Background Cystic echinococcosis (CE) is a zoonotic disease caused by the larval stage of the cestode Echinococcus granulosus sensu lato. We analyzed the proteomics of fertile and sterile Echinococcus granulosus cysts, in order to investigate the cause of sterility and provide a theoretical basis for identifying drug targets. Methods Three fertile and three sterile cysts were collected from yaks. After removing hydatid fluid, protoscoleces, and host granulomatous tissue, a TMT-based quantitative proteomic analysis was performed with the sterile cyst group as the control.The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE(1) partner repository with the dataset identifier PXD082611. Results A total of 1,695 proteins were identified. Among them, 119 proteins were differentially expressed, with 51 upregulated and 68 downregulated in fertile cysts. GO analysis revealed that these differential expressed proteins (DEPs) were primarily enriched in the following categories: for molecular function, binding (46.30%), catalytic activity (22.22%), structural molecule activity (11.11%), and molecular function regulator (5.56%); for cellular component, protein-containing complex (16.67%), cellular anatomical entity (28.52%), intracellular (27.04%), and cell (27.78%); for biological process, metabolic process (11.14%), cellular process (15.78%), biological regulation (10.90%), and cellular component organization or biogenesis (9.51%). KEGG pathway analysis indicated significant enrichment of these DEPs in the Ribosome and Glycosylphosphatidylinositol (GPI)-anchor biosynthesis pathways. The highly expressed PIG-M may participate in stress resistance, maintenance of germinal layer integrity, and immune evasion in E. granulosus . Conclusion The highly active protein synthesis pathway in fertile cysts supports their development, and GPI-anchored proteins may be key molecules for immune evasion in fertile cysts. These findings elucidate the protein differences between fertile and sterile cysts and provide valuable candidate targets for anti-hydatid research, and offer a reference for future control strategies against Echinococcus . Data are available via ProteomeXchange with identifier PXD082611.

Leaf rust is a highly destructive foliar disease in wheat which causes major constraints in wheat production worldwide. In this study, we conducted a comprehensive assessment of adult plant resistance to leaf rust in 590 accessions from the advanced backcross-nested association mapping plus inter-crossed (AB-NAMIC) population. We used 660K genotype data to perform a genome-wide association study (GWAS), which identified significant quantitative trait loci (QTLs) on chromosomes 1B, 2A, 2B, and 7D, and then focused on the candidate gene TaRLK-1B on chromosome 1B. A cleaved amplified polymorphic sequence (CAPS) marker developed based on TaRLK-1B haplotypes could effectively differentiate between resistant and susceptible varieties. This gene encodes a membrane-localized leucine-rich repeat receptor-like kinase (LRR-RLK) that is upregulated in response to the fungal infection that causes leaf rust. Targeted knockout of TaRLK-1B in wheat led to reduced resistance to leaf rust, underscoring its essential role as a positive regulator of the defense against this disease. We propose that TaRLK-1B interacts with the receptor-like cytoplasmic kinase TaRLCK1B, potentially facilitating immune signal transduction. Our findings also demonstrate that pyramiding minor effect QTLs significantly increases resistance to leaf rust. This study provides novel insights into rust resistance genes and valuable QTL information, which can improve marker-assisted wheat breeding efforts.

Abstract Background The Three-River-Source Region of the Qinghai-Tibet Plateau is a hyperendemic focus for echinococcosis, with Echinococcus granulosus, E. multilocularis, and E. shiquicus circulating between definitive canid hosts (dogs and foxes) and intermediate hosts (livestock and rodents). However, the extent of environmental contamination by Echinococcus eggs remains understudied and poses significant risks to human and animal health. Methods From 2019 to 2021, we collected 631 canid fecal samples (296 from dogs and 335 from foxes) and 398 adjacent soil samples across endemic counties in the Three-River-Source Region of the Qinghai-Tibet Plateau. Multiplex real-time PCR was employed to detect Echinococcus species DNA in feces and soil samples. Results The overall Echinococcus prevalence in canid feces was 7.13% (45/631), with 3.01% in E. multilocularis (19/631), 2.06% in E. granulosus (13/631), and 2.06% in E. shiquicus (13/631). Foxes presented increased E. multilocularis (3.88%, 13/335) and E. shiquicus (2.69%, 9/335) infections, whereas dogs presented increased E. granulosus prevalence (2.70%, 8/296). Soil contamination with Echinococcus species was detected in 2.51% (10/398) of the samples. The primary contaminants were E. multilocularis and E. shiquicus (1.01% each, 4/398), whereas E. granulosus was less frequent (0.50%, 2/398). Moreover, the soil near fox feces was contaminated with both E. multilocularis and E. shiquicus, whereas the dog-associated soil was contaminated with all three species. Conclusions This study suggests widespread environmental deposition of Echinococcus eggs on the Qinghai-Tibet Plateau, driven by canid defecation. If these eggs remain viable, their persistence in soil would indicate a potential zoonotic transmission risk, highlighting the need for integrated control strategies targeting both domestic and wild canids. Graphical abstract

