady intrinsic water use efficiency (Wi). This was mainly due to rapid stomatal responses, with the majority of sorghum accessions responding within ≤5 min. The maintenance of the ratio of the concentration of CO2 inside the leaf (Ci) and the surrounding atmospheric concentration (Ca) over a large range of accessions suggests high stomatal sensitivity to changes in Ci, that could underlie the rapid gs responses and extremely close relationship between A and gs under both dynamic and steady-state conditions. Therefore, sorghum represents a prime candidate for uncovering the elusive mechanisms that coordinate A and gs, and such information could be used to design crops with high A without incurring significant water losses and eroding Wi., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Enhancing crop water use efficiency (WUE) is a key target trait for climatic resilience and expanding cultivation on marginal lands. Engineering lower stomatal density to reduce stomatal conductance (gs) has improved WUE in multiple C3 crop species. However, reducing gs in C3 species often reduces photosynthetic carbon gain. A different response is expected in C4 plants because they possess specialized anatomy and biochemistry which concentrates CO2 at the site of fixation. This modifies the relationship of photosynthesis (AN) with intracellular CO2 concentration (ci), such that photosynthesis is CO2 saturated and reductions in gs are unlikely to limit AN. To test this hypothesis, genetic strategies were investigated to reduce stomatal density in the C4 crop sorghum. Constitutive expression of a synthetic epidermal patterning factor (EPF) transgenic allele in sorghum led to reduced stomatal densities, reduced gs, reduced plant water use, and avoidance of stress during a period of water deprivation. In addition, moderate reduction in stomatal density did not increase stomatal limitation to AN. However, these positive outcomes were associated with negative pleiotropic effects on reproductive development and photosynthetic capacity. Avoiding pleiotropy by targeting expression of the transgene to specific tissues could provide a pathway to improved agronomic outcomes., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Phytoremediation of cadmium (Cd)-polluted soil by using sweet sorghum displays a tremendous potential as it is a fast-growing, high biomass and Cd tolerant energy plant. Previous study has demonstrated SbNRT1.1B expression change is in accordance with enhanced Cd accumulation by external nitrate supply in sweet sorghum. Nevertheless, underlying mechanism of SbNRT1.1B response to Cd stress is still elusive. SbNRT1.1B exhibited a positive response to Cd stress in sweet sorghum. Overexpressing SbNRT1.1B increased primary root length, shoot fresh weight, nitrate and chlorophyll concentrations compared with Col-0 under Cd stress, while complementary SbNRT1.1B rescued these decreased values in mutant chl1-5. Cd concentrations in overexpressing SbNRT1.1B, complementary SbNRT1.1B and Col-0 lines were 3.2-4.1, 2.5-3.1 and 1.2-2.1 folds of that in chl1-5. Consistent with Cd concentrations, non-protein thiol (NPT), reduced glutathione (GSH) and phytochelatins (PCs) concentrations as well as the related genes expression levels showed the same trends under Cd stress. GSH biosynthesis inhibitor failed to reverse the patterns of GSH-dependent PCs concentrations changes in different lines, suggesting that SbNRT1.1B plays an upstream role in GSH-dependent PCs biosynthesis under Cd treatment. Altogether, SbNRT1.1B enhances nitrate concentrations contributing to increased chlorophyll concentrations and GSH-dependent PCs metabolites biosynthesis, thereby improving growth and Cd concentrations in plants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Backgrounds: Phosphorus is a critical nutrient in agriculture, influencing plant growth and nutritional quality., Objectives: This study, uniquely designed to investigate the effects of phosphorus (P) fertilization levels, sorghum cultivars, and growing locations on phytochemical content and antioxidant activity in sorghum grains, employed four sorghum cultivars (Hakeka, P954063, Tabat, and Tetron) grown under three P levels (0P, 1P, 2P) in two locations (Gezira and White Nile) in Sudan., Methods: In this study, four sorghum cultivars were grown in two distinct locations in Sudan, employing a split-plot design with three (P) fertilization levels. P was applied as triple super phosphate directly with the seeds, and additional fertilization included urea applied in two split doses. At physiological maturity, representative sorghum panicles were harvested, processed, and analyzed for bioactive compounds and antioxidant activities using standard extraction and quantification techniques such as Folin-Ciocalteu for phenolics and colorimetric flavonoid assays. Antioxidant activities were assessed through various assays, including DPPH and FRAP. Statistical analyses were performed using a three-way ANOVA to examine the effects of cultivar, P level, and location on the measured parameters, supplemented by multivariate analysis to further elucidate the interactions between these factors., Results: Significant interactions (p<0.001) were observed among cultivars, P levels, and locations for total phenolic content (TPC), total flavonoid content (TFC), carotenoids, tannins, and various antioxidant activity measures (DPPH, FRAP, ABTS, TRP, H2O2). P fertilization significantly increased all measured phytochemicals and antioxidant activities compared to non-treated cultivars, except for H2O2, which decreased with P application. Among cultivars, Hakeka consistently exhibited the highest TFC, carotenoid content, and antioxidant activities (DPPH, FRAP, TRP, ABTS), particularly at the 2P level. P954063 showed the highest TPC and tannin concentrations. Tetron generally had the lowest phytochemical and antioxidant levels. White Nile showed higher TPC, carotenoids, DPPH, FRAP, TRP, and ABTS levels, while Gezira had higher TFC, tannins, and H2O2 concentrations. The impact of phosphorus fertilization often varies between locations. Strong positive correlations were found between TPC and all antioxidant assays (r = 0.68-0.90) and total carotenoids and antioxidant activities (r = 0.73-0.93)., Conclusions: This study recommended cultivating the Tabat variety with 2P doses in Gezira. In addition, the Hakeka cultivar showed the highest increases in total flavonoid content, carotenoids, and antioxidant activities, particularly under the highest P level (2P). The findings highlight that P plays a critical role in enhancing sorghum's nutritional and health-promoting qualities, which are essential for leveraging this staple crop for food and nutrition security strategies in semi-arid regions., Competing Interests: The authors declare that they have no conflict of interests., (Copyright: © 2024 Elsafy et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Rising temperatures due to the current climate crisis will soon have devastating impacts on crop performance and resilience. In particular, CO2 assimilation is dramatically limited at high temperatures. CO2 assimilation is accomplished by rubisco, which is inhibited by the binding of inhibitory sugar phosphates to its active site. Plants therefore utilize the essential chaperone rubisco activase (RCA) to remove these inhibitors and enable continued CO2 fixation. However, RCA does not function at moderately high temperatures (42°C), resulting in impaired rubisco activity and reduced CO2 assimilation. We set out to understand temperature-dependent RCA regulation in four different C4 plants, with a focus on the crop plants maize (two cultivars) and sorghum, as well as the model grass Setaria viridis (setaria) using gas exchange measurements, which confirm that CO2 assimilation is limited by carboxylation in these organisms at high temperatures (42°C). All three species express distinct complements of RCA isoforms and each species alters the isoform and proteoform abundances in response to heat; however, the changes are species-specific. We also examine whether the heat-mediated inactivation of RCA is due to biochemical regulation rather than simple thermal denaturation. We reveal that biochemical regulation affects RCA function differently in different C4 species, and differences are apparent even between different cultivars of the same species. Our results suggest that each grass evolved different strategies to maintain RCA function during stress and we conclude that a successful engineering approach aimed at improving carbon capture in C4 grasses will need to accommodate these individual regulatory mechanisms., (© 2024 The Author(s).)

Cuticular wax (CW) is the first defensive barrier of plants that forms a waterproof barrier, protects the plant from desiccation, and defends against insects, pathogens, and UV radiation. Sorghum, an important grass crop with high heat and drought tolerance, exhibits a much higher wax load than other grasses and the model plant Arabidopsis. In this study, we explored the regulation of sorghum CW biosynthesis using a bloomless mutant. The CW on leaf sheaths of the bloomless 41 (bm41) mutant showed significantly reduced very long-chain fatty acids (VLCFAs), triterpenoids, alcohols, and other wax components, with an overall 86% decrease in total wax content compared with the wild type. Notably, the 28-carbon and 30-carbon VLCFAs were decreased in the mutants. Using bulk segregant analysis, we identified the causal gene of the bloomless phenotype as a leucine-rich repeat transmembrane protein kinase. Transcriptome analysis of the wild-type and bm41 mutant leaf sheaths revealed BM41 as a positive regulator of lipid biosynthesis and steroid metabolism. BM41 may regulate CW biosynthesis by regulating the expression of the gene encoding 3-ketoacyl-CoA synthase 6. Identification of BM41 as a new regulator of CW biosynthesis provides fundamental knowledge for improving grass crops' heat and drought tolerance by increasing CW., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Sorghum ( Sorghum bicolor ) is a significant crop serving food, energy, feed, and industrial raw materials, featuring extensive growth adaptability and diverse utility values. Despite the achievements in sorghum breeding in the last decades, conventional breeding methods still confront challenges such as lengthy breeding cycles, low efficiency, and complex genetic backgrounds. With the rapid advancement of molecular biology, genetics, and bioinformatics, molecular breeding has carved new pathways for enhancing sorghum yield and quality. This article reviews the molecular basic research progress in the key agronomic and adaptive traits of sorghum, including grain yield, grain quality, flowering time, plant height, tillering, stress resistance, and male sterility, and discusses future research priorities, offering novel insights and approaches for sorghum breeding.

