Table of Content

01 August 2026, Volume 59 Issue 15

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Functional Characterization and Molecular Marker Development of the Wheat Transcription Factor Gene TaWRKY65-3A

ZHANG YanFei, WU ZiYue, XUE ZhiWei, MA DongYun, KANG GuoZhang, FENG Wei, XIE YingXin, JING RuiLian, WANG WanZhang, MAO XinGuo, WANG ChenYang

Scientia Agricultura Sinica. 2026, 59(15):  3237-3251.  doi:10.3864/j.issn.0578-1752.2026.15.001

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【Objective】WRKY transcription factors are the plant-specific transcription factor families, and play a crucial role in plant growth, development, nutrient uptake, and response to biotic and abiotic stresses. Characterized the function of the wheat transcription factor gene TaWRKY65-3A and developed the molecular markers, providing genetic resources for wheat germplasm improvement through molecular breeding in this research.【Method】The sequence of TaWRKY65-3A was cloned from the cDNA of wheat variety Hanxuan 10. The domains were analyzed via the SMART website. The expression patterns of TaWRKY65-3A gene were analyzed by qRT-PCR under phytohormone ABA treatments, PEG and NaCl stresses. Predicted the polymorphisms of TaWRKY65-3A through the Wheat Genome Variation Joint Database, and validated them by using 21 wheat germplasms with rich genetic diversity and further developed molecular markers. Association analysis between TaWRKY65-3A haplotypes and phenotypic traits was carried out in a natural population consisting of 323 wheat accessions. The haplotype of 189 modern wheat cultivars from different released years were analyzed, aiming to clarify the selection trends of superior haplotypes during wheat breeding in China.【Result】TaWRKY65-3A is 1 998 bp in full length, comprising 2 exons and 1 intron, and harbors the characteristic WRKY domain and a C₂H₂-type zinc finger motif typical of the WRKY transcription factor family. The promoter region of TaWRKY65-3A contains a variety of cis-acting elements, including hormone-responsive elements, (e.g., to abscisic acid and methyl jasmonate) and stress-responsive elements (e.g., to drought and low temperature). Moreover, the expressions of TaWRKY65-3A was influenced by ABA, PEG and NaCl stresses. Six SNPs were identified in the TaWRKY65-3A region, forming three haplotypes: Hap-3A-1, Hap-3A-2, and Hap-3A-3. dCAPS markers were developed at positions of 1 265 bp (G/C) in the coding sequence and 1 658 bp (T/C) in the non-coding sequence, which were significantly associated with thousand-kernel weight, yield per plant, spikelet number per spike, and kernel number per spike across multiple environments. Notably, the haplotype Hap-3A-2 was demonstrated to be significantly correlated with higher spikelet number per spike, a great kernel number per spike, a higher thousand-kernel weight, and a higher yield per plant in multiple environments, including drought, high temperature, and the combined stress of drought and high temperature, and it was positively selected during wheat breeding in China.【Conclusion】TaWRKY65-3A gene responded to phytohormones ABA, PEG and NaCl stress, and were significantly correlated with spikelet number per spike, kernel number per spike, thousand-kernel weight and yield per plant under various environments such as drought, high temperature, and the combined stress of drought and high temperature. Hap-3A-2 is the favorable haplotype with higher spikelet number per spike, more kernel number per spike, higher thousand-kernel weight, and higher yield per plant. The molecular markers of dCAPS developed in this study provide valuable tools for marker-assisted breeding, facilitating the selection of germplasm with superior yield-related traits.

Functional Identification and Breeding Evolution of the Key Flowering Gene GhSUVs in Upland Cotton

WEI JIAZHI, ZHANG WENJIAO, ZHAO YAN, LI YING, JU JISHENG, YANG JUNNING, FENG YUYU, MA XIONGFENG, SU JUNJI, WANG CAIXIANG

Scientia Agricultura Sinica. 2026, 59(15):  3252-3266.  doi:10.3864/j.issn.0578-1752.2026.15.002

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【Objective】Early maturity is an important target trait in cotton genetic improvement in China, and flowering time is a key indicator of earliness. H3K9 histone lysine methyltransferases (SUVs) are involved in plant development and flowering regulation in model plants such as Arabidopsis thaliana and rice. This study aimed to investigate the role of the SUV gene family in flowering regulation in upland cotton and to identify elite allelic variations of key members. The findings provide valuable genetic resources for breeding early-maturing cotton cultivars.【Method】Protein sequences of the SUV family from Arabidopsis were used as reference sequences. HMMER, MEME, and TBtools were employed to identify and analyze SUV family members in upland cotton at the whole-genome level. Candidate genes involved in flowering regulation were screened through integrated RNA-seq and RT-qPCR analyses. Virus-induced gene silencing (VIGS) was used to validate the functions of candidate genes in flowering regulation. Resequencing data from 419 upland cotton accessions were analyzed to identify SNPs within GhSUV1, which were further validated by Sanger sequencing. Allele frequencies and genetic differentiation were examined across different cotton ecological regions.【Result】A total of 200 SUV homologous proteins were identified in nine higher plant species, including rice. Phylogenetic analysis divided them into SUVH and SUVR clades, indicating a relatively conserved evolutionary pattern. In upland cotton, 28 GhSUV genes were identified. Their expansion was mainly driven by polyploidization events. Most collinear gene pairs showed Ka/Ks<1, suggesting strong purifying selection. Gene structure and motif analyses revealed high similarity among members within the same clade. Promoter analysis showed that GhSUV genes are enriched in light-responsive and hormone-responsive cis-elements, indicating their involvement in multiple developmental regulatory pathways. RNA-seq and RT-qPCR analyses showed that GhSUV1 and GhSUV13 were expressed at higher levels in early-maturing materials. They were also highly expressed in leaves and certain floral organs. Functional validation using VIGS demonstrated that silencing GhSUV1 delayed squaring and flowering by 5.6 and 6.0 days, respectively. Silencing GhSUV13 delayed these stages by 5.8 and 6.8 days compared with the control. Moreover, suppression of GhSUV1 and GhSUV13 significantly reduced the expression of flowering-related genes GhSOC1, GhAP1, GhFT, and GhCAL, while significantly increasing GhSVP expression. These results indicate that both genes participate in flowering regulation. Sequence analysis identified a SNP in the exon region of GhSUV1 that was significantly associated with flowering time. The early-flowering GG allele increased in frequency with latitude. Genetic differentiation index and nucleotide diversity analyses revealed significant differences in the GhSUV1 genomic region between early- and late-maturing cultivars, suggesting that this locus has undergone breeding selection.【Conclusion】GhSUV1 and GhSUV13 are involved in flowering regulation in upland cotton. The elite allele of GhSUV1 is closely associated with early flowering and has been subject to breeding selection. These findings provide important genetic resources for the improvement of early-maturing cotton varieties.

