Table of Content

01 July 2026, Volume 59 Issue 13

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Molecular Detection of Disease Resistance Genes in Wheat Varieties Approved in Jiangsu Province

TAN YaNing, ZHANG BaoLong, CAO AiZhong, REN RunSheng

Scientia Agricultura Sinica. 2026, 59(13):  2763-2775.  doi:10.3864/j.issn.0578-1752.2026.13.001

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【Objective】Wheat is a crucial food crop in China, and its production security is closely tied to national food security. Jiangsu Province, a major wheat-producing region in eastern China, frequently suffers from diseases outbreaks due to its humid climate. Fusarium head blight (FHB), Stripe rust, and Powdery mildew are the primary diseases causing severe yield losses and quality degradation. Breeding and deploying multi-resistant varieties are the most economical and effective strategy for disease control. This study aimed to evaluate the comprehensive resistance of wheat varieties approved in Jiangsu over the past decade to these three diseases and to clarify the distribution and pyramiding of corresponding resistance genes using molecular markers, thereby providing a scientific basis and elite parental materials for disease resistance breeding in the region.【Method】Based on the approval data of 185 wheat varieties released in Jiangsu from 2014 to 2023, statistical analyses were conducted on their resistance levels to FHB, Stripe rust, and Powdery mildew. Varieties exhibiting resistance to at least two of these diseases were identified as “multi-resistant”. From 49 multi-resistant varieties, 44 germplasm accessions were selected for molecular detection. Specific markers were used to detect the FHB resistance gene Fhb1, the Powdery mildew resistance gene Pm21, and the Stripe rust resistance genes Yr5, Yr9, Yr15, Yr17, Yr18, Yr26, and Yr30. Experimental procedures included genomic DNA extraction using the CTAB method, optimized PCR systems and protocols, and the inclusion of known positive controls to ensure detection accuracy.【Result】According to the approval data, 53.52%, 43.92%, and 9.72% of the tested varieties showed resistance to Fusarium head blight, Stripe rust, and Powdery mildew, respectively. Five varieties (Ninghongmai 618, Yanmai 1902, Zhongyanmai 238, Yanmai 1903, and Ningmaizi 166) demonstrated resistance to all three diseases, and 49 varieties were resistant to at least two. Molecular marker detection indicated that all 44 multi-resistant accessions carried multiple resistance genes. Pm21, Yr5, Yr17, Yr18, and Yr26 were detected in all materials. Fhb1 was present in 84.1% of accessions, Yr15 in 93.2%, and Yr30 in 88.6%, while Yr9 was found in only three accessions. Notably, the varieties Xumai DH9 and Xumai 100 carried all nine tested resistance genes, demonstrating outstanding genetic potential for disease resistance and highlighting their suitability as ideal parental lines for multi-resistance breeding.【Conclusion】This study confirmed that the approved wheat varieties in Jiangsu Province have rich and unique gene resources for disease resistance through systematic molecular detection. It shows that the wheat varieties approved in Jiangsu province have high breeding and utilization potential. Their potential disease-resistant gene pool has not been fully developed. Based on this study, the screened wheat varieties can serve as a reserve germplasm bank, providing strong genetic support for future multi-resistance breeding in Jiangsu and East China, and alleviating the current challenge of limited sources of disease-resistant varieties.

Development and Application of InDel Marker Detection Technology for Field Identification of Maize Inbred Lines

WANG Rui, HU LiPing, ZHAO Wei, LIU ZhiHao, ZHANG MingQi, QING XiangYu, XU LiWen, HUO YongXue, GE JianRong, TIAN HongLi, YI HongMei, LIU YaWei, JIANG Bin, WU MingSheng, KUANG Meng, WANG FengGe

Scientia Agricultura Sinica. 2026, 59(13):  2776-2788.  doi:10.3864/j.issn.0578-1752.2026.13.002

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【Objective】Variety identification is vital for the security of agricultural production and integrity of seed markets. Current molecular approaches for identification of crop varieties predominantly require specialized laboratory equipment, resulting in relatively slow turnaround times and high costs. The Maize Point-of-Genuineness (M-POG) identification kit aims to extend variety identification from the laboratory to the field, address the lack of rapid on-site detection methods, and support the establishment of a rapid, efficient, and traceable molecular detection system for the modern seed industry. 【Method】Candidate InDel loci from the MaizeIDP50K chip were screened using a stepwise dichotomous partitioning method in combination with a dynamic optimization algorithm. Nine maize elite inbred lines and three closely related maize varieties were used to design dominant and co-dominant primers, which were evaluated by conducting quantitative real-time PCR and fluorescence capillary electrophoresis. An optimal combination algorithm was used to screen and determine the core set of loci. Based on the cycle threshold (Ct) value for the dominant primer for the core markers, a barcode conversion mechanism suitable for field detection was established. The detection thresholds were set using the core loci fingerprints of 2 270 maize inbred lines. The consistency and single sample detection efficiency were evaluated by comparison with the traditional method for variety identification using simple sequence repeat (SSR) markers, thereby verifying the accuracy and practicability of the M-POG method. 【Result】Eighty InDel candidate loci from the chip were screened for which 76 pairs of dominant primers were designed. Among these loci, 31 high quality loci showed clear qPCR amplification curves, high repeatability, and uniform amplification efficiency. Ultimately, 16 highly discriminative core loci were identified, achieving a variety recognition rate of 95.83% across the 2 270 inbred lines. The fingerprint detection threshold for each locus was calculated; loci with Ct<28 were classified as dominant type (coded as 1), and those with Ct>30 were classified as recessive type (coded as 0), enabling generation of a DNA fingerprinting for M-POG detection. Pairwise comparisons among the 2 270 inbred lines revealed that 98.29% of the pairs of lines differed at four or more loci. A comparative analysis between the M-POG method and the SSR molecular marker method, conducted using elite inbred lines and closely related varieties, showed that the samples were consistently discriminated by both methods in 90.91% of the comparisons. This result demonstrated the reliability of the M-POG method. Thus, the M-POG approach enables efficient screening of genetically dissimilar inbred lines, thereby substantially reducing the number of samples requiring further laboratory analysis. The complete field detection process required only 42 min, approximately one-quarter of the time required by standard identification procedures. 【Conclusion】Sixteen core InDel loci were selected for the M-POG method, achieving a variety recognition rate exceeding 95%. Based on the InDel on-site detection results, the minimum number of differential loci required for variety discrimination was determined. Development of the M-POG kit enables rapid and accurate identification of maize inbred lines in the field.