Melophagus ovinus (sheep keds) is a major ectoparasite and pathogen vector in the Qinghai-Tibet Plateau. To investigate the prevalence and genetic diversity of zoonotic Anaplasma spp. carried by M. ovinus, a total of 1000 M. ovinus samples were collected from five counties in Qinghai during 2024–2025. Nested PCR and genetic analysis showed that the carriage rate of Anaplasma ovis (A. ovis) was 27.3 %, with a total of 29 identified haplotypes. The genetic diversity exhibited a typical pattern of “low Hd + low Pi + dominant haplotype predominance” (Hd = 0.450, Pi = 0.004 06). The dominant haplotype AoH3 was widely distributed, and all A. ovis isolates clustered with the strains from Siberia and Tibet. A. bovis was only detected (0.9 %) in Huzhu county (Hd=1.000, Pi=0.09148). This study indicates that M. ovinus may serve as a natural carrier of A. ovis, highlighting the public health risk of zoonotic Anaplasma spp. to local herdsmen and high-risk populations in the Qinghai-Tibet Plateau.

Ruminal acidosis, particularly subacute ruminal acidosis, remains a major metabolic disorder that compromises animal health, production efficiency, and the sustainability of intensive ruminant systems. Although traditionally defined by reduced ruminal pH, it is increasingly recognized as a multidimensional disorder involving microbial ecological destabilization, disrupted metabolic cross-feeding, impaired epithelial barrier function, and dysregulated host inflammatory responses. This review synthesizes current knowledge of the microbial and metabolic mechanisms underlying acute and subacute ruminal acidosis and highlights processes that extend beyond pH depression alone. High-concentrate feeding shifts the balance among amylolytic and lactate-producing microorganisms, lactate-utilizing populations, and fibrolytic guilds, thereby promoting organic acid accumulation, reducing functional redundancy, and weakening microbial resilience. Concurrent increases in volatile fatty acids, lactate, lipopolysaccharide, histamine, and other microbially derived bioactive compounds increase epithelial acid load, disrupt tight-junction integrity, and facilitate inflammatory signaling. The principal novelty of this review is the integration of microbial functional guilds, metabolic cross-feeding, ecological resilience, epithelial barrier dysfunction, and host inflammation into a unified microbiota–metabolism–barrier–inflammation framework linking dietary perturbation with microbial dysfunction and host pathology. We further critically evaluate nutritional regulation, buffering agents, probiotics, yeast-derived products, postbiotics, and plant bioactive compounds according to their capacity to restore microbial function rather than merely correct ruminal pH. Additionally, this review may support multidimensional risk assessment, guide targeted intervention, and facilitate the integration of continuous ruminal monitoring with precision nutrition for earlier prediction and individualized prevention of ruminal acidosis.

Ovine theileriosis is a tick-borne hemoparasitic disease caused by protozoan parasites of the genus Theileria, which poses a substantial threat to sheep and goats. The disease is prevalent in northwestern China, yet studies on the diversity of its pathogens in Qinghai Province remain limited. From 2014 to 2025, 1062 blood samples were collected from sheep in 11 counties of Qinghai Province, and the Theileria spp. were investigated by PCR targeting the 18S rRNA gene. Genetic diversity analysis was performed to assess sequence variations and phylogenetic relationships. The results show that 424 samples tested positive for Theileria spp., with an overall infection rate of 39.92%, including T. uilenbergi 26.74%, T. luwenshuni 22.41%, T. ovis 18.17%, T. capreoli 0.56%, and Theileria sp. OT3 0.09%. The co-infection rate was 22.69%. Six haplotypes were detected in both T. uilenbergi and T. ovis, and fifteen in T. luwenshuni. Hap 1 was the dominant haplotype for all three species, with respective proportions of 73.08%, 77.06% and 82.17%. Phylogenetic analysis showed the strains were most closely related to those from Hunan, Turkey, Shaanxi, etc. These findings reveal a high diversity of Theileria species in Qinghai sheep, which provides crucial epidemiological insights into the transmission dynamics of ovine theileriosis in Qinghai and vital support for developing effective control strategies.