Regulatory Mechanisms of the Biosynthesis and Metabolism of Dhurrin and the Innovation Strategies for Sorghum Germplasm with Non- or Low- Dhurrin Content

LÜ XiaoSong, LAI ShangKun, ZHU Chao, LIU Wei, XIA QianHao, JIANG Lei, LI GuiYing, ZHANG ZhiJin, ZHU Li

Scientia Agricultura Sinica. 2026, 59(15):  3267-3282.  doi:10.3864/j.issn.0578-1752.2026.15.003

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Sorghum bicolor (L.) Moench, as the fifth most important cereal crop globally, plays an irreplaceable strategic role in ensuring food security, supporting feed production, and promoting the development of the brewing industry. Dhurrin, as cyanogenic glycoside unique to sorghum, exhibits a dual role in plant defense and industrial applications. On one hand, as a key stress-responsive metabolite, dhurrin enhances plant resilience against adverse stresses such as drought, salinity, and pests and diseases. On the other hand, upon tissue damage or during processing, dhurrin can be hydrolyzed by β-glucosidase to release toxic hydrogen cyanide (HCN), posing potential risks of livestock health and brewing quality. This paper systematically reviews the spatiotemporal distribution characteristics of dhurrin, elucidates its metabolic network (encompassing biosynthesis, activation, detoxification, and recycling) and regulatory mechanisms. Research indicates that the biosynthesis of dhurrin mainly relies on the sequential catalysis of cytochrome P450 enzyme family (CYP79A1 and CYP71E1), with the glycosylation reaction catalyzed by glycosyltransferases such as UGT85B1, and is completed through the cooperative reaction of multi-enzyme complexes; its activation process is closely related to the degree of plant tissue damage, and the hydrolysis action of β-glucosidase is a key step. In the field of animal feed utilization, residual dhurrin in insufficiently processed sorghum stalks or silage may induce HCN poisoning in animals, limiting its safety as feedstock. During the brewing process, cyanide released from dhurrin degradation can further react to form the carcinogen ethyl carbamate (EC), which poses a potential threat to the food safety of Baijiu (Chinese liquor). In response to the aforementioned risks, this paper summarizes multiple detoxification strategies, including physical pretreatment, enzyme inhibitors, and microbial degradation. More importantly, based on a deep understanding of the dhurrin metabolic pathway, this paper further proposes a feasible approach for developing non- or low-dhurrin sorghum germplasm through molecular breeding. By utilizing gene-editing technologies such as CRISPR/Cas9 to precisely knock out key biosynthetic genes, thereby reducing risks at the source. This research provides comprehensive theoretical support and practical guidance for the targeted regulation of metabolism, safe processing, and efficient utilization of dhurrin in sorghum, which is of great significance for promoting the green and sustainable development of the entire sorghum industry chain.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

Regulation of Wheat Lodging Resistance Through Culm Structure Optimization and Synergistic by Uniconazole and Prohexadione Calcium

WANG Qi, GUO ShuangQi, MEI LanXin, WU ChengDong, ZHENG ChangHong, ZHOU Qin, CAI Jian, WANG Xiao, Huang Mei, ZHONG YingXin, JIANG Dong

Scientia Agricultura Sinica. 2026, 59(15):  3283-3301.  doi:10.3864/j.issn.0578-1752.2026.15.004

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【Objective】The mechanical physicochemical properties of wheat culms are key intrinsic determinants of lodging resistance. This study aimed to determine the relationships between plant growth regulator-induced changes in culm physicochemical traits and improvements in mechanical properties.【Method】Two wheat varieties with different lodging resistance, Yangmai 22 and Ningmai 13, were selected. The experiment included four foliar spray treatments: water (control), uniconazole(20 mg·L^-1), prohexadione- calcium(4.5 g·L^-1) and a combination of uniconazole and prohexadione-calcium, applied at key growth stages. Culm morphological characteristics, mechanical traits, structural carbohydrate content, and yield components were systematically measured to analyze the regulatory effects of different treatments on culm lodging resistance and grain yield.【Result】Plant growth regulator treatments significantly reduced plant height and center of gravity height, shortened basal the first and second internodes, while increasing basal internode culm diameter, culm wall thickness, dry matter accumulation, and culm filling degree, with increased cell wall structural carbohydrate content. In 2022-2023, lignin content in the second basal internode at maturity stage of Yangmai 22 and Ningmai 13 increased by 21.17%-25.52% and 5.87%-8.61%, respectively. This optimization of morphology and matter accumulation further improved culm mechanical properties, as evidenced by significantly increased breaking force, section modulus, and breaking moment of the second basal internode, along with significantly reduced lodging index and markedly improved field lodging conditions. In 2023-2024, field lodging rate at maturity stage decreased by 7.81%, 8.94%, and 11.95%, respectively. Correlation analysis showed that breaking strength and breaking moment were significantly positively correlated with section modulus and culm wall thickness, while significantly negatively correlated with bending stress, center of gravity height, and basal internode length. The lodging index was significantly negatively correlated with breaking strength, breaking moment, section modulus, culm diameter, and wall thickness of the basal internodes, but significantly positively correlated with bending stress, plant height, center of gravity height, and internode length. In terms of yield, foliar application of uniconazole, prohexadione-calcium, and their combination increased grain yield. In 2022-2023, grain yield increased by 4.03%-28.40% in Yangmai 22 and by 9.35%-22.48% in Ningmai 13 compared with the control, respectively. In 2023-2024, spray treatments of Yangmai 22 increased by 15.71%, 14.97%, and 12.33% compared with the control, respectively.【Conclusion】Foliar application of uniconazole, prohexadione-calcium, and their combination significantly enhanced wheat culm lodging resistance by optimizing the morphological structure of basal internodes, including reducing plant height, shortening internodes, and increasing culm diameter; promoting dry matter accumulation, culm filling, and lignin deposition; and improving culm mechanical properties. These treatments also maintained or increased grain yield.

