ArchiveFasciclin-like arabinogalactan protein (FLA), a class of highly glycosylated proteins, are widely involved in plant growth, development and stress responses, yet their functional mechanisms in wheat remain unclear. To investigate the members and biological roles of FLA in wheat, this study conducted genome-wide identification and systematic analysis of the wheat FLA family using bioinformatics approaches. The RNA-Seq and qRT-PCR were employed to examine the expression patterns of TaFLAs across tissues and under abiotic stresses, while haplotype analysis was performed to explore the association between candidate genes and kernel weight-related traits. A total of 71 TaFLAs were identified in wheat, distributed across 17 chromosomes and designated TaFLA1 to TaFLA71. Phylogenetic analysis classified these genes into eight subfamilies, with members within the same subfamily exhibiting conserved gene structures and motifs, suggesting functional similarity. Intraspecific synteny analysis revealed 85 segmental duplication pairs (30 interchromosomal and 55 intrachromosomal duplications), while interspecific synteny highlighted closer evolutionary relationships between TaFLAs and BdFLAs from Brachypodium distachyon. The RNA-Seq and qRT-PCR results demonstrated widespread expression of TaFLAs across tissues and under stress conditions. Notably, TaFLA15, TaFLA34 and TaFLA62 showed high expression in spikes and grains, while TaFLA5, TaFLA15, TaFLA34 and TaFLA62 were upregulated under PEG 6000 treatment; TaFLA5 and TaFLA34 were induced by NaCl stress. A SNP molecular marker developed for TaFLA62 revealed, combined association analysis and geographical-temporal distribution studies, that the TaFLA62-HapⅡ haplotype significantly enhances kernel weight and has undergone positive selection during wheat breeding. Overall, the gene structure of wheat TaFLAs is similar, and most of them can respond to adversity stress, and its TaFLA62-apⅡ haplotype can be used to improve wheat yield traits.
Somatic embryogenesis receptor-like kinases (SERK) can sense and transmit extracellular signals which located on the cell membrane, play an important role in plant growth, development, and stress response. We identified SERKs in barley in order to explore the expression pattern of HvSERKs under drought and salt stress. We used bioinformatics methods to identify and analyze the HvSERKs from whole genome of barley with SERKs sequence of Arabidopsis as reference sequence, and analyzed their expression under drought and salt stress by qRT-PCR. The results in this article showed that eight HvSERKs locate in the barley genome, three of which were on 7H chromosome. The phylogenetic tree constructed with SERKs in Arabidopsis, wheat, rice, maize, rape, potato and barley showed that HvSERK1, HvSERK3, HvSERK6 and HvSERK8 of barley formed a separate branch. Gene structure and conserved protein motif analysis showed that HvSERKs were highly similar and conserved. The HvSERKs were located in the plasma membrane except for HvSERK7 which in the chloroplast, and secondary structures were mainly random coils. Under drought and salt condition, the expression level of HvSERKs except for HvSERK4 which is down-regulated in leaves with salt treatment is up-regulated. Additionally, HvSERKs genes are highly expressed in mature embryos compared to immature embryos. This study identified and analyzed the HvSERKs gene family in barley, and found that they respond to drought and salt stress, with the development regulation of embryos and spikes.
N6-methyladenosine (m6A), the most abundant epitranscriptomic modification in eukaryotic RNA, regulates plant growth, development, and stress responses through a dynamic “writer-eraser-reader” regulatory mechanism. Among these regulators, ECT family proteins act as m6A reader by recognizing m6A-modified sites to participate in RNA metabolism regulation. To investigate whether TaECT3 acts as an m6A reader protein in stress responses, this study cloned the coding sequence (CDS) of TaECT3 gene from a wheat cultivar Fielder and conducted comprehensive bioinformatics analyses. Results revealed that the CDS of TaECT3 spans 1 854 bp, encoding a 617-amino acid protein containing the characteristic YTH domain conserved in ECT family members. The TaECT3 protein exhibits a molecular weight of 66.93 kDa and a predicted isoelectric point of 6.48, demonstrating intrinsic hydrophilicity and thermodynamic stability. Secondary structure prediction revealed a predominance of random coils (79.74%) and α-helices (13.45%), while tertiary structure modeling confirmed the presence of a typical m6A-binding pocket. Phylogenetic analysis revealed that TaECT3 exhibits close evolutionary relationship to the ECT proteins of HvECT3, SbECT3, and BdDF3A from barley, sorghum, and brachypodium, respectively. Electrophoretic mobility shift assay (EMSA) confirmed that recombinant TaECT3 specifically binds m6A-modified RNA. Subcellular localization analysis suggested its potential membrane localization. qRT-PCR revealed significant regulation of TaECT3 expression in wheat cultivar Fielder under high-temperature (37 ℃, 6 h) and PEG8000-simulated drought (15%, 6 h) stress conditions, compared to the control. This study demonstrates TaECT3 functions as an m6A reader and participates in response to heat and drought stresses in wheat.
