Wheat Spikes Research Articles

Fusarium head blight resistance measured by type II inoculation method among UK wheat cultivars and investigations into the relationship with anthers trapped and anthers retained

AbstractThere is a paucity of information on the relationship between anthers and Type II expression of Fusarium Head Blight resistance (measured by point inoculation of a spikelet and observing disease spread over the wheat spike). Twenty‐nine UK commercial wheat varieties were evaluated for the expression of Type II resistance. Anther extrusion was measured by counting the number of spikelets with anthers trapped between the lemma and palea (anthers trapped). In five spikelets where trapped anthers were not observed, the number of spikelets with anthers contained within the spikelet was counted (anthers retained). There was variation in expression in the expression of Type II resistance (p < .001), anthers trapped (p < .001) and anthers retained (p < .001). Anthers trapped and anthers retained were not correlated with each other, indicating both should be considered independently. Later maturity correlated with great resistance in the AHDB Type I resistance ratings but to greater susceptibility to point inoculations (Type II). Consistent with previous literature higher numbers of anthers trapped to increase susceptibility to FHB Type I. However, the relationships were reversed for the expression of resistance measured by point inoculation within the spikelet, where anthers trapped between the lemma and palea reduced susceptibility to Type II point inoculation (p < .01). This suggests a benefit of releasing some pollen from within the spikelet for this inoculation method. Similarly, anthers retained contributed to increased Type II susceptibility (p < .05), potentially providing substrates within the spikelet inoculation site aiding fungal growth, but correlated to greater Type I resistance (p < .05) indicating the anthers are hidden from initial infection on the outside of the spikelet, particularly under low infection pressure. While preliminary, this work suggests that variation in anthers trapped, anthers retained and maturity date may increase Type II expression of resistance and could relate to increased Type I FHB susceptibility.

基于轻量级深度学习网络RepYOLO模型的麦穗实时检测

Real-time detection has become an essential component in intelligent agriculture and industry. In this paper, a real-time wheat spike detection method based on the lightweight deep learning network RepYOLO is proposed. Addressing the small and densely packed phenotype characteristics of wheat spikes, the channel attention mechanism module CBAM from the traditional YOLOv4 algorithm is introduced and multiple convolutional kernels are merged using a structural reparameterization method. Additionally, the ATSS algorithm is incorporated to enhance the accuracy of object detection. These approaches significantly reduce the model size, improve the inference speed, and lower the memory access cost. To validate the effectiveness of the model, it is trained and tested on a large dataset of diverse wheat spike images representing various phenotypes. The experimental results demonstrate that the RepYOLO algorithm achieves an average accuracy of 98.42% with a detection speed of 8.2 FPS. On the Jetson Nano platform, the inference speed reaches 34.20 ms. Consequently, the proposed model effectively reduces the memory access cost of deep learning networks without compromising accuracy and successfully improves the utilization of CPU/MCU limited performance.

Mining of QTLs for Spring Bread Wheat Spike Productivity by Comparing Spring Wheat Cultivars Released in Different Decades of the Last Century.

Genome-wide association studies (GWAS) are among the genetic tools for the mining of genomic loci associated with useful agronomic traits. The study enabled us to find new genetic markers associated with grain yield as well as quality. The sample under study consisted of spring wheat cultivars developed in different decades of the last century. A panel of 186 accessions was evaluated at VIR's experiment station in Pushkin across a 3-year period of field trials. In total, 24 SNPs associated with six productivity characteristics were revealed. Along with detecting significant markers for each year of the field study, meta-analyses were conducted. Loci associated with useful yield-related agronomic characteristics were detected on chromosomes 4A, 5A, 6A, 6B, and 7B. In addition to previously described regions, novel loci associated with grain yield and quality were identified during the study. We presume that the utilization of contrast cultivars which originated in different breeding periods allowed us to identify new markers associated with useful agronomic characteristics.

Reduction of Fusarium head blight and trichothecene contamination in transgenic wheat expressing Fusarium graminearum trichothecene 3-O-acetyltransferase.