Response of Dry Matter Accumulation and Yield Stability in Intercropped Maize to Biochar Application Under Nitrogen Reduction

WANG XiaoLi, FAN ZhiLong, NAN YunYou, CHEN GuiPing, JIANG LongXiu, YIN Wen, HU FaLong, CHAI Qiang

Scientia Agricultura Sinica. 2026, 59(15):  3302-3314.  doi:10.3864/j.issn.0578-1752.2026.15.005

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【Objective】In the arid irrigated areas of northwest China, monoculture cropping systems, high dependence on nitrogen fertilizer, and inadequate farmland protection have led to degraded soil quality and reduced farmland productivity, severely constraining crop yield improvement and sustainable production. This study integrated intercropping with leguminous crop, nitrogen reduction, and biochar application to investigate their synergistic effects on maize dry matter accumulation and distribution characteristics, maize yield, and yield stability, aiming to provide a theoretical basis and technical support for establishing a green and efficient sustainable maize production model in this region.【Method】The experiment was conducted from 2022 to 2024 at the Oasis Agriculture Comprehensive Experimental Station of Gansu Agricultural University. A split-split-plot field experiment was employed, with the main plots assigned to cropping patterns (maize ǁ fresh-edible pea, IM; sole maize, SM), the subplots to nitrogen application levels (conventional rate N₁, 360 kg·hm^-2; reduced nitrogen by 30% N₂, 250 kg·hm^-2), and the sub-subplots to biochar treatments (application, C, 15 t·hm^-2; no application, B). Indicators including maize dry matter accumulation and distribution, grain yield, and yield stability were systematically measured.【Result】Compared with monocropping, the intercropping pattern significantly increased maize grain yield, dry matter accumulation, and yield stability. Reducing nitrogen by 30% significantly decreased maize dry matter accumulation, maximum growth rate, ear dry matter allocation ratio, and grain yield, but biochar application effectively mitigated these negative effects. Biochar increased the maximum aboveground dry matter growth rate by 6.7%-33.7% for maize ǁ fresh-edible pea and by 9.4%-24.1% for sole maize, respectively. Under intercropping conditions, the treatment with 30% nitrogen reduction combined with biochar application (IMN₂C) showed no significant differences in grain yield, dry matter accumulation dynamics, and yield stability compared to the full nitrogen rate treatment (IMN₁C) and was significantly superior to other treatment combinations, and compared to the conventional nitrogen application without biochar in monocropped maize (SMN₁B), this treatment increased grain yield by 17.0%-19.2%. The underlying mechanism primarily involved biochar promoting maize root growth by increasing soil organic carbon and total nitrogen content, optimizing the root-to-shoot ratio, thereby ensuring efficient dry matter accumulation throughout the entire growth period. Particularly during the critical grain-filling stage, it maintained a high growth rate and promoted the preferential allocation of dry matter to the ears.【Conclusion】In the arid irrigated areas of northwest China, the integration of maize fresh-edible pea intercropping and biochar application, along with a 30% reduction in nitrogen fertilizer input, can effectively maintain maize yield and yield stability by improving soil fertility, promoting root development, and optimizing dry matter partitioning. This approach represents a feasible agronomic strategy for achieving synergistic reduction of chemical nitrogen fertilizer use and stable maize production in the region.

Comprehensive Evaluation of Defoliation Ability of Different Sugarcane Varieties/Lines and Analysis of Cell Morphology and Physiological Differences in Leaf Sheath Abscission Zone

DING Qian, HE LiLian, QIAN ZhenFeng, WU QiBin, ZHAO ChangZu, LIN PeiXia, NONG WeiYou, WANG YuJie, ZHANG YuanYuan, QUE YouXiong, LI FuSheng

Scientia Agricultura Sinica. 2026, 59(15):  3315-3327.  doi:10.3864/j.issn.0578-1752.2026.15.006

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【Objective】This study aimed to evaluate the defoliation ability of 56 sugarcane varieties/lines, to identity the germplasm with easy defoliation, and to elucidate the cellular morphological and physiological basis of sugarcane defoliation, so as to provide a theoretical basis for exploring the regulatory mechanism of sugarcane defoliation.【Method】The natural defoliation number, total number of nodes, natural defoliation rate, leaf sheath inclination angle (+8, +9, +10 leaf position) and defoliation force of 56 sugarcane varieties/lines were systematically investigated, and defoliation ability was comprehensively evaluated by entropy weight method and hierarchical cluster analysis. The base of the leaf sheath abscission zone at +2, +5 and +8 leaf positions of the easy-to-defoliate sugarcane variety XTT20 and the hard-to-defoliate sugarcane variety YAU11-818 were analyzed by combining paraffin sections,enzyme activities and endogenous hormone content the cell morphology and physiological basis of sugarcane defoliation.【Result】Entropy weight analysis showed that the comprehensive score of defoliation of 56 sugarcane varieties/lines ranged from 0.001 to 0.978. Systematic cluster analysis divided the 56 sugarcane varieties/lines into 3 groups, including 3 easily-defoliation varieties (XTT20, XTT22, YAU01-106), 7 difficult-to-defoliation lines, and 46 medium defoliation varieties/lines, accounting for 5.36%, 12.50% and 82.14% of the total number of varieties/lines, respectively. Compared with the difficult-to-defoliation line YAU11-818, the easily-defoliation variety XTT20 had a higher natural defoliation rate, larger leaf sheath inclination angle and smaller defoliation force; The cells in the leaf sheath abscission zone of the +8 leaf position of the defoliant varieties were more loosely arranged, the cell density was relatively low, and the cell degradation was larger, the percentage of aerated tissue area was significantly higher than that of the difficult-to-defoliation line (14.54% increase), and the difference between different leaf positions was significant (+8 leaf position increased by 16.97% compared to +5 leaf position). In addition, the polygalacturonase activity, cellulase activity and abscisic acid (ABA) content in +8 leaf sheath of XTT20 were significantly higher than those of YAU11-818. On the contrary, the content of indole-3-acetic acid (IAA) was higher in the difficult-to-defoliation line YAU11-818.【Conclusion】The degree of abscission zone and defoliation ability of +8 leaf sheaths at the maturity stage of sugarcane may be related to the activity of polygalacturonase, cellulase activity, peroxidase activity, ethylene synthesis precursors, abscisic acid, auxin content, etc., which can provide a reference for the identification of defoliation of sugarcane varieties.

