To obtain the superior genotype of HvHinb-1 gene related to the hardness of hulless barley grains, two hulless barley varieties with significant differences in hardness were seleeted: Suma (hardness value: 14.66±1.55) and Nanmulin (hardness value: 21.42±1.83). The HvHinb-1 gene sequence through PCR were doned to analyzed the sequence characteristics, protein physicochemical properties, and expression patterns of different genotypes, and explored their roles in the formation of hulless barley grain hardness. The distribution and genotype of HvHinb-1 in 21 hulless barley varieties with different hardness were analyzed, and the expression of HvHinb-1 in different stages of grain development (early, middle and late milk ripening) was detected by real-time quantitative PCR (qRT-PCR).The results showed that the HvHinb-1 was cloned from both the Suma and Nanmulin, with a sequence identity of 98.64%, and two amino acid differences. HvHinb-1 is a hydrophilic basic protein with an isoelectric point of 8.69 and a molecular weight of 16.12 to 16.14 kDa, primarily composed of α-helices and random coils. Phylogenetic analysis indicated that hulless barley HvHinb-1 has the closest genetic relationship to wheat. In the preliminary identification of 12 samples of low-hardness hulless barley (hardness value range: 9.04-15.04), the amino acid sequence at the mutation sites 78 and 94 was H, and in 9 samples of high-hardness hulless barley (hardness value range: 18.93-22.76), the amino acid sequence at the mutation sites 78 and 94 was Q. qRT-PCR analysis showed that in early milk ripening grains, the expression level of HvHinb-1 was significantly higher in Suma than that in Nanmulin, and in mid-milk ripening, it was extremely higher in Nanmulin than in Suma, suggesting that this period may be crucial for grain hardness formation.

To achieve rapid pre-harvest prediction of grain protein content (GPC) in winter wheat, this study developed an indirect prediction method integrating multispectral vegetation indices and texture features. Based on nitrogen fertilization experiments and spectral data collected from 2022 to 2024, texture parameters were extracted using Gray-level co-occurrence matrix (GLCM). Leaf nitrogen content (LNC) served as the critical intermediary to link spectral models with GPC, establishing a multispectral-based prediction model for winter wheat GPC. The results demonstrated that during the early grain-filling stage, vegetation indices combined with soil-background-filtered texture features achieved optimal LNC estimation. The exponential model y=0.281exp (0.097x), showed superior performance (r²=0.787, RMSE=0.221 g·kg^-1); significant correlations were observed between LNC and GPC across key growth stages, with correlation coefficients of 0.780(anthesis), 0.810 (early grain-filling), 0.704(mid-grain-filling), and 0.714(late grain-filling); the exponential model y=7.160exp (0.018x), developed using early grain-filling stage data, achieved the highest GPC prediction accuracy [r²=0.697, RMSE=0.096 g· (100 g)^-1]. The established early grain-filling stage GPC prediction model enhances field management optimization, enables grain quality classification, and provides technical support for applying remote sensing technology in high-quality wheat production and precision agriculture.

To clarify the physiological mechanisms by which exogenous brassinolide (Brassinolide, BR) regulates floret degeneration and grain formation in wheat, a spring wheat cultivar Yangmai 15 was used as the material. During the booting stage, plants were sprayed with 0, 0.1, 0.2, or 0.4 mg·L^-1 BR. The results showed that compared with un-spraying BR, spraying BR significantly reduced the floret degeneration rate and increased the number of grains per spike and grain weight. The 0.2 mg·L^-1 BR treatment was the most effective, increasing grains per spike by 4.76 and grain weight per spike by 7.58%. Spraying BR signifficant improved the chlorophyll content, maximum photochemical efficiency, and net photosynthetic rate from 5 to 25 days after spraying. Spraying BR promoted the accumulation of photosynthetic assimilates and their translocation to the spike. 10 days after spraying, sucrose content in the spike increased by 10.47% under 0.2 mg·L^-1 BR; sucrose synthase activity also increased, accompanied by upregulated expression of key sucrose-metabolism genes, including TaSUT1, TaSUS1, TaCWI, and TaAGPL1. Spraying BR modulated endogenous hormone balance in the spike: the 0.2 mg·L^-1 BR treatment increased gibberellin content by 13.42%, decreased abscisic acid content by 34.86%, and significantly raised the IAA/ABA and GA₃/ABA ratios by 37.10% and 42.71%, respectively. Spraying BR improved endosperm development. After anthesis, the relative areas of amyloplasts and protein bodies in endosperm cells under 0.2 mg·L^-1 BR increased significantly with 27.12% and 18.98% respectively. In conclusion, exogenous BR effectively reduced floret degeneration and promoted grain formation by enhancing photosynthetic capacity, optimizing assimilate partitioning, and regulating sucrose metabolism and hormonal homeostasis. This led to higher grains per spike and grain weight per spike, and ultimately increased yield. Under the experimental conditions, the optimal BR concentration was 0.2 mg·L^-1.