Fusarium graminearum, the causal agent of Fusarium head blight (FHB), produces various mycotoxins that contaminate wheat grains and cause profound health problems in humans and animals. Deoxynivalenol (DON) is the most common trichothecene found in contaminated grains. Our previous study showed that Arabidopsis-expressing F. graminearum trichothecene 3-O-acetyltransferase (FgTRI101) converted DON to 3-acetyldeoxynivalenol (3-ADON) and excreted it outside of Arabidopsis cells. To determine if wheat can convert and excrete 3-ADON and reduce FHB and DON contamination, FgTRI101 was cloned and introduced into wheat cv Bobwhite. Four independent transgenic lines containing FgTRI101 were identified. Gene expression studies showed that FgTRI101 was highly expressed in wheat leaf and spike tissues in the transgenic line FgTri101-1606. The seedlings of two FgTri101 transgenic wheat lines (FgTri101-1606 and 1651) grew significantly longer roots than the controls on media containing 5 µg/mL DON; however, the 3-ADON conversion and excretion was detected inconsistently in the seedlings of FgTri101-1606. Further analyses did not detect 3-ADON or other possible DON-related products in FgTri101-1606 seedlings after adding deuterium-labeled DON into the growth media. FgTri101-transgenic wheat plants showed significantly enhanced FHB resistance and lower DON content after they were infected with F. graminearum, but 3-ADON was not detected. Our study suggests that it is promising to utilize FgTRI101, a gene that the fungus uses for self-protection, for managing FHB and mycotoxin in wheat production.

A rapid, low-cost wheat spike grain segmentation and counting system based on deep learning and image processing

Spike grain number is a crucial parameter when estimating wheat yield. However, most methods predominantly focus on a single period, lacking universality. To achieve rapid and intelligent wheat spike grain counting from the filling stage to the mature stage, this study used different smartphones to obtain wheat ear images of different varieties and constructs the spike grain dataset required for deep learning segmentation model through image normalization and data enhancement. Then we propose a segmentation model called Multi-Attention TransUNet (MATransUNet) and a spike grain counting model (SGCountM) to achieve wheat spike grain counting. Our results indicate that, comparing with other models, MATransUNet achieves the best segmentation performance with strong robustness and generalization capabilities, achieving a mean intersection over union(mIoU) of 85.93%. Through the comparative analysis with manual counting results, our Method I, doubling the number of grains on one side of the ears, results in a root mean squared error (RMSE) of 4.38 and a coefficient of determination(R2) of 0.85; while the Method II, adding the grains on both sides of the ears, yields a RMSE of 3.13 and a R2 of 0.93. The counting accuracy significantly improves compared to not using the SGCountM. Moreover, transfer learning notably enhances the accuracy of the segmentation and counting model, enabling accurate spike grain counting from the filling stage to the mature stage. Finally, we integrate MATransUNet and SGCountM to design and implement a wheat spike grain segmentation and counting system (WeChat mini program). In practical tests, the mini program can effectively, accurately segments and counts wheat spike grain, demonstrating its feasibility and effectiveness. This research provides valuable insights into the intelligent application of wheat spike grain counting.

Dissecting the molecular basis of spike traits by integrating gene regulatory networks and genetic variation in wheat

Spike architecture influences both grain weight and grain number per spike, which are the two major components of grain yield in bread wheat (Triticum aestivum L.). However, the complex wheat genome and the influence of various environmental factors pose challenges in mapping the causal genes that affect spike traits. Here, we systematically identified genes involved in spike trait formation by integrating information on genomic variation and gene regulatory networks controlling young spike development in wheat. We identified 170 loci that are responsible for variations in spike length, spikelet number per spike, and grain number per spike through genome-wide association study and meta-QTL analyses. We constructed gene regulatory networks for young inflorescences at the double ridge stage and the floret primordium stage, in which the spikelet meristem and the floret meristem are predominant, respectively, by integrating transcriptome, histone modification, chromatin accessibility, eQTL, and protein–protein interactome data. From these networks, we identified 169 hub genes located in 76 of the 170 QTL regions whose polymorphisms are significantly associated with variation in spike traits. The functions of TaZF-B1, VRT-B2, and TaSPL15-A/D in establishment of wheat spike architecture were verified. This study provides valuable molecular resources for understanding spike traits and demonstrates that combining genetic analysis and developmental regulatory networks is a robust approach for dissection of complex traits.

Bronchoscopic removal of an aspirated vegetal foreign body in a small Münsterländer

Foreign bodies are common in pets and may cause vague clinical signs. One of the locations a foreign body can be found is the bronchial region. The foreign body itself, but also secondary inflammation and infection can lead to severe tissue reactions. Bronchoscopy is a useful diagnostic imaging tool, due to its diagnostic ability and therapeutic possibilities to directly remove foreign bodies. In the present case, a small Münsterländer was presented with a one-week existing productive cough. The dog inhaled a wheat spike into the airways, subsequently causing the development of secondary inflammation and infection. Through bronchoscopy, the foreign body was located in the left mainstem bronchus and removed successfully with a special forceps.