Wheat is a major staple crop for over one-third of the world's population, crucial for global food security, economic stability and cultural traditions. Recently, single-cell and spatial omics approaches have transformed biological discovery, primarily in medical and animal sciences, and they are now beginning to be applied in plant research. Here we summarize the technical innovations and feasibility of spatial omics applications in wheat research, particularly for understanding developmental and environmental responses, thereby potentially enhancing wheat breeding. We highlight how these tools can reveal spatial and temporal patterns in gene expression, cellular heterogeneity and tissue organization in wheat. Furthermore, we propose developing a spatially resolved single-cell atlas of wheat across its life cycle to facilitate breakthroughs in basic research and potential applications in breeding. To achieve these goals, we advocate for a Wheat Spatial Omics Consortium to foster worldwide collaboration for overcoming barriers and developing sustainable and climate-resilient wheat.

Wheat grain weight and flour quality largely depend on starch biosynthesis, yet the mechanisms by which transcription factors coordinate this process remain poorly understood. In this study, using an integrative strategy that combines genome-wide association analysis with yeast two-hybrid library screening, we identify TaNF-YC10 , a Nuclear Factor Y transcription factor, as a positive regulator of starch accumulation in the wheat endosperm. Loss of TaNF-YC10 reduces starch content and alters starch granule size distribution, whereas overexpression enhances starch accumulation and increases grain weight. TaNF-YC10 binds and activates core starch biosynthetic-related genes, including AGPL1 , GBSS1 , YUC11 , and NF-YB7 , and forms higher-order transcriptional complexes with TaNF-YB1 and TabHLH95 to coordinate multiple regulatory pathways. TaNF-YC10-A1 - Hap2 is associated with higher starch content and thousand grain weight and has been selected during wheat breeding in China. Collectively, our findings establish TaNF-YC10 as a pivotal transcriptional hub in starch regulation and highlight its potential as a target for genetic improvement of grain yield in wheat.

Fascioliasis, a globally prevalent zoonosis, severely threatens public health and livestock security. Current diagnostic approaches, hindered by the need for sophisticated instrumentation and specialized expertise, are inadequate for on-site surveillance in resource-constrained settings. This study developed a rapid, visual detection assay for Fasciola hepatica via recombinase-aided amplification (RAA) integrated with CRISPR/Cas12b, addressing critical equipment and operational constraints. Targeting a specific mitochondrial DNA fragment of F. hepatica, recombinant plasmid standards were constructed, RAA primers and sgRNA optimized, and three detection modalities (real-time fluorescence, UV lamp, test strip) integrated. Clinical validation against PCR demonstrated 45 min turnaround time, F. hepatica-specific positivity, and real-time fluorescence sensitivity of 2.6 copies/μL. Results showed high concordance with PCR and qPCR, with substantially reduced assay duration and streamlined workflow. This highly sensitive, specific, multi-visualized method overcomes limitations of conventional techniques, offering an efficient, field-deployable tool for fascioliasis surveillance and control in grassroots and pastoral regions.

Grain weight is primarily determined by the accumulation of storage compounds during grain filling, and elucidation of the underlying regulatory networks will help yield improvement. In this study, we identified TaPBF1-5A, a Dof-type transcription factor, and showed that it is a key positive regulator of grain development and thousand-grain weight (TGW). Knockout of TaPBF1 caused a significant reduction in both starch content and TGW, and altered SSP composition, particularly changes in glutenin subunits and gliadin fractions. We demonstrated that TaPBF1-5A directly activates starch biosynthesis genes including SSIIa-7D, GBSSI-7D, and SBEI-7D, and regulates the key regulatory factor TaNAC019-3B. Protein interaction assays revealed that TaPBF1-5A forms a nuclear complex with TaSPA-1B, which exerts differential regulatory effects on SSP-related genes, including Glu-1By and Gli-γ-7700. Furthermore, we identified a favourable haplotype, TaPBF1-5A-Hap1, associated with higher TGW and present evidence of positive selection during wheat breeding. Collectively, this work reveals that TaPBF1-5A promotes wheat grain weight mainly by regulating starch biosynthesis and provides a diagnostic marker for breeding.