Thioredoxins are a class of redox regulatory proteins widely present in plants, which play crucial roles in plant growth and development. To investigate the biological functions of the wheat thioredoxin TaTrxh10, we systematically analyzed its sequence characteristics, three-dimensional structure, and gene expression patterns. The protein was expressed in a prokaryotic system, and its disulfide bond reduction activity was validated in vitro. Sequence analysis revealed that wheat TaTrxh10 comprises three family members encoded by three homoeologous genes: TaTrxh10-A, TaTrxh10-B, and TaTrxh10-D. All three TaTrxh10 family members consist of 143 amino acids, with a sequence identity of 94.4%, and contain the classic thioredoxin conserved domain and catalytic active center (WCGPC). Protein structural analysis demonstrated that TaTrxh10 can fold into a sandwich-like spatial structure composed of five β-sheets and four α-helices. Gene expression analysis indicated that the three homoeologous genes of TaTrxh10 are expressed at very low levels in roots, stems, and leaves, and at relatively lower levels in the ovary and mature seeds; however, they are highly expressed in mature pollens. The thioredoxin reduction activity assay using DTNB demonstrated that all three family members of TaTrxh10 possess the ability to catalyze disulfide bond reduction, but their reduction activities varied among the members. In reaction systems with lower thioredoxin concentrations, TaTrxh10-D exhibited the highest catalytic activity, followed by TaTrxh10-B. These results indicate that wheat TaTrxh10 represents a class of active redox regulatory proteins specifically expressed in pollen, which provides important insights for further exploration of redox-dependent developmental regulatory mechanisms in wheat.
This study aimed to investigate the regulatory role of cell division cytokinin in wheat root morphological development and to identify key functional genes involved in root system architecture regulation. The seedlings of a wheat variety Jinmai 47 were treated with exogenous 0.05 μmol·L-1 6-benzylaminopurine (6-BA) to systematically analyze its effects on root phenotypic traits. The genotypic sensitivity of 110 wheat varieties (lines) to 6-BA was evaluated. Furthermore, the TaCKX1B gene was obtained through homology-based cloning, and its function was validated via heterologous overexpression in tobacco. The results showed that the treatment with 0.05 μmol·L-1 6-BA significantly inhibited primary root elongation while promoting root hair initiation and elongation. Considerable genetic variation in sensitivity to 6-BA was observed among wheat genotypes, with sensitivity indices predominantly ranging from 40% to 60%. Materials with low sensitivity at the seedling stage exhibited more developed root systems at the grain-filling stage. The TaCKX1B gene was located on chromosome 1B, with a coding sequence length of 1 569 bp. Heterologous overexpression of TaCKX1B in tobacco resulted in approximately a two-fold increase in root biomass in T1 transgenic plants compared to wild-type controls. In conclusion, the TaCKX1B gene plays a positive regulatory role in plant root development.
Wheat germplasms are the important foundation for wheat breeding. In this study, the genetic diversity was analyzed for 40 wheat germplasms from Pingdingshan region using SSR molecular markers. Results showed that the percentage of polymorphic bands for SSR primers ranges from 35% to 100%, with an average of 74.5%. A total of 164 polymorphic bands were obtained by 29 pairs of SSR primers, with an average of 5.60 bands for a single pair of SSR primers. The effective number of alleles, Nei's gene diversity index, and Shannon's information index of SSR primers ranged from 1.17 to 1.66, from 0.11 to 0.38, and from 0.17 to 0.55, respectively, with the average values of 1.43, 0.26, and 0.39, respectively. The genetic similarity coefficient between different wheat germplasms is distributed between 0.55 and 0.98, with an average of 0.77. Based on the genetic similarity coefficient, the 40 wheat germplasms can be divided into five groups. Of them, group I contained three wheat germplasms; group Ⅱ contained eleven wheat germplasms; group Ⅲ contained five wheat germplasms; and group IV contained three wheat germplasms. Group Ⅴ contained the largest number (eighteen) of wheat germplasms, accounting for 45.0% of the total wheat germplasms. The result of principal component analysis was basically consistent with the results of cluster analysis. In addition, DNA fingerprints of the 40 wheat germplasms were constructed with SSR primers.