Graphene Quantum Dots (GQD)-Mediated dsRNA Delivery for the Control of Fusarium Head Blight Disease in Wheat.

Graphene quantum dots (GQDs), a class of fluorescent carbon materials, have displayed significant potential in various fields such as energy devices, catalysis, sensing, bioimaging, and drug delivery. Because of their extremely small size, generally less than 100 nm, they also have tremendous potential in plant science research, especially for the delivery of nucleic acids, breaking the barrier of cell walls. In this study, we synthesized GQDs with a size range of 2-5 nm, characterized them, and surface-functionalized them with branched polyethylenimine (bPEI). We then used the surface-functionalized GQDs as carriers to deliver double-stranded RNA (dsRNA) that target two growth-and-development-related genes in Fusarium graminearum─the causative organism of the Fusarium head blight disease of wheat. The successful binding of dsRNA to GQDs-bPEIs was demonstrated through gel-shifting assays, showcasing the potential for efficient dsRNA delivery. We designed dsRNAs targeting the MGV1 and RAS1 genes of F. graminearum by using the pssRNAit pipeline, ensuring high specificity and no off-target effects. The coding sequences of the designed dsRAS1 and dsMGV1 were cloned into the L4440 vector and transformed into the Escherichia coli HT115 strain for dsRNA production. Fungal culture analysis revealed that the inclusion of dsRNAs in potato dextrose agar (PDA) media effectively slowed down the growth. Exogenous spraying experiments both in plate cultures and in intact wheat spikes demonstrated that the dsRNA:GQDs-bPEIs treatment was more effective in restricting fungal mycelium growth or the number of infected spikelets compared to naked dsRNA treatment. Our study demonstrates the promising potential of graphene quantum dots as carriers for dsRNA-based fungal disease management in wheat and other crops.

Phylogenetic, metabolic and pathogenic characteristics of Alternaria alternata strains from wheat in China

AbstractAlternaria is an important toxigenic fungal genus endangering the quality of cereal grains. No data are available in the literature regarding the composition of Alternaria species in wheat from the main wheat‐cultivating regions of China. In this study, a total of 282 strains were isolated from wheat kernels collected in various provinces. Phylogenetic characterization was performed using two informative loci, ASA10 and ASA19, demonstrating that all strains belonged to A. alternata. A low molecular diversity was observed, which did not appear to be influenced by geographic origin. The secondary metabolite profile of the strains was evaluated in rice culture. Altertoxin I was synthesized at a high concentration by the majority of the strains, while the accumulation of altenuene was lower in both frequency and level than that of alternariol and its derivative. A. alternata did not cause readily observable symptoms on wheat spikes after spraying of the conidia at anthesis, but a higher quantity of altertoxin I and Alternaria DNA were detected in inoculated grains. Artificial inoculations of apple fruit showed that all examined strains were capable of causing typical rot symptoms and producing all four toxins to a certain degree. The widespread occurrence of A. alternata on wheat grains and its ability to synthesize various toxic metabolites may pose a serious hazard for both growers and consumers.

Systematic identification of wheat spike developmental regulators by integrated multi-omics, transcriptional network, GWAS, and genetic analyses

The spike architecture of wheat plays a crucial role in determining grain number, making it a key trait for optimization in wheat breeding programs. In this study, we used a multi-omic approach to analyze the transcriptome and epigenome profiles of the young spike at eight developmental stages, revealing coordinated changes in chromatin accessibility and H3K27me3 abundance during the flowering transition. We constructed a core transcriptional regulatory network (TRN) that drives wheat spike formation and experimentally validated a multi-layer regulatory module involving TaSPL15, TaAGLG1, and TaFUL2. By integrating the TRN with genome-wide association studies, we identified 227 transcription factors, including 42 with known functions and 185 with unknown functions. Further investigation of 61 novel transcription factors using multiple homozygous mutant lines revealed 36 transcription factors that regulate spike architecture or flowering time, such as TaMYC2-A1, TaMYB30-A1, and TaWRKY37-A1. Of particular interest, TaMYB30-A1, downstream of and repressed by WFZP, was found to regulate fertile spikelet number. Notably, the excellent haplotype of TaMYB30-A1, which contains a C allele at the WFZP binding site, was enriched during wheat breeding improvement in China, leading to improved agronomic traits. Finally, we constructed a free and open access Wheat Spike Multi-Omic Database (http://39.98.48.156:8800/#/). Our study identifies novel and high-confidence regulators and offers an effective strategy for dissecting the genetic basis of wheat spike development, with practical value for wheat breeding.