Wheat leaf rust, caused by Puccinia triticina, poses a significant threat to global wheat production. MicroRNAs (miRNAs) are critical regulators of plant growth, development and stress responses; however, their role in wheat resistance to leaf rust remains poorly understood. In this study, we identified the miR171a-TaSCL6-1 module as a key regulator of leaf rust resistance in wheat. Expression of miR171a was upregulated in wheat seedlings following pathogen inoculation. Transgenic assays, including overexpression and short tandem target mimic approaches, demonstrated that miR171a negatively regulates resistance to leaf rust. Degradome sequencing confirmed TaSCL6-1 as the primary target of miR171a, and both overexpression and knockout of TaSCL6-1 established its role as a positive regulator of resistance. Transcriptome analysis revealed that TaSCL6-1 upregulates defence-related genes encoding peroxidases and transcription factors. TaSCL6-1 can activate the transcription of downstream genes by binding to the GAA motif in the promoters, and silencing of downstream genes (TaPOD2, TaPOD35 and TaERF114) increased wheat susceptibility to leaf rust. Furthermore, miR164-targeted TaNAC21/22 acts as an upstream regulator of MIR171a by binding to its promoter and activating its expression. Overexpression of TaNAC21/22 increased susceptibility to leaf rust, whereas transient silencing of TaNAC21/22 or overexpression of miR164 enhanced seedling resistance to leaf rust. Our findings elucidate the role of the miR171a-TaSCL6-1 module in regulating wheat defence against leaf rust, providing potential genetic targets for breeding resistant wheat varieties.

Protein malnutrition remains a major global health challenge, particularly in regions where cereal grains dominate daily diets and access to diverse protein sources is limited. Cereals such as rice, wheat and maize provide most of the world's calories, yet their grain proteins are often low in essential amino acids and poorly balanced for human nutrition. Improving both the quantity and quality of cereal protein therefore represents a critical opportunity to enhance human health while reducing reliance on environmentally intensive animal-based foods. In this Review, we synthesize recent advances in understanding how grain protein content and composition are regulated in cereals, and why protein enhancement has historically been constrained by trade-offs with starch accumulation and yield. We discuss how domestication and modern breeding reshaped carbon and nitrogen allocation in cereal grains, creating a starch-dominant optimum that limits protein concentration. Drawing on genetic studies from rice, maize and wheat, we highlight emerging strategies that improve nitrogen acquisition, amino acid transport, storage protein composition and endosperm buffering capacity, enabling partial decoupling of protein accumulation from yield penalties. Finally, we place cereal protein biofortification within a broader nutritional and environmental context. Enhancing protein density and amino acid balance in staple cereals can improve dietary adequacy for vulnerable populations while lowering greenhouse gas emissions per unit of nutrition. Together, these insights position cereal protein biofortification as a scalable and equitable pathway towards healthier diets and more sustainable food systems under global climate and population pressures.

RING finger E3 ligases are critical in regulating plant growth, development, and environmental responses; however, their role in wheat salt tolerance remains poorly understood. In this study, we identify TaGW2L, a GW2-like RING E3 ubiquitin ligase, as a novel regulator of salt tolerance in wheat. Expression analysis shows that TaGW2L homoeologs are downregulated in response to NaCl and ABA treatments. Overexpression of TaGW2L-7A resulted in a salt-sensitive phenotype, characterized by reduced survival rates, elevated reactive oxygen species (ROS) production, and decreased chlorophyll content under salt stress. Conversely, TaGW2L knockout lines exhibited enhanced salt tolerance, with improved survival rates, higher chlorophyll content, and lower ROS levels compared to wild-type plants. Transcriptome analysis revealed that TaGW2L knockout leads to the upregulation of salt-responsive genes, particularly those involved in ABA and ROS pathways. Ubiquitinome and biochemical analyses demonstrated that TaGW2L negatively regulates salt tolerance by mediating the ubiquitination and degradation of TaHMT1, a positive regulator of salt stress responses. Loss-of-function mutations in TaHMT1-5A impaired salt tolerance in wheat. This study underscores the critical role of the ubiquitin-proteasome system in wheat's salt stress response and offers new insights into potential targets for improving salt tolerance in wheat.

