Scientia Agricultura Sinica

Screening of Wheat Varieties with Low Nitrogen Tolerance and Genome-Wide Association Studies of Low Nitrogen Stress Tolerance Index

LI Ning, GAO LiFeng, HUANG Xin, SHI HuaWei, YANG JinWen, SHI YuGang, CHEN Ming, JIA JiZeng, SUN DaiZhen

Scientia Agricultura Sinica. 2025, 58(13): 2487-2503. doi:10.3864/j.issn.0578-1752.2025.13.001

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【Objective】 The excessive application of nitrogen fertilizers has led to ecological pollution and waste of agricultural resources. Developing nitrogen-efficient wheat varieties and improving nitrogen use efficiency are effective approaches for achieving sustainable agricultural development and environmental protection. Screening low-nitrogen-tolerant germplasm resources and identifying genetic loci and candidate genes associated with low-nitrogen tolerance can provide materials and theoretical foundations for breeding nitrogen-efficient wheat varieties. 【Method】 A natural population consisting of 389 wheat varieties was cultivated under high-nitrogen (HN) and low-nitrogen (LN) treatments in 10 field environments. Grain yield per plant (GYP) was measured to calculate the stress tolerance index (STI), thereby enabling the classification of varieties with differential low-nitrogen tolerance. Genome-wide association studies (GWAS) were conducted using 660K SNP array genotyping data to identify stable genetic loci associated with low-nitrogen tolerance. Candidate genes were prioritized through haplotype analysis, expression profiling, and functional annotation. 【Result】 Twelve wheat varieties with strong low-nitrogen tolerance were identified, including Zhongluo 08-1, Jimai 15, Jinghua 2, Kehong 1, Mianyang 19, Jimai 22, Zhenmai 4, Yumai 35, Fengkang 7, Mianyang 11, Jinmai 31, and Lumai 5. Fourteen loci significantly associated with STI were detected, among which four (qSTI1A.1, qSTI3B, qSTI6A, and qSTI7A.2) overlapped with previously reported low-nitrogen tolerance or yield-related QTLs. Notably, qSTI3B-replicated across three environments-was identified as a key locus governing low-nitrogen tolerance. Functional annotation revealed that its candidate gene, TraesCS3B02G042400, encodes an AP2/EREBP (APETALA2/ethylene-responsive element-binding protein) transcription factor. Haplotype analysis showed significant STI divergence among varieties carrying distinct haplotypes, while expression levels of TraesCS3B02G042400 exhibited nitrogen dose-responsive upregulation. 【Conclusion】 Twelve wheat varieties with strong low-nitrogen tolerance were screened. A stable genetic locus, qSTI3B, and a candidate gene, TraesCS3B02G042400, associated with low-nitrogen tolerance were identified.

Characterization of Maize Germplasm Resistance to Common Smut and Analysis of Physiological Differences

LI XiangYu, LIU JianZhuo, HU DanDan, LIU GengYu, CHEN LiangYu, LI Bing, DU WanLi, SONG Bo

Scientia Agricultura Sinica. 2025, 58(13): 2504-2521. doi:10.3864/j.issn.0578-1752.2025.13.002

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【Objective】 Common smut (Mycosarcoma maydis) is a major fungal disease affecting maize production in China. This study aimed to screen for resistant germplasm resources and analyze their physiological and biochemical responses to pathogen infection, providing scientific support for disease resistance breeding. 【Method】 A total of 425 maize germplasm resources were selected and systematically screened for their resistance to common smut. Sugar metabolism, oxidative stress responses, and photosynthetic parameters were analyzed to identify high-resistant, moderate-resistant, and susceptible inbred lines. WGA-AF488/PI staining was used to analyze the hyphal proliferation to reveal the disease resistance traits of different inbred lines. 【Result】 The study found significant annual differences in the disease index and incidence of common smut in 2021 and 2022, primarily influenced by temperature and precipitation. Principal component analysis showed that the disease index on day 4, 8, and 12 post-inoculation was a key indicator of disease severity, while the disease incidence at the grain filling stage (R2) and wax ripening stage (R4) revealed differences across growth stages. Additionally, 6 high-resistance inbred lines (e.g., Q319), 67 medium-resistance inbred lines (e.g., D599), and 171 susceptible inbred lines (e.g., M407) were identified. Sugar metabolism analysis showed significant differences in sucrose, fructose, and glucose contents across inbred lines with different disease resistance, indicating the critical role of sugar metabolism in the competition between the pathogen and the host. Oxidative stress analysis revealed significant increases in H₂O₂ and O^2- contents post-infection, with the high-resistant inbred line Q319 exhibiting the strongest OH^- scavenging capacity. SOD and POD activities were significantly increased on days 4 and 8 post-inoculation, with the SOD activity of D599 increasing by 114.98% on day 8 and the SOD activity of Q319 increasing by 96.08%. On day 12, the POD activity of D599 and Q319 increased by 164.27% and 160.91%, respectively, indicating strong antioxidant defense capabilities in resistant materials. WGA staining showed that hyphal extension was limited in Q319, primarily concentrated near the vascular bundles, suggesting strong cell wall defense. D599 exhibited intermediate hyphal expansion speed and range, representing moderate defense capacity, while M407 displayed extensive hyphal spread with large intercellular infection, indicating weak cell wall defense. Post-inoculation, the net photosynthetic rate of Q319, D599, and M407 decreased by 52.5%, 52.8%, and 100.2%, respectively, compared to the control group, with significant reductions in photosynthetic pigment content; however, the decline decreased from 4 to 12 d. 【Conclusion】 This study reveals significant differences in sugar metabolism, oxidative stress responses, antioxidant capacity, and photosynthesis among different maize inbred lines under Mycosarcoma maydis infection. The high-resistance inbred line Q319 exhibits strong disease resistance through low sucrose, high fructose and glucose metabolism, along with higher chlorophyll and carotenoid content and efficient antioxidant ability. The high-susceptible inbred line M407 shows increased sucrose accumulation and insufficient antioxidant response, resulting in significantly decreased net photosynthetic rate and transpiration rate, leading to increased disease susceptibility. The medium-resistance inbred line D599 displays intermediate photosynthetic function and pigment accumulation, with disease resistance between Q319 and M407.