To investigate the mechanisms underlying the formation of quality traits in different wheat types, this study utilized nine wheat varieties recently approved for cultivation in the Huang-Huai-Hai wheat-growing region. These included a strong-gluten wheat cultivar Weilong 169, a purple wheat cultivar Jimai 8, a medium-gluten wheat cultivar Xinong 100, along with the national standard control variety Zhoumai 36. Grain nutritional quality, processing quality, high-molecular-weight glutenin subunit (HMW-GS) composition, and quality-related gene differences were analyzed. The results showed that Weilong 169 exhibited the highest gliadin-to-glutenin ratio (1.84), while Xinong 112 had the highest glutenin macropolymer (GMP) content (11.51%). Among the varieties, Xinong 136 displayed the highest total starch content (80.69%); Xinong 106 had the highest amylose content (39.55%), and Xinong 100 had the highest amylopectin content (43.18%). All medium-gluten wheat varieties (except Jimai 8) exhibited B-type starch content above 87.64%. Protein percentage area, junction count, and junction density in Xinong 112 were significantly higher than those in Weilong 169 (P<0.05). Jimai 8 showed the highest total sulfhydryl and disulfide bond contents (20.01 μmol·g-1 and 7.24 μmol·g-1, respectively). Furthermore, Xinong 109 and Xinong 136 had the lowest loss tangent values. For HMW-GS, high-quality subunits were detected at the Glu-A1, Glu-B1 and Glu-D1 loci in all varieties except Xinong 100, Xinong 109 and Xinong 112. The number of superior allelic variations among the tested varieties ranged from six to ten. In conclusion, significant differences in quality traits were observed among the wheat varieties. Medium-gluten wheat varieties, such as Xinong 100, demonstrated superior performance across various nutritional and processing quality indicators. Strong-gluten wheat (Weilong 169) and purple wheat (Jimai 8) possessed high-quality subunit combinations (Ax1, Bx7+By8, Dx5+Dy10) and the largest number of superior allelic variations, reflecting rich genetic diversity in quality-related traits.
The carotenoid content in wheat kernels serves as a crucial determinant of color appearance and commercial value, representing an important indicator in wheat breeding. While conventional detection methods (ultraviolet spectrophotometry, thin-layer chromatography, and high-performance liquid chromatography) demonstrate accuracy, their operational costs and time requirements prove prohibitive for large-scale applications. This study provided a rapid non-destructive prediction method through the integration of near-infrared spectroscopy (NIRS) with chemometric techniques. We systematically evaluated: (1) three dataset partitioning methods hold-out method (HOM), K-fold cross-validation (KFCV), time series split (TSS); (2) four spectral preprocessing techniques [Savitzky-Golay smoothing (SG), multiplicative scatter correction (MSC), standard normal variate transformation (SNV), trend correction (TC)], three feature selection methods were obtained by fusing variance threshold feature selection (VTFS), SelectKBest (SKB), and recursive feature elimination (RFE) with principal component analysis (PCA), respectively. Subsequent modeling employed partial least squares regression (PLSR), support vector machine regression (SVR), and gradient boosting regression (GBR) were used to build different prediction models. The results showed that HOM achieved optimal dataset partitioning, SG preprocessing provided superior spectral enhancement, and PCA-SKB feature fusion delivered maximum information retention. The optimized was PCA-SKB-GBR. In the validation set and prediction set, R2 were 0.99 and 0.89 with RMSE of 0.04 and 0.34 μg·g-1, respectively. The residual prediction deviation (RPD) reached 3.01. Therefore, based on the hold-out method of sample set, SG spectral pretreatment and PCA-SKB feature selection method, the PCA-SKB-GBR model can be used to predict the carotenoid content of wheat grain quickly and efficiently.