Whole Genome Association Study of Protein and Threonine Content in Mung Bean Seeds and Development of KASP Molecular Marker

CHEN ShengQi, LIU JinYang, HUANG JiaLi, LIU XiaoCen, ZHAO ShuoQian, LIN Yun, XUE ChenChen, YAN Qiang, CHEN JingBin, WU RanRan, CHEN Xin, CHENG JinPing, YUAN XingXing

Scientia Agricultura Sinica. 2026, 59(13):  2789-2801.  doi:10.3864/j.issn.0578-1752.2026.13.003

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【Objective】Protein and threonine are the core nutrients of mung bean seeds, which have a critical impact on the nutritional quality of mung beans. Among them, mung bean protein is a high-quality protein source for vegetarians due to its easy digestion, absorption, and rich biological activity, which is of great value to human health. This study aims to explore genetic loci and candidate genes significantly associated with the protein content and relative content of threonine in mung bean seeds, providing a theoretical basis for genetic improvement of protein and threonine content in mung bean seeds. 【Method】This study used Kjeldahl nitrogen determination method and liquid chromatography-mass spectrometry to determine the protein content and relative content of threonine in 279 mung bean varieties planted in 2022. The general linear model of Tassel5 software was used for genome-wide association study to explore genetic loci related to protein content and relative content of threonine in mung bean seeds. By combining linkage disequilibrium analysis, transcriptome comparative analysis, and comparative genomics, key candidate genes related to nitrogen metabolism were identified. 【Result】Six key loci located on chromosome 3 were identified, which are related to the protein content and relative content of threonine in mung bean seeds, and can explain 6.06% to 12.20% of phenotypic variation. These 6 loci are closely linked and can be mainly divided into two haplotypes, with Hap2 being the dominant haplotype. Then, KASP molecular marker was designed for the key candidate locus SLG03_966208 (A/G), which can successfully classify 80 mung bean varieties into three genotypes: AA, GG, and AG, with a detection accuracy rate of 87.5%. The key candidate gene EVM0000757 was identified within the locus linkage region, and the homologous gene AT5G65750 in Arabidopsis was significantly associated with nitrogen metabolism. Its gene expression was significantly different between high and low protein varieties (P=1.84E-03, |log₂FC|=1.39), making it a reliable candidate gene.【Conclusion】279 mung bean varieties were identified for their grain protein content and relative content of threonine. Six SNP loci were detected within the 939 296-1 039 749 bp region of chromosome 3. A KASP molecular marker for the SLG03_966208 (A/G) locus was developed, which can distinguish the high and low levels of protein and threonine content in mung bean grains. A key candidate gene related to nitrogen metabolism, EVM0000757, was identified.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

Effects of Potassium Dihydrogen Phosphate on Membrane System Damage, Defense Mechanisms and Yield in Peanut Under Intercropping with Maize

MA PengLong, WANG MeiWen, LI XingZhu, SHANG WanTing, LIU Dan, MA XinRou, WANG Jing, JIANG ChunJi, ZHAO XinHua, WANG XiaoGuang

Scientia Agricultura Sinica. 2026, 59(13):  2802-2814.  doi:10.3864/j.issn.0578-1752.2026.13.004

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【Objective】Maize-peanut intercropping system can enhance resource utilization efficiency and yield. However, the yield of peanuts in the edge and middle rows decreases after the flower-pegging stage due to the shading effect of maize. Foliar spraying of potassium dihydrogen phosphate can effectively alleviate this impact. This study aimed to elucidate the regulatory effects of foliar potassium dihydrogen phosphate application on membrane system damage, defense capacity, and yield in intercropped peanut systems, for providing a theoretical basis for high-yield cultivation management of maize-peanut intercropping system.【Method】Using the maize variety Liangyu 99 and the peanut variety Huayu 22 as experimental materials, a maize:peanut with 2:4 intercropping system was employed. During the flower-pegging stage, foliar applications of a 0.5% potassium dihydrogen phosphate solution were administered three times at a rate of 730 kg·hm^-2 per application, to investigate the effects of foliar potassium dihydrogen phosphate application on peanut membrane system damage, osmoregulatory capacity, antioxidant enzyme activity, and yield.【Result】In maize-peanut intercropping system, foliar application of potassium dihydrogen phosphate significantly reduced both the relative electrical conductivity and malondialdehyde content in peanuts compared with no treatment. During the pod-filling stage, relative electrical conductivity values in sole cropping plots, middle rows of intercropping plots, and edge rows of intercropping plots decreased by 15.56%, 35.29%, and 43.37%, respectively, compared with untreated controls, respectively. The trend in malondialdehyde levels mirrored that of relative electrical conductivity: sole cropping < intercropping middle rows < intercropping edge rows. Foliar application also markedly increased soluble protein, soluble sugar, and free proline content in peanuts while enhancing antioxidant enzyme activity, with edge rows demonstrating significantly superior effects to middle rows, which in turn outperformed sole cropping. Consequently, foliar application substantially improved individual peanut plants' fruit number, full fruit weight per plant, and total weight per hundred fruits compared with control conditions, with yield increases 7.24%-18.86% in sole cropping, 14.48%-20.02% in middle rows, and 16.75%-33.13% in edge rows. There were significantly greater yield improvements in intercropping system than in sole cropping.【Conclusion】During the flower-pegging stage, foliar applications of a 0.5% potassium dihydrogen phosphate solution could help mitigate leaf membrane system damage caused by peanut shading in maize, enhance its osmoregulatory capacity and antioxidant enzyme activity, increase both the number and weight of full fruits per plant as well as the total weight per hundred fruits, thereby improving peanut yield under the maize-peanut intercropping system.

Analysis of Physiological and Metabolic Response Mechanisms in Maize Varieties with Different Salt Tolerances Under Salt Stress

MA XiaoYing, QU XiaoYu, LI Tao, LIU WeiFan, LIU Hao, MA FengLan, WAN MengHu, WU Na, LIU JiLi

Scientia Agricultura Sinica. 2026, 59(13):  2815-2827.  doi:10.3864/j.issn.0578-1752.2026.13.005

