Background: Leaf angle is a key determinant of plant architecture and yield under high-density planting. Loss of the ligule reduces leaf angle, offering a strategy for breeding compact maize. This study aims to identify the genetic basis of the liguleless mutant jd30 and develop a functional marker for breeding. Methods: The jd30 mutant was phenotypically characterized and compared with the wild type (WT). Genetic analysis was performed using F2 populations. Map-based cloning was conducted with simple sequence repeat (SSR) and insertion/deletion (InDel) markers. Candidate genes were annotated and sequenced. Allelism tests were performed by crossing jd30 with other lg1 mutants. A Cleaved Amplified Polymorphic Sequence (CAPS) marker was developed based on the causal mutation. Results: The jd30 mutant showed complete ligule loss from the V2 stage onward and had a significantly smaller leaf angle than the wild type. The phenotype is controlled by a single recessive nuclear gene, mapped to a 123-kb interval on chromosome 2 containing four candidate genes. A single-base cytosine (C) deletion at nucleotide 522 of ZmLG1 caused a frameshift and premature stop codon. Two other lg1 mutants, 20292 and 29163, carried distinct mutations—a C insertion and multiple base substitutions, respectively. Allelism tests confirmed jd30 as a novel allele of ZmLG1. A co-dominant CAPS marker, M5, was developed to distinguish wild-type and mutant alleles. Conclusions: This study identifies a novel loss-of-function allele of ZmLG1, designated ZmLG1-1, and the M5 marker, providing valuable genetic resources for modifying leaf architecture and improving maize plant compactness.

Wheat is one of the most important food crops in the world, and grain number per spike (GNS) is one of the most important factors affecting wheat grain yield. Therefore, identifying genes controlling GNS is important for wheat production. However, wheat is an allohexaploid species and has a highly complex genome, and the isolation of wheat genes is challenging. In this study, we identified a candidate gene, TraesCS4B02G047100 (TaRLK-4B), associated with GNS using both quantitative trait locus (QTL) mapping and genome-wide association study (GWAS) based on RNA-Seq. We obtained three homozygous mutant lines, bb-1, bb-2 and bb-3, using the CRISPR/Cas9 gene editing system. Compared with Fielder (wild type, WT), the three mutant lines exhibited significant reductions in GNS, total spikelets per spike (TSS) and spike length (SL). These results indicated that TaRLK-4B positively regulates GNS and its related traits TSS and SL. The TaRLK-4B gene contains two exons and one intron and belongs to the largest subfamily of LRR-RLKs. We performed RNA-Seq analysis using spikes from the WT and bb-2 mutant line at the tillering stage. A total of 1,193 differentially expressed genes (DEGs) were identified, including previously cloned TaSPL17 homologous genes in the three subgenomes and their orthologous gene OsIPA1 (OsSPL14) in rice. Combining RNA-Seq data of TaRLK-4B and DAP-Seq data of the TaSPL17-7D, we identified 136 overlapping genes which are likely downstream targets of TaSPL17-7D. Therefore, we hypothesized that TaRLK-4B represents a novel putative component of the TaSPL17-centered regulatory network. In addition, haplotype analysis revealed that Haplotype 2 (Hap2) is a favorable haplotype for increasing GNS, but the thousand-grain weight (TGW) was not significantly different between Haplotype 1 (Hap1) and Hap2.

IntroductionWheat is a major global food crop, and improving its yield is essential for food security. Identifying genetic loci and candidate genes associated with yield-related traits is crucial for molecular breeding.MethodsA panel of 768 common wheat varieties from domestic and international sources was evaluated over two consecutive years (2024 and 2025) in Tai’an. Three growth-stage traits, eight agronomic traits, and three seedling-stage traits, including root length, seedling weight, and seedling height, were measured. Genotyping was performed using a 55K SNP array, and a genome-wide association study was conducted using a linear mixed model.ResultsPopulation structure and kinship analyses classified the varieties into primitive landraces and modern cultivars. A total of 49,768 high-quality SNPs were identified, covering 82.73% of the genome with an average marker density of 0.29 Mb. Among them, 17,293, 18,313, and 14,162 SNPs were located on the A, B, and D subgenomes, respectively. Association analysis revealed 544 SNP loci significantly associated with yield-related traits across 21 chromosomes, of which 49 were consistently detected in both years. Additionally, 60 loci were associated with two or more traits, indicating potential pleiotropic effects, and were distributed on 16 chromosomes (excluding 3D, 4B, 4D, 7A, and 7B).DiscussionThis study identifies SNP markers associated with key traits across growth stages in common wheat and provides a basis for the discovery of candidate genes and favorable alleles. These findings may facilitate the utilization of elite genetic resources for yield improvement in wheat breeding.