PLANT PROTECTION

Establishment and Application of RPA-CRISPR/Cas12a-based Visual Detection of Botrytis cinerea

ZHOU YuXin, SHEN YongJun, ZHU GuangXue, CHAI ALi, XIE XueWen, LI Lei, FAN TengFei, SUN XianHua, XIANG Sheng, LI BaoJu, SHI YanXia

Scientia Agricultura Sinica. 2026, 59(15):  3328-3339.  doi:10.3864/j.issn.0578-1752.2026.15.007

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【Objective】Botrytis cinerea, the causal agent of gray mold in facility vegetables, is characterized by a wide host range and rapid post-infection spread, often leading to severe economic losses. This paper aims to establish an early and precise detection method for gray mold before its visible symptoms appear, and to provide technical support for the early monitoring and targeted control of gray mold in facility vegetable cultivation.【Method】A conserved gene sequence (BC1G_05726) from B. cinerea strain B05.10 was used as the target. Specific recombinase polymerase isothermal amplification (RPA) primers and CRISPR/Cas12a crRNAs were designed and screened. Key parameters including reaction time and the concentration ratio of Cas12a to crRNA were systematically optimized to establish a rapid visual detection method combining RPA and CRISPR/Cas12a. Fluorescence signals were visualized using a portable blue light transilluminator. The specificity, sensitivity, and applicability of the developed method were evaluated using complex environmental samples (infested soil, seeds, symptomatic plants, and air samples).【Result】The optimal crRNA (crRNA-1) and primer pair (RPA-bc-F/R) were selected. The optimized detection conditions were as follows: RPA reaction at 37 ℃ for 20 min, followed by CRISPR/Cas12a reaction with final concentrations of 200 nmol·L^-1 for Cas12a and 50 nmol·L^-1 for crRNA for another 20 min. The entire detection process was completed within 40 min. The sensitivity assay revealed a detection limit of 10^-4 ng·μL^-1 of genomic DNA of B. cinerea, which was 100-fold more sensitive than conventional PCR. Specificity testing showed that only B. cinerea produced positive fluorescence signals, while none of the other 14 tested fungal and bacterial species or the negative control generated any signal, indicating excellent specificity of the assay. In application tests, the method successfully detected B. cinerea strains from different hosts. After artificial inoculation on cucumber leaves, the pathogen was stably detected as early as 1 d post-inoculation, preceding symptom appearance. Dynamic monitoring of air samples in the facility environment demonstrated that B. cinerea could be detected in the air at 60 h post-inoculation, at which time typical disease lesions had not yet developed on the plants, validating the early warning capability of this method.【Conclusion】The RPA-CRISPR/Cas12a-based visual detection method established in this study enables rapid, sensitive, and specific detection of B. cinerea under isothermal conditions without requiring complex instrumentation, and the results can be read by the naked eye. This method is suitable for on-site detection of the pathogen in soil, seed, plant, and air samples.

Sexual Dimorphism in Phototactic Behavior and Dynamic Transcriptional Regulation of Opsins in Myllocerinus aurolineatus

FU NanXia, YUAN JinKang, LUO ZongXiu, LI ZhaoQun, BIAN Lei, XIU ChunLi, ZHOU Li, CHEN ZongMao, CAI XiaoMing

Scientia Agricultura Sinica. 2026, 59(15):  3340-3351.  doi:10.3864/j.issn.0578-1752.2026.15.008

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【Background】Phototaxis is widely utilized to monitor and control pest populations in agricultural practices. Tea weevil (Myllocerinus aurolineatus) is a server regional outbreak tea pest in Chinese tea plantations, and its control currently relies heavily on chemical pesticides. Previous studies have shown that adults of M. aurolineatus exhibit strong phototaxis at night.【Objective】To lay basis for developing light-based management strategies, this study clarified the effects of different light spectra on the phototactic behavior of male and female M. aurolineatus, and further revealed the regulatory effects of different light stimuli on the transcriptional expression of their opsin genes.【Method】Firstly, to reveal the phototactic characteristic of this pest, phototaxis of female and male adult M. aurolineatus to eight types of monochromatic light was tested through behavioral choice experiments. Subsequently, bioinformatic analysis was performed to identify opsin genes in M. aurolineatus, and a phylogenetic tree of identified opsins along with those from other insect species was constructed using the neighbor-joining method. Finally, the spatiotemporal expression profiles of opsin genes were examined across different developmental stages, under circadian rhythm, and various light stimuli using quantitative real-time PCR (RT-qPCR).【Result】Behavioral assays revealed that violet light elicited the highest attractiveness to males and was the second preferred wavelength for females, surpassed only by red light. In contrast, the blue light was the least attractive to both sexes. Moreover, the phototaxis rates of males to yellow, green and blue light were significantly higher than those of females. Bioinformatic analysis showed M. aurolineatus contains only two opsin genes, a long-wavelength-sensitive opsin (Ma-LW) and an ultraviolet-sensitive opsin (Ma-UV); no blue-light-sensitive opsin gene was detected, which was in accordance with the lowest phototactic rate to blue light observed in behavioral assays. Spatiotemporal expression profiling showed that both Ma-LW and Ma-UV were highly expressed in adults, with no significant difference throughout the light-dark cycle. Furthermore, dark adaptation up-regulated both Ma-LW and Ma-UV expression in M. aurolineatus adults, while subsequent short-term light stimuli (red, blue and violet) significantly reversed this up-regulation. Among these, red light exerted significantly stronger suppressive effect on Ma-LW expressional abundance than blue or violet light, while no significant difference in suppressive effect on Ma-UV expression was detected among the three lights.【Conclusion】Sexual dimorphism is evident in the phototactic behavior of adult M. aurolineatus, and exposure to different light spectra can down-regulate opsin gene transcription in dark-adapted M. aurolineatus, these findings contribute to the development of light-based M. aurolineatus management strategies.

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

Effects of Cover Crop Species and Incorporation Ratios on Soil Fertility and Economic Returns in Salinized Cotton Fields

WANG YuXia, LUO ChuXin, LI ZiShuang, DU MengYang, LI HongJie, ZHOU XiaoLin, LIU BaiLin, CAO WeiDong, LI ZiZhong

Scientia Agricultura Sinica. 2026, 59(15):  3352-3373.  doi:10.3864/j.issn.0578-1752.2026.15.009