Comprehensive Evaluation of Potato Flour Quality and Variety Screening

ZHAO TianTian, YUAN JianLong, ZHUO FengQi, TANG ZhenSan, XU Jie, ZHANG Feng

Scientia Agricultura Sinica. 2025, 58(13): 2522-2537. doi:10.3864/j.issn.0578-1752.2025.13.003

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【Objective】 This study analyzes the quality characteristics of potato flour from different potato varieties (lines), aiming to identify key indicators for selecting suitable varieties for potato flour processing and to provide a theoretical basis for breeding high-quality processing varieties. 【Method】 Thirty-two domestically and internationally cultivated potato varieties (lines) were employed as experimental materials, with the variety Atlantic as the control. The contents of dry matter, starch, amylose, reducing sugars, ascorbic acid, citric acid, chlorogenic acid, and glycoalkaloids in the tubers were measured, along with the quality and physicochemical properties of the potato flour, including moisture, ash, crude starch, crude fat, solubility, swelling capacity, water-holding capacity, oil-holding capacity, blue value, freeze-thaw stability, whiteness, brightness, and thermal characteristics. After principal component analysis, cluster analysis, correlation analysis, and stepwise regression analysis, the differences in tuber and potato flour quality among the various varieties (lines) were compared, and representative indicators and superior processing quality varieties were identified. 【Result】 Among the thirty-two varieties, the tuber dry matter content ranged from 17.14% to 25.58% FW, starch content ranged from 11.34% to 19.52% FW, amylose content ranged from 15.24% to 24.44% FW of the starch content, reducing sugars content ranged from 0.10% to 1.07% FW, ascorbic acid content ranged from 0.51 to 4.10 mg·g^-1 DW, citric acid content ranged from 9.60 to 28.42 mg·g^-1 DW, chlorogenic acid content ranged from 121.32 to 2 558.36 mg·kg^-1 DW, and glycoalkaloids content ranged from 25.63 to 882.90 mg·kg^-1 DW. The potato flour moisture content ranged from 2.05 to 7.21 g·100 g^-1, ash content ranged from 2.00 to 5.34 g·100 g^-1, crude starch content ranged from 44.50 to 77.50 g·100 g^-1, crude fat content ranged from 0.10 to 0.93 g·100 g^-1, solubility ranged from 14.11% to 25.17%, swelling capacity ranged from 12.75% to 17.71%, water-holding capacity ranged from 2.76 to 3.79 g·g^-1, oil-holding capacity ranged 2.37 to 3.30 g·g^-1, blue value ranged from 6.35 to 22.52, bleeding rate ranged from 23.96% to 54.34%, whiteness ranged from 11.47 to 31.00, brightness ranged from 75.92 to 88.03, onset temperature ranged from 60.70 to 70.04 ℃, peak temperature ranged from 64.84 to 74.06 ℃, crystallization temperature ranged from 69.37 to 78.15 ℃, and pasting enthalpy ranged from 2.36 to 4.92 J·g^-1. Significant correlations were found among various indicators, including agronomic traits of tubers and the quality and physicochemical properties of potato flour. Notably, there are an extremely significant negative correlation among swelling capacity, bleeding rate, To, Tp, and Tc. A comprehensive evaluation model for potato flour quality was developed: Y=0.612+0.352×Dry matter (%)-0.344×Bleeding rate (%)-0.233×Glycoalkaloid content (μg·g^-1)-0.238×Peak temperature (℃)+0.152×Crude starch (g·100 g^-1) (R²=0.952, P<0.01). 【Conclusion】 The content of dry matter, glycoalkaloid, crude starch, freeze-thaw stability, and peak temperature are key indicators for evaluating and selecting potatoes for flour processing. Gannongshu No.7 and Zhongshu No.49 are excellent varieties for potato flour processing.

Multi-Objective Optimization of Stable Yield and Emission Reduction of Dryland Spring Wheat Based on DNDC and NSGA-Ⅲ. Coupling Model

CAO JingWen, NIE ZhiGang, LI Guang, YANG Jie

Scientia Agricultura Sinica. 2025, 58(13): 2538-2551. doi:10.3864/j.issn.0578-1752.2025.13.004

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【Objective】 In response to growing food demand and ecological sustainability requirements, this study explored the comprehensive impact of integrated irrigation and fertilization management on spring wheat yield, soil CO₂ and N₂O emissions fluxes in arid areas of Northwest China, with the aim of identifying optimal irrigation and fertilization strategies to achieve coordinated development of agricultural production and environmental benefits. 【Method】 Based on the DNDC agricultural ecosystem simulation model, using field trial data from Anjiapo Village, Fengxiang Town, Dingxi City, Gansu Province, from 2021 to 2023, the model was calibrated and validated. Different fertilization levels (0-400 kg·hm^-2) and irrigation levels (0-300 mm). The model simulates the response patterns of wheat growth dynamics and soil greenhouse gas (CO₂ and N₂O) emission fluxes under different irrigation and fertilization management measures. Combining the NSGA-III multi-objective optimization algorithm, a multi-objective optimization framework was established with three objective functions: “maximizing crop yield”, “minimizing soil CO₂ emissions flux”, and “minimizing soil N₂O emissions flux” as the three objective functions, achieving synergistic optimization between wheat yield enhancement and soil greenhouse gas emission reduction, and determining the optimal management scheme that balances yield and environmental benefits. 【Result】 The DNDC model effectively simulates spring wheat yield and soil greenhouse gas emission fluxes. Under four fertilization gradient treatments, the normalized root mean square error (NRMSE) for yield, soil CO₂ emissions, and N₂O emissions over three years was 17.4%-18.8%, 7.62%-11.41%, and 9.19%-12.47%, respectively. Under two irrigation treatments, the normalized root mean square error NRMSE for yield over three years was 13.3%-17.2%. The optimized irrigation and fertilization rates indicate that when fertilization is controlled at 150-180 kg·hm^-2 and irrigation volume is 110-150 mm, wheat yield can be increased to 2 088.48 kg·hm^-2, while soil CO₂ emissions flux is controlled at 4 998.87-5 011.5 kg·hm^-2 per year, and soil N₂O emission flux is controlled at 4.06-4.14 kg·hm^-2 per year. 【Conclusion】 Coupling the DNDC model with the NSGA-III algorithm enables the simultaneous optimization of spring wheat yield and soil greenhouse gas emissions fluxes in dryland areas. When the irrigation amount is set between 110-150 mm and nitrogen application rate between 150-180 kg·hm^-2, it is possible to maintain stable yields while effectively controlling soil CO₂ and N₂O emission fluxes. This provides a scientific basis for achieving both yield stability and emission reduction in dryland spring wheat systems in central Gansu.