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【Objective】As a major food crop in China, maize (Zea mays L.) is relatively sensitive to salt stress, which significantly inhibits its photosynthesis and causes metabolic disorders. Therefore, this study utilized maize varieties with different levels of salt tolerance to systematically investigate their photosynthetic physiology and metabolic responses under salt stress, with the aim of elucidating the underlying causes of inter-varietal differences in salt tolerance at the physiological and metabolic levels. 【Method】A pot experiment was conducted using salt-tolerant maize varieties Yinyu 238 (YY238) and Heyu 157 (HY157), and the salt-sensitive variety Xianyu 335 (XY335). Three salt stress concentration gradients were established: 0 mmol NaCl·L^-1 (CK), 120 mmol NaCl·L^-1 (S1), and 240 mmol NaCl·L^-1 (S2). Stress treatment was applied at the six-leaf stage of the maize seedlings. Samples were taken at 24 hours after stress treatment to measure the net photosynthetic rate (P_n), photosynthetic performance index (PI), ion content (K⁺, and Na⁺), and the content of malondialdehyde (MDA), proline (Pro), abscisic acid (ABA), and jasmonic acid (JA). Non-targeted metabolomics technology was used to analyze leaf metabolites. Differential metabolites were screened based on a variable importance in projection (VIP) > 1 and P<0.05, and pathway enrichment analysis was performed using the KEGG database. 【Result】Salt stress significantly reduced both P_n and PI. Compared with CK, the salt-sensitive variety XY335 exhibited a maximum P_n reduction of 39.8% under S2, while the salt-tolerant variety YY238 showed a relatively smaller reduction of 31.9%. In terms of ion homeostasis, salt stress significantly decreased the shoot K⁺/Na⁺ ratio, with XY335 experiencing a reduction of 76.4%, significantly higher than the 74.6% reduction in YY238. Salt-tolerant varieties demonstrated stronger osmotic adjustment capacity, with YY238 showing a 199.2% increase in Pro accumulation. In contrast, the salt-sensitive variety XY335 suffered more severe oxidative damage, with a 172.1% increase in MDA content. Hormonal responses indicated that XY335 had a higher increase in ABA (59.3%), while YY238 showed the largest increase in JA under S2, reaching 56.3%. Metabolomic analysis revealed that salt stress induced variety-specific metabolic reprogramming. The metabolic disorder in the salt-sensitive variety XY335 worsened with increasing stress intensity, with the number of differential metabolites surging to 127. In contrast, salt-tolerant varieties exhibited a stronger ability to maintain metabolic homeostasis. Salt-tolerant varieties specifically accumulated stress resistance-related metabolites: YY238 significantly upregulated amino acid derivatives and organic acids, including Nα-methylhistidine and trans-aconitate; HY157 specifically accumulated amino acids and their derivatives, such as γ-aminobutyric acid and L-glutamine, as well as phenolic acids like salicylic acid. In comparison, the salt-sensitive variety XY335 primarily accumulated nucleic acid metabolites related to energy metabolism, such as β-guanidinopropionic acid, and organic acids like isonicotinic acid. KEGG pathway enrichment analysis revealed that salt-tolerant varieties were commonly enriched in pathways, such as phenylpropanoid biosynthesis and ABC transporters. Additionally, YY238 specifically activated the biosynthesis of flavonoids and flavonols, while HY157 was significantly enriched in alanine, aspartate, and glutamate metabolism pathways. 【Conclusion】In summary, salt stress enhanced the adaptability of salt-tolerant maize varieties by maintaining photosynthetic performance and ion homeostasis, mitigating oxidative damage, and activating specific metabolic pathways. Salt-tolerant varieties maintained higher photosynthetic efficiency through precise ion compartmentalization and alleviate oxidative damage by rapidly accumulating osmotic adjustment substances and effectively scavenging reactive oxygen species. At the metabolic level, salt-tolerant varieties specifically activated pathways, such as phenylpropanoid biosynthesis and amino acid metabolism, providing the key material and signaling foundations for their salt tolerance.

PLANT PROTECTION

The Role of Iron in Enhancing Rice Blast Resistance Induced by Endophytic Streptomyces hyaluromycini OsiPR-1

ZHAO XingYu, GAO Yan, TANG XinKe, FENG YinChen, CHEN SiYing, GU TianYu, PENG JiaShi

Scientia Agricultura Sinica. 2026, 59(13):  2828-2840.  doi:10.3864/j.issn.0578-1752.2026.13.006

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【Objective】 Iron (Fe) is widely involved in critical life processes such as plant growth, development, and immune responses. Endophytes colonizing plant tissues have evolved efficient mechanisms for Fe uptake and homeostasis regulation, thereby positively influencing host disease resistance. This study aims to explore rice endophytic resources and investigate the impact of Fe nutrition on their ability to induce host resistance, and to provide a theoretical basis for green management of plant diseases.【Method】Endophytic bacteria were isolated from rice leaves using surface disinfection combined with the streak plate method. Target strains with antagonistic activity against Magnaporthe oryzae were screened using the dual culture method. The selected strain was identified through morphological observation and phylogenetic analysis based on 16S rRNA and multi-gene sequences. Its siderophore-producing ability was analyzed using CAS assay medium and by amplifying genes related to siderophore biosynthesis. Three culture conditions (normal Fe, Fe-deficient, and high Fe) were established to assess the reactive oxygen species (ROS) burst, Fe accumulation, and expression levels of Fe metabolism-related genes in rice leaves after spraying with the target strain. The efficacy of the endophyte in inducing rice resistance to rice blast under different Fe regimes was evaluated.【Result】An endophytic actinomycete strain, OsiPR-1, with stable antagonistic activity against M. oryzae, was isolated from rice and identified as Streptomyces hyaluromycini. This strain exhibited a strong ability to produce siderophores, with peak production (60.2%) observed on the 6th day under Fe-deficient conditions, and three siderophore biosynthesis gene clusters were detected in its genome. Reinoculation tests showed that the strain’s colonization and its induction of blast resistance were closely related to environmental Fe levels. Under normal Fe conditions, OsiPR-1 induced early accumulation of Fe and ROS in rice stomata to trigger immune responses, and subsequently regulated Fe homeostasis restoration via secreted siderophores to establish a stable symbiotic relationship, significantly reducing rice blast lesion length by 74.8%. Under high Fe conditions, Fe and ROS accumulation induced by OsiPR-1 appeared transiently in leaf veins only at 4 h post-treatment and then rapidly disappeared. Although the strain proliferated substantially, the symbiotic relationship was unstable, resulting in a lesion length reduction of only 64.5%. Under Fe-deficient conditions, Fe and ROS accumulation in stomata was weak. However, genes involved in Fe transport (OsIRT1), Fe homeostasis (OsFRO2), and lipoxygenase catalysis (OsLOX2) were significantly up-regulated (7.7-10.3 folds). The drastic fluctuation in Fe homeostasis and the subsequent intense immune response inhibited endophyte colonization, hindering the establishment of symbiosis and leading to a sharp decline in disease control efficacy.【Conclusion】Normal Fe supply is a critical prerequisite for S. hyaluromycini OsiPR-1 to enhance rice resistance against blast through a siderophore-mediated immune induction mechanism. This study provides an important theoretical foundation for developing efficient biocontrol agents against rice blast based on precise regulation of Fe nutrition and offers new insights into the role of Fe nutrition in plant-endophyte interactions.

Biological Activity and Mechanism of 2-Octyl-3-isothiazolinone Against Alternaria alternata Causing Tobacco Brown Spot

GUO FuQiang, PENG ZhiXin, LI DeRui, LIN MuSen, XU LiFeng, ZHANG ChengSheng

Scientia Agricultura Sinica. 2026, 59(13):  2841-2852.  doi:10.3864/j.issn.0578-1752.2026.13.007

