With the passage of time and the advancement of technology, crop breeding has gone through generations from 1.0 to 4.0 and is now moving towards generation 5.0. Although the 3.0 and 4.0 generations of breeding have received extensive attention, only hybrid breeding of the 2.0 generation can enable the parents to achieve genome-wide recombination, resulting in a large number of complex and unpredictable interactions within and between genes, which may be the basis for the emergence of breakthrough traits. Thus hybrid breeding still holds an important position. However, at present, taking rice as an example, the hybrid breeding operations carried out by the majority of breeders may still have issues that need improvement in terms of scientificity and efficiency. In light of the current situation, in order to select high-yielding, high-quality, and multi-resistant varieties, and to overcome the homogenization of varieties, hybrid rice breeding should pay attention to the following aspects. Firstly, the breeding goals should be combined with the local natural conditions and effectively coordinate the combination of advantageous traits. Only in this way can the high-yield, high-quality and highly-resistant high-level goals be achieved, so as to break through the homogenization of varieties. Secondly, because the F₁ generation combines the superior traits of both parents and has certain hybrid vigor, it may be the best-performing generation of the same combination. If F₁ performs poorly overall, it is difficult for its offspring to produce the expected types that meet the breeding goals. Therefore, this generation should be selected as a key generation, which is conducive to significantly improving the efficiency of breeding. Thirdly, in the early stage of breeding, the main task is to promote generations. To enhance the breeding efficiency, direct seeding should be adopted, which can save land and resources. During the breeding process, the current generation should be combined with the early-generation tests to increase predictability and further eliminate combinations to improve the breeding efficiency. Fourth, during the high-generation selection process, after field selecting, the panicle traits of the combinations should be further compared indoors to select the optimal combination, so as to achieve the best from the best. Finally, the intelligent varieties of the 5.0 generation of breeding are those that can adapt to the ecological and biological factors of the wide range of environments, and can meet the production needs with wide adaptability. Due to the complexity of the environmental conditions for crop growth, it is necessary to conduct extensive and long-term identification of the varieties to achieve the breeding goals. In conclusion, by optimizing the field operations and selection techniques in hybrid breeding, the breeding efficiency will be significantly enhanced, laying the foundation for the selection of breakthrough varieties.

【Objective】 Rapid on-site screening of genetically modified (GM) crops is crucial for effective biosafety regulation. To overcome the limitations of current detection methods, such as equipment dependency and operational complexity, this study developed a closed-tube detection system by integrating recombinase polymerase amplification (RPA) with split DNAzyme (MNAzyme). The system enables rapid, sensitive, and on-site detection of the GM soybean event DBN9004, supporting regulatory compliance and industrial safety management. 【Method】 Using GM soybean DBN9004 and its non-GM counterpart Jack as experimental materials, we firstly identified event-specific sequences for target detection through bioinformatics analysis. Then a recombinant plasmid (9004P) was constructed as a standard template. An asymmetric RPA system was designed to efficiently amplify the target sequence while generating abundant single-stranded DNA (ssDNA) products for MNAzyme activation. Critical reaction parameters were systematically optimized, including reaction temperature (35-60 ℃), probe concentration (125-1 000 nmol·L^-1), and RPA primer ratios (10 000﹕10 000 nmol·L^-1-10 000﹕31.25 nmol·L^-1). Sensitivity assessment was evaluated using gradient-diluted plasmids (8×10^-1-8×10⁵ copies/μL), while specificity evaluation was verified against ten GM crop lines (GTS40-3-2, ZH10-6, etc.). Field samples (n=13) were tested and compared with qPCR results. 【Result】 The method demonstrated exceptional sensitivity (8 copies/reaction), good repeatability (RSD=4.44%) and reproducibility (RSD=5.75%), absolute specificity for DBN9004 with no cross-reactivity against ten prevalent GM soybean varieties. Field testing demonstrated perfect concordance (100%) with qPCR results (n=13). 【Conclusion】 This study implemented an asymmetric RPA strategy to efficiently generate target-specific ssDNA amplicons. The resulting ssDNA products demonstrate specific binding affinity for pre-engineered split DNAzyme subunits (A/B), triggering their activation and subsequent continuous cleavage of fluorophore-quencher labeled substrate probes. Leveraging this molecular mechanism, we established a novel RPA-MNAzyme integrated platform for rapid and reliable detection of genetically modified soybean event DBN9004. By combining asymmetric RPA with MNAzyme cascade amplification, the method achieves dual-specificity recognition and signal enhancement. The closed-tube design prevents aerosol contamination, while the dual-mode output system accommodates both laboratory and on-site screening needs.