Effects of Reducing Nitrogen Application on Maize Agronomic Traits, Grain Yield and Quality Under Green Manure Returning to Field System in Arid Areas

WANG PengFei, YU AiZhong, WANG Feng, WANG YuLong, LÜ HanQiang, SHANG YongPan, Yin Bo, LIU YaLong, ZHANG DongLing, HUO JianZhe, JIANG KeQiang, PANG XiaoNeng

Scientia Agricultura Sinica. 2025, 58(13): 2552-2563. doi:10.3864/j.issn.0578-1752.2025.13.005

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【Objective】 In order to seek a reasonable nitrogen application system suitable for high-yield and high-quality production of maize in arid areas, this study focused on the effects of reducing nitrogen application under the condition of returning green manure to the field on maize grain yield and quality, with a view to providing a theoretical basis for the regional development of nitrogen-saving, efficiency-enhancing, high-yield and high-quality agricultural production models. 【Method】 Field experiment were conducted at the Wuwei Oasis Agricultural Experiment Station from 2020 to 2022, and different nitrogen reduction rates were investigated under the condition of green manure returned to the field (Green manure retention with nitrogen reduction 0%, N100, 360 kg N·hm^-2; Green manure retention with nitrogen reduction 10%, N90, 324 kg N·hm^-2; Green manure retention with nitrogen reduction 20%, N80, 288 kg N·hm^-2; Green manure retention with nitrogen reduction 30%, N70, 252 kg N·hm^-2; and Green manure retention with nitrogen reduction 40%, N60, 216 kg N·hm^-2) on maize agronomic traits, yield components and grain quality. 【Result】 Compared with the N100 treatment, the N80 treatment ensured that maize plant height, stem diameters, and dry weight did not decrease, while the N70 treatment reduced them by 8.8%, 11.3%, and 16.4%, respectively, and the N60 treatment reduced them by 10.6%, 12.9%, and 21.3%, respectively. Among the yield components, there was no significant difference in maize bare top length among treatments; ear length, row number per ear, kernel number per row, and ear numbers under N100, N90, and N80 treatments were no significant difference too, but they were all significantly higher than those under N70 and N60 treatments. In addition, N80 treatment had the advantage of ensuring that the maize grain protein content was not reduced and fat content was increased while ensuring that maize yield was not reduced. And this system helped to reduce the amount of nitrogen accumulation in maize stems and leaves, and increased the amount of nitrogen accumulation in maize grain. Mantel test analysis revealed that maize grain yield, protein content, and fat content exhibited significant positive correlations (P<0.05, Mantel’s r>0.25) with nitrogen application rate, grain nitrogen accumulation, plant height, stem diameter, dry weight, ear length, and row number per ear. However, no significant correlations were observed with ear height, bare top length, or kernels number per row. Grey correlation analysis further showed that the ear numbers, plant dry weight and row number per ear were the key factors affecting maize kernel yield. 【Conclusion】 In the context of sustainable agriculture, reducing nitrogen by 20% under the condition of returning green manure to the field (288 kg N·hm^-2) could be used as a reasonable nitrogen management system for developing nitrogen-saving and input-reducing, high-yield and high-quality agriculture in arid areas.

Effects of Co-Ridge Planting on the Distribution Characteristics of Soil Available Phosphorus and the Absorption and Utilization of Phosphorus by Crops in Maize||Peanut

LIANG Na, WANG JiangTao, WANG YingChao, ZHENG Bin, WANG XiaoXiao, LIU Juan, LIU Ling, FU GuoZhan, JIAO NianYuan

Scientia Agricultura Sinica. 2025, 58(13): 2564-2577. doi:10.3864/j.issn.0578-1752.2025.13.006

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【Objective】 Co-ridge planting can further enhance the yield advantage of maize (Zea mays L.) and peanut (Arachis hypogaea L.) intercropping (maize||peanut). This study aimed to explore the distribution characteristics of available phosphorus in maize||peanut soil and the characteristics of phosphorus absorption and utilization in crops under the co-ridge planting, which could provide theoretical and technical basis for sustainable high-yield cultivation of maize||peanut. 【Method】 The experiment was conducted in the farm of Henan University of Science and Technology from 2023 to 2024, using a field randomized block experiment. Under the conditions of no phosphorus (P₀) and 180 kg P₂O₅·hm^-2 (P₁₈₀), the effects of co-ridge planting of maize and peanut intercropping (RIC) on the distribution characteristics of soil available phosphorus in 0-60 cm soil layer as well as the absorption and utilization of phosphorus by crops in maize||peanut were studied with flat planting of maize and peanut intercropping (FIC) as the control. 【Result】 In the horizontal direction, the distribution of soil available phosphorus in 0-60 cm soil layer under FIC treatment exhibited “ ” (gentle) characteristics, while under RIC treatment, it showed “ ” (ridge) characteristics, under P₁₈₀ the “low-high-low” “ ” (ridge) characteristics was more prominent. Compared with FIC, RIC was beneficial to increase the soil available phosphorus content in the 0-40 cm soil layer within the planting unit, which significantly increased by 18.38%-21.29% under P₁₈₀. Compared with FIC, RIC alleviated the interspecific phosphorus nutrition competition, increased the phosphorus content of intercropping maize and peanut, significantly increased the phosphorus accumulation, and promoted the phosphorus distribution to maize grain and peanut seed. Compared with FIC, RIC significantly increased the phosphorus absorption and yield of intercropping maize, intercropping peanut, and intercropping system, within the increase range of phosphorus absorption being 18.83%-32.62%, 24.08%-41.78% and 21.06%-37.14%, respectively (P<0.05). Compared with P₀, P₁₈₀ could further increase soil available phosphorus content and phosphorus content in maize and peanut, promote phosphorus absorption in intercropping system, and significantly increase maize and peanut yield and intercropping advantage. 【Conclusion】 The co-ridge planting could further improve the yield advantage of maize||peanut compared with flat planting. The key lied in the fact that it improved the available phosphorus distribution in 0-60 cm soil and increased the available phosphorus content in the 0-40 cm soil layer of the planting unit, alleviated the interspecific phosphorus nutrition competition, and promoted the absorption of phosphorus in maize and peanut and its distribution to seed. Phosphorus application had a significant positive regulation effect.