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【Objective】The objective of this study is to clarify the inhibitory effect of 2-octyl-3-isothiazolinone, a metabolite from the co-culture of Trichoderma asperellum HG1 and Bacillus subtilis Tpb55, on Alternaria alternata (the pathogen causing tobacco brown spot) and its impact on the physiological metabolism of the pathogen, and to provide a theoretical basis for the application of this natural product in the management of plant fungal diseases.【Method】The antifungal activities of eight secondary metabolites from the co-culture against A. alternata were assessed using the mycelial growth rate method. The toxicity of 2-octyl-3-isothiazolinone was evaluated, and its EC₅₀ value was calculated. Field trials were conducted to determine its control efficacy against tobacco brown spot. Transcriptome sequencing was employed to analyze the gene expression profiles of A. alternata following treatment with 2-octyl-3-isothiazolinone. The impact on energy metabolism was assessed by measuring ATP content and ATPase activity. Propidium iodide (PI) staining and reactive oxygen species (ROS) detection were used to evaluate cell membrane integrity and oxidative stress levels, respectively.【Result】At a concentration of 10 μg·mL^-1, 2-octyl-3-isothiazolinone exhibited 100% inhibition against A. alternata, which was significantly superior to that of the positive control dimethachlon (27.60%). In the field experiment, the control efficacies at concentrations of 100 and 200 μg·mL^-1 were 80.80% and 84.33%, respectively, which were equivalent to that of the positive control (40% dimethachlon diluted 400-fold, 80.15%), and there was no toxic effect on tobacco itself. Further studies showed that 2-octyl-3-isothiazolinone exerted its antifungal effect through a dual mechanism: on the one hand, it significantly inhibited the key enzymes of glycolysis and the tricarboxylic acid cycle, blocking the energy metabolism of the pathogen; on the other hand, the expression levels of genes related to ROS scavenging in the pathogen (AALT_g9032, AALT_g9033) were significantly down-regulated, eventually leading to the massive accumulation of ROS, causing oxidative damage and destroying the integrity and biological function of the cell membrane. Further verification demonstrated that 2-octyl-3-isothiazolinone significantly reduced ATP content and ATPase activity, confirming disruption of energy metabolism. PI staining revealed concentration-dependent damage to cell membrane integrity, while DCFH-DA fluorescence staining indicated substantial intracellular ROS accumulation, triggering oxidative stress and exacerbating cellular damage. These results were consistent with the transcriptomic findings.【Conclusion】As a green pesticide with broad application prospects, 2-octyl-3-isothiazolinone interferes with the normal life activities of A. alternata through the synergistic effect of energy blockage-oxidative damage. This study provides a theoretical basis and application support for its use as a new antifungal agent to control tobacco brown spot.

Biocontrol Mechanism of Bacillus-Derived Lipopeptides Through Targeted Reduction of Pathogenicity in Ralstonia solanacearum

LIU YuFan, CHEN Zheng, CHEN DeJu, LIU Xin, XIAO RongFeng, WANG JiePing, WANG Xun, LIU Bo, HE Jin, CHEN MeiChun

Scientia Agricultura Sinica. 2026, 59(13):  2853-2866.  doi:10.3864/j.issn.0578-1752.2026.13.008

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【Objective】Bacterial wilt caused by Ralstonia solanacearum is a highly destructive soil-borne disease. Lipopeptides produced by Bacillus species are promising biocontrol agents. However, whether lipopeptides can exert biocontrol effects by attenuating the pathogenicity of R. solanacearum remains unclear. This study aims to investigate the mechanism by which lipopeptides target and reduce the pathogenicity of R. solanacearum, and to provide a theoretical basis for developing novel biocontrol agents based on pathogenicity regulation.【Method】The minimum inhibitory concentration (MIC) of lipopeptides was determined using the broth dilution method. The control efficacy of lipopeptides against bacterial wilt was evaluated through pot experiments. Defense-related enzyme activities including superoxide dismutase (SOD), catalase (CAT), polyphenol oxidase (PPO), and peroxidase (POD), as well as the contents of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) in tomato plants were determined using commercial assay kits. Combined with scanning electron microscopy, semi-solid plate motility test, and transcriptome sequencing, the effects of lipopeptides on the morphology, motility, and gene expression of R. solanacearum were systematically analyzed.【Result】The MIC of lipopeptides from Bacillus sp. FJAT-2349 was 0.1875 mg·mL^-1. At a sub-inhibitory concentration (0.075 mg·mL^-1), the incidence of tomato bacterial wilt decreased by 29.2% compared to the R. solanacearum control group. No significant changes were observed in the activities of SOD, CAT, PPO, POD, or the contents of MDA and H₂O₂ in tomato leaves, indicating that this concentration of lipopeptides failed to induce systemic resistance in tomato plants. SEM analysis showed that lipopeptide-treated R. solanacearum cells exhibited an elongated rod-shaped morphology with a length approximately three times that of normal cells, suggesting impaired cell division. Motility assays showed that bacterial movement was reduced by approximately 36%. Transcriptomic analysis revealed significant down-regulation of genes associated with cell division (ZapE), type III secretion system (T3SS) regulation (hrpB1, hrpB2), flagellar transcriptional regulator (flhC) and genes encoding virulence effector proteins. The gene expressions of cell division-related enzyme (ftsK), flagellin (fliC), histidine utilization-related enzyme (hipO), and RNA polymerase β' subunit (rpoC) were up-regulated. KEGG enrichment analysis indicated that differentially expressed genes were significantly enriched in pathways related to metabolism (energy metabolism, amino acid metabolism, and biosynthesis of secondary metabolites), genetic information processing (translation and RNA processing and metabolism), and environmental information processing (ABC protein transport), suggesting that lipopeptide treatment interferes with energy metabolism and substance transport, disrupts protein synthesis and turnover, reprograms the metabolic activity of R. solanacearum, and thereby inhibits its growth and reproduction.【Conclusion】Bacillus sp. FJAT-2349 lipopeptides at sub-inhibitory concentration exert their biocontrol effect primarily through a dual-core mechanism: blocking cell division progression and suppressing T3SS virulence, which synergistically interfere with pathogen energy metabolism and substance transport, disrupt protein synthesis and turnover, and reduce pathogen motility, thereby attenuating the pathogenicity of R. solanacearum.

SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT

The Influence of Straw Returning to the Field on the Soil Agglomeration Structure and Carbon Sequestration Effect of Meadows

HUANG YiWen, SUI ShiJiang, HE ZhiGang, YE Xin, XU JiaYi, ZOU XiaoJin, GONG Liang

Scientia Agricultura Sinica. 2026, 59(13):  2867-2877.  doi:10.3864/j.issn.0578-1752.2026.13.009