Drought is the primary agrometeorological disaster affecting spring wheat growth in the Hexi Corridor. Accurately identifying its drought characteristics is crucial for field management and disaster prevention and mitigation in local spring wheat cultivation. Based on the daily meteorological data from Wuwei Meteorological Station and the growth stage, soil relative humidity (R_sm) and yield data of spring wheat from the Agrometeorological Experiment Station during 1995-2023, the effective precipitation (P_e), crop water requirement (ET_c), and crop water deficit index (CWDI) during the growth periods of spring wheat were calculated, and regression analysis, Spearman correlation analysis, and Mann-Kendall (M-K) detection were employed to reveal the water supply-demand dynamics and drought characteristics of spring wheat. The results showed that the ET_c during the growth period of spring wheat in the Hexi Corridor significantly exceeded P_e during 1995-2023. Under non-irrigation conditions, severe water imbalance occurred, with the peak water deficit reached 58.7 mm during jointing-heading stage. Drought frequency analysis showed frequent drought occurrences across all growth stages without irrigation, particularly during the jointing-heading stage, where the frequency of severe drought reached 86%. Under irrigation, only the sowing-three leaf stage exhibited a severe drought frequency of 55%, but the actual yield reduction rate was only 2.7%-9.4%. The interannual fluctuations of CWDI were significant between 1995 and 2023, under non-irrigation conditions, continuous drought persisted throughout the growth periods, with CWDI reaching 85% during the heading-grain filling stage. The CWDI showed an extremely significant upward trend at 9.5% a^-1 (P<0.01), with an abrupt change occurring in 2014-2015. Under irrigation, the CWDI showed a monomodal pattern, with an overall increase rate slowing down to 1.4% a^-1, however significantly downward trend at 52.0% a^-1 (P<0.05) during three leaf-jointing stage. Although CWDI reached 81% during sowing-three leaf stage, winter irrigation ensured soil moisture, which basically met crop requirements. Climate change intensified water stress on spring wheat. Adopting precision irrigation technologies such as drip irrigation during the critical water requirement stages of spring wheat can effectively mitigate the adverse impacts of climate change induced spring droughts and late spring early summer droughts on crops. This approach ensures normal growth and development of spring wheat and stabilized yields.

NF-YB transcription factors play key regulatory roles in plant growth, development, and stress responses. In this study, HMMER and BLAST were used to identify members of the wheat NF-YB gene family, and their physicochemical properties, phylogenetic relationships, and gene structures were analyzed. Using wheat varieties including Lantian 134, Longzimai 1, and Chang 6878, the expression pattern of TaNF-YB-2 at the seedling stage under drought and salt stress was examined. A total of 10 TaNF-YB family members were identified in the wheat genome and sequentially named TaNF-YB-1 to TaNF-YB-10. These proteins were unstable and acidic. The TaNF-YB members in common wheat showed high similarity to their homologs in spelt wheat, indicating evolutionary conservation. TaNF-YBs genes were mainly located on homologous groups 1 and 4, with five-gene pairs showing collinearity. Cis-element analysis suggested that TaNF-YB gene family are involved in stress responses. Under drought and salt stress, the expression of TaNF-YB-2 increased in both roots and leaves. Under drought stress, the relative expression levels of TaNF-YB-2 in three drought-resistant varieties (Lantian 134, Longzimai 1, and Chang 6878) were consistently significantly higher than in two drought-sensitive varieties (Xikemai 518 and Lanhangxuan 121). Under salt stress, the relative expression levels of TaNF-YB-2 in the three salt-tolerant varieties were significantly higher than those in the salt-sensitive materials—in leaves after 4 h and in roots after 6 h of treatment.