Screening of Target Genes Downstream of VviERF045, a Transcription Factor Associated with Gray Mold Resistance in Vitis vinifera

ZHAO YuLei, XIN JiaLu, LI ChengNan, LI Shan, XIE XuFei, YIN Xiao

Scientia Agricultura Sinica. 2025, 58(13): 2578-2590. doi:10.3864/j.issn.0578-1752.2025.13.007

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【Background】 Gray mold is an important fungal disease that seriously endangers the grapevine (Vitis vinifera) industry worldwide. As an important transcription factor family in plants, ethylene-responsive factor (AP2/ERF) family plays a key role in regulating plant growth and development and stress response. 【Objective】 The study aimed to elucidate the molecular mechanisms by which the grapevine ethylene-responsive factor VviERF045 mediates defense against Botrytis cinerea. Through prediction and functional analysis of its downstream target genes, this research provides new insights into the transcriptional regulatory network associated with grapevine resistance to B. cinerea, establishing a foundation for breeding disease-resistant cultivars. 【Method】 Grapevine leaves were inoculated with B. cinerea using both agar disc and spore suspension methods. The expression profiles of VviERF045 and its predicted targets were evaluated via quantitative real-time PCR (qRT-PCR). Promoter cis-acting elements were analyzed using PlantCARE, while phylogenetic relationships and sequence alignments of VviERF045 were assessed using MEGA 7 and DNAMAN. Functional validation was conducted through Agrobacterium-mediated transient overexpression of VviERF045 in ‘Cabernet Sauvignon’ leaves. To further explore its regulatory landscape, DNA affinity purification sequencing (DAP-seq) was performed on ‘Pinot Noir’ leaves at 12 hours post-inoculation (hpi) with B. cinerea. 【Result】 Transient overexpression of VviERF045 significantly enhanced resistance to gray mold in ‘Cabernet Sauvignon’ leaves. DAP-seq analysis was performed on V. vinifera ‘Pinot Noir’ leaves at 12 hpi. Comparative analysis of peaks between two experimental replicates identified 51 806 consensus peaks. Subsequent genomic annotation revealed these peaks were predominantly located within ± 2 kb regions flanking promoter-transcription start sites (TSS). The top 2 000 most statistically significant peaks were selected for functional characterization through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Through integrated analysis of potential target genes and peak calling, three high-confidence candidate target genes (TCP8, SAP5, and bHLH48) were identified, all showing significant binding peaks in their promoter regions. Subsequent qRT-PCR validation confirmed the transcriptional upregulation of these genes, suggesting their cooperative involvement with VviERF045 in the grapevine’s response to B. cinerea infection.【Conclusion】VviERF045 functions as a positive regulator of grapevine defense against B. cinerea, likely by activating key stress-responsive genes such as TCP8, SAP5, and bHLH48. These results provide mechanistic insights into the pathogen-responsive transcriptional network in grapevine and identify potential molecular targets for breeding resistant V. vinifera cultivars.

Insecticidal Active Component Identification of Camellia oleifera Shell Against Mythimna separata and Its Action Mechanism

ZHAN Li, LIANG ZongSuo, YU Jing, LU Jun, LIANG Qian

Scientia Agricultura Sinica. 2025, 58(13): 2591-2603. doi:10.3864/j.issn.0578-1752.2025.13.008

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【Objective】 The objective of this study is to clarify the insecticidal active component and mechanism of Camellia oleifera shell on Mythimna separate, and to provide a basis for the development of botanical insecticides for M. separata. 【Method】 The insecticidal activity of petroleum ether, chloroform, ethyl acetate, and aqueous extracts of C. oleifera shell against 3rd instar larvae of M. separata was evaluated using the immersion and leaf-dipping methods. The petroleum ether extract was subjected to silica gel column chromatography for separation, followed by gas chromatography-mass spectrometry (GC-MS) analysis to identify the active components. Furthermore, a non-targeted metabolomics approach was employed to annotate, screen, and analyze differential metabolites in M. separata. 【Result】 The petroleum ether extract of C. oleifera shell exhibited the highest contact toxicity against M. separata, with LC₅₀values of 9.53, 8.00, 7.51, and 7.02 mg·mL^-1 on 1, 3, 5, and 7 d, respectively. The petroleum ether extract was separated by silica gel column chromatography to obtain 20 fractions, among which fraction 6 showed the best contact toxicity against M. separata, with LC₅₀ values of 3.99, 3.85, 3.72, and 3.72 mg·mL^-1 on 1, 3, 5, and 7 d, respectively. Further separation of fraction 6 yielded 21 subfractions, with subfraction 6.4 demonstrating the highest contact toxicity. GC-MS analysis identified hexadecane (19.33%), 1-hexadecene (14.21%), octacosane (4.03%), octadecane (9.24%), dichlorochrysanthemic acid (6.73%), 1-octadecene (7.27%), 1-tetracosanol (3.87%) and isophytol (5.92%) as the main compounds. Among these, dichlorochrysanthemic acid exhibited the strongest contact toxicity against M. separata, with LC₅₀ values of 91.62, 55.61, 34.94, and 24.43 μg·mL^-1 on 1, 3, 5, and 7 d, respectively. Its insecticidal efficacy was comparable to that of the positive control, pyrethrin. Non-targeted metabolomics analysis identified six differential metabolites, among which piceid, cannabinoid, 5-fluoro-cannabinoid metabolite, and caproylglycine were significantly upregulated, while 4-ethyl-2,6-dihydroxyphenyl sulfate and N-pentacosanoylglycine were significantly downregulated. These differential metabolites were primarily enriched in four metabolic pathways: cofactor biosynthesis, tryptophan metabolism, niacin and nicotinamide metabolism, ubiquinone and terpenoid-quinone biosynthesis. 【Conclusion】 Dichlorochrysanthemic acid from C. oleifera shell extract exhibits insecticidal activity against M. separata, providing a new avenue for the biological control of M. separata and the high-value utilization of C. oleifera shell.