GWAS of 17 spike traits in 246 wheat varieties identifies three novel QTLs, 70 candidate genes, and seven validated KASP markers for spike architecture. Spike morphology is a critical determinant of wheat yield, influencing grain formation and photoassimilate partitioning. To dissect its genetic basis, we conducted a genome-wide association study (GWAS) on 17 spike-related traits. A natural population comprising 246 wheat varieties was evaluated in five field environments over two growing seasons. Genotyping was performed using a 60 K SNP array, yielding 36,591 high-quality markers (36,554 SNPs and 37 InDels) for analysis. GWAS identified 1,246 significant markers, including 139 with pleiotropic effects (associated with ≥ 2 traits) and 26 that were stable across multiple environments (traits detected in ≥ 3 environments). These stable markers delineated 16 quantitative trait loci (QTLs) for eight key traits, such as spike length, grain dimensions, and spikelet density. Among these, we discovered three novel QTLs: one for spike length and two for total spikelet number per spike. Gene annotation within the QTL intervals yielded 98 candidate genes. Subsequent expression analysis refined this set to 70 high-confidence candidates, five of which are known wheat genes with potential novel roles in spike development. To facilitate breeding, we developed seven Kompetitive Allele-Specific PCR (KASP) markers and subsequently validated them in an independent cohort: five for spike length and two for spikelet density. This study provides novel genetic insights and practical molecular tools, facilitating marker-assisted selection for improved wheat spike architecture and yield.

AIMS:Excessive NO3--N and NO2--N accumulation threatens aquaculture water quality, highlighting the need for aerobic microorganisms capable of converting dissolved inorganic nitrogen into biomass-associated organic nitrogen. This study aimed to characterize the assimilatory NO3-/NO2- reduction mechanism of Priestia megaterium BZ-95 and to evaluate its potential for aquaculture wastewater treatment. METHODS AND RESULTS:A NO3--reducing bacterium, P. megaterium BZ-95, was investigated using whole-genome analysis, gene expression profiling, and nitrogen-removal assays. Genome analysis revealed two nitrogen-assimilation modules, namely a NO3- assimilation cluster (nasC-nirB_2-narT) and a nitrite assimilation operon (nirB_1-nirD-nirC-cysG), where as no denitrification or DNRA genes were detected. In NO3--N medium, NO3--N decreased from 72.59 to 8.07 mg/l within 24 h, accompanied by transient NH4+ accumulation. In NO2--N medium, NO2--N decreased from 70.00 to 0.009 mg/l within 21 h, with no detectable NH4+ accumulation. In mixed-N medium, NO3--N and NO2--N decreased from 30.00 to 3.33 mg/l and from 30.00 to 0.99 mg/l, respectively, within 24 h. Gene expression analysis showed that NO3- mainly induced the nasC-nirB_2-narT module, whereas NO2- strongly induced the nirB_1-nirD-nirC-cysG module. In addition, the mutant BZ-95D4 showed a 61.08% higher NO3--N removal capacity than the parental strain BZ-95. CONCLUSIONS:Priestia megaterium BZ-95 possesses a dual-module assimilatory nitrogen-reduction system with distinct responses to NO3- and NO2-. These findings improve understanding of microbial nitrate assimilation and indicate that BZ-95, particularly the improved strain BZ-95D4, is a promising candidate for aquaculture wastewater treatment.