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【Objective】This study aimed to explore the effects of different straw return methods on the distribution and stability of soil aggregates, as well as the organic carbon components and carbon sequestration effect in meadow soil, so as to provide the theoretical and practical basis for improving soil aggregate structure and enhancing carbon sequestration potential.【Method】Based on an 11-year long-term field experiment, the treatments were no straw returning (CK), full no-tillage with straw mulching (SNT), half no-tillage with straw mulching (NT), full annual incorporation of straw (S1), and full biennial incorporation of straw (S2). 【Result】Compared with the CK treatment, the SNT treatment significantly increased soil organic carbon content in the 0-40 cm soil layer by 2.5% to 6.6%. Straw returning could increase the content of active carbon components (easily oxidized organic carbon and microbial biomass carbon) in the 0-40 cm soil layer to varying degrees. Among them, the S1 treatment significantly increased the easily oxidized organic carbon content in the 0-40 cm soil layer from 15.4% to 20.1%, and the NT treatment significantly increased the microbial biomass carbon content in the 0-40 cm soil layer from16.0% to 17.6%. The SNT treatment significantly increased the easily oxidized organic carbon and microbial biomass carbon content in the 0-40 cm soil layer, with increases of 53.6% to 54.1% and 34.6% to 57.2%, respectively. Compared with the CK treatment, the S2 treatment significantly increased the percentage of water-stable macroaggregates (>0.25 mm) in the 0-20 cm soil layer and improved the distribution of aggregates of different sizes. The content of 3-5 mm and 0.5-1 mm aggregates increased significantly by 16.8% and 49.0%, respectively. The S1 treatment and NT treatment significantly increased the percentage of water-stable macroaggregates (>0.25 mm) in the 20-40 cm soil layer. The content of >5 mm aggregates in the S1 treatment increased significantly by 10.3%, and the content of 3-5 mm aggregates in the NT treatment increased significantly by 25.7%. The content of easily oxidized organic carbon in the surface soil was significantly positively correlated with the percentage of >0.25 mm macroaggregates. The effects of different straw returning methods on the content of easily oxidizable organic carbon in each particle size aggregate were different. The SNT treatment and the S1 treatment significantly increased the content of easily oxidizable organic carbon in each particle size aggregate. Among them, the SNT treatment increased the content by 9.5%-15.9% in the 0-20 cm soil layer and by 9.1%-23.9% in the 20-40 cm soil layer, while the S1 treatment increased the content by 12.7%-23.2% in the 0-20 cm soil layer and by 14.8%-19.4% in the 20-40 cm soil layer.【Conclusion】Different straw returning methods could increase the soil organic carbon content and the content of active carbon components in the 0-40 cm soil layer to varying degrees, and improve the distribution of aggregate size classes. Among them, full amount no-tillage and straw mulching returning was more conducive to increasing soil organic carbon, easily oxidized organic carbon and microbial biomass carbon content, and promoting the formation of easily oxidized organic carbon in aggregates to enhance soil carbon sequestration potential.

Effect of Nitrogen Addition on Soil Microorganisms in Leymus secalinus Grassland in Northern Shanxi

JIA HuiLi, ZHU YiChen, CHEN SiXue, LIU Miao, DONG KuanHu, ZHAO Xiang, SHI YongHong

Scientia Agricultura Sinica. 2026, 59(13):  2878-2893.  doi:10.3864/j.issn.0578-1752.2026.13.010

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【Objective】This study aimed to investigate the effects of different nitrogen addition levels on soil microbial community structure, diversity and nitrogen cycling functional genes in Leymus secalinus grassland of northern Shanxi, which held the profound significance for understanding nutrient cycling in grassland ecosystems under the context of nitrogen deposition.【Method】This experiment was conducted based on the multi-level nitrogen (N) addition platform established in 2017 at the Youyu Loess Plateau Grassland Ecosystem Research Station, Shanxi Province. Eight treatments were set up: N0, N1, N2, N4, N8, N16, N24, and N32, corresponding to nitrogen application rates of 0, 1, 2, 4, 8, 16, 24, and 32 g N·m^-2·a^-1, respectively. Utilizing metagenomic technology, soil microorganisms were studied under different levels of nitrogen addition. 【Result】 (1) Nitrogen addition significantly altered the soil bacterial and fungal community structures. It increased the relative abundance of Acidobacteria and Thermodesulfovibrionales in the bacterial community while reducing that of Nitrospirales. In the fungal community, nitrogen addition increased the relative abundance of Ascomycota but decreased that of Mucoromycota and Basidiomycota. (2) Nitrogen addition significantly affected bacterial community structures richness, but had no significant effect on diversity, with no significant impact on the richness and diversity of fungal communities. (3) Microbial differential groups enriched with different levels of nitrogen addition were different. (4) Nitrogen addition significantly increased the relative abundances of nitrification genes amoABC and hao; the relative abundances of denitrification genes nirK and nirS significantly increased under the N4 treatment, while norBC relative abundances significantly increased under N1 and N24 treatments; the relative abundance of dissimilatory nitrate reduction genes nirBD and nrfAH increased significantly under the N16-N32 treatments, while the relative abundance of napAB and narGHI decreased significantly under the N1, N2, N4, N16 and N24 treatments; the relative abundances of nitrogen fixation genes nifDKH and vnfDKGH significantly increased under the N32 treatment (5) With the increase in nitrogen addition levels, soil water content (SWC) and pH decreased significantly, while total carbon (TC) and total nitrogen (TN) showed a trend of first increasing and then decreasing. Similarly, microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN) also exhibited a trend of first increasing and then decreasing. These changes in soil factors significantly affected the composition of the bacterial community structure, among which SWC was the most significant factor. (6) The genes involved in the process of dissimilatory nitrate reduction to ammonium were significantly negatively correlated with SWC and pH. The genes involved in the process of assimilatory nitrate reduction to ammonium were significantly positively correlated with MBC and MBN, nitrogen fixation process genes were significantly positively correlated with SWC, nitrification process genes were significantly negatively correlated with soil moisture, and were significantly positively correlated with NO₃^--N.【Conclusion】 In summary, nitrogen addition significantly affected the structure of soil microbial communities by influencing soil water content (SWC) and pH, which notably impacted the relative abundances of genes involved in nitrification, dissimilatory nitrate reduction to ammonium, nitrogen fixation, and denitrification processes, thereby altering the progression of soil nitrogen cycling. Soil microorganisms play a pivotal role in nutrient cycling. Further investigating the effects of nitrogen addition on soil microorganisms not only provided a theoretical basis for the rational management of Leymus secalinus grasslands in northern Shanxi but also held significant implications for the restoration of ecosystem functions in these grasslands.

Preparation and Application of Surfactant-Modified Castor Oil-Based Polyurethane-Coated Urea

LIAN SiRong, TANG JianWei, WANG YongJie, HUANG JingLong, WANG ChengQian, LI Rui, MAO KeLu, WANG Xu, YUE YanJun, LIU Yong, WANG BaoMing, LIU PengFei

Scientia Agricultura Sinica. 2026, 59(13):  2894-2905.  doi:10.3864/j.issn.0578-1752.2026.13.011