Soil Enzyme Activities and Their Stoichiometry Under Prolonged Rice Cultivation

ZHANG XinYao, WANG Ping, LIU YaLong, WANG JingKuan

Scientia Agricultura Sinica. 2025, 58(13): 2604-2613. doi:10.3864/j.issn.0578-1752.2025.13.009

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【Objective】 The theoretical evaluation of ecological stoichiometry of soil microbial metabolism and soil nutrient restriction in paddy soil was carried out, so as to provide the support for understanding the nutrient cycle of rice ecosystem. 【Method】 In this study, a chronosequence of paddy soils with 1 000 years was collected and analyzed soil enzyme activity and enzyme stoichiometry ratio with prolonged rice cultivation and clarified the influencing factors. 【Result】 The invertase activity increased with the chronosequence, while the activities of α-1,4-glucosidase, β-1,4-glucosidase, β-D-cellobiohydrolase and β-1,4-xylosidase enzymes decreased significantly at the early stage of chronosequence (50 years), and then increased significantly and remained stable. The dynamic of urease was similar to invertase, while the activities of n-acetyl-β-glucosaminidase were similar to most of the enzymes concerned in the carbon cycle. The C:N ratio of soil enzymes was the lowest at 1.28 when the soil was not cultivated (0 year), while the C:P and N:P ratios of soil enzymes were the highest at 1.77 and 1.38 at the same time, respectively. Generally, the C:N and C:P ratios of soil enzymes were all greater than 1 in all rice-growing years, while the N﹕P ratio of soil enzymes was less than 1 except for the uncultivated soil (0 year). 【Conclusion】 According to the comprehensive stoichiometric analysis (soil enzyme activity ratio, vector length and angle) could be seen that the soil microorganisms in the marsh before reclamation were mainly limited by carbon, while the soil microorganisms in the early stage (50 years) of rice cultivation after reclamation were mainly limited by phosphorus, and with the chronosequence, the soil microorganisms were changed to be limited by nitrogen. The research results could provide a theoretical basis for understanding the biogeochemical cycling mechanisms mediated by microorganisms in paddy ecosystems, as well as guiding soil nutrient management and ecological sustainable development.

The Characteristics of Ammonia Volatilization and Crop Yield Under Legume-Wheat Rotation System in Fluvo-Aquic Soil in Northern Henan Province

WANG ShiJi, LI YueYing, CHEN Chen, JIANG GuiYing, LIU ChaoLin, ZHU ChangWei, YANG Jin, WANG MengRu, JIE XiaoLei, LIU Fang, LIU ShiLiang

Scientia Agricultura Sinica. 2025, 58(13): 2614-2629. doi:10.3864/j.issn.0578-1752.2025.13.010

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【Objective】 Based on the long-term rotation experiment, this study explored the characteristics of ammonia volatilization and crop yield change in Fluvo-aquic soil. 【Method】 The long-term experiment started in 2016, and this study carried out 2023-2024. A randomized block design with five crop rotation treatments were set as: (1) continuous wheat-maize (WMWM); (2) continuous wheat-peanut (WPWP); (3) continuous wheat-soybean (WSWS); (4) 1-year wheat-maize + 1-year wheat-soybean (WMWS); (5) 1-year wheat-maize + 1-year wheat-summer peanut (WMWP). The ammonia volatilization rate (AVR) and cumulative volatilization (ACV), the content of different nitrogen forms in soil, nitrogen content in crop, and crop yield were measured and analyzed. 【Result】 The ACV in summer-autumn season was higher than that in wheat season, and the AVR in both seasons concentrated in 1st-9th day after fertilization. Under the same fertilization condition, at the summer- autumn season, the AVR peak in WMWM treatment was the highest one with 4.15 kg·hm^-2·d^-1. At the wheat season, the lowest AVR of wheat base fertilizer was observed under WMWS (0.77 kg·hm^-2·d^-1), and the lowest AVR after topdressing fertilizer was under WSWS (0.40 kg·hm^-2·d^-1). The lowest ACV was under WMWP and WMWS during both summer-autumn and wheat seasons, which decreased by 18.7% and 14.8%, 12.5% and 28.6%, respectively, compared with WMWM. In the 0-20 cm soil layer, the content of ammonium, nitrate, and total nitrogen under WPWP and WSWS were significantly higher than those under WMWM during the two seasons. Compared with WMWM, the content of nitrogen in wheat grains under WPWP and WSWS were increased by 12.9% and 17.2%, respectively, and the similar trend was observed in grain nitrogen accumulation, with 135 kg·hm^-2 and 137 kg·hm^-2, respectively. The maize equivalent yield (MEY equal to price yield) under WPWP treatment was significantly higher than other treatments, which was 41.7% higher than that under WMWM treatment. Compare with WMWM, the panicle number (7.0% and 3.5%), kernel number per spike (20.7% and 15.9%), thousand grain weight (5.4% and 4.1%) and yield (10.8% and 10.9%) were enhanced under WPWP and WSWS treatment. During the whole rotation cycle, crops absorbed nitrogen accounted for 43.8%-56.3%, and ammonia volatilization accounted for 1.9%-4.5%. The highest crop absorbed proportion was under WMWP, while the lowest proportion ammonia volatilization was under WSWS. 【Conclusion】 In the Fluvo-aquic soil area of northern Henan Province, the rotation of wheat and legume crops improved the soil nutrient, reduced the soil ammonia volatilization, increased the nitrogen content and wheat yield, which was recommended as a suitable rotation mode in this area.