Nile tilapia (Oreochromis niloticus) mono-sex male culture is widely adopted due to the faster growth rate of males. High-temperature-induced sex reversal is an effective sex control technology. However, the cellular and molecular mechanisms underlying spermatogenesis in high-temperature-induced XX pseudo-males remain unclear. In this study, we performed single-nucleus RNA sequencing to compare testicular transcriptomes between genetic XY males and XX pseudo-males. We obtained high-quality transcriptional profiles of 6175 cells from genetic males and 6395 cells from pseudo-males, which were annotated into 13 cell types including 4 germ cell subtypes and 8 somatic cell subtypes. While genetic males and pseudo-males shared an identical repertoire of testicular cell types, significant transcriptomic differences were observed in both germ cells and the somatic niche. Specifically, spermatogonia from pseudo-males exhibited dysregulated expression of apoptosis- and proliferation-related genes, which likely contributes to the reduced spermatogonial numbers reported in previous studies. In contrast, pseudo-male spermatocytes showed substantially elevated expression of cell cycle (ccni, ccnd2) and meiosis (sycp1, sycp3) genes, which may enable them to maintain normal sperm production. Furthermore, Leydig cells from pseudo-males showed elevated expression of steroidogenic genes (cyp17a1, hsd11b2, star2, etc.) throughout testicular development. Sertoli cells showed downregulation of male-biased genes (gsdf, stat3 and jarid2b) and upregulation of female-biased genes (nr5a2, ctnnb1 and wt1a), suggesting that a dynamic balance between TGF-β and Wnt/β-catenin signaling is key to temperature-induced sex reversal. These findings provide new insights into the molecular basis of spermatogenesis in sex-reversed fish, which will facilitate the optimization of high-temperature-induced sex reversal technologies in aquaculture.

As a staple cereal crop cultivated worldwide,common wheat (Triticum aestivum L.) accounts for approximately 35％ of global dietary energy requirements (Sharma and Sharma 2025). However,agricultural yields are currently under significant pressure due to the combined impacts of anthropogenic climate change and geopolitical conflicts (Ortiz-Bobea et al. 2021;Hultgren et al. 2025),which threatens global food security. These challenges necessitate the urgent development of molecular breeding approaches to improve wheat production sustainably.

IntroductionThe success of allopolyploids is partly attributable to their ability to resolve genomic conflicts during the early generations following allopolyploidization. Codon usage patterns represent important genomic signatures, and codon usage bias can influence gene expression and cellular function through multiple processes. However, little is known about how subgenomic conflicts in codon usage are resolved during allopolyploidization.MethodsWe characterized changes in codon usage from the wild progenitors of bread wheat to modern bread wheat. In addition, we generated full-length transcriptomes of Aegilops tauschii (male parent), Triticum durum (female parent), their triploid hybrids, and spontaneously doubled allohexaploid wheat to investigate changes in codon usage associated with hybridization and genome doubling.ResultsThe evolutionarily divergent subgenomes of bread wheat exhibited similar codon usage patterns, whereas the codon usage patterns of bread wheat differed markedly from those of its wild progenitors, which tended to have higher GC content at the third codon position (GC3). These differences were associated with asymmetric changes in subgenomic GC3 during the transition from the wild progenitors to bread wheat. Changes in GC3 were also associated with sharp decreases in the average number of transcripts from GC3-poor genes. Analysis of the synthetic allopolyploidization system further revealed distinct changes in codon usage during successive stages of bread wheat allopolyploidization, including hybridization and genome doubling. In particular, genome doubling was accompanied by an increase in GC3, contributing to the GC3-rich codon usage bias observed in bread wheat.DiscussionThese findings reveal dynamic remodeling and convergence of codon usage patterns during wheat allopolyploidization and suggest that hybridization and genome doubling contribute differently to this process. Our results provide new insights into the evolution of codon usage landscapes in allopolyploids and may improve our understanding of subgenomic accommodation following allopolyploidization.