ABSTRACT Pre‐harvest sprouting (PHS) poses a major threat to wheat yield and quality, yet the genetic basis of seed dormancy underlying PHS resistance remains poorly understood. Here, through integrated genome‐wide association and transcriptomic analyses, we identify TaMYB7‐A1 as a key regulator of seed dormancy and PHS resistance. TaMYB7‐A1 encodes an R2R3‐MYB transcription factor that directly activates TaABI5 to modulate abscisic acid (ABA) signaling and indirectly fine‐tunes ABA–gibberellin (GA) homeostasis to enforce dormancy. Evolutionary and haplotype analyses revealed that the superior allele, TaMYB7‐A1Hap−1, originated from wild einkorn and was introgressed into domesticated emmer and subsequently into modern bread wheat. A miniature inverted‑repeat transposable element (MITE) insertion in its promoter substantially elevates TaMYB7‐A1 expression by increasing chromatin accessibility and facilitating the recruitment of upstream regulators, while two key amino acid substitutions (Gly23 and Gly92) strengthen its DNA‐binding and transcriptional activation capacity. Combinations of promoter and coding‐region variants generate graded PHS resistance across haplotypes, mirroring local adaptation to harvest‑season precipitation. Introgression of TaMYB7‐A1Hap−1 into modern cultivar enhances PHS resistance without yield penalties. These findings elucidate a molecular and evolutionary framework for precipitation‐driven adaptation and provide a valuable genetic target for developing climate‐resilient wheat varieties.

Starch is the primary storage compound in wheat grains and is essential for both flour quality and grain weight. In this study, we identified TaMYB44, an R2R3-MYB transcription factor gene that controls starch content in wheat grains, through a genome-wide association study. The TaMYB44 homoeologs were predominantly expressed in developing grains, with peak levels observed 10 days after pollination. Functional analyses revealed that TaMYB44 acts as a negative regulator of starch synthesis in the endosperm, limiting grain size by repressing starch synthesis-related genes and modulating secondary metabolism. Knockout mutants of TaMYB44 exhibited significantly increased starch accumulation, larger grain size, and improved yield stability across diverse growing environments. Furthermore, we discovered that TaWDR1 interacts with TaMYB44, alleviating its repressive effects to restore starch synthesis and enhance grain weight. Notably, the functions of MYB44 appear to be partially conserved between wheat and rice, underscoring its potential as a target for genetic improvement. Our findings offer valuable insights into the transcriptional regulation of starch synthesis and provide genetic resources for enhancing grain yield in wheat and rice.

Dogs act as natural reservoirs of a large number of zoonotic pathogens, including the intestinal Cryptosporidium spp. and Giardia duodenalis, the most relevant protozoan species causing gastrointestinal diseases worldwide. An epidemiological study aiming to assess the prevalence and molecular diversity of Cryptosporidium and G. duodenalis was conducted in the Qinghai Tibetan Plateau area (QTPA), Northwest China. A total of 217 dog faecal samples were collected from sheep farms, dog farms and pet hospitals. The species/genotypes of Cryptosporidium and G. duodenalis isolated from dogs were identified by the PCR-based method targeting the partial 18S ribosomal RNA gene for Cryptosporidium and beta-giardin gene for G. duodenalis. The results of Cryptosporidium spp. and G. duodenalis infections in dogs showed an overall prevalence of 2.8% (6/217) and 6.9% (15/217), respectively. No Cryptosporidium and G. duodenalis co-infections were observed. The PCR-sequence results confirmed detection of C. parvum (n=3), C. canis (n=2), C. andersoni (n=1) and G. duodenalis assemblages B (n=9), C (n=3) and D (n=3) in dogs from the QTPA. The results of the present study demonstrated that dogs in the QTPA are commonly exposed to Cryptosporidium spp. and G. duodenalis. This may indicate that dogs are potential sources of infection of Cryptosporidium spp. and G. duodenalis transmitting to humans and animals.