【Objective】 The upper reaches of the Yangtze River represent one of China’s major rapeseed-producing regions, playing a pivotal role in ensuring the national supply of edible vegetable oil and improving the self-sufficiency rate of oil crops. However, the region is characterized by complex and variable climatic conditions, and traditional genetic improvement evaluation methods are highly susceptible to environmental interference, making it difficult to accurately track trends in varietal genetic potential. This study aimed to establish a method for tracing genetic improvement using annually top-yielding lines as representatives, thereby systematically revealing the genetic progress trajectory and agronomic trait evolution of rapeseed lines in the upper reaches of the Yangtze River from 2004 to 2023. The goal was to clarify the synergistic regulatory mechanisms of key agronomic traits in high-yield lines and provide theoretical support for high-yield and stable-yield rapeseed breeding. 【Method】 Annual top-yielding lines from the National Winter Rapeseed Regional Trials conducted in the upper reaches of the Yangtze River from 2004 to 2023 were selected as the study objects. A mixed linear model was used to separate environmental effects via the best linear unbiased prediction (BLUP) model. Linear regression, Pearson’s correlation analysis, standardized path analysis, and principal component analysis (PCA) were integrated to comprehensively assess yield genetic progress trends and trait variation patterns over the past two decades, and to construct a multi-trait synergistic regulation network. 【Result】 Both actual yield and BLUP-based yield of the rapeseed lines showed a significant upward trend from 2004 to 2023. The improvement of traits in high-yield lines exhibited clear stage-specific changes: from 2004-2013, breeding strategies emphasized compact plant type, with significant reductions in branching number, siliques per plant, and whole growth period; from 2014-2023, strategies shifted toward seed number type, with marked increases in siliques per plant and thousand-seed weight. Correlation and path analyses revealed that yield per plant is the core direct driving factor for enhancing population yield, while silique number per plant and branching number primarily contribute to population yield through indirect pathways via their effects on yield per plant. PCA revealed that the first five principal components all had eigenvalues greater than 1, cumulatively explaining 76% of the total variance. The PC1 axis, predominantly characterized by structural traits such as silique number per plant, branching number, and plant height, accounted for 29% of the variance, representing the primary dimension underlying inter-varietal differentiation, indicating a breeding trend from “single-trait breakthroughs” to “multi-factor synergy”. 【Conclusion】 The breeding focus for rapeseed in the upper Yangtze River has shifted from early-stage optimization of plant architecture toward late-stage enhancement of seed number. The study identified a high-yield model centered on yield per plant, supported by the coordinated improvement of branching number and siliques per plant, with balanced allocation to seed size traits. The lack of promotion area for high-yield varieties in the upper reaches of the Yangtze River from 2004 to 2013 indicates that the breeding direction in this region should comprehensively consider oil production and lodging resistance, in order to achieve sustained break throughs in rapeseed yield under different ecological and management conditions in the upper reaches of the Yangtze River.