Identification of high carotenoid germplasm is crucial to assist breeders in provitamin-A biofortification of sorghum (Sorghum bicolor [L.] Moench). High-performance liquid chromatography is the gold standard for carotenoid quantification, however, it is not feasible for large scale phenotyping due to its high cost and low throughput. In this study, we tested the feasibility of using grain color as a high-throughput method of carotenoid biofortification breeding. We hypothesized that visual, color-based selection can be an effective strategy to identify high-carotenoid accessions. Yellow grain had significantly higher carotenoid content than red, brown, and white grain. The degree of yellowness could distinguish the presence or absence of carotenoids, but could not distinguish carotenoid concentrations within yellow-only accessions. The degree of luminosity of the grain, however, was able to better predict carotenoid concentrations within yellow-only accessions. Genome-wide association studies identified significant marker-trait associations for qualitative and quantitative grain color traits and carotenoid concentrations near carotenoid pathway genes-ZEP, PDS, CYP97A, NCED, CCD, and LycE-three of which were common between grain color and carotenoid traits. These findings suggest that using grain color as a method for screening germplasm may be an effective high-throughput selection tool for prebreeding and early-stage breeding in carotenoid biofortification., (© 2024. The Author(s).)

Plant biomass can significantly contribute to alternative energy sources. Sorghum bicolor is a promising plant for producing energy, but is susceptible to iron deficiency, which inhibits its cultivation in iron-limiting calcareous soils. The molecular basis for the susceptibility of sorghum to iron deficiency remains unclear. Here, we explored the sorghum genome to identify genes involved in iron uptake and translocation. Iron deficiency-responsive gene expression was comparable to that in other graminaceous plants. A nicotianamine synthase gene, SbNAS1, was induced in response to iron deficiency, and SbNAS1 showed enzyme activity. Sorghum secreted 2'-deoxymugineic acid and other phytosiderophores under iron deficiency, but their levels were relatively low. Intercropping of sorghum with barley or rice rescued iron deficiency symptoms of sorghum. To produce bioengineered sorghum with enhanced tolerance to iron deficiency, we introduced four cassettes into sorghum: 35S promoter-OsIRO2 for activation of iron acquisition-related gene expression, SbIRT1 promoter-Refre1/372 for enhanced ferric-chelate reductase activity, and barley IDS3, and HvNAS1 genomic fragments for enhanced production of phytosiderophores and nicotianamine. The resultant single sorghum line exhibited enhanced secretion of phytosiderophores, increased ferric-chelate reductase activity, and improved iron uptake and leaf greenness compared with non-transformants under iron-limiting conditions. Similar traits were also conferred to rice by introducing the four cassettes. Moreover, these rice lines showed similar or better tolerance in calcareous soils and increased grain iron accumulation compared with previous rice lines carrying two or three comparable cassettes. These results provide a molecular basis for the bioengineering of sorghum tolerant of low iron availability in calcareous soils., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

In this study, the effects on silage performance and microbial community of sorghum straw treated with the addition of enzymes (cellulase (CE), xylanase (XE)) and extrusion puffing technology, combined with SEM, XRD, and FTIR techniques, were thoroughly investigated. The results showed that the enzyme preparations, especially xylanase, significantly improved the nutritional value and fermentation efficiency of straw and enhanced the silage effect. Extruding significantly changes the surface structure of the straw, increasing the surface area and porosity, and promoting the attachment of microorganisms. This study not only optimized the sorghum straw silage performance but also provided technical support for the efficient use of straw resources, which is of great significance for the sustainable development of animal husbandry and the resource utilization of agricultural waste., (© 2024. The Author(s).)

Interactions between plants and soil microbial communities that benefit plant growth and enhance nutrient acquisition are driven by the selective release of metabolites from plant roots, or root exudation. To investigate these plant-microbe interactions, we developed a photoaffinity probe based on sorgoleone ( so rgoleone d iazirine a lkyne for p hoto a ffinity l abeling, SoDA-PAL), a hydrophobic secondary metabolite and allelochemical produced in Sorghum bicolor root exudates. We applied SoDA-PAL to the identification of sorgoleone-binding proteins in Acinetobacter pittii SO1, a potential plant growth-promoting microbe isolated from sorghum rhizosphere soil. Competitive photoaffinity labeling of A. pittii whole cell lysates with SoDA-PAL identified 137 statistically enriched proteins, including putative transporters, transcriptional regulators, and a subset of proteins with predicted enzymatic functions. We performed computational protein modeling and docking with sorgoleone to prioritize candidates for experimental validation and then confirmed binding of sorgoleone to four of these proteins in vitro : the α/β fold hydrolase SrgB (OH685&#95;09420), a fumarylacetoacetase (OH685&#95;02300), a lysophospholipase (OH685&#95;14215), and an unannotated hypothetical protein (OH685&#95;18625). Our application of this specialized sorgoleone-based probe coupled with structural bioinformatics streamlines the identification of microbial proteins involved in metabolite recognition, metabolism, and toxicity, widening our understanding of the range of cellular pathways that can be affected by a plant secondary metabolite.IMPORTANCEHere, we demonstrate that a photoaffinity-based chemical probe modeled after sorgoleone, an important secondary metabolite released by sorghum roots, can be used to identify microbial proteins that directly interact with sorgoleone. We applied this probe to the sorghum-associated bacterium Acinetobacter pittii and showed that probe labeling is dose-dependent and sensitive to competition with purified sorgoleone. Coupling the probe with proteomics and computational analysis facilitated the identification of putative sorgoleone binders, including a protein implicated in a conserved pathway essential for sorgoleone catabolism. We anticipate that discoveries seeded by this workflow will expand our understanding of the molecular mechanisms by which specific metabolites in root exudates shape the sorghum rhizosphere microbiome., Competing Interests: The authors declare no conflict of interest.

Sustainably grown biomass is a promising alternative to produce fuels and chemicals and reduce the dependency on fossil energy sources. However, the efficient conversion of lignocellulosic biomass into biofuels and bioproducts often requires extensive testing of components and reaction conditions used in the pretreatment, saccharification, and bioconversion steps. This restriction can result in a significant and unwieldy number of combinations of biomass types, solvents, microbial strains, and operational parameters that need to be characterized, turning these efforts into a daunting and time-consuming task. Here we developed a high-throughput feedstocks-to-fuels screening platform to address these challenges. The result is a miniaturized semi-automated platform that leverages the capabilities of a solid handling robot, a liquid handling robot, analytical instruments, and a centralized data repository, adapted to operate as an ionic-liquid-based biomass conversion pipeline. The pipeline was tested by using sorghum as feedstock, the biocompatible ionic liquid cholinium phosphate as pretreatment solvent, a "one-pot" process configuration that does not require ionic liquid removal after pretreatment, and an engineered strain of the yeast Rhodosporidium toruloides that produces the jet-fuel precursor bisabolene as a conversion microbe. By the simultaneous processing of 48 samples, we show that this configuration and reaction conditions result in sugar yields (~70%) and bisabolene titers (~1500 mg/L) that are comparable to the efficiencies observed at larger scales but require only a fraction of the time. We expect that this Feedstocks-to-Fuels pipeline will become an effective tool to screen thousands of bioenergy crop and feedstock samples and assist process optimization efforts and the development of predictive deconstruction approaches., Competing Interests: B.A.S. has a financial interest in Caribou Biofuels, Illium Technologies, and Erg Bio. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Pidatala et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

While considerable knowledge exists about the enzymes pivotal for C 4 photosynthesis, much less is known about the cis- regulation important for specifying their expression in distinct cell types. Here, we use single-cell-indexed ATAC-seq to identify cell-type-specific accessible chromatin regions (ACRs) associated with C 4 enzymes for five different grass species. This study spans four C 4 species, covering three distinct photosynthetic subtypes: Zea mays and Sorghum bicolor (NADP-dependent malic enzyme), Panicum miliaceum (NAD-dependent malic enzyme), Urochloa fusca (phosphoenolpyruvate carboxykinase), along with the C 3 outgroup Oryza sativa . We studied the cis- regulatory landscape of enzymes essential across all C 4 species and those unique to C 4 subtypes, measuring cell-type-specific biases for C 4 enzymes using chromatin accessibility data. Integrating these data with phylogenetics revealed diverse co-option of gene family members between species, showcasing the various paths of C 4 evolution. Besides promoter proximal ACRs, we found that, on average, C 4 genes have two to three distal cell-type-specific ACRs, highlighting the complexity and divergent nature of C 4 evolution. Examining the evolutionary history of these cell-type-specific ACRs revealed a spectrum of conserved and novel ACRs, even among closely related species, indicating ongoing evolution of cis -regulation at these C 4 loci. This study illuminates the dynamic and complex nature of cis -regulatory elements evolution in C 4 photosynthesis, particularly highlighting the intricate cis- regulatory evolution of key loci. Our findings offer a valuable resource for future investigations, potentially aiding in the optimization of C 3 crop performance under changing climatic conditions., Competing Interests: Competing interests statement:R.J.S. is a co-founder of REquest Genomics, LLC, a company that provides epigenomic services.