To investigate freshwater snail distribution, which are intermediate hosts of Fasciola hepatica (Linnaeus, 1758), in pastoral areas around Qinghai Lake and provide a scientific basis for fascioliasis control in Qinghai Province of Qinghai-Tibet Plateau, 234 snail samples were collected from Gangcha and Haiyan Counties. After preliminary screening by morphology, species confirmation and phylogenetic analysis were performed using cytochrome c oxidase I (cox1) sequencing. All 234 samples passed quality control, and five species were identified: Radix auricularia (28.6%, 67/234) (Linnaeus, 1758), Tibetoradix kruglovi (27.4%, 64/234) (Vinarski, 2022), Orientogalba ollula (28.2%, 66/234) (Gould, 1859), Orientogalba viridis (6.0%, 14/234) (Quoy, 1833), and Gyraulus chinensis (9.8%, 23/234) (Dunker, 1848). Notably, T. kruglovi (an endemic species specific to the Qinghai-Tibet Plateau) was dominant in high-altitude Gangcha (ca. 3,300 m), in striking contrast to the multispecies assemblage in Haiyan (ca. 3,000 m), suggesting possibly altitude-driven geographic isolation. Haplotype network analysis indicated recent demographic expansions in T. kruglovi, O. ollula, and G. chinensis. This study represents the first basin-wide investigation of freshwater snail biodiversity and genetic structure around Qinghai Lake, providing critical data to inform targeted fascioliasis control strategies.

As the second important staple crop next to rice in China, common wheat (Triticum aestivum) plays a decisive role in national food security. Wild and semi-wild relatives of wheat provide abundant genetic resources for wheat genetic improvement. In China, wheat wide hybridization and chromosome engineering breeding initiated in the 1950s and developed into a well-defined theoretical and technical system over the next three decades through learning, exploration and practice. Subsequently, the technological innovation in alien chromatin identification and the isolation and analysis of alien resistance genes sponsored by continuous national projects have significantly enhanced China's impact on the world in this field. Eminent scientists such as Professor Li Zhensheng, who was awarded the Medal of the Republic before the National Day in 2024, have made outstanding contributions to the establishment and development of the research in this area in China. This article reviews the history of wheat wide hybridization and chromosome engineering breeding in China, aiming to honor the senior scientists and inspire future researchers to work hard in germplasm innovation and alien gene transfer, cloning and utilization in breeding.

Twenty-six QTLs associated with yield-related traits in wheat were identified through joint linkage and association analysis, with TraesCS5A03G0002500 being selected as a candidate gene for QGl.caas-5A.1. As a major staple crop worldwide, continuously increasing wheat yield is crucial for ensuring food security. Wheat yield is influenced by multiple traits, and elucidating the genetic basis of yield-related traits lays a foundation for future gene cloning and molecular mechanism studies. In this study, a recombinant inbred line (RIL) population derived from 292 lines of Hengguan 35/Zhoumai 18 was genotyped with the Affymetrix wheat 660 K SNP array. Combined with the phenotype of the RIL population in 13 environments, linkage analysis of six yield-related traits including plant height, grain number per spike, thousand-grain weight, grain length, grain width, and grain thickness was conducted. A total of 262 quantitative trait locus (QTLs) (logarithm of odds [LOD] > 3) were identified across 21 chromosomes, in which 50 QTLs were repeatedly detected in more than three environments. Numerous QTLs harbored cloned genes and overlapped with those reported in previous studies. Subsequently, joint analysis of genome-wide association study (GWAS) data from the advanced backcross-nested association mapping plus inter-crossed (AB-NAMIC) population and QTLs identified in the RIL population revealed 26 overlapping genomic regions. Notably, the QGl.caas-5A.1 associated with grain length on chromosome 5A was detected in both the RIL and AB-NAMIC populations, and TraesCS5A03G0002500 was selected as a candidate gene. A kompetitive allele-specific PCR (KASP) marker based on a variant [A/G] in TraesCS5A03G0002500 was developed and validated in a natural population containing 350 accessions. Taken together, these results provide valuable information for fine mapping and cloning of yield-related wheat genes in the future.