【Objective】 Sodium nitroprusside (SNP), as an exogenous nitric oxide (NO) donor, plays a critical regulatory role in plant stress responses. This study aimed to investigate the effects of exogenous SNP on sugar metabolism in rice seedlings under alkaline stress, for providing a theoretical basis for elucidating the mechanism of SNP enhancing rice alkali tolerance. 【Method】 Seedlings of rice cultivars Zhonghua 11 and Ningjing 52 were used as test materials. Four treatments were applied: control (CK), CK+SNP (50 µmol·L^-1), alkaline stress (AS, a mixed alkaline solution containing NaHCO₃ and Na₂CO₃ at a 1:1 ratio, pH 9.55, 20 mmol·L^-1), and AS+SNP (20 mmol·L^-1 AS+50 µmol·L^-1 SNP). The effects of these treatments on sugar metabolism in the seedlings of the two rice cultivars were analyzed. 【Result】 Compared with AS treatment, AS+SNP treatment significantly increased rice seedling height, root length, fresh weight, and dry weight of rice seedlings. AS+SNP treatment upregulated the expression of chlorophyll synthase-related genes (OsChlH, OsCAO1, and OsCAO2), thereby increasing chlorophyll content and promoting photosynthesis. Concurrently, it upregulated the expression of the sucrose invertase gene OsNIN1, leading to decreased fructose and sucrose content but increased glucose content, which facilitated glycolysis and the tricarboxylic acid cycle, promoting the accumulation of malate and citrate to maintain energy balance. Additionally, SNP induced the expression of sucrose phosphate synthase OsSPS1, gibberellin synthase-related genes (OsGA3ox2, and OsGA20ox1), and proline synthesis gene (OsP5CS), increasing the levels of gibberellin (GA), abscisic acid (ABA), proline (Pro), and soluble sugar. This regulation helped maintain osmotic and hormonal balance and promoted sucrose transport from source to sink tissues. 【Conclusion】 Exogenous SNP application enhanced rice seedling tolerance to alkaline stress by upregulating chlorophyll biosynthesis genes, promoting chlorophyll accumulation, and sustaining photosynthetic efficiency, thereby facilitating glucose synthesis. Moreover, SNP activated glycolysis and the tricarboxylic acid cycle, directing sugar metabolites toward malate and citrate production, thus promoting organic acid accumulation. Additionally, exogenous SNP modulated osmotic regulation and GA/ABA hormonal balance, facilitating sucrose transport and sugar metabolic pathway activation, and ensuring efficient energy metabolism. These mechanisms collectively improved rice seedling resilience under alkaline stress.

【Objective】 To address the scientific challenges of limited yield potential, inefficient resource use efficiency, and constrained economic benefits in equidistant maize planting in the Northwest oasis irrigation zones of China, this study investigated the effects of wide-narrow row planting patterns on maize density tolerance and yield potential. 【Method】 This experiment began in 2017 and the data were collected from 2019 to 2021, using a split-plot design. The main plot treatments comprised three line spacing configurations characterized by alternating wide-narrow line spacing: L1 (7:3 ratio, 56 cm wide line spacing: 24 cm narrow line spacing), L2 (6:4 ratio, 48 cm wide line spacing: 32 cm narrow line spacing), and L3 (5:5 ratio: 40 cm wide line spacing, 40 cm narrow line spacing). The split-plot treatments were four maize planting densities: D1 (8.25×10⁴ plants/hm², local conventional planting density), D2 (9.00×10⁴ plants/hm², increased density by 9.1%), D3 (9.75×10⁴ plants/hm², increased density by 18.2%), and D4 (10.50×10⁴ plants/hm², increased density by 27.3%). The effects of these spacing arrangements under increasing planting density on maize photosynthetic characteristics, yield, resource utilization, soil nitrogen content, and economic benefits were investigated. 【Result】 Both wide-narrow line spacing planting and increased maize planting density enhanced photosynthetic characteristics, yield, and resource utilization efficiency of maize in the Hexi Oasis Irrigation District. Compared with wide-narrow line spacing ratio of 5:5 treatment, wide-narrow line spacing ratio of 7:3 treatment increased maize average leaf area index, light interception score at the big flare stage, grain yield, light use efficiency, water use efficiency, and nitrogen use efficiency by 9.7%, 7.1%, 8.8%, 8.2%, 12.7%, and 14.1%, respectively. Increased density by 9.1% and 18.2% treatments increased maize average leaf area index by 11.4% and 15.7%, light interception score at the big flare stage by 7.4% and 10.1%, grain yield by 9.6% and 11.3%, light use efficiency by 4.0% and 6.1%, and water use efficiency by 10.2% and 20.5% than that with local traditional planting density, respectively. Wide-narrow line spacing ratio of 7:3 with an 18.2% density increase demonstrated significant potential for comprehensive improvement. Compared with wide-narrow line spacing ratio of 5:5 and traditional planting density, wide-narrow line spacing ratio of 7:3 and increased density by 18.2% increased average leaf area index by 28.4%, enhanced light interception score at big flare and grain filling stages by 22.0% and 17.1%, respectively, and raised grain yield, biomass, and harvest index by 22.2%, 13.1%, and 8.0%, respectively. Wide-narrow line spacing ratio of 7:3 and increased density by 18.2% also improved average leaf area index, water use efficiency, and nitrogen use efficiency by 13.5%, 39.6%, and 24.0%, respectively, while reducing soil total nitrogen, ammonium nitrogen, and nitrate nitrogen contents by 11.2%, 18.0%, and 16.8% compared with wide-narrow line spacing ratio of 5:5 and traditional planting density, respectively. Additionally, wide-narrow line spacing ratio of 7:3 and increased density by 18.2% treatment increased net profit and the ratio of benefit and cost by 50.1% and 23.3%, compared with wide-narrow line spacing ratio of 5:5 and traditional planting density, respectively. 【Conclusion】 The wide-narrow line spacing ratio cropping pattern of 7:3 enhanced the comprehensive effects of densely planted maize by improving photosynthetic characteristics, resource use efficiency, and soil nitrogen supply.