In order to clarify the effect of the new carbon-based blended fertilizer (CBF) on wheat following rice stubble, using F1 (24-12-8), F2 (24-10-10), F3 (18-10-7) and F4 (15-10-10) as materials, four application patterns were designed under similar fertilizer cost conditions: CBF applied once pre sowing (M1), CBF applied twice (M2), and CBF applied twice along with urea supplemented pre-sowing (M3) or at jointing stage (M4). Using urea four times application as control (CK), the effects of different treatments on wheat population structure, leaf senescence, yield component, nitrogen efficiency, and economic benefit were studied. The results showed that M1 had the lowest spike number at maturity stage and the percentage of ear bearing tiller, as well as the largest decrease of LAI and SPAD value at milking stage, resulting in a decrease rate in yield of 7.96%-12.91%, a decrease in apparent nitrogen use efficiency of 0.93-3.52 percentage points, and a decrease in net benefit of 2 012.49-3 070.72 yuan·hm-2 compared with CK. Compared with M1, M2, M3 and M4 promoted spike number, kernel number per spike and 1 000-grain weight, and increased grain yield by 12.60%, 19.20% and 16.46%, respectively. M3 was more conducive to improving tillering and matter accumulation at the early stage of wheat, promoting seedling strength, maintaining higher population LAI and flag leaf SPAD value, and increasing dry matter and nitrogen accumulation at maturity stage, which increased yield and apparent nitrogen use efficiency by 2.23%-12.19% and 3.62-12.48 percentage points compared with CK, respectively. From the perspective of economic benefit, M3 achieved the highest output value and saved twice labor input of topdressing, and thus improved net benefit, compared with CK in F1, F2 and F3, which achieved the highest net benefit in F2, with an increase of 1 399.65 yuan·hm-2 compared with CK. In different fertilizer formulations, F3 and F4 increased apparent nitrogen use efficiency but failed to reach the expected yield and benefits under the similar fertilizer costs. F2 showed higher yield and benefits in M2, M3, and M4 compared with other formula fertilizers. In summary, among different formulations of CBF, F2 applied twice along with urea supplement pre-sowing is the most beneficial fertilizer method to synergistically improve wheat yield, economic benefit and apparent nitrogen use efficiency, which can promote the high-yield and high-efficiency cultivation of wheat following rice stubble.
To clarify the regulatory effects of phosphorus application on drought tolerance during the development of wheat floret, a multi-spike variety Yumai 49-198 (V1) and a large-spike variety Zhoumai 22 (V2) were used as experimental materials. A pot experiment was conducted under two soil water conditions: severe drought (35%-45% of field capacity, W0) and normal moisture (75%-85% of field capacity, W1), and two phosphorus application levels (no phosphorus application, P0; 0.134 g·kg-1 phosphorus application, P1). The effects of phosphorus application on the number of fertile florets and grain number per spike during wheat floret development, as well as the photosynthetic characteristics, osmotic regulation, and antioxidant system of the flag leaf were investigated. The results showed that compared with W1, the number of fertile florets and grains per spike in the two wheat varieties under W0 condition were significantly reduced; the relative chlorophyll content (SPAD), net photosynthetic rate (Pn), maximum quantum efficiency of PSⅡ (Fv/Fm) of flag leaf, and plant phosphorus content decreased significantly; the content of soluble protein (SP), proline (PRO), and malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) and catalase (CAT) in the flag leaf were significantly increased. Compared with P0, P1 significantly increased the number of fertile florets and grain number per spike of the two varieties, with an increase rate of 5.74%-22.99%, 9.90%-17.48% for V1 and 12.16%-15.63%, 7.61%-27.45% for V2, respectively; P1 effectively increased the SPAD, Pn, Fv/Fm, SP content, PRO content, SOD and CAT activities in flag leaf, and decreased the MDA of flag leaf. The differences were significant under W0 condition, and the regulatory effect on V2 was greater than that on V1. In this study, phosphorus application significantly enhanced the drought tolerance of wheat plants, improved the photosynthetic assimilation, provided sufficient nutritional security for the formation and development of fertile florets into grains, thus achieving stable yield with increased grains, and the regulatory effect of which on the large-spike variety was higher than that of the multi-spike variety.