Advantageous spike-to-stem competition for assimilates contributes to the reduction in grain number loss in wheat spikes under water deficit stress

This study was conducted to investigate the mechanisms of assimilate distribution and sugar metabolism in spike-stem that inhibit the formation of fertile florets and grains per spike in winter wheat under pre-reproductive drought stress. Two winter wheat cultivars, CH58 (relatively strongly drought tolerant) and LH6 (relatively weakly drought tolerant), were subjected to successive soil drought treatments from jointing to heading during the 2020–2022 growing seasons. The results showed that pre-reproductive drought stress intensified the degradation and abortion of wheat florets. Compared to CH58, the decrease in the number of fertile florets and grains per spike of LH6 under drought stress increased by an average of 5.3%−8.0% and 8.3%−9.0%, respectively. Drought significantly inhibited the distribution of 13C-photosynthates in wheat spikes (15.7%−24.7%) and stems (8.5%−11.7%) during the booting stage. The number of differentially expressed genes enriched in starch and sucrose metabolic pathways was much higher in the spike and stem of LH6 than in those of CH58. Drought significantly reduced sucrose and hexose in young spikes but increased hexose and fructan concentrations in stems. Compared to LH6, the higher invertase activity and accompanying high expression of sugar transporter protein (STP) in CH58 spikes under drought stress contributed to the utilization of sucrose in young spikes. Additionally, under severe drought, the higher fructan concentration and expression of sugar transport proteins (SWEET and SUT) in the stem of CH58 improved the ability of the stem to transport assimilates to young spikes and thus alleviated the loss of grain number per spike. Exogenous spermidine optimized hexose and sucrose allocation in young wheat spikes and stems after drought stress, thereby increasing the number of fertile florets under drought stress. Overall, the ability of the dominant spike to utilize sugar and the ability to compete for assimilates between the spike and stem contribute to the resistance to drought stress-induced grain reduction in the spike. Exogenous chemicals can regulate the number of fertile florets through this pathway, thereby promoting the formation of wheat grain number.

Image-based classification of wheat spikes by glume pubescence using convolutional neural networks.

Pubescence is an important phenotypic trait observed in both vegetative and generative plant organs. Pubescent plants demonstrate increased resistance to various environmental stresses such as drought, low temperatures, and pests. It serves as a significant morphological marker and aids in selecting stress-resistant cultivars, particularly in wheat. In wheat, pubescence is visible on leaves, leaf sheath, glumes and nodes. Regarding glumes, the presence of pubescence plays a pivotal role in its classification. It supplements other spike characteristics, aiding in distinguishing between different varieties within the wheat species. The determination of pubescence typically involves visual analysis by an expert. However, methods without the use of binocular loupe tend to be subjective, while employing additional equipment is labor-intensive. This paper proposes an integrated approach to determine glume pubescence presence in spike images captured under laboratory conditions using a digital camera and convolutional neural networks. Initially, image segmentation is conducted to extract the contour of the spike body, followed by cropping of the spike images to an equal size. These images are then classified based on glume pubescence (pubescent/glabrous) using various convolutional neural network architectures (Resnet-18, EfficientNet-B0, and EfficientNet-B1). The networks were trained and tested on a dataset comprising 9,719 spike images. For segmentation, the U-Net model with EfficientNet-B1 encoder was chosen, achieving the segmentation accuracy IoU = 0.947 for the spike body and 0.777 for awns. The classification model for glume pubescence with the highest performance utilized the EfficientNet-B1 architecture. On the test sample, the model exhibited prediction accuracy parameters of F1 = 0.85 and AUC = 0.96, while on the holdout sample it showed F1 = 0.84 and AUC = 0.89. Additionally, the study investigated the relationship between image scale, artificial distortions, and model prediction performance, revealing that higher magnification and smaller distortions yielded a more accurate prediction of glume pubescence.

Dual RNA-sequencing of Fusarium head blight resistance in winter wheat.