Cadmium (Cd) pollution in the soil is a serious environmental issue worldwide. Phytoextraction of Cd-polluted soil is a cost-effective, sustainable and environmentally-friendly strategy. Agricultural fertilizer management is beneficial for promoting the Cd phytoremediation efficiency. Potassium (K) is the nutrient required in the largest amount cation by plants. Sweet sorghum exhibits a substantial phytoremediation potential of Cd-polluted soil. Clarifying the mechanism of K-mediated Cd accumulation in sweet sorghum is imperative. Sweet sorghum plants were grown hydroponically with an extra K supply in the presence or absence of Cd treatment. An extra K application significantly increased plant growth under non-Cd addition, while K lost the profitable effect under Cd stress. K supplementation remarkably enhanced Cd concentrations and Cd accumulation in shoots and roots of sweet sorghum. Transcriptome analysis demonstrated that zinc ion transport, cysteine and methionine metabolism, flavonoid biosynthesis and phenylpropanoid biosynthesis pathways might contribute to the increased Cd accumulation as affected by an extra K supply. Furthermore, SbZIP9, SbSTP8, SbYS1, SbMAG and SbFOMT-like were targeted as they closely correlated with both plant growth and Cd stress in sweet sorghum. SbFOMT-like showed an independent pathway, while SbZIP9, SbSTP8, SbYS1 and SbMAG displayed positive correlations mutually. Notably, SbZIP9 and SbFOMT-like were highly expressed when compared with other target genes. Taken together, SbZIP9 and SbFOMT-like were upregulated and downregulated by an extra K supply under Cd stress, suggesting that SbZIP9 and SbFOMT-like enhances and declines Cd accumulation as regulated by K addition in sweet sorghum respectively., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Sorghum (Sorghumbicolor L.), a model for C 4 grass and an emerging biofuel crop, is known for its robust tolerance to low input field. However, the focus on enhancing nitrogen use efficiency (NUE) in sorghum under low nitrogen (N) conditions has been limited. This study conducted hydroponic experiments and field trials with two sorghum inbred lines, contrasting in their N efficiency: the N-efficient (398B) and the N-inefficient (CS3541) inbred lines. The aim was to analyze the key factors influencing NUE by integrating phenotypic, physiological, and multi-omics approaches under N deficiency conditions. The field experiments revealed that 398B displayed superior NUE and yield performance compared to CS3541. In hydroponic experiments, the growth of 398B outperformed CS3541 following N deficiency, attributing to its higher photosynthetic and sustaining activity of N metabolism-related enzymes. Genomic and transcriptomic integration highlighted fewer genomic diversities and alterations in global gene expression in 398B, which were likely contributor to its high NUE. Additionally, co-expression network analysis suggested the involvement of key genes which impact N uptake efficiency (NUpE) and N utilization efficiency (NUtE) in both lines, such as an N transporter, Sobic.003G371000.v3.2leaf(NPF5.10) and a transcription factor, Sobic.002G202800.v3.2leaf(WRKY) in bolstering NUE under low-N stress. The findings collectively suggested that 398B achieved higher NUpE and NUtE, effectively coordinating photosynthesis and N metabolism to enhance NUE. The candidate genes regulating N uptake and utilization efficiencies could provide valuable insights for developing sorghum breeds with improved NUE, contributing to sustainable agricultural practices and bioenergy crop development., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

In this study, phenolic profile/content was analyzed by high-resolution untargeted metabolomics after short germination (72 h) and seedling growth (144 h), using three sorghum genotypes varying in tannin content (IS 29569, Macia and IS 30400). In vitro antioxidant capacity and phenolic bioaccessibility were determined by microplate-based and INFOGEST methods, respectively. A total of 58 % annotated compounds were found in all genotypes; and phenolic acids and flavonoids represent more than 80 % of sorghum total abundance. PCA analysis showed higher phenolic variability in germination times (72 %) than genotypes (51 %). Germination reduced total ion abundance (-7 %) and free:bound phenolic compounds ratio (2.4-1.1), but antioxidant capacity remained constant. These results indicate the cell matrix-phenolic decomplexation, with the free compounds were quickly consumed after radicle emergence. Germination increased phenolic bioaccessibility (mainly in oral phase) but reduces flavonoids contents in gastric/intestinal digestion steps. This work can stimulate seed germination as a viable option for sorghum-based foods development, with improved nutritional and bioactive properties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Cover cropping is an agricultural practice that uses secondary crops to support the growth of primary crops through various mechanisms including erosion control, weed suppression, nutrient management, and enhanced biodiversity. Cover crops may elicit some of these ecosystem services through chemical interactions with the soil microbiome via root exudation, or the release of plant metabolites from roots. Phytohormones are one metabolite type exuded by plants that activate the rhizosphere microbiome, yet managing this chemical interaction remains an untapped mechanism for optimizing plant-soil-microbiome interactions. Currently, there is limited understanding on the diversity of cover crop phytohormone root exudation patterns and our aim was to understand how phytochemical signals selectively enrich specific microbial taxa and functionalities in agricultural soils., Results: Here, we link variability in cover crop root exudate composition to changes in soil microbiome functionality. Exudate chemical profiles from 4 cover crop species (Sorghum bicolor, Vicia villosa, Brassica napus, and Secale cereal) were used as the chemical inputs to decipher microbial responses. These distinct exudate profiles, along with a no exudate control, were amended to agricultural soil microcosms with microbial responses tracked over time using metabolomes and genome-resolved metatranscriptomes. Our findings illustrated microbial metabolic patterns were unique in response to cover crop exudate inputs over time, particularly by sorghum and cereal rye amended microcosms. In these microcosms, we identify novel microbial members (at the genera and family level) who produced IAA and GA 4 over time. Additionally, we identified cover crop exudates exclusively enriched for bacterial nitrite oxidizers, while control microcosms were discriminated for nitrogen transport, mineralization, and assimilation, highlighting distinct changes in microbial nitrogen cycling in response to chemical inputs., Conclusions: We highlight that root exudate amendments alter microbial community function (i.e., N cycling) and microbial phytohormone metabolisms, particularly in response to root exudates isolated from cereal rye and sorghum plants. Additionally, we constructed a soil microbial genomic catalog of microorganisms responding to commonly used cover crops, a public resource for agriculturally relevant microbes. Many of our exudate-stimulated microorganisms are representatives from poorly characterized or novel taxa, revealing the yet to be discovered metabolic reservoir harbored in agricultural soils. Our findings emphasize the tractability of high-resolution multi-omics approaches to investigate processes relevant for agricultural soils, opening the possibility of targeting specific soil biogeochemical outcomes through biological precision agricultural practices that use cover crops and the microbiome as levers for enhanced crop production. Video Abstract., (© 2024. The Author(s).)

A fundamental assumption in plant science posits that leaf air spaces remain vapor saturated, leading to the predominant view that stomata alone control leaf water loss. This concept has been pivotal in photosynthesis and water-use efficiency research. However, recent evidence has refuted this longstanding assumption by providing evidence of unsaturation in the leaf air space of C 3 plants under relatively mild vapor pressure deficit (VPD) stress. This phenomenon represents a nonstomatal mechanism restricting water loss from the mesophyll. The potential ubiquity and physiological implications of this phenomenon, its driving mechanisms in different plant species and habitats, and its interaction with other ecological adaptations have. In this context, C 4 plants spark particular interest for their importance as crops, bundle sheath cells' unique anatomical characteristics and specialized functions, and notably higher water-use efficiency relative to C 3 plants. Here, we confirm reduced relative humidities in the substomatal cavity of the C 4 plants maize, sorghum, and proso millet down to 80% under mild VPD stress. We demonstrate the critical role of nonstomatal control in these plants, indicating that the role of the CO 2 concentration mechanism in CO 2 management at a high VPD may have been overestimated. Our findings offer a mechanistic reconciliation between discrepancies in CO 2 and VPD responses reported in C 4 species. They also reveal that nonstomatal control is integral to maintaining an advantageous microclimate of relatively higher CO 2 concentrations in the mesophyll air space of C 4 plants for carbon fixation, proving vital when these plants face VPD stress., Competing Interests: Competing interests statement:The authors declare no competing interest.