Thousand-kernel weight is one of the key yield components in modern wheat breeding where significant progress has been made through genetic improvement. It is crucial to conduct in-depth research on the important genes controlling wheat kernel weight and their functions. In this study, molecular markers for five genes related to wheat kernel weight (TaSus2-2B, TaGW8-B1, TaCwi-A1, TaGS-D1, and TaGW2-6A) were used to test 263 new wheat varieties (lines) collected from the 2023-2024 national trials in the Huanghuai wheat region. Among these varieties (lines), all kernel weight-related genes existed in combinations. There were nine types of combinations for the five prior haplotypes, specifically: TaGW8-B1a+TaGW2-6A-A, TaSus2-2Ba+TaGW8-B1a+TaGW2-6A-A, TaGW8-B1a+TaGS-D1a+TaCwi-A1a, TaGW8-B1a+TaGS-D1a+TaGW2-6A-A, TaGW8-B1a+TaCwi-A1a+TaGW2-6A-A, TaSus2-2Ba+TaGW8-B1a+TaGS-D1a+TaGW2-6A-A, TaGW8-B1a+TaGS-D1a+TaCwi-A1a+TaGW2-6A-A, TaSus2-2Ba+TaGW8-B1a+TaCwi-A1a+TaGW2-6A-A, and TaSus2-2Ba+TaGW8-B1a+TaGS-D1a+TaCwi-A1a+TaGW2-6A-A. Among these combinations, the TaSus2-2Ba+TaGW8-B1a+TaGW2-6A-A combination had the lowest frequency at 1.90%, while the TaGW8-B1a+TaGS-D1a+TaCwi-A1a+TaGW2-6A-A combination had the highest frequency at 31.56%. Further evaluation and analysis of kernel traits in 2023 in Zhengzhou and Zhumadian and in 2024 in Xinxiang revealed that among these nine gene combination types, the TaGW8-B1a+TaGW2-6A-A combination exhibited a relatively weak effect on increasing wheat kernel weight, whereas the combination type with five pyramided superior kernel weight-related alleles(TaSus2-2Ba+TaGW8-B1a+TaGS-D1a+TaCwi-A1a+TaGW2-6A-A) showed a relatively strong effect on improving wheat kernel weight. The results of this study indicated that increasing the number of superior kernel weight-related genes(haplotypes) could effectively enhance the kernel weight of wheat varieties in the Huanghuai wheat region.

To investigate the molecular mechanism of high-temperature all-stage stripe rust resistance in Xiaoyan 6, an E3 ubiquitin ligase gene, TaPUB23, was identified based on the previous transcriptome sequencing results. This gene, which was significantly upregulated under both stripe rust infection and high-temperature conditions, has a full-length sequence of 1,272 bp, encoding 423 amino acids, and is localized in the cell membrane, cytoplasm, and nucleus. Virus-induced transient overexpression of TaPUB23 significantly reduced the stripe rust resistance of Xiaoyan 6 plant. Based on the yeast two hybrid (Y2H) system, we identified the interaction target TaDJA7 of TaPUB23 and demonstrated that TaPUB23 ubiquitinated TaDJA7. The interaction between TaPUB23 and TaDJA7 was further validated through luciferase complementation assays (LCA), bimolecular fluorescence complementation (BiFC), and Pull-down assays. These findings reveal that the E3 ubiquitin ligase TaPUB23 regulates high-temperature all-stage stripe rust resistance in wheat through ubiquitination of TaDJA7.

The aleurone layer of blue-grained wheat is rich in anthocyanins, which offer numerous health benefits to humans. To elucidate the molecular mechanisms underlying anthocyanin synthesis and regulation in the aleurone layer, this study compared the blue-grained wheat material E10-2B with its white-grained counterpart E10-2W using agronomic trait evaluation, molecular markers, liquid array technology, and transcriptome analysis. The results showed that compared to E10-2W, E10-2B exhibited a 15.31% reduction in plant height, indicating a dwarfing trait. However, its panicle length, 1 000-grain weight, and grain length decreased by 17.93%, 5.99%, and 4.40%, respectively. Molecular marker and liquid chip analyses revealed that E10-2B contains the complete genome of common wheat, along with the 4E chromosome from Thinopyrum ponticum. The grain color of E10-2B undergoes a transformation between 20 and 25 days post-anthesis, with 25 days post-anthesis identified as the critical time point for anthocyanin accumulation. Transcriptome sequencing analysis identified 748 and 5 246 differentially expressed genes at 20 and 25 days post-flowering, respectively. KEGG enrichment analysis indicated that the differentially expressed genes at 25 days post-flowering were significantly enriched in pathways related to phenylpropanoid, flavonoid, and anthocyanin biosynthesis, suggesting activation of the anthocyanin synthesis pathway at this stage. To further identify key genes in the anthocyanin synthesis pathway, we analyzed transcriptome data from the blue-grained wheat material Blue 1 and its white-grained control White 1, available in public databases. Comparative analysis focused on the expression patterns of anthocyanin synthesis-related genes in the two blue-grained materials (E10-2B and Blue 1). A total of 77 and 21 up-regulated anthocyanin synthesis-related genes were identified in E10-2B and Blue 1, respectively, including chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), and MYB transcription factors. Notably, 15 genes showed shared up-regulation in both blue-grained materials, accounting for 19.48% of the up-regulated genes in E10-2B. Among these, three flavonoid 3′,5′-hydroxylase genes (F3′5′H) and three flavonoid 3-O-glucosyltransferase genes (UFGT) exhibited high expression levels in both materials. These co-upregulated genes likely play a pivotal role in the differential anthocyanin accumulation between blue and white wheat.