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【Objective】Surfactants affect the properties and controlled-release performance of coated fertilizers by influencing the structure of polyurethane film materials based on castor oil. This study offered a theoretical support for the practical use of surfactant-modified plant oil-based polyurethane coated urea in agriculture through a methodical examination of the properties of coated urea, surfactant-modified castor oil-based polyurethane film, and field tests. 【Method】Using surfactant modification, castor oil-based polyurethane-coated urea (BPCU) was created. The surfactant modification mechanism was examined, and FT-IR was used to corroborate the polyurethane synthesis. The effects of surfactants on the hydrophobic properties of membrane materials were investigated. In Zhaodi Village, Xinxiang, Henan Province, a field experiment was carried out using homemade slow-release fertilizer to reduce fertilizer application by 20% while using regular urea as the control. The impact of BPCU on summer maize yield was examined, and Zhongjinyu 2513 was the test crop. 【Result】By improving the compatibility of the polyurethane matrix and creating a pseudo-crosslinking effect, the surfactant could make the high hydroxyl value castor oil-based polyol and PM-200 more evenly distribution on the coating film's surface. Additionally, the cross-linking density was further enhanced by lowering the energy consumption of the interfacial reaction between the high hydroxyl value castor oil-based polyol and PM-200 to form polyurethane. The improved membrane material had a decreased residual carbon content, according to the results. The improved membrane shell's water contact angle was 105.5°, a 13.0 % increase from the unmodified membrane shell's 93.4°. The modified membrane material had a 7-day water absorption rate of 1.64%, a porosity of 1.14%, and a swelling degree of 1.18%. The 7-day water absorption rate, porosity, and swelling degree were all reduced by 76.8%, 76.7%, and 25.3%, respectively, when compared with the unmodified membrane material. The slow-release duration of BPCU was extended from 70 to 83 days at 0.5% surfactant addition, which was 18.6% longer than that of castor oil-based polyurethane-coated urea (PCU). With a 20% decrease and a 5% coating, the corn yield of the BPCU fertilizer was 11 908.1 kg·hm^-2, a 22.0% increase over the maize yield of 9 757.2 kg·hm^-2 following the application of regular urea. In contrast, the thousand-grain weight likewise rose by 16.9%. The yield and thousand-grain weight of maize were significantly impacted by the application of BPCU. It mostly influenced the thousand-grain weight of maize, which in turn impacted its yield. 【Conclusion】It was demonstrated from aspects such as nitrogen release period and summer maize yield that the performance of BPCU was superior to that of the unmodified PCU. The application of BPCU could be regarded as an effective way to reduce fertilizer application and increase efficiency. The creation of novel coated controlled-release fertilizers was supported theoretically and technically by this study.

HORTICULTURE

Character Analysis and Evaluation of Different Pepper Varieties in Plateau Solar Greenhouses

WANG GuangSheng, DAWAPUCHI, CIWANG, PUQIONG, NIMASANGZHU

Scientia Agricultura Sinica. 2026, 59(13):  2906-2918.  doi:10.3864/j.issn.0578-1752.2026.13.012

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【Objective】Under plateau solar greenhouse conditions, the agronomic traits of roots, stems, leaves and fruits of different pepper varieties were analyzed, the relationship between each trait and the yield of a single pepper plant was explored, and a comprehensive evaluation was made, so as to provide theoretical and practical reference for improving cultivation techniques, introducing excellent varieties (lines), and enriching local resources. 【Method】Descriptive statistics, entropy weight method, path, correlation and other multivariate analysis methods are used to make judgments, and the membership function method and cluster analysis are used for comprehensive evaluation to clarify the main traits that affect the yield of a single plant and the pepper varieties suitable for regional ecological conditions. 【Result】There are certain differences between different organs of each variety. The coefficient of variation of the four root traits ranges from 7.70% to 16.20%, the variation coefficient of the five stem traits ranges from 6.99% to 16.50%, and the variation coefficient of the three leaf traits ranges from 11.50% to 15.90%. The coefficients of variation of the four fruit traits ranged from 15.90% to 26.50%. The coefficients of variation of single fruit quality, number of fruits per plant, and yield per plant were 39.60%, 44.20%, and 19.50% respectively. The entropy weight method combined with path analysis showed that the direct effects affecting the yield of a single plant were ordered from large to small as leaves > fruits > stems > roots. Correlation analysis showed that in root organs, the number of roots was significantly positively correlated with yield per plant; in leaf organs, leaf length, leaf width, and petiole length were all highly significantly positively correlated with yield per plant. Principal component analysis simplified 19 agronomic traits into 6 principal components, with a cumulative contribution rate of 81.631%, and established a comprehensive evaluation model D=0.435μ₁+0.222μ₂+0.112μ₃+0.092μ₄+0.072μ₅+0.066μ₆, three varieties with excellent comprehensive scores, Zhongjiao 506, Zhongjiao Y213, and Zhongjiao Y216, were screened out. When promoting varieties, the local preference for screw peppers should be taken into consideration, and Zhongjiao 129, Meiluo 28, and Zhongjiao 209 were listed as targets for promotion and application. Based on the characteristics of each organ and D value, 30 peppers were divided into 5 groups. The differences in leaves and fruits between each category were significant. TypeⅠ has a compact plant type, firm fruits, and numerous fruits; TypeⅡ has shorter leaves and petioles, and overall, the plant height is shorter and the yield per plant is lower; the fruits of typeⅢ are short and thick, and the quality of single fruit is the highest; the fruits of typeⅣ are short and thick, and the quality of single fruit is lower than that of typeⅢ; the number of typeⅤ is the largest, and the D value is in the middle position, showing a more balanced performance. 【Conclusion】The types of introduced resources are relatively rich. Under plateau solar greenhouse conditions, leaf traits and the number of roots are the direct factors that affect the yield of a single pepper plant. Based on the characteristics of each organ, principal component analysis and membership function method can be used to comprehensively evaluate the introduced pepper varieties and screen out excellent varieties. Leaf and fruit traits can be used as the main basis to distinguish taxa and enhance the utilization of different resources.

Analysis of Sugar and Organic Acid Components and Related Metabolic Gene Expression During Fruit Development in 14 Blueberry Cultivars

HUANG MengChi, CHEN LiNa, GAO DengTao, LIU Li, HAN JingYi, DUAN ShuYun, LIU ZhaoNing, LI HaoXian, LU ZhenHua

Scientia Agricultura Sinica. 2026, 59(13):  2919-2932.  doi:10.3864/j.issn.0578-1752.2026.13.013

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【Objective】To characterize the dynamic changes in sugar and organic acid composition across three key developmental stages in 14 major blueberry cultivars currently cultivated in China, which differ markedly in ripening period and flavor profile, and to elucidate the relationship between PEPC and organic acid metabolism in blueberry fruit, thereby providing theoretical support for blueberry production and flavor quality improvement.【Method】Fruits of 14 blueberry cultivars at three key ripening stages (green, pink, and blue) were used as materials. Soluble sugar and organic acid components were determined by HPLC, and the expression patterns of the PEPC gene were analyzed by qRT-PCR.【Result】During fruit ripening, the accumulation dynamics of TA and TSS varied among different blueberry cultivars and could be classified into three acid-accumulation patterns (continuous decline, stable-decline, and increase-decline) and three sugar-accumulation patterns (continuous increase, stable- increase, and decline-increase). Glucose and fructose were the major soluble sugar components in blueberry fruits, whereas citric acid, quinic acid, and malic acid were the predominant organic acids. Among them, quinic acid exhibited a relatively high coefficient of variation. Cluster analysis based on sugar and organic acid composition divided the 14 cultivars into four groups. In four cultivars, the major organic acids were citric acid, quinic acid, and malic acid; seven cultivars were dominated by citric acid and malic acid; and one cultivar was characterized primarily by citric acid and quinic acid. In these 12 cultivars, the proportions of glucose and fructose were relatively balanced. In contrast, the remaining two cultivars showed a clear predominance of quinic acid, with fructose content significantly higher than that of glucose. Total sugar content in blueberry fruits was significantly positively correlated with glucose and fructose contents, while total organic acid content was extremely significantly positively correlated with the contents of citric acid, quinic acid, oxalic acid, and lactic acid. In the three blueberry cultivars representing different acid-accumulation patterns, PEPC expression increased from the green fruit stage to the pink fruit stage, but was downregulated from the pink fruit stage to the blue fruit stage. During this latter period, the contents of malic acid and citric acid in the fruits also decreased correspondingly.【Conclusion】During blueberry fruit development, the variation patterns of TA and TSS were closely associated with cultivar characteristics. The major soluble sugar components were glucose and fructose, while the predominant organic acids were citric acid, quinic acid, and malic acid. Based on organic acid composition, the cultivars could be categorized into two types: high-citric-acid and high-quinic-acid types. In three blueberry cultivars representing different acid-accumulation patterns, PEPC expression was downregulated from the pink fruit stage to the blue fruit stage, which was consistent with the declining trends of malic acid and citric acid contents, indicating that PEPC is involved in the metabolism of organic acids in blueberry fruits.