Sorghum, a globally grown gluten-free cereal, is used mainly as an animal feed in developed countries regardless of its potential for human consumption. In this study, we utilized nontargeted lipidomics to thoroughly analyze, compare, and characterize whole-grain lipids in six sorghum cultivars (cv) grown in a single field trial in Australia: Buster, Bazley, Cracker, Liberty, MR43, and Tiger. In total, 194 lipid molecular species representing five major lipid classes were identified. Multivariate analysis unveiled distinct lipid profiles among the cultivars. The most distinct lipid profile belonged to cv. MR43. The lower ω-6 to ω-3 ratio and optimal P/S ratio in cv. Bazley reflect this as a valuable source of balanced essential fatty acids in the diet. The novel bioactive lipids known as FAHFAs (fatty acid esters of hydroxy fatty acids) were identified and characterized in sorghum grains. These findings further emphasize the potential of whole-grain sorghum as a basis for new health-promoting food products.

The utilization of biomass ash in sustainable agriculture and increasing its fertilizing efficiency by biological agents, potentially sequestering CO 2 , have become important issues for the global economy. The aim of this paper was to investigate the effects of ash from sorghum ( Sorghum bicolor L. Moench) and Jerusalem artichoke ( Helianthus tuberosus L.) biomass, a biogas plant digestate, and a Spirodela polyrhiza extract, acting alone or synergistically, on soil fertility and the development, health and physiological properties of sorghum plants. The results show novel information concerning differences in the composition and impact of ash, depending on its origin, soil properties and sorghum plant development. Sorghum ash was more effective than that from Jerusalem artichoke. Ash used alone and preferably acting synergistically with the digestate and Spirodela polyrhiza extract greatly increased soil fertility and the growth, biomass yield and health of sorghum plants. These improvements were associated with an increased chlorophyll content in leaves, better gas exchange (photosynthesis, transpiration, stomatal conductance), greater enzyme activity (acid and alkaline phosphatase, RNase, and total dehydrogenase), and a higher biomass energy value. The developed treatments improved environmental conditions by replacing synthetic fertilizers, increasing the sequestration of CO 2 , solving the ash storage problem, reducing the need for pesticides, and enabling a closed circulation of nutrients between plant and soil, maintaining high soil fertility.

Photosynthetic organisms must cope with rapid fluctuations in light intensity. Nonphotochemical quenching (NPQ) enables the dissipation of excess light energy as heat under high light conditions, whereas its relaxation under low light maximizes photosynthetic productivity. We quantified variation in NPQ kinetics across a large sorghum (Sorghum bicolor) association panel in four environments, uncovering significant genetic control for NPQ. A genome-wide association study (GWAS) confidently identified three unique regions in the sorghum genome associated with NPQ and suggestive associations in an additional 61 regions. We detected strong signals from the sorghum ortholog of Arabidopsis thaliana Suppressor Of Variegation 3 (SVR3) involved in plastid-nucleus signaling. By integrating GWAS results for NPQ across maize (Zea mays) and sorghum-association panels, we identified a second gene, Non-yellowing 1 (NYE1), originally studied by Gregor Mendel in pea (Pisum sativum) and involved in the degradation of photosynthetic pigments in light-harvesting complexes. Analysis of nye1 insertion alleles in A. thaliana confirmed the effect of this gene on NPQ kinetics in eudicots. We extended our comparative genomics GWAS framework across the entire maize and sorghum genomes, identifying four additional loci involved in NPQ kinetics. These results provide a baseline for increasing the accuracy and speed of candidate gene identification for GWAS in species with high linkage disequilibrium., (© 2024 The Author(s). The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Eco-friendly and sustainable practices must be followed while using the right plants and microbes to remove harmful heavy metals from the soil. The goal of the current study was to ascertain how effectively sorghum plants removed cadmium (Cd) from the soil using polyamines and mycorrhiza. Plant-biochemicals such as free amino acids, ascorbic acids, anthocyanin, proline, and catalase, APX, peroxidase activities were considered as markers in this study which revealed the adverse plant growth performance under 70 and 150 ppm of Cd concentration (w/w) after 30,60, and 90 days of treatment. The plants showed a mitigating effect against high Cd-concentration with exogenous use of mycorrhiza and putrescine. The treatment T17 (mycorrhiza +5 mM putrescine) showed a substantial decrease in the content of total free amino acid, ascorbic acid, catalase, APX, peroxidase by 228.36%, 39.79%, 59.06%, 182.79% 106.97%, respectively after 90 days as compared to T12 (150 ppm Cd). Anthocyanin content was negatively correlated (-0.503, -0.556, and -0.613) at p < 0.01 with other studied markers, with an increase by 10.52% in T17 treated plant as compared to T12. The concentration of Cd in root increased by 49.6% (141 ppm) and decreased in the shoot by 71% (17.8 ppm) in T17 treated plant as compared to T12 after 90 days. The application of mycorrhiza and putrescine significantly increased BCF (>1) and decreased TF (<1) for Cd translocation. The administration of mycorrhiza and putrescine boosted the Cd removal efficiency of sorghum plants, according to FTIR, XRD, and DSC analysis. As a result, this study demonstrates novel approaches for induced phytoremediation activity of plants via mycorrhiza and putrescine augmentation, which can be a promising option for efficient bioremediation in contaminated sites., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Drought stress strongly affects crop yield. Although knowledge of long non-coding RNAs (lncRNAs) has been updated continuously and rapidly, information about lncRNAs in drought resistance regulation is extremely limited in sorghum. Here, lncRNA-sequencing was performed with seedlings of a sorghum cultivar (Jinza29) under three water control treatments to investigate the mechanism of lncRNAs responsible for drought resistance in sorghum. A total of 377 differentially expressed lncRNAs (DElncRNAs) were identified. We also predicted 4322 and 2827 transcripts as potential cis-target and trans-target genes for drought-responsive lncRNAs, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that those target genes exhibited marked enrichment into "oxidoreductase activity", "signal transducer activity", "DNA repair", "photosynthesis", "glutathione metabolism", and "phenylpropanoid biosynthesis" and other terms associated with abiotic stress resistance. Moreover, several lncRNAs were estimated to modulate the expression of other genes related to stress response and photosynthetic carbon metabolism. Additionally, we found 107 DElncRNAs that might be candidate target mimics for 56 miRNAs. LncRNAs play important roles in drought adaptation of sorghum through interacting with protein-encoding genes. The obtained results provided novel insights into the biological characteristics of lncRNAs and offered potential regulatory factors for genetically enhancing drought resistance in sorghum., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Salt stress is one of the significant environmental stresses that severely affect plant growth and development. Here, we report quantitative N-glycoproteomics characterization of differential N-glycosylation in Sorghum bicolor under low, median and high salinity stress. 21,621 intact N-glycopeptides coming from the combination of 127 N-glycan structures on 6574 N-glycosites from 5321 proteins were identified; differential N-glycosylation was observed for 682 N-glycoproteins which are mainly involved in the pathways of biosynthesis of secondary metabolites, biosynthesis of amino acids and several metabolic pathways. 41 N-glycan structures modifying on 338 N-glycopeptides from 122 glycoproteins were co-quantified and deregulated under at least one salt stress, including enzymes of energy production and carbohydrate metabolisms, cell wall organization related proteins, glycosyltransferases and so on. Intriguingly, with increasing salt concentration, there was an increase in the percentage of complex N-glycans on the altered N-glycopeptides. Furthermore, the observation of glycoproteins with distinct salt sensitivity is noteworthy, particularly the upregulated hyposensitive glycoproteins that predominantly undergo complex N-glycan modification. This is the first N-glycoproteome description of salt stress response at the intact N-glycopeptide level in sorghum and a further validation of data reported here would likely provide deeper insights into the stress physiology of this important crop plant., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Sorghum seeds can discolor during storage. Treatment of seeds with citric acid improves sensory quality and antioxidant activity. This study compared the differences in phenotypic and antioxidant activity between citric acid-treated and water-treated sorghum seeds. The study used transcriptomics and metabolomics approaches to investigate the regulatory mechanisms. The ∆a, ∆b and ∆l values of citric acid-treated sorghum seeds significantly increased after 6 months of storage. The SOD, POD and CAT enzyme activities of the citric acid-treated group were 1.94, 1.91 and 2.45 times higher than those of the control, respectively. The joint transcriptome and metabolome analysis showed that the citric acid-induced changes were mainly focused on the flavonoid biosynthetic pathway. Citric acid treatment up-regulated CHS, ANR, MYB and bHLH genes and promoted flavonoid accumulation. In conclusion, citric acid treatment promotes flavonoid accumulation, delays sorghum seed discoloration, and enhances antioxidant activity and storage life., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Zhenhua Li reports financial support, administrative support, article publishing charges, equipment, drugs, or supplies, and travel were provided by Agriculture of Guizhou Province and the Key Laboratory of Molecular Breeding of Grain and Oil Crops of Guizhou Province. Zhenhua Li reports a relationship with Agriculture of Guizhou Province and the Key Laboratory of Molecular Breeding of Grain and Oil Crops of Guizhou Province. that includes: funding grants, non-financial support, speaking and lecture fees, and travel reimbursement. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Colletotrichum boninense is the main pathogenic fungus causing leaf spot disease in Sorghum sudangrass hybrids , which seriously impairs its quality and yield. In order to find an efficient and green means of control, this study used the agar disk diffusion method to screen for a fungicide with the strongest inhibitory effect on C. boninense from among several bacteria, fungi, and chemicals. Then, the changes in the plant's antioxidant system and metabolic levels after treatment were used to compare the three means of control. The lowest inhibitory concentration of Zalfexam was 10 mg/mL, at which point C. boninense did not grow, and the inhibition rates of Bacillus velezensis (X7) and Trichoderma harzianum were 33.87-51.85% and 77.86-80.56%, respectively. Superoxide dismutase (SOD) and chitinase were up-regulated 2.43 and 1.24 folds in the Trichoderma harzianum group (M group) and SOD activity was up-regulated 2.2 folds in the Bacillus velezensis group (X7 group) compared to the control group (CK group). SOD, peroxidase (POD), and chitinase activities were elevated in the Zalfexam group (HX group). The differential metabolites in different treatment groups were mainly enriched in amino acid metabolism and production, flavonoid production, and lipid metabolism pathways. Compared with the diseased plants (ZB group), the M, X7, HX, and CK groups were co-enriched in the tryptophan metabolic pathway and glutamate-arginine metabolic pathway, and only the CK group showed a down-regulation of the metabolites in the two common pathways, while the metabolites of the common pathways were up-regulated in the M, X7, and HX groups. In addition, the salicylic acid-jasmonic acid pathway and ascorbic acid-glutathione, which were unique to the M group, played an important role in helping Sorghum sudangrass hybrids to acquire systemic resistance against stress. This study fills the gap in the control of Colletotrichum boninene , which causes leaf spot disease in Sorghum sudangrass hybrids . This paper represents the first reported case of biological control for leaf spot disease in Sorghum sudangrass hybrids and provides a reference for the control of leaf spot disease in Sorghum sudangrass hybrids as well as other crops infected with Colletotrichum boninense .