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【Objective】Aiming at the soil degradation problems (including low organic matter content, nutrient imbalance, and secondary salinization) caused by long-term continuous cropping of cotton fields in northern Shandong, this study aimed to identify the suitable catch crop species and optimal incorporation ratio for salinized cotton fields in this region, so as to provide the technical support for ecological restoration and green production of cotton fields.【Method】The experiment was conducted at the experimental demonstration base in Wucheng County, Dezhou City, Shandong Province from 2022 to 2023. A split-plot design was adopted, with the main plots being three catch crop species: Orychophragmus violaceus, Vicia villosa Roth, and Lolium perenne L., and the subplots being four incorporation ratios: 100%, 70%, 30%, and 0. Winter fallow bare land was used as the control (CK). The biomass and nutrient accumulation of catch crops were systematically determined before incorporation, and soil physicochemical properties (such as available phosphorus, available potassium, alkali-hydrolyzable nitrogen, organic matter, pH, and electrical conductivity), microbial biomass carbon and nitrogen, and activities of key soil enzymes (such as urease, sucrase, and catalase) were dynamically monitored at the seedling, flowering, and harvest stages of cotton. The comprehensive soil fertility index (IFI) was calculated using the fuzzy mathematics membership function combined with principal component analysis, and a comprehensive evaluation was conducted by integrating cotton yield and annual economic benefits.【Result】(1) There were significant differences in soil improvement functions among different catch crops (P<0.05): Vicia villosa Roth had the highest total nitrogen accumulation (70.4 kg·hm^-2), showing the best performance in improving soil nitrogen supply and comprehensive fertility; Lolium perenne L. had the highest fresh weight (31 951.4 kg·hm^-2), dry weight (6 058.0 kg·hm^-2), and total carbon accumulation (3 129.9 kg·hm^-2), with significant advantages in carbon input and biomass accumulation; Orychophragmus violaceus had stable nutrient release and balanced overall performance. (2) The incorporation ratio significantly regulated soil indicators (P<0.05), and the 100% incorporation treatment performed the best. Compared with CK, it reduced soil electrical conductivity by 32.7% on average, increased organic matter by 15.9%, and enhanced the activities of urease, sucrase, and catalase by 142.3%, 46.4%, and 54.6%, respectively. (3) There were growth stage differences between soil physicochemical properties and biological activities: organic matter was the core for driving factor at the cotton seedling stage (significantly positively correlated with microbial biomass carbon and nitrogen, and enzyme activities, P<0.05); the inhibitory effect of electrical conductivity was significantly enhanced at the harvest stage (correlation coefficients -0.5--0.9). (4) The IFI under 100% incorporation treatment of Vicia villosa Roth was the highest (0.76), with a cotton yield of 5 263.6 kg·hm^-2 and a net income of 21 477.0 yuan/hm², an increase of 50.0% compared with CK; although 100% incorporation of Lolium perenne L. increased soil carbon storage, it significantly inhibited cotton growth, and the net income decreased by 24.0% compared with CK.【Conclusion】100% incorporation of Vicia villosa Roth was the optimal improvement model for slightly salinized cotton fields in northern Shandong, which could achieve the coordinated improvement of ecological and economic benefits. The matching mechanism between catch crop functional characteristics and incorporation ratio under this model provided a reference for the improvement of similar salinized farmland.

Regulation Effect of Biochar Combined with Slow-Release Fertilizer on Soil Water Use in Oasis Irrigation Area

WANG Shen, LI Pan, LIANG Miao, WANG LiPing, ZHANG DianKai, ZHAO LianHao, FAN ZhiLong, HU FaLong, NAN YunYou, YIN Wen, CHAI Qiang

Scientia Agricultura Sinica. 2026, 59(15):  3374-3386.  doi:10.3864/j.issn.0578-1752.2026.15.010

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【Objective】To address water scarcity in oasis irrigation areas and issues such as excessive nitrogen fertilizer application and low water use efficiency in maize production, this study investigated the effects of biochar combined with slow-release fertilizers on maize soil water use characteristics under reduced nitrogen application, so as to provide the practical evidence for high maize yields and efficient water resource utilization.【Method】A split-plot design was employed. The main plots featured soil conditioner (biochar (B)) and no conditioner (C). Split plots included two nitrogen fertilizer types: traditional chemical nitrogen fertilizer (T) and slow-release nitrogen fertilizer (S). Sub-split plots tested two nitrogen application rates: traditional nitrogen application rate (N2: 360 kg·hm^-2), and a 30% reduced traditional nitrogen application rate (N1: 252 kg·hm^-2). This study investigated the effects of biochar combined with slow-release fertilizer on maize yield, water consumption, and water use efficiency under reduced nitrogen application during the 2023-2024 growth season.【Result】Applying biochar with slow-release fertilizer to reduce nitrogen application (BSN1) increased pre-sowing soil water storage capacity in nitrogen-reduced maize and lowered maize water consumption throughout its entire growth stage. Regarding pre-sowing soil water storage, biochar increased by 5.3%-6.5% compared with no conditioner; no significant differences were observed between slow-release fertilizer and traditional chemical nitrogen fertilizer, nor between reducted nitrogen application and traditional nitrogen application rates (P>0.05). Compared with traditional chemical nitrogen fertilizer application rate without soil conditioner (CTN2), BSN1 increased pre-sowing soil water storage by 11.1%-12.5%. Biochar reduced water consumption during the maize sowing to jointing stage and the big flare to flowering stage by 6.4%-6.7% and 4.6%-5.5%, respectively, though the evapotranspiration modulus showed no significant differences. Slow-release fertilizer reduced maize water consumption from sowing to maturity by 8.1%-10.0%, but only lowered the evapotranspiration modulus during the sowing to flowering stage. Reduced nitrogen application decreased water consumption during the big flare stage to flowering stage by 6.3%-7.5%, with no significant difference in the evapotranspiration modulus. Comprehensive analysis of soil conditioner, nitrogen fertilizer type, and nitrogen application rate showed that BSN1 reduced maize water consumption by 21.4%-29.5% and the evapotranspiration modulus by 6.3%-7.6% compared with CTN2 during the sowing to flowering stage, water consumption decreased by 8.0%-9.9% during flowering to maturity stage, while the evapotranspiration modulus showed an upward trend. The combination of biochar and slow-release fertilizer effectively coordinated water supply and demand during different growth stages of nitrogen-reduced maize. Consequently, maize under BSN1 exhibited a 13.6%-14.8% reduction in total water consumption compared with CTN2, with grain yield and water use efficiency increasing by 8.6%-9.1% and 24.1%-24.7%, respectively.【Conclusion】The application of biochar combined with slow-release fertilizer and reduced nitrogen rates significantly increased maize grain yield and water use efficiency compared with traditional chemical nitrogen fertilizers without soil conditioners at traditional nitrogen application levels. This approach served as a viable technology for nitrogen-saving, yield-enhancing, and water-efficient maize production in the Hexi Oasis irrigation areas.