FOOD SCIENCE AND ENGINEERING

Differential Absorption of Components in Buckwheat Polyphenols via MKN28/Caco-2 Continuous Transport Model and Their Lipid- Lowering Effects

YAO YiJun, CHEN Yu, JU XingRong, WANG LiFeng

Scientia Agricultura Sinica. 2026, 59(13):  2933-2945.  doi:10.3864/j.issn.0578-1752.2026.13.014

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【Background】 Polyphenolic compounds exhibit pronounced lipid-lowering effects, and their absorption in the human gastrointestinal tract varies considerably, which critically influences their bioavailability. Buckwheat, a natural food being rich in polyphenols, has been suggested to possess unique lipid-lowering potential. 【Objective】 This study aimed to investigate the differential absorption of buckwheat polyphenols using an MKN28/Caco-2 sequential transport model, and to systematically evaluate their lipid-lowering effects and underlying mechanisms through cellular assays and gut microbiota fermentation experiments.【Method】 Buckwheat polyphenol extract was characterized by ultra-high-performance liquid chromatography coupled with quadrupole Orbitrap mass spectrometry (UHPLC-Q-Orbitrap MS). Based on the MKN28/Caco-2 transport model, the extract was fractionated into readily absorbable and poorly absorbable components. The lipid-lowering effects of the absorbable fraction were assessed in adipocytes using Oil Red O staining and intracellular lipid quantification. In addition, the impact of the non-absorbable fraction on gut microbiota composition was evaluated in vitro fermentation.【Result】 Component analysis revealed that there are 23 kinds of polyphenolic compounds rutin and protocatechuic acid exhibited relatively high signal intensities. The absorbable fraction significantly reduced lipid accumulation in adipocytes, with triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) levels in 3T3-L1 cells decreased by 17.8%, 28.6%, and 15.2%, respectively. Furthermore, in vitro fermentation demonstrated that the poorly absorbable fraction (BWP-AP) modulated gut microbiota composition by increasing the relative abundance of Bacteroides and Bifidobacterium, while decreasing Escherichia-Shigella and Alistipes.【Conclusion】 Buckwheat polyphenol extract contained 23 identified phenolic compounds. The readily absorbable fraction exerted significant intracellular lipid-lowering effects, whereas the poorly absorbable fraction regulated metabolic homeostasis via modulation of gut microbiota. These findings revealed a dual-pathway mechanism involving direct host absorption and microbiota-mediated regulation, providing a theoretical basis for the development of functional foods based on buckwheat polyphenols.

Elevation-Related Variation and Key Environmental Drivers of Fresh Bamboo Shoot Quality in Chimonobambusa utilis from Jinfo Mountain in China

ZENG QingPing, LIU Han, NIE TangJie, JIANG XuanBin, LI HongYan, TANG JiaJia, SHEN Ling, CHEN LiJie, LUO XiaoGang, XIE ChengWei, TONG Long

Scientia Agricultura Sinica. 2026, 59(13):  2946-2961.  doi:10.3864/j.issn.0578-1752.2026.13.015

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【Objective】 This study aimed to reveal the variation patterns of Chimonobambusa utilis fresh shoot quality along an altitudinal gradient and to identify the main soil nutrient driving factors, providing a theoretical basis for the precise cultivation and efficient utilization of high-quality C. utilis shoots.【Method】Based on the distribution pattern and sprouting regularity of C. utilis in the nature reserve, five standard sample plots were established at altitudes of 1 850, 1 950, 2 050, and 2 150 m, respectively. The nutritional components, amino acid composition, flavor substances, and texture of fresh shoots at different altitudes were systematically determined, and the nutrient status of the understory soil was simultaneously analyzed. Redundancy analysis (RDA), linear regression, and structural equation modeling (SEM) were used to comprehensively analyze the altitudinal differences in fresh shoot quality and its main soil factors.【Result】Among basic nutrients and mineral elements, there were no significant differences (P>0.05) in protein, crude fat, soluble sugar, vitamin C, amino acid-state nitrogen, reducing sugar, triterpenoids, ash, dietary fiber, sodium, magnesium, calcium, iron, zinc, and selenium among different altitudes. However, potassium content increased significantly with increasing altitude (P<0.05), and the potassium content of fresh bamboo shoots at 2 150 m was 1.50 times higher than that at 1 850 m. There were no significant differences in total amino acids and the content of 13 amino acids among different altitudes (P>0.05), but serine, alanine, and tyrosine were sensitive to altitude. Specifically, serine in fresh bamboo shoots at 2 150 m, 2 050 m, and 1 950 m was significantly higher than that at 1 850 m; alanine in fresh bamboo shoots at 1 950 m was significantly higher than that at 1 850 m; tyrosine in fresh bamboo shoots at 2 050 m was significantly higher than that at 1 850 m (P<0.05). Among the taste and texture indicators of fresh bamboo shoots, there were no significant differences in cellulose, lignin, oxalic acid, total flavonoids, umami, bitterness, and tasteless amino acids among different altitudes (P>0.05). However, tannin and sweet amino acids were significantly affected by altitude (P<0.05). The highest hardness of fresh bamboo shoots was at 1 850 m, and the highest crispness was at 2 150 m. Regression analysis showed that potassium and serine content exhibited a positive linear relationship with increasing altitude, while tannin content exhibited a negative linear relationship (R²=0.50). The RDA results indicated that soil pH, SOC (soil organic carbon), and TN (total nitrogen) were the primary soil nutrient factors driving the differentiation of fresh bamboo shoot quality. The SEM results further revealed that altitude indirectly regulated the nutritional and flavor quality of fresh bamboo shoots by promoting TN and SOC accumulation and decreasing soil pH. 【Conclusion】High altitude was conducive to the accumulation of potassium and sweet amino acids while reducing tannins, thus improving flavor. TN, SOC, and pH were the main driving factors for the differentiation of fresh bamboo shoot quality, and altitude indirectly affected the quality of fresh bamboo shoots by regulating soil nutrient content.