Plant cell culture technology helps to obtain natural plant-derived metabolites. The callus of sorghum, a prominent cereal crop, possesses various metabolites with potential health benefits. However, the epigenetic mechanism regulating metabolic biosynthetic capabilities in sorghum remains unknown. Therefore, we conducted N6-methyladenine (6mA) methylome analysis using transcriptome profiling and metabolome analysis to investigate the role of 6mA alterations in two calluses having different biosynthetic capacities, which were derived from immature sorghum embryos. Our findings indicate that the 6mA upregulation within gene bodies is crucial in transcriptional activity potentially mediated by the DNA demethylase SbALKBH1. Furthermore, 6mA was significantly enriched in genes involved in the biosynthesis of flavonoids and isoflavonoids. This could serve as a novel source of bioactive compounds for human health. Thus, 6mA could play an essential role in flavonoid biosynthesis in the sorghum callus.

Cereal grains play an important role in human health as a source of macro- and micronutrients, besides phytochemicals. The metabolite diversity was investigated in cereal crops and their milling fractions by untargeted metabolomics ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) of 69 samples: 7 species (barley, oat, pearl millet, rye, sorghum, triticale, and wheat), 23 genotypes, and 4 milling fractions (husk, bran, flour, and wholegrain). Samples were also analyzed by in vitro antioxidant activity. UHPLC-MS/MS signals were processed using XCMS, and metabolite annotation was based on SIRIUS and GNPS libraries. Bran and husk showed the highest antioxidant capacity and phenolic content/diversity. The major metabolite classes were phenolic acids, flavonoids, fatty acyls, and organic acids. Sorghum, millet, barley, and oats showed distinct metabolite profiles, especially related to the bran fraction. Molecular networking and chemometrics provided a comprehensive insight into the metabolic profiling of cereal crops, unveiling the potential of coproducts and super cereals such as sorghum and millet as sources of polyphenols.

Sorghum ratooning, a time and labor-saving cultivation practice, is increasingly being adopted by farmers in Southwest China as an alternative. Efficient N fertilizer management is critical for economical production of sorghum and the long-term protection of the environment. To investigate the impact of N management on grain yield and nitrogen use efficiencies (NUEs) of ratoon sorghum system, a three-year field experiment was conducted for Jinyunuo3 (a hybrid cultivar) and Guojiaohong1 (an inbred cultivar) using 12 combinations of N rates and splitting ratios. The results showed that increasing N rate and splitting application times led to improvements in various growth parameters such as dry matter weight, crop growth rate (CGR), leaf area index (LAI), and photosynthetic potential (PP). The main, ratoon, and annual yields increased with N rate increase, but there was no significant difference between 225 and 150 kg N ha -1 in the ratoon and annual yields. Splitting the application of N fertilizer enhanced grain yield compared to a single dose application method, especially three-split applications yielded higher than two-split applications. Compared with N rates of 225 and 150 kg ha -1 , N rate of 75 kg ha -1 increased apparent recovery rate of applied nitrogen (RE N ), agronomic efficiency of applied nitrogen (AE N ), and partial factor productivity from applied nitrogen (PFP N ) in both main season and whole year. But through splitting application methods at high N rates could achieve similar or even higher levels of NUEs compared to all applied as basal fertilizer at low N rates. Therefore, it could be recommended that applying 150 kg N ha -1 with a basal-jointing-heading fertilizer ratio of 2:4:4 represented an efficient N management practice to synchronously obtain high grain yield and NUEs in ratoon sorghum system in Southwest China., (© 2024. The Author(s).)

The pollen wall protects pollen during dispersal and is critical for pollination recognition. In the Poaceae family, the pollen exine stereostructure exhibits a high degree of conservation with similar patterns across species. However, there remains controversy regarding the conservation of key factors involved in its formation among various Poaceae species. EPAD1 , as a gene specific to the Poaceae family, and its orthologous genes play a conserved role in pollen wall formation in wheat and rice. However, they do not appear to have significant functions in maize. To further confirm the conserved function of EPAD1 in Poaceae, we performed an analysis on four EPAD1 orthologs from two distinct sub-clades within the Poaceae family. The two functional redundant barley EPAD1 genes ( HvEPAD1 and HvEPAD2 ) from the BOP clade, along with the single copy of sorghum ( SbEPAD1 ) and millet ( SiEPAD1 ) from the PACMAD clade were examined. The CRISPR-Cas9-generated mutants all exhibited defects in pollen wall formation, consistent with previous findings on EPAD1 in rice and wheat. Interestingly, in barley, hvepad2 single mutant also showed apical spikelets abortion, aligning with a decreased expression level of HvEPAD1 and HvEPAD2 from the apical to the bottom of the spike. Our finding provides evidence that EPAD1 orthologs contribute to Poaceae specific pollen exine pattern formation via maintaining primexine integrity despite potential variations in copy numbers across different species.

Water scarcity is a major environmental constraint on plant growth in arid regions. Soluble sugars and amino acids are essential osmolytes for plants to cope with osmotic stresses. Sweet sorghum is an important bioenergy crop and forage with strong adaptabilities to adverse environments; however, the accumulation pattern and biosynthesis basis of soluble sugars and amino acids in this species under osmotic stresses remain elusive. Here, we investigated the physiological responses of a sweet sorghum cultivar to PEG-induced osmotic stresses, analyzed differentially accumulated soluble sugars and amino acids after 20% PEG treatment using metabolome profiling, and identified key genes involved in the biosynthesis pathways of soluble sugars and amino acids using transcriptome sequencing. The results showed that the growth and photosynthesis of sweet sorghum seedlings were significantly inhibited by more than 20% PEG. After PEG treatments, the leaf osmotic adjustment ability was strengthened, while the contents of major inorganic osmolytes, including K + and NO 3 - , remained stable. After 20% PEG treatment, a total of 119 and 188 differentially accumulated metabolites were identified in the stems and leaves, respectively, and the accumulations of soluble sugars such as raffinose, trehalose, glucose, sucrose, and melibiose, as well as amino acids such as proline, leucine, valine, serine, and arginine were significantly increased, suggesting that these metabolites should play key roles in osmotic adjustment of sweet sorghum. The transcriptome sequencing identified 1711 and 4978 DEGs in the stems, as well as 2061 and 6596 DEGs in the leaves after 20% PEG treatment for 6 and 48 h, respectively, among which the expressions of genes involved in biosynthesis pathways of sucrose (such as SUS1 , SUS2 , etc.), trehalose (including TPS6 ), raffinose (such as RAFS2 and GOLS2 , etc.), proline (such as P5CS2 and P5CR ), leucine and valine (including BCAT2 ), and arginine (such as ASS and ASL ) were significantly upregulated. These genes should be responsible for the large accumulation of soluble sugars and amino acids under osmotic stresses. This study deepens our understanding of the important roles of individual soluble sugars and amino acids in the adaptation of sweet sorghum to water scarcity.