Effects of Potassium Application on Root Rhizosphere Microbial Community Changes and Growth of Sugarcane

ZHAO Yong, ZHANG ZhongFu, WANG YuTong, AI Jing, LIU JiaYong, WU JianMing, DENG Jun, ZHANG YueBin

Scientia Agricultura Sinica. 2025, 58(13): 2630-2644. doi:10.3864/j.issn.0578-1752.2025.13.011

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【Objective】 This study aimed to explore the characteristics of changes in the bacterial and fungal communities and functions in the rhizosphere soil and their influences on sugarcane growth during the tillering stage under different potassium application levels, so as to provide a theoretical basis and production guidance for potassium application during the tillering stage of sugarcane. 【Method】 Using the new variety YZ1696 as the experimental material, a three-year continuous field trial was conducted during the sugarcane tillering stage, applying five different potassium fertilizer levels (0, 75, 150, 225, and 300 kg·hm^-2) to investigate the effects on the growth rate of sugarcane, the structure and function of the rhizosphere soil bacterial and fungal community, and the sucrose yield of sugarcane. 【Result】 (1) Within two months after potassium application during the tillering stage, the absolute growth rate of sugarcane initially increased and then decreased as the potassium application level increased. After three consecutive years of targeted application of different potassium fertilizer treatments, the growth performance of new plantings, one-year-old ratoons, and two-year-old ratoons was consistent, showing that both the sugarcane yield and sucrose content first increased and then decreased with increasing potassium fertilization gradient, and the growth rate, yield, and sucrose content of sugarcane were the optimal under a potassium application of 150 kg·hm^-2. (2) Different potassium application treatments had significantly influenced the abundance of bacteria and fungi in the rhizosphere soil and the composition of the microbial community and diversity. The differences in bacterial abundance mainly concentrated in Proteobacteria, Acidobacteriota, Actinobacteriota, Nitrospirota, and Firmicutes, while the differences in fungal abundance mainly concentrated in Basidiomycota, Ascomycota, and Chytridiomycota. The structure and diversity of the rhizosphere bacterial and fungal communities exhibited a trend of decreasing first and then increasing with the increase in potassium application level, and a fertilization inflection point was at a potassium application of 150 kg·hm^-2. (3) Correlation analysis between the abundance of the main microbial communities and the sucrose yield of sugarcane under different potassium application treatments indicated that the abundances of bacterial Actinobacteriota and fungal Proteobacteria, Basidiomycota, Ascomycota, and Chytridiomycota were significantly correlated with the sucrose yield of sugarcane. These microorganisms played positive roles in nitrogen fixation, organic matter decomposition, nutrient cycling, and inhibition of pathogenic bacteria in the soil. 【Conclusion】 Appropriate potassium application during the tillering stage could improve the structure and function of the rhizosphere soil microbial community, thereby promoting sugarcane growth. However, different potassium application levels had varying effects on sugarcane yield, sucrose content, the functional microbial communities of bacteria and fungi, among which the potassium application level of 150 kg·hm^-2 had the most significant effect on optimizing the structure of the soil microbial community and enhancing the sucrose yield of sugarcane.

Multi-Omics Analysis Reveals the Changes of Monoterpenes and Anthocyanins Accumulation During Veraison in Red Muscat-Type Grape

WANG HuiLing, ZHANG YingYing, YAN AiLing, WANG XiaoYue, LIU ZhenHua, REN JianCheng, XU HaiYing, SUN Lei

Scientia Agricultura Sinica. 2025, 58(13): 2645-2662. doi:10.3864/j.issn.0578-1752.2025.13.012

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【Objective】 The accumulation of anthocyanins and monoterpenes during grape berry color change at both metabolic and transcriptional levels were analyzed, to explore the spatio-temporal synthesis mechanisms of anthocyanins and monoterpenes, and provide a theoretical basis for the regulation of anthocyanin and monoterpene synthesis in table grape. 【Method】 The grape berries of Ruiduhongyu were used as materials, sampling was started at 5 days before the veraison and continued until 40 days after the initial color change. The contents of total soluble solids and titratable acid in the berry samples were determined by conventional methods; The changes in monoterpene components and content in berries were determined with headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPEME-GC-MS); The content of total flavonoids and total anthocyanins were detected using a spectrophotometer; The expression changes of key genes involved in monoterpenes and anthocyanins synthesis were analyzed by transcriptome sequencing and real-time fluorescence quantitative PCR. 【Result】 With the advancement of the coloration process, the main components of 25 free and bound monoterpenes in the Ruiduhongyu berries fluctuated. Most of the free monoterpenes began to be synthesized in large quantities from the 20th day of coloration and reached the highest level at the 35-40 th day of coloration. The content of bound monoterpenes reached the highest level at 30 days after coloration. The content of bound monoterpenes was higher than that of free monoterpenes. Flavonoids were synthesized and accumulated in large quantities before the fruit coloration; the synthesis of anthocyanins was initiated along with the fruit coloration and reached the highest level at 20 days after coloration, followed by a slight decrease. Based on transcriptome sequencing, a total of 5 836 differentially expressed genes were identified, and the number of differentially expressed genes varied significantly among different developmental stages. The differentially expressed genes were enriched in the pathways of phenylalanine synthesis, flavonoid synthesis, and monoterpene synthesis. Among them, 14 differentially expressed genes were related to the monoterpene synthesis pathway, and 11 were related to the anthocyanin synthesis pathway. The expression patterns of these genes were consistent with the accumulation of monoterpenes and anthocyanins. Further correlation analysis screened out 24 transcription factors that were significantly correlated with the expression of multiple genes involved in the monoterpene and anthocyanin synthesis pathways. 【Conclusion】 The synthesis of anthocyanins in red Muscat type grape berries initiates earlier than that of the aroma compound monoterpenes, and the synthesis of the two types of compounds is regulated in a spatio-temporal pattern. The accumulation of monoterpenes and anthocyanins is closely related to the expression of multiple key enzyme genes in their synthesis pathways, and their synthesis is regulated at the transcriptional level of various genes.