Fusarium head blight (FHB) is a devastating fungal disease responsible for significant yield losses in wheat and other cereal crops across the globe. FHB infection of wheat spikes results in grain contamination with mycotoxins, reducing both grain quality and yield. Breeding strategies have resulted in the production of FHB-resistant cultivars, however, the underlying molecular mechanisms of resistance in the majority of these cultivars are still poorly understood. To improve our understanding of FHB-resistance, we performed a transcriptomic analysis of FHB-resistant AC Emerson, FHB-moderately resistant AC Morley, and FHB-susceptible CDC Falcon in response to Fusarium graminearum. Wheat spikelets located directly below the point of inoculation were collected at 7-days post inoculation (dpi), where dual RNA-sequencing was performed to explore differential expression patterns between wheat cultivars in addition to the challenging pathogen. Differential expression analysis revealed distinct defense responses within FHB-resistant cultivars including the enrichment of physical defense through the lignin biosynthesis pathway, and DON detoxification through the activity of UDP-glycosyltransferases. Nucleotide sequence variants were also identified broadly between these cultivars with several variants being identified within differentially expressed putative defense genes. Further, F. graminearum demonstrated differential expression of mycotoxin biosynthesis pathways during infection, leading to the identification of putative pathogenicity factors.

Correlação entre resistência à brusone em cultivares de trigo e taxa de esporulação de conídios de Pyricularia oryzae Triticum

ABSTRACT: The use of resistant wheat cultivars is a fundamental strategy to minimize the damages caused by blast, a disease caused by the fungus Pyricularia oryzae Triticum (PoT). The objetive of this stydy was to (a) evaluate the reaction to blast of Brazilian wheat cultivars and (b) determine whether there is correlation between severity of symptoms on wheat spikes and “sporulation rate of PoT conidia per gram of wheat spike rachis” (Rscon). Plants of 16 wheat cultivars were grown in greenhouse until flowering (Zadoks stage 65), when their spikes were inoculated with a suspension formed by mixing the conidia of three PoT isolates. The evaluated variables were blast severity on spikes at 5, 7 and 11 days after inoculation (dai) and Rscon. Rachis were evaluated individually to determine the Rscon. Correlation analyzes were carried out between blast severity means on spikes of cultivars at 5, 7 and 11 dai and Rscon. The cultivars TBIO Mestre, TBIO Aton, ORS 1401, ORS 1403, ORS Destak, ORS Feroz, and CD 116 stood out for being classified in the statistical groups with the highest resistance to blast for the four variables considered in the study. The correlation between blast severity on spikes at 5, 7 and 11 dai and Rscon is at most moderate.

WheatNet: Attentional Path Aggregation Feature Pyramid Network for Precise Detection and Counting of Dense and Arbitrary-Oriented Wheat Spikes

WheatNet: Attentional Path Aggregation Feature Pyramid Network for Precise Detection and Counting of Dense and Arbitrary-Oriented Wheat Spikes

From Fields to Pixels: UAV Multispectral and Field-Captured RGB Imaging for High-Throughput Wheat Spike and Kernel Counting

From Fields to Pixels: UAV Multispectral and Field-Captured RGB Imaging for High-Throughput Wheat Spike and Kernel Counting

The Wheat Endophyte Epicoccum layuense J4-3 Inhibits Fusarium graminearum and Enhances Plant Growth.

Fungal endophytes are well-known for their ability to promote plant growth and hinder fungal diseases, including Fusarium head blight (FHB) caused by Fusarium graminearum. This study aimed to characterize the biocontrol efficacy of strain J4-3 isolated from the stem of symptomless wheat collected from Heilongjiang Province, China. It was identified as Epicoccum layuense using morphological characteristics and phylogenetic analysis of the rDNA internal transcribed spacer (ITS) and beta-tubulin (TUB). In a dual culture assay, strain J4-3 significantly inhibited the mycelial growth of F. graminearum strain PH-1 and other fungal pathogens. In addition, wheat coleoptile tests showed that lesion symptoms caused by F. graminearum were significantly reduced in wheat seedlings treated with hyphal fragment suspensions of strain J4-3 compared to the controls. Under field conditions, applying spore suspensions and culture filtrates of strain J4-3 with conidial suspensions of F. graminearum on wheat spikes resulted in the significant biocontrol efficacy of FHB. In addition, wheat seedlings previously treated with spore suspensions of strain J4-3 before sowing successfully resulted in FHB reduction after the application of conidial suspensions of F. graminearum at anthesis. More importantly, wheat seedlings treated with hyphal fragments and spore suspensions of strain J4-3 showed significant increases in wheat growth compared to the controls under greenhouse and field conditions. Overall, these findings suggest that E. layuense J4-3 could be a promising biocontrol agent (BCA) against F. graminearum, causing FHB and a growth-promoting fungus in wheat.