HORTICULTURE

Screening Evaluation and Physiological Difference Analysis of Kiwifruit Germplasm Resources for Tolerance to Combined Stress of Iron Deficiency and Waterlogging

FENG YaQing, SUN YaHan, ZHOU QiaoHui, LIN Peng, LI JiaYi, WANG HuiShen, LIU Tao, LIU ZhanDe, WANG NanNan

Scientia Agricultura Sinica. 2026, 59(15):  3387-3399.  doi:10.3864/j.issn.0578-1752.2026.15.011

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【Objective】To screen kiwifruit germplasm with tolerance to combined stress of iron (Fe) deficiency and waterlogging, and to clarify physiological and biochemical differences under single or combined stresses of the two factors, so as to provide a scientific basis and genetic resources for the breeding of kiwifruit germplasm tolerant to Fe deficiency and waterlogging.【Method】Seventeen tissue-cultured kiwifruit genotypes belonging to four species, namely Actinidia chinensis (Ac), A. macrosperma (Am), A. polygama (Ap) and A. valvata (Av), were used to determine plant growth parameters, leaf net photosynthetic rate (Pn), and biomass in various parts for assessing the tolerance differences under combined stress of Fe deficiency and waterlogging. Moreover, two tolerance-contrasting genotypes (Av1 and Av5) from the same kiwifruit species were analyzed for the similarities and differences in physiological responses to combined stress.【Result】Using the weight-based membership function method and cluster analysis, the combined tolerance of the seventeen kiwifruit genotypes was classified into three groups: High tolerant, including Av1, Av2, Am1, Av3, Av4, Am2, Ap1, Am3, and Av5; Medium tolerant, including Acd1, XX, Ap2, YX, Acd2, and HWD; And low tolerant, including QH and QM. Across kiwifruit species, the greatest tolerance was observed in Av and Am, followed by Ap, and then Ac. The superior tolerance of Av and Am was related to their excellent aboveground performance, including plant height, new leaf number, leaf area, leaf length and width, fresh weight of aboveground parts and whole plant; and that of Ap was associated with its good root performance, including root volume and root fresh and dry weight. Combined stress enhanced proton extrusion, ferric chelate reductase activity and Fe concentration in roots of both genotypes. However, in new leaves, combined stress decreased Fe concentration in Av5 but increased that in Av1, which contributed to more positive root physiological regulation in Av5. Both waterlogging and combined stresses increased root porosity of both genotypes. In Av1, combined stress increased cortex proportion and decreased the ratio of the endodermis to phloem of roots and stems. Additionally, combined stress increased C/N in various plant organs, particularly for Av1.【Conclusion】The tolerance in combined Fe deficiency and waterlogging stress differed substantially among four kiwifruit species or different genotypes within the same species. Compared with Av5, Av1 is better able to uptake and transport Fe, to adjust anatomical structures of roots and stems, and to raise C/N in various plant organs, thereby performing better in tolerance to combined stress of Fe deficiency and waterlogging.

Effects of Pneumatic Atomization Spraying on Pesticide Deposition Efficiency and Control Efficacy Against Postharvest Gray Mold in Cut Roses

LI Lei, YANG MingShan, HUA WeiHan, LIU Xin, SHAO Qing, CHEN Rui, WANG WenSheng, GAO JunPing, SUN XiaoMing

Scientia Agricultura Sinica. 2026, 59(15):  3400-3411.  doi:10.3864/j.issn.0578-1752.2026.15.012

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【Objective】To address the problems of pesticide waste, cross-infection, and low efficiency associated with traditional postharvest gray mold control in cut roses, this study aims to explore and establish an efficient and low-pesticide-consumption chemical control technique.【Method】The droplet size and density of pesticide were determined using the water-sensitive paper method to optimize the operational parameters for pneumatic atomization spraying. Three main cut rose cultivars, ‘Peach Avalanche’ ‘Nightingale’ and ‘Jumilia’, were used as test materials to systematically compare four pesticide application methods. The treatments included: (1) pneumatic atomization spraying at 0.40 MPa, (2) hydraulic electric spraying at 0.25 MPa, (3) hydraulic manual spraying at 0.16 MPa, and (4) traditional flower head dipping. Their effects on pesticide deposition parameters and gray mold control efficacy were evaluated. Pesticide deposition parameters were determined using the allura red tracer method and the water-sensitive paper method. Two-level multiple linear regression models of deposition and control efficacy were constructed to quantitatively analyze the relationships among application parameters, deposition efficiency and control efficacy, and to explore the internal mechanism underlying the differences in control efficacy among different treatments.【Result】(1) Spray pressure and target distance significantly affected droplet characteristics during pneumatic atomization spraying. The volume median diameter (VMD) was the smallest and droplet density the highest at 0.4 MPa and 40 cm target distance, representing the optimal application parameter combination. (2) The droplet volume diameter produced by pneumatic atomization spraying was 87.7 μm, the droplet density was 332.6 droplets/cm², and the uniformity (CV=8.85%) was significantly better than that of hydraulic electric spraying and hydraulic manual spraying. (3) The pesticide deposition rate of pneumatic atomization spraying reached 67.6%, which was equivalent to 89.0% of that of flower head dipping (76.0%). This value was significantly higher than that obtained with hydraulic electric spraying (33.7%) and hydraulic manual spraying (24.7%). (4) After 96 h of simulated postharvest cold-chain transportation, the gray mold incidence of ‘Peach Avalanche’ treated with pneumatic atomization spraying was only 5.56%, with a control efficacy of 82.18%, which did not differ significantly from that of traditional flower head dipping, but was significantly superior to that of conventional hydraulic spraying methods. Notably, the pesticide dosage per stem was merely 0.27 mL, representing an 84.66% reduction compared with that of flower head dipping (1.76 mL/stem). On the 8th day of vase life, the survival rate of cut flowers treated with pneumatic atomization spraying ranged from 60% to 70%, comparable to that of traditional flower head dipping. Two-level multiple linear regression models were constructed for droplet parameters-deposition efficiency (adjusted R²=0.964) and deposition efficiency- control efficacy (adjusted R²=0.838). The results confirmed that pneumatic atomization spraying, with its high droplet density and superior deposition performance, achieved optimal control efficacy. (5) Verification tests on ‘Nightingale’ and ‘Jumilia’ confirmed that pneumatic atomization spraying achieved stable control efficacy across cultivars with differing resistance levels, exhibiting no significant difference from traditional flower head dipping.【Conclusion】Pneumatic atomization spraying is a postharvest gray mold control technology with the advantages of high efficiency, pesticide use reduction and cost savings, which can provide important support for improving the quality and efficiency of the cut rose industry.