To provide a reference for breeding high-yield, high-quality, and green wheat varieties in the Huainan region of Jiangsu, this study conducted a systematic analysis of agronomic traits (growth period, plant height, spike number, grains per spike, thousand-kernel weight, and disease resistance) and quality traits of 116 wheat varieties approved in Jiangsu from 2010 to 2024. The pedigree information of these varieties was traced, and their genetic relationships were studied using phylogenetic and cluster analysis. The results showed that agronomic traits and quality of the approved wheat varieties in the Huainan region from 2010 to 2024 exhibited significant improvement trends. The average growth period of varieties was 207 days, a decrease of 3 to 5 days compared to early-maturity varieties; plant height remained stable at 80-88 cm, averaging 84.51 cm. Among the three yield components, spike number per hm² increased by 4.31%, thousand-kernel weight improved by 9.64%, and grains per spike remained relatively stable. Quality improvements shifted toward medium to strong gluten, with a notable increase in medium to strong gluten wheat varieties. In terms of disease resistance, varieties maintained moderate resistance to fusarium head blight, but showed enhanced resistance to powdery mildew and yellow mosaic virus disease. However, varieties resistant to sharp eyespot remained relatively scarce. Genetic relationship analysis revealed that the genetic similarity coefficients of parentage (COP) for 6 670 pairs among the 116 varieties ranged from 0 to 0.66, with an average COP of 0.32. Cluster analysis based on phylogenetic relationships divided the tested wheat varieties into four groups, with key parental varieties such as Ningmai 9, Ningmai 13, Yangmai 158, Zhenmai 9, and Zhenmai 168 forming three major core clusters, accounting for 86.21% of the approved varieties. The study indicated that while yield and quality traits of varieties in the Huainan region of Jiangsu have significantly improved, but their genetic foundation remains relatively narrow. The repeated utilization of key parental varieties like Ningmai 9, Yangmai 158, and Zhenmai 9, along with limited incorporation of new external parents, has led to insufficient genetic diversity in the developed varieties of the Huainan region in Jiangsu.

Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, is a devastating foliar disease that significantly threatens wheat production. In this study, molecular marker and Sanger sequencing were adopted to analyse the composition or sequence variation of 17 cloned or common powdery mildew (Pm) resistance genes in 40 wheat varieties mainly from Yuncheng city. Molecular markers were used to identify the 15 Pm loci, and the results showed that five loci (Pm2, Pm6, Pm8, Pm30 and Pm52) were identified as single gene or gene pyramiding. The frequency of Pm2, Pm6, Pm8, and Pm30 in the tested varieties was 35%, 20%, 20%, and 10%, respectively. Five varieties (Yunmai 2008, Jimai 22, Zhongmai 30, GA20015, and Liangxing 99) have both Pm2 and Pm6, and combination of Pm8 and Pm30 was only detected in Yunhei 1619 and Jinzimai 33. To analyse the variation of partial resistance gene Pm46, a pair of specific primers was designed to amplify the full-length genomic sequence of Pm46 in the 40 varieties. A predicted 4 708 bp DNA fragment was obtained in 34 varieties whereas no amplicon was obtained from the remaining six colored wheat varieties. By marker analysis, the six varieties all carry 4Ag (4E) chromosome of blue-grained wheat, which is the donor of ThMYC4E. Pm46 gene was sequenced in the 34 varieties and 3 haplotypes were identified, all of which encode susceptible proteins. Additionally, allelic variation analysis of Pm5 gene in the tested materials revealed that only Linnuo 178 carries the resistant allele Pm5b, while the resistant allele Pm5e was not detected. The phenotypic identification results show that germplasm carrying the Pm6 and germplasm carrying the blue-grained wheat 4Ag chromosome exhibit good resistance to the local powdery mildew races in Yuncheng, which can be further used for future wheat genetic improvement.