Nocardia seriolae is a prevalent opportunistic pathogen that primarily affects immunocompromised or surface-damaged fish, and it can result in persistent, progressive infections with high mortality rates. However, the proliferation dynamics of N. seriolae during chronic infection and the immune responses of host macrophages remain poorly understood. In this study, we established an in vivo infection model using fluorescently labeled N. seriolae in northern snakehead. Cryosectioning and semi-quantitative PCR analyses revealed that N. seriolae progressively expanded within host tissues, colonizing target organs within 2-3 days via hematogenous dissemination and subsequently inducing granuloma formation. Complete bacterial clearance was observed in advanced granulomas, and this was accompanied by extensive macrophage apoptosis within the lesions. Antibodies against macrophage-expressed gene 1 (Mpeg1) and mannose receptor C1 (Mrc1) were used to label total macrophages and M2 macrophage-polarized subsets, respectively. Immunofluorescence analysis demonstrated that during granuloma maturation, M2 macrophages were localized to the granuloma periphery, and M1 macrophages were in close contact with pathogens in the granuloma core; this spatial arrangement likely contributed to the containment and elimination of bacteria. Our study is the first to demonstrate tissue-specific N. seriolae proliferation in fish and changes in macrophage subsets during infection. These findings provide important insights into the interactions between N. seriolae and its host and have implications for the development of targeted therapies for nocardial infections.

While intensive aquaculture has developed rapidly, the consequent buildup of nitrogenous compounds, poses a critical threat to aquatic organisms. Microbial degradation offers an environmentally sustainable solution. We investigated the metabolic regulatory capacity of Priestia megaterium BZ-95 under four nitrogen regimes-ammonium (NH4+-N), nitrite (NO2--N), nitrate (NO3--N), and a mixture of them (Mix)-using comparative transcriptomics. We revealed that BZ-95 in NH4+-N activated a direct assimilation program prioritizing branched-chain amino acid biosynthesis. Conversely, under nitrate, BZ-95 enhanced membrane transport and 2-oxocarboxylic acid metabolism to facilitate the rapid incorporation of nitrate-derived ammonium into biomass. Nitrite stress triggered a coordinated response involving the assimilatory nir module (nirC-nirB-nirD) and enhanced energy metabolism to meet the heightened demand for reducing power during its rapid reduction. Under mixed nitrogen sources, BZ-95 established a highly synergistic carbon-nitrogen network, simultaneously processing multiple nitrogen inputs without a hierarchical preference, highlighting its remarkable metabolic plasticity. Intersection analysis defined a refined core of 692 nitrite-specific DEGs and revealed broad transcriptional activation under nitrite stress. Analysis of the NO2--specific core identified enhanced transmembrane transport capacity, coupled with auxiliary metabolic tuning, as central adaptive strategies for nitrite processing. Collectively, these findings provide crucial insights into the molecular basis of nitrogen coordination in P. megaterium BZ-95.

Hepcidin is an antimicrobial peptide involved in innate immunity, and the addition of hepcidin or probiotics expressing hepcidin to feed can significantly enhance the disease resistance of farmed animals. Lactobacillus plantarum has the advantages of stable colonization in the animal gut, promotion of intestinal health, and efficient expression of foreign proteins. Previously, we have used L. plantarum (LP37) as a feed additive to promote the growth of Micropterus salmoides and Oreochromis mossambicus. The aim of this study is to construct a recombinant LP37 expressing hepcidin and evaluate its role in improving antibacterial ability and growth promotion in M. salmoides. Based on the expression vector pSIP403, we used green fluorescent protein (GFP) as a reporter gene to compare the effects of four promoters on the expression levels of foreign proteins and selected the promoter TUFA with the highest expression level. By introducing the ribosomal binding site (RBS) and secretory signal peptide sequence (SP-PGH), we constructed a novel vector capable of efficiently expressing M. salmoides hepcidin (MsHep) extracellularly. Transformation of the novel vector into LP37 to obtain the recombinant strain: LP37-MsHep. This recombinant LP37-MsHep demonstrated considerable antibacterial activity against Aeromonas hydrophila and several other microbes. The dietary supplementation of recombinant LP37-MsHep for 60 days significantly enhanced the survival rates of M. salmoides and reduced tissue bacterial load in M. salmoides post infection with A. hydrophila, and regulated gene expression of FPN1, IL-10, TNF- α , and MHC-II involved in iron metabolism and immunity. Similar to LP37, dietary supplementation of recombinant LP37-MsHep also significantly improved the final body weight (FBW), weight gain ratio (WGR), and specific growth rate (SGR) and reduced the feed coefficient rate (FCR). Collectively, the recombinant LP37-MsHep is capable of significantly improving growth performance and disease resistance of M. salmoides, presenting it as a promising green feed additive in aquaculture with broad application potential.