Identification of the VcTPS/VcTPP Gene Family in Blueberries and Functional Analysis of VcTPS24 in Floral Induction

WU XinLiang, WU HuiLing, ZHANG SuiLin, CAO Man, FENG Xin, ZHOU BingJie, HOU ZhiXia

Scientia Agricultura Sinica. 2025, 58(13): 2663-2681. doi:10.3864/j.issn.0578-1752.2025.13.013

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【Objective】 Trehalose-6-phosphate synthase (TPS), as a key enzyme in the trehalose synthesis pathway, is involved in regulating various developmental process such as germination, plant growth, flowering, and stress resistance. In this study, genes encoding TPS and TPP in the blueberry trehalose synthesis pathway were identified. We analyzed the molecular mechanism of VcTPS24 in regulating floral induction through molecular biology methods, laying the foundation for revealing the sugar signaling pathway regulating blueberry floral induction. 【Method】 Bioinformatics methods was used to analyze the number of amino acid, gene structure, conserved motifs, promoter sequences, and subcellular localization of VcTPS/VcTPP family members in the trehalose metabolism pathway of blueberries (Vaccinium corymbosum L.). The expression characteristics of VcTPS class I genes during blueberry floral differentiation were analyzed through qRT-PCR experiments. We analyzed the expression of VcTPS24 in blueberries under hormone and low temperature stress by exogenous application of gibberellin, auxin and other hormones, and 4 ℃ low temperature treatment. The regulatory effects of VcSOC1 and VcSVP on the VcTPS24 promoter were analyzed by dual luciferase assay. We validated the function of VcTPS24 in regulating growth and floral induction by overexpressing it in tobacco. 【Result】 A total of 28 VcTPS and 19 VcTPP family members were identified from blueberry, which can be divided into two subfamilies. VcTPS family members encoding proteins ranging from 465 to 1 382 amino acids, with an average isoelectric point (pI) of 6.02. Most members have 3 exon structures. VcTPP family members encoding proteins ranging from 241 to 434 amino acids, with an average isoelectric point (pI) of 7.48. Most members have 11 exon structures. Cis acting elements such as G-Box, P-Box, LTR, and circadian, which are involved in light, hormone, stress response, and regulation of plant growth, were found in the promoter sequences of VcTPS/VcTPP family members. Members of the VcTPS/VcTPP family are mostly located in chloroplasts and vacuoles. The expression level of VcTPS class I genes significantly increased during blueberry floral induction, and the expression level of VcTPS24 was significantly higher through the entire period of flower bud differentiation than in the undifferentiated group. Subcellular localization assay showed that the expression of VcTPS24 can be significantly upregulated in blueberry seedlings by exogenous IAA, GA₃, SA, MeJA, and low temperature stress. The dual luciferase assay showed that the promoter activity of VcTPS24 was significantly increased by VcSOC1, while it was significantly decreased by VcSVP. VcTPS24 is located in the nucleus and cytoplasm. Under the condition of no exogenous sucrose supply, the germination and radicle elongation of tobacco seeds overexpressing VcTPS24 were promoted, but excessive sucrose hindered the growth of tobacco seedlings overexpressing VcTPS24. The expression level of VcTPS24 was upregulated under short day condition. Under both long and short day conditions, the relative expression levels of NtLFY, NtFUL, and NtSOC1 in overexpressing VcTPS24 tobacco lines were all increased to varying degrees, and the formation time of their inflorescences was significantly earlier than that of the wild type. This indicates that VcTPS24 is involved in the regulation of floral induction. 【Conclusion】 A total of 28 blueberry VcTPS and 19 VcTPP family members were identified, and the VcTPS class I gene played an important role during blueberry floral induction. The expression of VcTPS24 was influenced by exogenous hormones, and the overexpression of VcTPS24 can promote tobacco flowering, which may be related to the upregulated expression of flowering genes.

Research Progress on the Function of Bovine Thyroid Gene and Its Correlation with Environmental Adaptability

LI Hao, TIAN YuYang, ZHANG ZiMing, CAO YiFan, CIREN LuoBu, NIMA CangJue, DANZENG LuoSang, LEI ChuZhao, BASANG ZhuZha, CHEN NingBo

Scientia Agricultura Sinica. 2025, 58(13): 2682-2692. doi:10.3864/j.issn.0578-1752.2025.13.014