ANIMAL SCIENCE·VETERINARY SCIENCE

Transcription Factor ZBTB6 Activates the Transcription and Function of NORHA in Sow Granulosa Cells

SUN ZhenYu, CUI YiGe, LI YuQi, WANG SiQi, DU Xing, LI QiFa

Scientia Agricultura Sinica. 2026, 59(13):  2962-2974.  doi:10.3864/j.issn.0578-1752.2026.13.016

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【Objective】 This study was aimed to characterize the distal core promoter of the porcine lncRNA NORHA gene, and explore the mechanism by which the transcription factor (TF) ZBTB6 activated its transcription and function, so as to provide a basis for analyzing the transcriptional regulatory network of NORHA. 【Method】 The sequence of the distal core promoter of the NORHA gene in Yorkshire pigs was obtained by PCR amplification and sequencing. Bioinformatic method was used to predict the TF-binding sites (TFBSs) and to characterize the ZBTB6 encoding sequence. A ZBTB6 overexpression vector was constructed by restriction enzymatic digestion, and its overexpression efficiency in sow granulosa cells (sGCs) was verified by qPCR and western blot. The effects of ZBTB6 on the activity of the distal core promoter and transcription of NORHA in sGCs were determined by luciferase assay and qPCR. The binding of ZBTB6 to the ZBTB6-binding site (ZBS) in the distal core promoter in sGCs was detected by chromatin immunoprecipitation (ChIP) and DNA sequencing. The concentrations of progesterone (P4) and estradiol (E2) in follicular fluid were measured by ELISA. ZBTB6 and NORHA levels in follicles, and BCL2 and BAX levels in ZBTB6-overexpressing sGCs were detected by qPCR. 【Result】 TFBSs of 255 TFs, including ZBTB6, were predicted in 526-bp of the distal core promoter of Yorkshire NORHA gene. The overexpression vector successfully achieved the overexpression of ZBTB6 levels in sGCs. The transcriptional levels of NORHA in sGCs were markedly upregulated in ZBTB6-overexpressing sGCs. Correlation analysis revealed that ZBTB6 and NORHA levels in follicles were markedly positively correlated. A ZBS motif was discovered at -2276/-2264 nt in the distal core promoter. Luciferase assay revealed that ZBTB6 activates the activity of the distal core promoter via the ZBS motif. ChIP and sequencing confirmed that ZBTB6 directly bound to the ZBS motif in the distal core promoter of the NORHA gene in sGCs. The ZBTB6 gene in pigs and other mammalian species was evolutionally conserved and all contained a conserved DNA-binding domain. ZBTB6 was markedly upregulated during sow follicular atresia and showed a marked positive correlation with the P4/E2 ratio, a marker for follicular atresia. Co-transfection experiments showed that overexpression of ZBTB6 markedly downregulated the anti-apoptotic gene BCL2 levels, upregulated the pro-apoptotic gene BAX levels, and downregulated the BCL2/BAX ratio, an apoptotic marker, whereas knockdown of NORHA reversed this. 【Conclusion】 ZBTB6 was a transcription activator of the NORHA gene in sGCs, which accelerated sGC apoptosis and follicular atresia by directly activating NORHA transcription.

Exploration of Candidate Genes Affecting Meat Quality of Jingyuan Chicken Based on Weighted Gene Co-expression Network Analysis

ZHAO Wei, TIAN JinLi, CHEN SiYu, YANG LiJuan, MA LiNa, ZHANG GuoJun, ZHANG Hu, HU WeiLong, YU HaiRong, ZHANG HaiBo, ZHANG Juan

Scientia Agricultura Sinica. 2026, 59(13):  2975-2992.  doi:10.3864/j.issn.0578-1752.2026.13.017

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【Background】The Jingyuan chicken, a local chicken breed in Ningxia, boasts advantages such as delicious meat quality and unique flavor. However, it faces challenges including slow growth rates and uneven intramuscular fat deposition. Previous studies have shown that adding 0.6% fresh corn extract (FCE) to the diet could significantly improve the meat quality of Jingyuan chickens, but the key candidate genes through which FCE affected meat quality remained unclear. 【Objective】This research aimed to delve deeper into the key candidate genes influenced by FCE affecting meat quality in Jingyuan chickens, thereby providing a scientific basis for improving meat quality and optimizing meat chicken feeding strategies.【Method】 In earlier studies, 120 Jingyuan chickens at 135 days old (1.5±0.2 kg), raised under the same conditions, were divided into four groups. The control group was fed a basic diet, while the experimental groups received diets supplemented with 0.3%, 0.6%, and 0.9% FCE, respectively. After 45 days of feeding, the chickens reached 180 days old, at which point breast muscle was collected to assess meat quality, determining that the optimal addition amount was 0.6% FCE. Based on the outcomes of previous research, serum and breast muscle lipid factors were measured for the control and 0.6% FCE groups. Weighted Gene Co-expression Network Analysis (WGCNA) was utilized to conduct an in-depth analysis of the transcriptome sequencing data of breast muscle, uncovering key candidate genes significantly associated with meat quality.【Result】 The serum levels of triglycerides (TG) and very-low-density lipoproteins (VLDL) in the 0.6% FCE group were significantly higher than those in the control group (P<0.05). The TG, total cholesterol (TC), and high-density lipoproteins (HDL) in the breast muscle tissue of the 0.6% FCE group were also found to be significantly elevated (P<0.05). Through WGCNA analysis, a total of 19 co-expression modules were identified, with six target modules significantly associated with meat quality being selected: purple, midnight blue, light green, cyan, grey60, and salmon (|R²| ≥ 0.5, P<0.5). KEGG pathway enrichment analysis and Gene Ontology (GO) functional enrichment analysis of the hub genes within these modules revealed significant enrichment of the hub genes in the purple module in pathways, such as insulin signaling and mitochondrial autophagy; the midnight blue module hub genes were enriched in pyrimidine and purine metabolism; the light green module hub genes were associated with protein processing in the endoplasmic reticulum and MAPK signaling pathway; the cyan module hub genes mainly enriched pathways related to cell adhesion molecules and phagosome processes; the grey60 module hub genes were linked to vascular smooth muscle contraction and actin cytoskeleton regulation; and the salmon module hub genes enriched in MAPK signaling and p53 signaling pathways. Through comparative analyses of hub genes and differently expressed genes (DEGs), ten overlapping genes were identified. Protein interaction and functional enrichment analyses identified CLDN18, NKX2-1, and TRIB1 as key candidate genes influencing meat quality. 【Conclusion】 This study ultimately identified key candidate genes affecting meat quality, namely CLDN18, NKX2-1, and TRIB1. The findings provided a reference for understanding the molecular mechanisms by which FCE regulates meat quality and offered a new perspective for the improvement of meat quality in local chicken breeds.