The evaluation of genetic diversity in germplasm resources is fundamental to crop breeding. A total of 183 oat germplasm resources were evaluated through field trials at Xinjin District and Shandan County, located in southern and northern China, respectively. Phenotypic and agronomic traits were assessed, including six qualitative and sixteen quantitative characteristics. Results revealed significant variation in panicle attitude and grain color, based on the statistical analysis using SPSS. Among the sixteen quantitative traits, coefficient of variation ranged from 4.92% to 48.02% with the second internode thickness exhibiting the highest genetic diversity index. Correlation analysis of sixteen quantitative traits was performed using R Studio, and the results indicated significant positive relationships between grain weight and several ear characteristics, including spikelet number, ear length, layer numbers, and grain numbers per ear. Principal component analysis categorized the sixteen quantitative phenotypic traits into five independent factors. The structural equation modeling using SPSS-AMOS indicated that ear characteristics showed strong direct contributions to grain weight, establishing it as a key indicator for future breeding efforts. The multiple correspondence analysis by R Studio suggested that a total of nineteen oat germplasm resources showed the grain and biomass production potential across both experimental regions.

Reducing plant height (PH) plays an important role in reducing lodging and increasing wheat yield. The potential of wheat yields has reached a plateau in recent years. However the discovery and application of new plant height genes could further increases wheat yield potential. This study located a wheat dwarfing gene downstream of Rht1 on chromosome 4B through QTL mapping, called the phosphatidylinositol 4-kinase alpha (PI4KA-4B). The TaPI4KA-4B gene is located on the cell membrane, and the mating based split-ubiquitin system assays (mbSUS) showed that this gene interacts with TaCSN7 (COP9 signalosome subunit 7). Furthermore, this gene was knocked out in the variety Fielder using the CRISPR/Cas9 gene editing technology. Two homozygous mutant genotypes, AAbbDD (-1 bp) and AAbbDD (-2 bp), were obtained by multi-generation hybridization. Compared with the wild type, the plant height of the mutant was significantly reduced by 6-8 cm, and the stem cell length was significantly shortened with irregular cell morphology. RNA-seq analysis of wild type fielder and mutant plants revealed that the differentially expressed genes (DEGs) were mainly involved in microtubule-based processes, macromolecule biosynthesis, carbohydrate metabolism, and cytoskeletal composition, implying that TaPI4KA-4B regulates plant height through modulating organic synthesis and cytoskeletal reorganization. These results indicate that TaPI4KA-4B is a novel Rht gene that controls plant height in wheat.

Numerous studies have reported a significant positive correlation between wheat yield and the quantity of wheat heads. However, collecting data on wheat heads in the field poses a challenge for several reasons, including the uncontrollable nature of the environment, inconsistent data quality, and ambiguous data truth. To address these challenges, we developed a simulation strategy to replicate the conditions of a real wheat field, which enabled the data collection process to be conducted indoors over a short period. After applying grayscale image processing to process the simulated wheat images, we trained and tested nine deep learning models: Faster-RCNN, YOLOv7, YOLOv8, CenterNet, SSD, RetinaNet, EfficientDet, Deformable-DETR and DINO. Our results indicated that YOLOv7 performed the best (R2 = 0.963, RMSE = 2.463). We then compared our model trained on simulated wheat data to a model trained on real wheat data (R2 = 0.963 vs 0.972, RMSE = 2.463 vs 2.692). We also achieved good model performance on five test sets: GWHD, SDAU2021-SDAU2024. The results demonstrated the efficacy of our simulation, which provides an efficient and convenient strategy for the precision agriculture community.