FOOD SCIENCE AND ENGINEERING

Flavonoid Composition and Gut Homeostasis-Modulatory Effects of Shatianyu (Citrus grandis L. Osbeck) Whole Fruit Fermented by Lactobacillus rhamnosus

MA XIAONI, WANG JIANMING, ZHANG MINGWEI, ZHANG LIN, LUO GUANGYI, ZHANG RUIFEN, DENG MEI

Scientia Agricultura Sinica. 2026, 59(15):  3412-3423.  doi:10.3864/j.issn.0578-1752.2026.15.013

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【Objective】This study aimed to elucidate the differences in flavonoid composition and gut homeostasis-regulating effects of Shatianyu (Citrus grandis L. Osbeck) whole fruit before and after debittering by Lactobacillus rhamnosus fermentation, thereby providing the theoretical foundation for the precise nutritional processing of Shatianyu.【Method】Shatianyu whole fruit powder (SWFP) and Lactobacillus rhamnosus-fermented Shatianyu whole fruit powder (F-SWFP) were used as test materials. High-performance liquid chromatography (HPLC) was adopted to analyze the differences in flavonoid composition and content between the above-mentioned fruit powder. Subsequently, SWFP and F-SWFP were subjected to in vitro fecal microbiota fermentation for 24 h, respectively. Gas chromatography (GC) and a lipopolysaccharide (LPS) enzyme-linked immunosorbent assay (ELISA) kit were employed to determine the content of short-chain fatty acids (SCFAs) and LPS in each fermentation system, respectively. 16S rRNA sequencing was adopted to determine the structure of fecal microbiota. The correlation between the relative abundance of fecal microbiota among SCFAs and LPS contents was revealed by Pearson correlation analysis.【Result】Eight monomeric flavonoids, namely neoeriocitrin, naringin, melitidin, cigranoside B, cigranoside A, cigranoside C, rhoifolin, and hesperidin, were detected in both SWFP and F-SWFP. Naringin, melitidin, and cigranoside B were the major flavonoid components, accounting for approximately 85% of the total content of detected flavonoids. Fermentation with Lactobacillus rhamnosus significantly reduced the total flavonoid content in SWFP, cigranoside B, neoeriocitrin, rhoifolin, hesperidin and melitidin were the main degraded flavonoids in F-SWFP, with the degradation rates ranging from 28.1% to 78.5%. After 24 h-fecal microbiota fermentation, both SWFP and F-SWFP significantly reduced the relative abundance ratio of Bacillota to Bacteroidota in fecal microbiota, with F-SWFP exhibiting a more pronounced effect. SWFP specifically induced the proliferation of Bifidobacterium and Mediterraneibacter, whereas F-SWFP not only specifically promoted the proliferation of Segatella and g_norank_f_Prevotellaceae, but also inhibited the growth of Collinsella, Blautia, Parabacteroides, and Dialister. 24 h-fecal microbiota fermentation of SWFP or F-SWFP all significantly increased acetic, propionic, and butyric acid content but reduced LPS content in the fermentation system. Notably, F-SWFP fermentation group showed higher SCFAs but lower LPS content than those of SWFP fermentation group, also, the butyric acid and LPS content in F-SWFP group was 1.2 and 0.7 times those of SWFP group, respectively. Correlation analysis revealed that the characteristic microbes in SWFP group were significantly positively correlated with SCFAs content, whereas those in F-SWFP group were not only positively correlated with SCFAs content, but also negatively correlated with LPS content.【Conclusion】Compared with SWFP, F-SWFP showed a lower total flavonoid content, but exhibited greater efficacy in promoting butyrate production while inhibiting LPS production by fecal microbiota. Given the key roles of butyrate and LPS in obesity-related metabolic diseases, F-SWFP serves as a functional ingredient for ameliorating obesity-related metabolic diseases and is suitable for use in the development of healthy foods.

Effects of Controlled Atmosphere and 1-MCP Treatment on Quality of Yuhuangli Plum During Refrigeration

LU XiaoFeng, HAO ShuoJie, WANG Yang, GAO Na, TONG Wei, TIAN RuiDong, SUN HaiLong, LIU Shuo

Scientia Agricultura Sinica. 2026, 59(15):  3424-3439.  doi:10.3864/j.issn.0578-1752.2026.15.014

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【Objective】This study aimed to investigate the effects of 1-methylcyclopropene (1-MCP) and controlled atmosphere (CA) storage on the preservation and internal quality of YuHuangli plums, to identify the optimal storage strategies, so as to provide the theoretical and technical references for industrial-scale storage.【Method】YuHuangli plums were subjected to four storage treatments: 1.0 µL·L^-1 1-MCP, CA storage with 3% O₂ + 1% CO₂, CA storage with 3% O₂ + 5% CO₂, and normal cold storage as a control (CK). Fruit samples were analyzed at 0, 30, 60, and 90 days of storage. External and internal quality indices—including peel color, texture, sugar-acid composition, respiration intensity, and aroma-were measured using a colorimeter, texture analyzer, gas chromatograph, high-performance liquid chromatograph, and gas chromatography-mass spectrometer.【Result】Compared with CK, both 1-MCP and CA treatments delayed the decline in fruit firmness and titratable acidity, maintained higher levels of soluble solids, vitamin C, and polyphenols, reduced respiration rates, and slowed the reduction of cohesiveness, springiness, gumminess, chewiness, rupture force, rupture deformation, yield force, and yield deformation, thus better preserving fruit texture. Both treatments also partially inhibited the degradation of sugar-acid components, with acidity being better maintained after 60 days. Peel color parameters (L*, a*, b*, and C*) declined during storage, with the fastest decrease under CK, followed by 3% O₂ + 5% CO₂, then 3% O₂ + 1% CO₂, and the slowest under 1-MCP treatment, significantly suppressing browning and darkening (P<0.05). After 60 days, 1-MCP exhibited the most pronounced preservation effect, followed by 3% O₂ + 1% CO₂ and 3% O₂ + 5% CO₂, while CK showed the greatest quality deterioration. At this stage, the flavor profile of YuHuangli plum was dominated by aldehydes, alcohols, and esters, with trans-2-hexenal and hexanal accounting for about 84% of the total relative content. Over time, the relative content of aldehydes and alcohols decreased in the 1-MCP and CA groups, whereas esters increased.【Conclusion】Both 1-MCP (1.0 µL·L^-1) and CA storage (3% O₂ + 1% CO₂) effectively preserved the quality of ‘YuHuangli’ plums during cold storage. Among them, 1-MCP provided the most comprehensive maintenance of overall fruit quality—including texture, color, and flavor—with the optimal preservation observed after 60 days.