Background: Solid-state fermentation (SSF) has been widely used in the processing of sorghum grain (SG) because it can produce products with improved sensory characteristics. To clarify the influence of different microbial strains on the SSF of SG, especially on the polyphenols content and composition, Lactiplantibacillus plantarum, Saccharomyces cerevisiae, Rhizopus oryzae, Aspergillus oryzae, and Neurospora sitophila were used separately and together for SSF of SG. Furthermore, the relationship between the dynamic changes in polyphenols and enzyme activity closely related to the metabolism of polyphenols has also been measured and analyzed. Microstructural changes observed after SSF provide a visual representation of the SSF on the SG., Results: After SSF, tannin content (TC) and free phenolic content (FPC) were decreased by 56.36% and 23.48%, respectively. Polyphenol oxidase, β-glucosidase and cellulase activities were increased 5.25, 3.27, and 45.57 times, respectively. TC and FPC were negatively correlated with cellulase activity. A positive correlation between FPC and xylanase activity after 30 h SSF became negative after 48 h SSF. The SG surface was fragmented and porous, reducing the blocking effect of cortex., Conclusion: Cellulase played a crucial role in promoting the degradation of tannin (antinutrient) and phenolic compounds. Xylanase continued to release flavonoids while microbial metabolism consumed them with the extension of SSF time. SSF is an effective way to improve the bioactivity and processing characteristics of SG. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Trait heritability and the response to selection depend on genetic variation, a prerequisite to developing sorghum varieties with desirable agronomic traits and high carbon sequestration for sustainable crop production and soil health. The present study aimed to assess the extent of genetic variability and associations among agronomic and carbon storage traits in selected sorghum genotypes to identify the best candidates for production or breeding. Fifty genotypes were evaluated at Ukulinga, Bethlehem and Silverton sites in South Africa during the 2022/23 growing season. The following agronomic and carbon storage traits were collected: days to 50% heading (DTH), days to 50% maturity (DTM), plant height (PH), total plant biomass (PB), shoot biomass (SB), root biomass (RB), root-to-shoot biomass ratio (RS), grain yield (GY), harvest index (HI), shoot carbon content (SCc), root carbon content (RCc), grain carbon content (GCc), total plant carbon stock (PCs), shoot carbon stock (SCs), root carbon stock (RCs), and root-to-shoot carbon stock ratio (RCs/SCs), and grain carbon stock (GCs). Higher genotypic coefficient of variations (GCVs) were recorded for GY at 45.92%, RB (39.24%), RCs/SCs (38.45), and RCs (34.62). Higher phenotypic coefficient of variations (PCVs) were recorded for PH (68.91%), followed by GY (51.8%), RB (50.51%), RS (41.96%), RCs/SCs (44.90%), and GCs (41.90%). High broad-sense heritability and genetic advance were recorded for HI (83.76 and 24.53%), GY (78.59 and 9.98%), PB (74.14 and 13.18%) and PCs (53.63 and 37.57%), respectively, suggesting a marked genetic contribution to the traits. Grain yield exhibited positive association with HI (r = 0.76; r = 0.79), DTH (r = 0.13; r = 0.31), PH (r = 0.1; r = 0.27), PB (r = 0.01; r = 0.02), RB (r = 0.05; r = 0.06) based on genotypic and phenotypic correlations, respectively. Further, the path analysis revealed significant positive direct effects of SB (0.607) and RB (0.456) on GY. The RS exerted a positive and significant indirect effect (0.229) on grain yield through SB. The study revealed that PB, SB, RB, RS, RCs, and RCs/SCs are the principal traits when selecting sorghum genotypes with high yield and carbon storage capacity., (© 2024. The Author(s).)

Heterosis is a crucial factor in enhancing crop yield, particularly in sorghum (Sorghum bicolor ). This research utilised six sorghum restorer lines, six sorghum sterile lines, and 36 hybrid combinations created through the NCII incomplete double-row hybridisation method. We evaluated the performance of F1 generation hybrids for leaf photosynthesis-related parameters, carbon metabolism-related enzymes, and their correlation with yield traits during the flowering stage. Results showed that hybrid sorghum exhibited significant high-parent heterosis in net photosynthetic rate (P n ), transpiration rate (T r ), stomatal conductance (G s ), apparent leaf meat conductance (AMC), ribulose-1,5-bisphosphate (RuBP) carboxylase, phosphoenolpyruvate (PEP) carboxylase, and sucrose phosphate synthase (SPS). Conversely, inter-cellular carbon dioxide concentration (C i ), instantaneous water uses efficiency (WUE), and sucrose synthase (SuSy) displayed mostly negative heterosis. Traits such as 1000-grain weight (TGW), grain weight per spike (GWPS), and dry matter content (DMC) exhibited significant high-parent heterosis, with TGW reaching the highest value of 82.54%. P n demonstrated positive correlations with T r , C i , G s , RuBP carboxylase, PEP carboxylase, GWPS, TGW, and DMC, suggesting that T r , C i , and G s could aid in identifying high-photosynthesis sorghum varieties. Concurrently, P n could help select carbon-efficient sorghum varieties due to its close relationship with yield. Overall, the F1 generation of sorghum hybrids displayed notable heterosis during anthesis. Combined with field performance, P n at athesis can serve as a valuable indicator for early prediction of the yield potential of the F1 generation of sorghum hybrids and for screening carbon-efficient sorghum varieties.

Background: Sorghum (Sorghum bicolor) is a promising opportunity crop for arid regions of Africa due to its high tolerance to drought and heat stresses. Screening for genetic variability in photosynthetic regulation under salt stress can help to identify target trait combinations essential for sorghum genetic improvement. The primary objective of this study was to identify reliable indicators of photosynthetic performance under salt stress for forage yield within a panel of 18 sorghum varieties from stage 1 (leaf 3) to stage 7 (late flowering to early silage maturity). We dissected the genetic diversity and variability in five stress-sensitive photosynthetic parameters: nonphotochemical chlorophyll fluorescence quenching (NPQ), the electron transport rate (ETR), the maximum potential quantum efficiency of photosystem II (F V /F M ), the CO 2 assimilation rate (A), and the photosynthetic performance based on absorption (PI ABS ). Further, we investigated potential genes for target phenotypes using a combined approach of bioinformatics, transcriptional analysis, and homologous overexpression., Results: The panel revealed polymorphism, two admixed subpopulations, and significant molecular variability between and within population. During the investigated development stages, the PI ABS varied dramatically and consistently amongst varieties. Under higher saline conditions, PI ABS also showed a significant positive connection with A and dry matter gain. Because PI ABS is a measure of plants' overall photosynthetic performance, it was applied to predict the salinity performance index (SPI). The SPI correlated positively with dry matter gain, demonstrating that PI ABS could be used as a reliable salt stress performance marker for forage sorghum. Eight rubisco large subunit genes were identified in-silico and validated using qPCR with variable expression across the varieties under saline conditions. Overexpression of Rubisco Large Subunit 8 increased PI ABS , altered the OJIP, and growth with an insignificant effect on A., Conclusions: These findings provide insights into strategies for enhancing the photosynthetic performance of sorghum under saline conditions for improved photosynthetic performance and potential dry matter yield. The integration of molecular approaches, guided by the identified genetic variability, holds promise for genetically breeding sorghum tailored to thrive in arid and saline environments, contributing to sustainable agricultural practices., (© 2024. The Author(s).)

Cereal seeds are vital for food, feed, and agricultural sustainability because they store and provide essential nutrients to human and animal food and feed systems. Unraveling molecular processes in seed development is crucial for enhancing cereal grain yield and quality. We analyze spatiotemporal transcriptome and metabolome profiles during sorghum seed development in the inbred line 'BTx623'. Morphological and molecular analyses identify the key stages of seed maturation, specifying starch biosynthesis onset at 5 days post-anthesis (dpa) and protein at 10 dpa. Transcriptome profiling from 1 to 25 dpa reveal dynamic gene expression pathways, shifting from cellular growth and embryo development (1-5 dpa) to cell division, fatty acid biosynthesis (5-25 dpa), and seed storage compounds synthesis in the endosperm (5-25 dpa). Network analysis identifies 361 and 207 hub genes linked to starch and protein synthesis in the endosperm, respectively, which will help breeders enhance sorghum grain quality. The availability of this data in the sorghum reference genome line establishes a baseline for future studies as new pangenomes emerge, which will consider copy number and presence-absence variation in functional food traits., (© 2024. The Author(s).)