Wheat (Triticum aestivum L.) is a vital staple crop globally, with its grain microelement content playing a crucial role in human nutrition and health. In this study, the concentrations of eight essential microelements (micronutrients and toxic elements): iron (Fe), manganese (Mn), copper (Cu), zinc (Zn), selenium (Se), chromium (Cr), cadmium (Cd), and arsenic (As), were quantified in 272 wheat varieties using inductively coupled plasma mass spectrometry (ICP-MS) under three different environments. A genome-wide association study (GWAS) was conducted using 176,357 molecular markers, comprising 163,223 single-nucleotide polymorphisms (SNPs) and 13,134 insertion-deletion (InDels) variants, identified through RNA sequencing. A total of 196 significant markers associated with microelement content traits were identified across 21 chromosomes in various environments. Of these, 14 significant markers consistently appeared across environments, forming 13 QTLs and linking to 45 candidate genes. Among these, 29 genes were homologs of known genes in Arabidopsis and rice, while 16 were novel candidates. Haplotype analysis indicated significant phenotypic variation in microelement accumulation, with TraesCS6A02G204300Hap2 notably enhancing iron content. This study provides valuable insights into the genetic architecture of microelement accumulation in wheat grains and introduces novel genetic resources for breeding wheat varieties aimed at improving micronutrient content and ensuring food safety.

Low temperature (LT) in spring has become one of the principal abiotic stresses that restrict the growth and development of wheat. Diverse analyses were performed to investigate the mechanism underlying the response of wheat grain development to LT stress during booting. These included morphological observation, measurements of starch synthase activity, and determination of amylose and amylopectin content of wheat grain after exposure to treatment with LT during booting. Additionally, proteomic analysis was performed using tandem mass tags (TMT). Results showed that the plumpness of wheat grains decreased after LT stress. Moreover, the activities of sucrose synthase (SuS, EC 2.4.1.13) and ADP-glucose pyrophosphorylase (AGPase, EC 2.7.7.27) exhibited a significant reduction, leading to a significant reduction in the contents of amylose and amylopectin. A total of 509 differentially expressed proteins (DEPs) were identified by proteomics analysis. The Gene Ontology (GO) enrichment analysis showed that the protein difference multiple in the nutritional repository activity was the largest among the molecular functions, and the up-regulated seed storage protein (SSP) played an active role in the response of grains to LT stress and subsequent damage. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that LT stress reduced the expression of DEPs such as sucrose phosphate synthase (SPS), glucose-1-phosphate adenylyltransferase (glgC), and β-fructofuranosidase (FFase) in sucrose and starch metabolic pathways, thus affecting the synthesis of grain starch. In addition, many heat shock proteins (HSPs) were found in the protein processing in endoplasmic reticulum pathways, which can resist some damage caused by LT stress. These findings provide a new theoretical foundation for elucidating the underlying mechanism governing wheat yield development after exposure to LT stress in spring.

Nicosulfuron, a widely utilized herbicide, is detrimental to some maize varieties due to their sensitivity. Developing tolerant varieties with resistance genes is an economical and effective way to alleviate phytotoxicity. In this study, map-based cloning revealed that the maize resistance gene to nicosulfuron is Zm00001eb214410 (CYP81A9), which encodes a cytochrome P450 monooxygenase. qRT- PCR results showed that CYP81A9 expression in the susceptible line JS188 was significantly reduced compared to the resistant line B73 during 0-192 hours following 80 mg/L nicosulfuron spraying. Meanwhile, a CYP81A9 overexpression line exhibited normal growth under a 20-fold nicosulfuron concentration (1600 mg/L), while the transgenic acceptor background material Zong31 did not survive. Correspondingly, silencing CYP81A9 through CRISPR/Cas9 mutagenesis and premature transcription termination mutant EMS4-06e182 resulted in the loss of nicosulfuron resistance in maize. Acetolactate Synthase (ALS), the target enzyme of nicosulfuron, exhibited significantly reduced activity in the roots, stems, and leaves of susceptible maize post-nicosulfuron spraying. The CYP81A9 expression in the susceptible material was positively correlated with ALS activity in vivo. Therefore, this study identified CYP81A9 as the key gene regulating nicosulfuron resistance in maize and discovered three distinct haplotypes of CYP81A9, thereby laying a solid foundation for further exploration of the underlying resistance mechanisms.