ANIMAL SCIENCE·VETERINARY SCIENCE

Effects of Grazing Intensities on Root Functional Traits in Plant Communities of Inner Mongolia Meadow Steppe

KONG MengQiao, LI Hui, LI ZiHao, PING XiaoYan, JIANG CuiXia, GUO HaoNan, YANG YiJing, XIN XiaoPing

Scientia Agricultura Sinica. 2026, 59(15):  3440-3451.  doi:10.3864/j.issn.0578-1752.2026.15.015

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【Objective】Roots are the most critical underground organs of plants. This study investigates the dynamic effects of grazing disturbance on the root functional traits of meadow steppe communities, aiming to clarify the mechanisms by which grazing affects the structure and function of grassland community roots. It provides insights into the regulatory mechanisms and survival strategies of grassland plants under grazing disturbance.【Method】The root functional traits including total root length, number of root tips, number of root forks, root average diameter, root surface area, and total root volume were measured using minirhizotron technology under six grazing intensity treatments: no grazing (G0), light grazing (G1), light moderate grazing (G2), moderate grazing (G3), heavy grazing (G4), and extreme grazing (G5). Each grazing intensity treatment was replicated with nine minirhizotron tubes.【Result】(1) Both below-ground biomass and root-soil ratio exhibited significant decreasing trends with increasing soil depth (P<0.05). The maximum below-ground biomass and root-soil ratio were observed in the 0-10 cm soil layer under G4 grazing treatment, but there were no significant differences in below-ground biomass among grazing treatments. (2) The root morphological characteristics of meadow steppe communities vary significantly with the progression of the growing season and are affected by grazing intensity, with total root length and number of root forks showing an increasing trend over the growing season, whereas both root average diameter and total root volume reached their maxima in July before subsequent declines. Under varying grazing intensity gradients, root traits exhibit distinct patterns: moderate grazing (G1-G3) promotes root growth in the early growing season, whereas heavy or extreme grazing increases total root length and number of root forks in the late growing season but still reduces root average diameter and total root volume. (3) Specific root length (SRL) and specific root area (SRA) showed an increasing trend as the growing season progressed, peaking in September, while root tissue density (RTD) fluctuated. From June to July, SRL and SRA were highest under G3 grazing intensity, while from August to September, SRL increased with grazing intensity, and SRA decreased. RTD was generally higher under G3-G5 intensities compared to G0-G2. (4) Over time, plant height and coverage increased seasonally and peaked in September, while root-shoot ratio (R:S) decreased. With increasing grazing intensity, both height and coverage significantly decreased significantly in each month, while R:S significantly increased. (5) SRL and SRA were significantly correlated with plant community characteristics, and total root length and RTD were strongly correlated with R:S. However, average root diameter and number of root tips showed no significant correlation with soil factors or plant community characteristics.【Conclusion】The response of root morphological traits to grazing intensity exhibited dynamic differences, reflecting trade-offs in resource acquisition and structural strength at different periods. The functional composition of plant communities was strongly correlated with root traits, warranting further research. This study provides valuable insights into the response mechanisms and adaptability of meadow steppe plant roots to grazing intensity and seasonal dynamics.

Establishment of an Infection Model in Pigs for Chinese Clinical Strains of Lawsonia intracellularis

XIE Rui, GU ZhengQuan, LIU SiWen, LUO YaJuan, CUI YuCong, HUA Lin, SONG WenBo, HUANG Chao, TANG XiBiao, XU GaoYuan, CHEN HuanChun, WU Bin, PENG Zhong

Scientia Agricultura Sinica. 2026, 59(15):  3452-3464.  doi:10.3864/j.issn.0578-1752.2026.15.016

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【Objective】This study aimed to establish a stable porcine proliferative enteropathy (PPE) model using Chinese isolates of Lawsonia intracellularis (LI) and clinical infected intestinal tissue homogenates from different sources, construct a systematic evaluation index system covering fecal bacterial shedding, fecal score, average daily gain, feed conversion ratio, serum antibody level and histopathological changes, and identify the optimal challenge dose and preferred strain, so as to provide the standardized model and data support for the research on LI pathogenic mechanism, vaccine efficacy evaluation and prevention and control strategies.【Method】Twenty-eight-day-old weaned piglets were selected and divided into a negative control group and multiple infection groups. In Experiment 1, the pathogenicity of four LI tissue homogenates (JX703, GX301, HN312, and LI-HuB23) was compared at the dose of 1×10⁹ LI per piglet. In Experiment 2, JX703 tissue homogenate with the strongest comprehensive pathogenicity and the in vitro cultured LI-HuB23 isolate were selected to orally inoculate 28-day-old piglets at gradient doses (10¹⁰ LI per pig, 10⁸ LI per pig, and 10⁶ LI per pig). The piglets were monitored for 28 days after inoculation, during which fecal bacterial shedding, fecal score, average daily gain, feed conversion ratio, serum antibody level and histopathological changes were observed and recorded.【Result】In Experiment 1, typical clinical symptoms and pathological manifestations of PPE were successfully reproduced in all infection groups inoculated with tissue homogenates of different LI strains. LI was detectable in feces from day 4 post-inoculation, and bacterial shedding persisted until day 28 post-inoculation. Diarrhea occurred in the infection groups from day 4 post-inoculation, with the most severe symptoms observed from day 19 to 21 post-inoculation. All infection groups showed lower average daily gain and higher feed conversion ratio compared with the negative control group. Serum antibody monitoring showed that seroconversion occurred from day 14 after the establishment of infection and lasted until day 28 post-inoculation. Pathological examination revealed thickened intestinal mucosal folds in the infection groups; hematoxylin-eosin (HE) staining showed intestinal crypt hyperplasia and decreased goblet cells, and LI was detected in the cytoplasm of intestinal crypt epithelial cells by immunohistochemical staining (IHC). Overall, JX703 tissue homogenate induced the most prominent symptoms with the strongest virulence. In Experiment 2, the results of gradient dose test showed that inoculation with LI-HuB23 in 10⁶ LI per piglet was sufficient to establish effective infection, accompanied by fecal bacterial shedding, diarrhea, and corresponding pathological changes. The severity of symptoms increased with the elevation of infection dose, and the 10¹⁰ LI per pig group exhibited the most typical patterns in terms of peak fecal scores, pathological scores, and reductions in average daily gain.【Conclusion】A PPE infection model was successfully established using Chinese LI isolates and field-derived infected tissue homogenates in this study. The selected evaluation indexes had an important reference value for related research, including fecal bacterial shedding, fecal score, average daily gain, feed conversion ratio, serum antibody level and histopathological changes. The infection model established with an inoculation dose of 10¹⁰ LI per pig presented more typical clinical and pathological characteristics of PPE.