Main Conclusion: Millets' protein studies are lagging behind those of major cereals. Current status and future insights into the investigation of millet proteins are discussed. Millets are important small-seeded cereals majorly grown and consumed by people in Asia and Africa and are considered crops of future food security. Although millets possess excellent climate resilience and nutrient supplementation properties, their research advancements have been lagging behind major cereals. Although considerable genomic resources have been developed in recent years, research on millet proteins and proteomes is currently limited, highlighting a need for further investigation in this area. This review provides the current status of protein research in millets and provides insights to understand protein responses for climate resilience and nutrient supplementation in millets. The reference proteome data is available for sorghum, foxtail millet, and proso millet to date; other millets, such as pearl millet, finger millet, barnyard millet, kodo millet, tef, and browntop millet, do not have any reference proteome data. Many studies were reported on stress-responsive protein identification in foxtail millet, with most studies on the identification of proteins under drought-stress conditions. Pearl millet has a few reports on protein identification under drought and saline stress. Finger millet is the only other millet to have a report on stress-responsive (drought) protein identification in the leaf. For protein localization studies, foxtail millet has a few reports. Sorghum has the highest number of 40 experimentally proven crystal structures, and other millets have fewer or no experimentally proven structures. Further proteomics studies will help dissect the specific proteins involved in climate resilience and nutrient supplementation and aid in breeding better crops to conserve food security., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The OVATE gene family plays an important role in regulating the development of plant organs and resisting stress, but its expression characteristics and functions in sorghum have not been revealed. In this study, we identified 26 OVATE genes in the sorghum BTx623 genome, which were divided into four groups and distributed unevenly across 9 chromosomes. Evolutionary analysis showed that after differentiation between sorghum and Arabidopsis, the OVATE gene family may have experienced unique expansion events, and all OVATE family members were negatively selected. Transcriptome sequencing and RT-qPCR results showed that OVATE genes in sorghum showed diverse expression characteristics, such as gene SORBl&#95;3001G468900 and SORBl&#95;3009G173400 were significantly expressed in seeds, while SORBI&#95;3005G042700 and SORBI&#95;3002G417700 were only highly expressed in L1. Meantime, in the promoter region, a large number of hormone-associated cis-acting elements were identified, and these results suggest that members of the OVATE gene family may be involved in regulating specific development of sorghum leaves and seeds. This study improves the understanding of the OVATE gene family of sorghum and provides important clues for further exploration of the function of the OVATE gene family., (© 2024. The Author(s).)

Over evolutionary time, plants have developed sophisticated regulatory mechanisms to adapt to fluctuating nitrogen (N) environments, ensuring that their growth is balanced with their responses to N stress. This study explored the potential of L-tryptophan (Trp) in regulating sorghum root growth under conditions of N limitation. Here, two distinct sorghum genotypes (low-N tolerance 398B and low-N sensitive CS3541) were utilized for investigating effect of low-N stress on root morphology and conducting a comparative transcriptomics analysis. Our foundings indicated that 398B exhibited longer roots, greater root dry weights, and a higher Trp content compared to CS3541 under low-N conditions. Furthermore, transcriptome analysis revealed substantial differences in gene expression profiles related to Trp pathway and carbon (C) and N metabolism pathways between the two genotypes. Additional experiments were conducted to assess the effects of exogenous Trp treatment on the interplay between sorghum root growth and low-N tolerance. Our observations showed that Trp-treated plants developed longer root and had elevated levels of Trp and IAA under low-N conditons. Concurrently, these plants demonstrated stronger physiological activities in C and N metabolism when subjected to low-N stress. These results underscored the pivotal role of Trp on root growth and low-N stress responses by balancing IAA levels and C and N metabolism. This study not only deepens our understanding of how plants maintain growth plasticity during environmental stress but also provides valuable insights into the availability of amino acid in crops, which could be instrumental in developing strategies for promoting crop resilience to N deficiency., Competing Interests: Declaration of competing interest The authors declare no conflict of interests., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Sorghum [Sorghum bicolor (L.) Moench] yield is limited by the coincidence of drought during its sensitive stages. The use of cerium oxide nanoparticles in agriculture is minimal despite its antioxidant properties. We hypothesize that drought-induced decreases in photosynthetic rate in sorghum may be associated with decreased tissue water content and organelle membrane damage. We aimed to quantify the impact of foliar application of nanoceria on transpiration rate, accumulation of compatible solutes, photosynthetic rate and reproductive success under drought stress in sorghum. In order to ascertain the mechanism by which nanoceria mitigate drought-induced inhibition of photosynthesis and reproductive success, experiments were undertaken in a factorial completely randomized design or split-plot design. Foliar spray of nanoceria under progressive soil drying conserved soil moisture by restricting the transpiration rate than water spray, indicating that nanoceria exerted strong stomatal control. Under drought stress at the seed development stage, foliar application of nanoceria at 25 mg L -1 significantly improved the photosynthetic rate (19%) compared to control by maintaining a higher tissue water content (18%) achieved by accumulating compatible solutes. The nanoceria-sprayed plants exhibited intact chloroplast and thylakoid membranes because of increased heme enzymes [catalase (53%) and peroxidase (45%)] activity, which helped in the reduction of hydrogen peroxide content (74%). Under drought, compared to water spray, nanoceria improved the seed-set percentage (24%) and individual seed mass (27%), eventually causing a higher seed yield. Thus, foliar application of nanoceria at 25 mg L -1 under drought can increase grain yield through increased photosynthesis and reproductive traits., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The effect on the morpho-physiological parameters and yield of sorghum cultivated in a greenhouse with reclaimed water (RW) and (dehydrated sludge (DS) obtained in a sewage treatment plant, was evaluated. Six treatments (T), with five repetitions each, were carried out in entirely randomized blocks. Water (W) was used in T1 (W) (control), T2 (W + NPK), and T3 (W + DS); RW was used in T4 (RW), T5 (RW + P), and T6 (RW + DS). The results showed that irrigation with only RW (T4), or W + DS (T3) was very suitable for the cultivation since an adequate nutritional supply was provided. The positive effects on the morpho-physiological parameters, plant height, stem diameter and stem length (in cm), were: T3 - 148.8, 1.50, and 103, respectively; T4 - 154, 1.70, and 107, respectively; and on the grain production in weight of 1000 seeds (g), and productivity in grains per plant: T3 - 6.97 and 1453, respectively; T4 - 6.81 and 1636, respectively. Both treatments showed for most of the parameters, no significant differences compared with those of T2 or T5 with supplementary fertilizers. A high production of metabolites (mg g -1 ) like free amino acids was also shown: T3 - 6.45; T4 - 8.43 and proline: T3 - 1.86; T4 - 1.77, known to be a good indication of a plant natural defence against stress conditions, and in soluble protein: T3 - 11.20; T4 - 13.51. Therefore, since the production of such grains with RW or DS can be environmentally and economically beneficial, their use is recommended for small and medium farmers in semiarid regions.

Sorghum ( Sorghum bicolor ), the fifth most important cereal crop globally, serves as a staple food, animal feed, and a bioenergy source. Paclobutrazol-Resistance ( PRE ) genes play a pivotal role in the response to environmental stress, yet the understanding of their involvement in pest resistance remains limited. In the present study, a total of seven SbPRE genes were found within the sorghum BTx623 genome. Subsequently, their genomic location was studied, and they were distributed on four chromosomes. An analysis of cis-acting elements in SbPRE promoters revealed that various elements were associated with hormones and stress responses. Expression pattern analysis showed differentially tissue-specific expression profiles among SbPRE genes. The expression of some SbPRE genes can be induced by abiotic stress and aphid treatments. Furthermore, through phytohormones and transgenic analyses, we demonstrated that SbPRE4 improves sorghum resistance to aphids by accumulating jasmonic acids (JAs) in transgenic Arabidopsis, giving insights into the molecular and biological function of atypical basic helix-loop-helix (bHLH) transcription factors in sorghum pest resistance.