In wheat ( Triticum aestivum L.), yield component traits (YCTs) are the most important yield traits. Only several genes for YCTs have been originally cloned. The efficient cloning of genes for YCTs directly from wheat remains a challenge. Here, we proposed a strategy for cloning genes from quantitative trait loci (QTLs) by sequencing of recombinant inbred lines (RILs) (QTL-Seq-RIL). Using the ‘TN18 × LM6’ RIL population as an example, we identified 138 candidate unigenes (CUGs) for YCTs from 77 stable QTLs. The average of CUGs per QTL was 1.8, which enabled us to confirm the CUGs directly. We have confirmed seven CUGs, TaIFABPL, TaDdRp, TaRLK, TaTD, TaTFC3, TaKMT and TaSPL17 , via the CRISPR/Cas9 system. Of these, six genes were found firstly to regulate YCTs in crops except for TaSPL17 . Five CUGs (include TaSPL17 ) for which orthologous genes have been cloned previously with the same or similar agronomic functions. It is to say, 11 CUGs were preliminarily validated using a single RIL population. QTL-Seq-RIL provides an efficient method for rapid gene cloning using existing RIL populations.### Competing Interest StatementThe authors have declared no competing interest.

Background: Sulfur (S) is a vital element for the normal growth and development of plants, performing crucial biological functions in various life processes. Methods: This study investigated thirteen S utilization efficiency (SUE)-related traits at the seedling stage of wheat using a recombinant inbred line (RIL) population. The quantitative trait loci (QTLs) were mapped by genetic mapping. Thirteen S utilization efficiency-related traits were investigated under two hydroponic culture trials with low S (0.1S, T1), moderate S (0.5S, T2), and high S (1.5S, T3) levels, using the wheat RILs. Results: A total of 170 QTLs for the thirteen traits in different treatment environments were identified. Among them, 89, 103, and 101 QTLs were found in T1, T2, and T3, respectively. A total of 63 QTLs were found in the multiple treatment environments, the other 107 QTLs only being detected in a single treatment environment. Among them, thirteen relatively high-frequency QTLs (RHF-QTLs) and eleven QTL clusters were found. Five (QSh-1D, QRn-1D, QSdw-1D, QTdw-1D, and QTsc-1D) and six (QRdw-6A, QSdw-6A, QTdw-6A, QRsc-6A, QSsc-6A, and QTsc-6A) RHF-QTLs were identified in QTL clusters C3 and C10, respectively. Conclusion: These thirteen RHF-QTLs and eleven QTL clusters are expected to apply to the molecular marker-assisted selection (MAS) of wheat.

The cuticular wax, acting as the ultimate defense barrier, is essential for the normal morphogenesis of plant organs. Despite this importance, the connection between wax composition and leaf development has not been thoroughly explored. In this study, we characterized a new maize mutant, ragged leaf4 (rgd4), which exhibits crinkled and ragged leaves starting from the sixth leaf stage. The phenotype of rgd4 is conferred by ZmCER1, which encoding an aldehyde decarbonylase involved in wax biosynthesis. ZmCER1 function deficient mutant displayed reduced cuticular wax density and disordered bulliform cells (BCs), while ZmCER1 overexpressing plants exhibited the opposite effects, indicating that ZmCER1 regulates cuticular wax biosynthesis and BCs development. Additionally, as the density of cuticular wax increased, the water loss rate of detached leaf decreases, suggesting that ZmCER1 is positively correlated with plant drought tolerance.