To clarify the influence of exogenous alpha-ketoglutaric acid (α-KG) spraying after anthesis on the photosynthesis of wheat flag leaves and the characteristics of grain filling, Shumai 830 and Shumai 1868, with significant differences in thousand-grain weight, were selected as materials. Using no spraying as the contrel (CK), 0.50, 0.75, and 1.00 g·L-1 concentrations of α-KG spraying at 0 d and 15 d after anthesis were set up. The differences in chlorophyll content and photosynthetic parameters of flag leaves, grain-filling parameters, and yield under different treatments were compared. The results indicated that compared with the CK, the α-KG treatment enhanced wheat photosynthesis and promoted grain filling, thereby increasing the grain weight of wheat. Among them, the treatment of 0.75 g·L-1 α-KG showed the best effects. Under the treatment with 0.75 g·L-1 of α-KG, the chlorophyll content at 21 d after anthesis and the net photosynthetic rate at 20 d after anthesis of Shumai 830 and Shumai 1868 were significantly higher than those of the CK, with an increase rate of 12.0% and 9.6% for chlorophyll content, and 11.1% and 6.6% for net photosynthetic rate, respectively. And the active grain-filling period, average grain-filling rate, and maximum theoretical thousand-grain weight of Shumai 830and Shumai 1868 were significantly higher than those of the CK, with an increase rate of 3.8%, 4.6%, and 7.9% for Shumai 830, and 1.6%, 4.9%, and 4.9% for Shumai 1868. After the treatment with 0.75 g·L-1 of α-KG, the thousand-grain weight of Shumai 830 and Shumai 1868 was increased by 3.8% and 2.7%, respectively, compared with CK, and the yield was increased by 6.2% and 7.9%, respectively, all showing significant difference from the CK. Thus, exogenous α-KG spraying after anthesis can promote photosynthesis and grain filling by increasing the chlorophyll content of wheat, and ultimately increase grain weight and yield.
To investigate the effects of different postponing nitrogen application treatments on oat grain yield and nitrogen accumulation and translocation, a randomized block design was adopted using two oat varieties Bayou 1 and Dingyou 8 as the experimental materials. Field trial were conducted in 2021 and 2022, and four nitrogen application treatments were set up under a total nitrogen rate of 100 kg·hm-2: all basal application (CK), 1/2 basal application + 1/2 tillering stage fertiliser (N1), 1/2 basal application + 1/2 jointing stage fertiliser (N2), 1/3 basal application + 1/3 tillering stage fertiliser + 1/3 jointing stage fertiliser (N3). The effects of different postponing nitrogen application treatments on yield of oats, dry matter accumulation and translocation in each organ, and nitrogen accumulation and translocation in each organ were determined and analysed. The results indicated that different postponing nitrogen application treatments had an extremely significant impact on oat yield. Compared with CK, the yield of both Bayou 1 and Dingyou 8 significantly increased in N3 treatment, with an average increase rate of 7.96%-8.82% over the two years. However, the yield of oats were reduced by 9.64%-4.50% and 4.12%-6.59% in N1 and N2 treatments respectively, N3 treatment enhanced the pre-anthesis dry matter and nitrogen translocation, post-anthesis dry matter, and nitrogen accumulation, with the increase rate of 21.83%-37.79%, 23.59%-30.19%, 7.58%-12.35%, and 8.48%-11.54%, respectively. Therefore, under a total nitrogen rate of 100 kg·hm-2, applying 1/3 basal application + 1/3 tillering stage fertiliser + 1/3 jointing stage fertiliser better coordinated the nitrogen accumulation and transport of oats, and improved the yield.