The study evaluated the effects of treating irrigation water with a coaxial flow variator (CFV) on the morpho-physiology of pot-cultivated test species, including cucumber (Cucumis sativus, CU), lettuce (Lactuca sativa, LE), and sorghum (Sorghum vulgare, SO), in early stages of growth. CFV caused a lower oxidation reduction potential (ORP), increased pH and flow resistance and inductance. It induced changes in the absorbance characteristics of water in specific spectral regions, likely associated with greater stretching and reduced bending vibrations compared to untreated water. While assimilation rate and photosynthetic efficiency were not significantly affected at 60 days after sowing, treated water increased the stomatal conductance to water vapour gsw (+79%) and the electron transport rate ETR (+10%) in CU, as well as the non-photochemical quenching NPQ (+33%) in SO. Treated water also reduced leaf temperature in all species (-0.86 °C on average). This translated into improved plant biomass (leaves: +34%; roots: +140%) and reduced leaf-to-root biomass ratio (-42%) in SO, allowing both faster aerial growth and soil colonization, which can be exploited to improve plant tolerance against abiotic stresses. In the C3 species CU and LE, plant biomass was instead reduced, although significantly in LE only, while the leaf-to-root biomass ratio was generally enhanced, a result likely profitable in the cultivation of leafy vegetables. This is a preliminary trial on the effects of functionalized water and much remains to be investigated in other physiological processes, plant species, and growth stages for the full exploitation of this water treatment in agronomy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

The present investigation explored the antifungal effectiveness of Trachyspermum ammi essential oil (TAEO) against Aspergillus flavus, aflatoxin B 1 (AFB 1 ) contamination, and its mechanism of action using biochemical and computational approaches. The GC-MS result revealed the chemical diversity of TAEO with the highest percentage of γ-terpinene (39 %). The TAEO exhibited minimum inhibitory concentration against A. flavus growth (0.5 µL/mL) and AFB 1 (0.4 µL/mL) with radical scavenging activity (IC 50  = 2.13 µL/mL). The mechanism of action of TAEO was associated with the alteration in plasma membrane functioning, antioxidative defense, and carbon source catabolism. The molecular dynamic result shows the multi-regime binding of γ-terpinene with the target proteins (Nor1, Omt1, and Vbs) of AFB 1 biosynthesis. Furthermore, TAEO exhibited remarkable in-situ protection of Sorghum bicolor seed samples against A. flavus and AFB 1 contamination and protected the nutritional deterioration. Hence, the study recommends TAEO as a natural antifungal agent for food protection against A. flavus mediated biodeterioration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: On tropical regions, phosphorus (P) fixation onto aluminum and iron oxides in soil clays restricts P diffusion from the soil to the root surface, limiting crop yields. While increased root surface area favors P uptake under low-P availability, the relationship between the three-dimensional arrangement of the root system and P efficiency remains elusive. Here, we simultaneously assessed allelic effects of loci associated with a variety of root and P efficiency traits, in addition to grain yield under low-P availability, using multi-trait genome-wide association. We also set out to establish the relationship between root architectural traits assessed in hydroponics and in a low-P soil. Our goal was to better understand the influence of root morphology and architecture in sorghum performance under low-P availability., Result: In general, the same alleles of associated SNPs increased root and P efficiency traits including grain yield in a low-P soil. We found that sorghum P efficiency relies on pleiotropic loci affecting root traits, which enhance grain yield under low-P availability. Root systems with enhanced surface area stemming from lateral root proliferation mostly up to 40 cm soil depth are important for sorghum adaptation to low-P soils, indicating that differences in root morphology leading to enhanced P uptake occur exactly in the soil layer where P is found at the highest concentration., Conclusion: Integrated QTLs detected in different mapping populations now provide a comprehensive molecular genetic framework for P efficiency studies in sorghum. This indicated extensive conservation of P efficiency QTL across populations and emphasized the terminal portion of chromosome 3 as an important region for P efficiency in sorghum. Increases in root surface area via enhancement of lateral root development is a relevant trait for sorghum low-P soil adaptation, impacting the overall architecture of the sorghum root system. In turn, particularly concerning the critical trait for water and nutrient uptake, root surface area, root system development in deeper soil layers does not occur at the expense of shallow rooting, which may be a key reason leading to the distinctive sorghum adaptation to tropical soils with multiple abiotic stresses including low P availability and drought., (© 2024. The Author(s).)

Drought stress is one of the most severe natural disasters in terms of its frequency, length, impact intensity, and associated losses, making it a significant threat to agricultural productivity. Sorghum ( Sorghum bicolor ), a C4 plant, shows a wide range of morphological, physiological, and biochemical adaptations in response to drought stress, paving the way for it to endure harsh environments. In arid environments, sorghum exhibits enhanced water uptake and reduced dissipation through its morphological activity, allowing it to withstand drought stress. Sorghum exhibits physiological and biochemical resistance to drought, primarily by adjusting its osmotic potential, scavenging reactive oxygen species, and changing the activities of its antioxidant enzymes. In addition, certain sorghum genes exhibit downregulation capabilities in response to drought stress. Therefore, in the current review, we explore drought tolerance in sorghum, encompassing its morphological characteristics and physiological mechanisms and the identification and selection of its functional genes. The use of modern biotechnological and molecular biological approaches to improving sorghum resistance is critical for selecting and breeding drought-tolerant sorghum varieties.

Background: The sorghum aphid Melanaphis sacchari (Zehntner) (Homoptera: Aphididae) is an important insect in the late growth phase of sorghum (Sorghum bicolor L.). However, the mechanisms of sorghum response to aphid infestation are unclear., Results: In this paper, the mechanisms of aphid resistance in different types of sorghum varieties were revealed by studying the epidermal cell structure and performing a transcriptome and metabolome association analysis of aphid-resistant and aphid-susceptible varieties. The epidermal cell results showed that the resistance of sorghum to aphids was positively correlated with epidermal cell regularity and negatively correlated with the intercellular space and leaf thickness. Transcriptome and metabolomic analyses showed that differentially expressed genes in the resistant variety HN16 and susceptible variety BTX623 were mainly enriched in the flavonoid biosynthesis pathway and differentially expressed metabolites were mainly related to isoflavonoid biosynthesis and flavonoid biosynthesis. The q-PCR results of key genes were consistent with the transcriptome expression results. Meanwhile, the metabolome test results showed that after aphidinfestation, naringenin and genistein were significantly upregulated in the aphid-resistant variety HN16 and aphid-susceptible variety BTX623 while luteolin was only significantly upregulated in BTX623. These results show that naringenin, genistein, and luteolin play important roles in plant resistance to aphid infestation. The results of exogenous spraying tests showed that a 1‰ concentration of naringenin and genistein is optimal for improving sorghum resistance to aphid feeding., Conclusions: In summary, the physical properties of the sorghum leaf structure related to aphid resistance were studied to provide a reference for the breeding of aphid-resistant varieties. The flavonoid biosynthesis pathway plays an important role in the response of sorghum aphids and represents an important basis for the biological control of these pests. The results of the spraying experiment provide insights for developing anti-aphid substances in the future., (© 2024. The Author(s).)

Introduction: Millets, owing to their rich nutritional and low-to-moderate glycemic index values, are termed superfoods; however, some anti-nutritional factors, such as tannins, limit the absorption of micro and macronutrients. Non-thermal processing technologies, such as fermentation, can improve nutrient content and reduce these anti-nutritional factors., Methods: The effect of a controlled submerged fermentation of whole grain sorghum, pearl millet, and dehusked Kodo millet using mixed lactic acid bacteria (LAB) culture in tofu whey-based media on the proximate, antioxidant, tannin content, vitamin B, amino acids profile and estimated glycemic index (eGI) of different millets were evaluated., Results: The protein content (2-12.5%), carbohydrate content (2-13.6%), antioxidant activity (3-49%), vitamin B complex, amino acid profile (89-90%), and eGI of whole grain sorghum, pearl millet, and dehusked Kodo millet improved due to LAB-assisted submerged fermentation. In contrast, fat (4-15%), ash (56-67%), crude fiber (5-34%), minerals, tannin and resistant starch content decreased due to LAB fermentation., Conclusion: Controlled LAB fermentation can improve the nutritional quality of sorghum and millets while reducing anti-nutritional factors. This non-thermal process can be adopted industrially to produce more palatable and nutritionally superior millet products., Competing Interests: Author Yogesh Sharma is employed by Waters India Pvt. Ltd., New Delhi. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Sorghum germplasm showed grain Fe and Zn genetic variability, but a few varieties were biofortified with these minerals. This work contributes to narrowing this gap. Fe and Zn concentrations along with 55,068 high-quality GBS SNP data from 140 sorghum accessions were used in this study. Both micronutrients exhibited good variability with respective ranges of 22.09-52.55 ppm and 17.92-43.16 ppm. Significant marker-trait associations were identified on chromosomes 1, 3, and 5. Two major effect SNPs (S01&#95;72265728 and S05&#95;58213541) explained 35% and 32% of Fe and Zn phenotypic variance, respectively. The SNP S01&#95;72265728 was identified in the cytochrome P450 gene and showed a positive effect on Fe accumulation in the kernel, while S05&#95;58213541 was intergenic near Sobic.005G134800 (zinc-binding ribosomal protein) and showed negative effect on Zn. Tissue-specific in silico expression analysis resulted in higher levels of Sobic.003G350800 gene product in several tissues such as leaf, root, flower, panicle, and stem. Sobic.005G188300 and Sobic.001G463800 were expressed moderately at grain maturity and anthesis in leaf, root, panicle, and seed tissues. The candidate genes expressed in leaves, stems, and grains will be targeted to improve grain and stover quality. The haplotypes identified will be useful in forward genetics breeding., (© 2024. The Author(s).)