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The thyroid gland is an important endocrine organ in cattle that plays a biological role by secreting thyroid hormone (TH), which promotes nutrient metabolism, growth and development, improves neural function; and regulates physiological functions. The multifunctionality of the thyroid gland is indispensable for maintaining a healthy body in animals and has a profound impact on their growth, development and environmental adaptation. The genetic background of Chinese yellow cattle is complex, and after long-term hybridization and selection in various regions, at least 58 local breeds have been formed. During the spread of Chinese yellow cattle, they faced diverse geographical and ecological environments, and their genomes continuously microevolved to adapt to different environments. Among these genes, thyroid-related genes may play important roles in adaptation. This article reviews the research progress on the regulatory functions of the thyroid and related genetic variations from the perspectives of animal physiology and genomics. First, the thyroid gland dynamically regulates TH levels through negative feedback regulation and then regulates animal development, metabolism, rhythm and reproductive function. The thyroid signal is highly sensitive to environmental influences and can respond accordingly, such as increasing heat production under cold conditions, reducing heat production under hot conditions, and regulating immunity and nerve cells to resist diseases and other adverse factors. The process from TH synthesis to its function is complex. Many genes are involved in the regulation of this process, and the genetic variation of some genes has an impact on the health and performance of animals. In recent years, research on the bovine genome has developed rapidly, with in-depth discussions on the origin, production, and stress resistance traits of cattle. With increasing research on the bovine genome, more new functions of bovine thyroid-related genes have been discovered. Thyroid-related genes affect key traits such as growth, metabolism, and environmental adaptation in cattle. For example, the DUOXA2 gene is involved in calcium regulation in dairy cows and in the high-altitude adaptation of Tibetan cattle, and its Leu20Pro variation has a potential impact on yellow cattle in the Qinghai‒Tibet Plateau. The TG gene is one of the key regulatory genes related to beef cattle production traits, with three SNPs associated with sexual maturity in bulls, which can significantly reduce the age of sexual maturity in bulls. Finally, this paper highlights the potential of genome resequencing technology and single-nucleus RNA and chromatin accessibility sequencing technology for exploring the genetic variations of thyroid related genes. The feasibility of these two technologies is demonstrated from aspects such as cost, functionality, and application examples. Genome resequencing can identify various types of variant sites and pinpoint key variants on the basis of selection signals. Single-nucleus RNA and chromatin accessibility sequencing technology can be used to investigate the gene expression differences within each cell under different environmental conditions and at various developmental stages, facilitating the verification of the mechanisms of action of key regulatory genes. Hence, delving into the functions of bovine thyroid-related genes and their genetic variations linked to growth and environmental adaptability is pivotal for revealing novel genes associated with important economic traits and stress resistance in cattle. This paper introduces the regulatory mechanism of the thyroid gland and the effects of thyroid-related genes on the growth, metabolism, biological rhythm regulation, reproductive function and environmental adaptation of livestock, and proposes new technologies that can be used for thyroid research in the future to provide a reference for future research on thyroid function and cattle breeding.

Co-Exposure of DON and PRRSV Induces Autophagy in PAM-KNU Cells Through the PI3K/AKT/mTOR Signaling Pathway

WANG LinYuan, XUAN JingYan, DU Yu, CHEN Tong, NIU RuiYan

Scientia Agricultura Sinica. 2025, 58(13): 2693-2706. doi:10.3864/j.issn.0578-1752.2025.13.015

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【Objective】 Environmental factor deoxynivalenol (DON) and infectious agent Porcine reproductive and respiratory syndrome virus (PRRSV), as two factors that cause serious harm to the pig industry in China, often occur simultaneously in clinical practice, but few studies have been conducted on their co-exposure, and their specific mechanisms are still unclear. The aim of this study was to study the effects of DON and PRRSV on host cell porcine alveolar macrophages (PAM-KNU), and to provide theoretical reference for the environmental requirements of pig industry. 【Method】 In this experiment, the PRRSV collected after amplification was diluted by gradient, and the lesion phenomenon (CPE) of Marc-145 cells infected with PRRSV was observed by microscope, and the titer of the virus was calculated. PAM-KNU cells were infected with PRRSV with MOI=1, and the expression level of PRRSV-N and virus titer of supernatant were detected by western blot at 0, 12, 24, 36, 48, 60 and 72 h, respectively, and the appropriate time was selected. At the selected time, the effects of DON concentrations of 0, 8, 16, 32, 64, 128, 256, 512, 1 024 and 2 048 nmol·L^-1 on the cell viability of PAM-KNU were detected by CCK-8 method. Therefore, the appropriate 4 DON concentrations were selected for follow-up tests. Four different concentrations of DON and PRRSV were exposed to PAM-KNU cells, and the protein expression of PRRSV-N was detected by western blot, and the final DON concentration was selected. On this basis, DON and PRRSV co-exposed cell models were established, which were divided into control group (group C), DON group (group D), PRRSV group (group V) and DON+PRRSV group (group VD). Autophagy in PAM-KNU cells was examined using electron microscopy. The levels of autophagy-related proteins LC3-II, Beclin1, p62, and ULK 1 were quantified by Western Blot and indirect immunofluorescence. Proteomic sequencing was conducted using Astral DIA technology, followed by KEGG pathway enrichment analysis of differentially expressed proteins. The expression of proteins involved in the PI3K/AKT/mTOR pathway was validated. 【Result】 By observing the CPE phenomenon, the titer of PRRSV is calculated to be 10^7.3 TCID₅₀/mL. The growth dynamic curve of PRRSV in PAM-KNU cells showed that the expression of PRRSV-N protein reached an exponential growth period at 24 h. The results of CCK-8 showed that DON concentration at 8, 16, 32 and 64 nmol·L^-1 had no significant effect on PAM-KNU cell viability. When the DON concentration is greater than or equal to 128 nmol·L^-1, it has a significant impact on cell viability, and the cell viability gradually decreases. Four of the DON concentrations (32, 128, 256, 512 nmol·L^-1) were exposed to PAM-KNU cells together with PRRSV. Western Blot results showed that when the DON concentration was 128 nmol·L^-1, the expression of PRRSV-N protein began to decrease significantly. On this basis, PAM-KNU cell model was established. The results of transmission electron microscopy indicated a decrease in the number of autophagosomes in the VD group compared to the D and V groups. Additionally, the expression levels of LC3-Ⅱ, Beclin1, p62, and ULK 1 were significantly reduced, suggesting that the level of autophagy was inhibited. Differential proteins were identified through proteomic sequencing and subsequently enriched in multiple pathways as determined by KEGG analysis. In this study, the PI3K/AKT/mTOR pathways were specifically validated. Western blot analysis revealed increased expression levels of p-PI3K/PI3K, p-AKT/AKT, and p-mTOR/mTOR, corroborating the findings from indirect immunofluorescence. 【Conclusion】 Co-exposure to DON and PRRSV significantly inhibited autophagy in PAM-KNU cells by upregulating the PI3K/AKT/mTOR pathway. This inhibition may result in the dysregulation of the host cell immune response, thereby affecting the host cell's ability to respond to pathogens.

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