In order to explore the adaptability of strong-gluten wheat under different soil types and the differences in yield and quality among varieties, the pot experiments were conducted with three strong-gluten wheat varieties Gaoyou 2018, Shiluan 02-1 and Shiyou 20 as materials in fluvo-aquic soil environment and black soil environment. The changes of flag leaf chlorophyll content, grain weight and nitrogen content of various organs of the strong-gluten wheat varieties from flowering stage to maturity stage under fluvo-aquic soil and black soil were compared. The similarities and differences of stem number per plant, yield and its components, grain protein and component content, and protein yield at maturity stage were compared; Since the flowering period began, the nitrogen content of various plant vegetative organs gradually decreased, while the nitrogen content of grains showed a trend of first decreasing and then increasing. The results showed that the chlorophyll content of wheat in the two soils increased first and then decreased from flowering to 25 days after flowering, and reached the maximum at 5 days after flowering. Compared with the fluvo-aquic soil, the leaf staying green time and grain filling time of wheat were prolonged under the conditions of black soil, and the duration of grain filling gradual increasing period of the three varieties was prolonged by an average of 3.0 days. The stem number per plant, effective spike number and 1 000-grain weight increased by 10.91%, 18.62% and 3.85%, respectively; the number of ineffective spikes decreased by 1.35%; the non-gluten protein content and gluten protein content in grain increased by 0.18% and 1.48%, respectively; and the protein yield increased by 0.51 g·pot-1. In summary, under the conditions of this experiment, black soil significantly promoted wheat tillering, protein content, and increased yield compared with fluvo-aquic soil.
To investigate the effects of sulfur fertilization on wheat yield and quality under different nitrogen levels in the lime concretion black soil area of northern Anhui Province, a field split-plot experiment was conducted in Suzhou, Anhui Province, from 2022 to 2024. The wheat varieties tested were Annongda 1216 and Huachengmai 1688. The main plots were based on a basal nitrogen application of 196 kg·hm-2, with two top-dressed nitrogen levels of 0 kg·hm-2 (N1) and 75 kg·hm-2 (N2) at the jointing stage. The sub-plot was set up with two sulfur levels of 0 kg·hm-2 (S1) and 18.29 kg·hm-2 (S2). The study compared and analyzed the effects of different treatments on wheat yield, quality, dry matter accumulation and translocation, and photosynthetic characteristics. The results showed that compared to the N1S1 treatment, the N2S2 treatment increased the average values of leaf area index (LAI), spike number, grain number per spike, grain yield, and protein content by 13.43%, 6.29%, 2.82%, 12.17%, and 1.07 percentage points, respectively. Under the same nitrogen application rate, Pn, Gs, and Tr in the S2 treatment were higher than those in S1, while Ci was lower. The post-flowering dry matter accumulation and grain dry matter distribution at maturity stage were on average 7.78% and 5.60% higher in S2, and the spike number, grain number per spike, and yield increased by 3.75%, 1.70%, and 6.14%, respectively. Under the same nitrogen-sulfur fertilization level, N2 resulted in higher tiller number and leaf area index than N1, with Pn, Gs, and Tr increasing by 5.67%, 16.62%, and 12.41%, respectively. The post-flowering dry matter accumulation and grain dry matter distribution at maturity stage were on average 8.47% and 6.23% higher, and the spike number, grain number per spike, yield, and protein content increased by 3.16%, 1.39%, 6.23%, and 2.05 percentage points, respectively. In conclusion, top-dressing nitrogen fertilizer and basal sulfur application can enhance wheat leaf area index, improve photosynthetic capacity, promote post-flowering dry matter accumulation, and ultimately increase wheat grain yield and protein content.
In order to investigate the control efficacy of the combination of Bacillus methylotrophicus Lw-6 and chemical fungicides against wheat stripe rust, Puccinia striiformis f. sp. tritici (Pst) urediniospores were used as the target fungus, and strain Lw-6 was screened through antagonistic tests in vitro. The chemical fungicide used in combination with Lw-6 was selected by compatibility testing, and the optimal concentration was selected by greenhouse pot tests. Finally, the control and growth promotion effects were determined by greenhouse pot tests and field test. The results showed that diniconazole had the best compatibility with strain Lw-6, and the combined application of 128 mg·L-1 Di and Lw-6 fermentation liquid achieved 92.78% and 86.22% control efficacy against wheat stripe rust in pot experiments and field trials, respectively, which were higher than the controlling effects of using individual diniconazole or Lw-6. In addition, the 1 000-grain weight and yield of wheat were significantly increased after compound treatment, and the contribution rate of plant protection of 6 500 mg·L-1 Lw-6 powder combined with 128 mg·L-1 diniconazole was 24.98%. In summary, the combination of Lw-6 and diniconazole can effectively control wheat stripe rust, and it has promising application potential.