Fusarium head blight in wheat: Impact of growing season, wheat variety and nitrogen fertilization under natural infection

AbstractThe fungal genus Fusarium encompasses a diverse group of species responsible for synthesizing mycotoxins, particularly deoxynivalenol, fumonisin, and zearalenone and inducing Fusarium head blight in wheat. The research was undertaken over a period of two consecutive growing seasons (2020 and 2021) on the premises and facilities of the Hungarian University of Agriculture and Life Sciences (MATE). The objective of this study was to investigate the impact of growing season, nitrogen fertilization, and wheat variety on Fusarium infection as well as mycotoxin contamination in wheat kernel. Zearalenone was absent throughout the course of the two growing seasons, whereas deoxynivalenol was found solely in 2020. The findings demonstrate that nitrogen fertilization failed to exhibit a statistically significant impact on both Fusarium infection and mycotoxin production. The impact of wheat variety on Fusarium infection and deoxynivalenol was not found to be statistically significant. However, it exerted a significant effect on fumonisin production. The growing season exerted a statistically significant impact on the incidence of Fusarium infection and the ensuing contamination with mycotoxins, attributable to augmented precipitation levels in 2021 compared to 2020, specifically during the flowering period when the spike of wheat is highly susceptible to Fusarium infection.

Low sucrose availability reduces basal spikelet fertility by inducing abscisic acid and jasmonic acid synthesis in wheat.

Within a spike of wheat, the central spikelets usually generate three to four fertile florets, while the basal spikelets generate zero to one fertile floret. The physiological and transcriptional mechanism behind the difference in fertility between the basal and central spikelets is unclear. This study reports a high temporal resolution investigation of transcriptomes, number and morphology of floret primordia, and physiological traits. The W6.5-W7.5 stage was regarded as the boundary to distinguish between fertile and abortive floret primordia; those floret primordia reaching the W6.5-W7.5 stage during the differentiation phase (3-9 d after terminal spikelet stage) usually developed into fertile florets in the next dimorphism phase (12-27 d after terminal spikelet stage), whereas the others aborted. The central spikelets had a greater number of fertile florets than the basal spikelets, which was associated with more floret primordia reaching the W6.5-W7.5 stage. Physiological and transcriptional results demonstrated that the central spikelets had a higher sucrose content and lower abscisic acid (ABA) and jasmonic acid (JA) accumulation than the basal spikelets due to down-regulation of genes involved in ABA and JA synthesis. Collectively, we propose a model in which ABA and JA accumulation is induced under limiting sucrose availability (basal spikelet) through the up-regulation of genes involved in ABA and JA synthesis; this leads to floret primordia in the basal spikelets failing to reach their fertile potential (W6.5-W7.5 stage) during the differentiation phase and then aborting. This fertility repression model may also regulate spikelet fertility in other cereal crops and potentially provides genetic resources to improve spikelet fertility.

Residual Effect of Biochar and Legumes on Soil Fertility, Yield and Yield Components of Wheat

Biochar and the use of legumes in cropping systems are considered sustainable approaches to boost crop yield and preserve soil fertility. In the current study, the effects of leftover biochar and previously planted legumes on wheat yield and soil N status were examined at various nitrogen (N) levels. The experiment included testing two levels of previously applied biochar (0 and 50 tons ha-1), three legumes under four levels of N (0, 60, 90, and 120 kg ha-1), cowpea (Vigna unguiculata) for fodder, Sesbenia (Sesbenia grandaflora) for green manuring, and mung bean (Vigna radiata) for grain. Results showed that biochar application enhanced wheat tiller m-2, spikes m-2, grains per spike, thousand grain weight, grain yield, biological yield, and soil total N status by 3%, 6.5%, 3.7%, 1.8%, 7.8%, 9.5%, and 11%, respectively. Moreover, applying nitrogen at a rate of 90 kg ha-1 increased the amount of wheat spike m-2 by 20%, grain spike-1 by 10%, grain yield by 70%, biological yield by 48%, harvest index by 27%, and the N content of the grain, straw, and soil by 13%, 14%, and 36% respectively. Meanwhile, 1000 grain weight resulted higher by 6.17%. Legumes that had been previously seeded outperformed fallow and increased spikes m-2, grain yield, biological yield, grain N content, and soil total N content by 8.2%, 11%, 6.78%, 25%, and 42%, respectively. It is determined that applying biochar to the summer gap left by legumes can increase soil fertility and wheat output.