【Objective】 The Huang-Huai-Hai region is a major production area for summer soybeans (Glycine max L.) in China. In this region, frequent wind, rain, and hail during the summer often cause damage to leave at the seedling stage, resulting in a reduction in leaf area. By using artificial defoliation, this study investigated the changes in photosynthetic characteristics, pod distribution, and yield formation of soybeans following leaf removal at the seedling stage, so as to provide support for the integrated application of stress-resistant and high-yield soybean cultivation techniques. 【Method】 Using 'Zhonghuang 301' as the experimental material, a two-year field experiment was conducted. Different defoliation treatments were established at the soybean V3 stage: CK (no defoliation), C1 (removal of 1 trifoliolate leaf), C2 (removal of 2 trifoliolate leaves), and C3 (removal of 3 trifoliolate leaves). These treatments simulated the reduction in leaf area caused by adverse weather conditions in seedlings to systematically analyze the effects of seedling defoliation on soybean yield and physiological characteristics. 【Result】 Defoliation at the seedling stage reduced soybean yield, and the degree of yield loss was positively correlated with defoliation intensity. The C1, C2, and C3 treatments resulted in average yield reductions of 5.4%, 10.2%, and 19.3% over two years, respectively. This was primarily attributed to an increase in the height of the lowest pod (which increased by 7.09%, 24.5%, and 42% compared with CK, respectively) and a decrease in the number of pods in the lower layer (0-30 cm) of the plant (which decreased by 13.6%, 33.8%, and 59.6%, respectively). The Leaf Area Index (LAI) decreased with increasing defoliation, leading to reduced aboveground dry matter accumulation; compared with CK, the accumulation under C1, C2, and C3 treatments at the R1 and R3 stages decreased by an average of 8.5%, 16.5%, and 37.1%, respectively. At the R8 stage, the grain dry weight for C1, C2, and C3 decreased by 12.1%, 24.3%, and 32.7% compared with CK, respectively. The photosynthetic characteristics analysis showed that there were no significant differences in photosynthetic indices between the C1/C2 treatments and the CK treatment at any growth stage. However, the C3 treatment significantly inhibited the net photosynthetic rate, relative chlorophyll content, and Photosystem II activity at the R1 stage, although its photosynthetic capacity recovered by the R3 stage. Nitrogen metabolism studies indicated that defoliation treatments reduced total aboveground nitrogen accumulation. Although the proportion of grain nitrogen reached 93.6% in the late reproductive stage, insufficient nitrogen supply from source organs limited sink capacity. At maturity stage, the difference in protein content between the treatment with the highest and the lowest values was only 1.27%. Principal Component Analysis (PCA) indicated that yield was positively correlated with LAI, photosynthetic rate, nitrogen accumulation, and lower-layer pod number, but negatively correlated with the height of the lowest pod. 【Conclusion】 Defoliation at the seedling stage exerts a significant negative impact on summer soybean yield, with the magnitude of yield loss increasing with defoliation intensity. Comprehensive analysis indicates that leaf area index (LAI), photosynthetic performance, nitrogen accumulation, and basal pod number serve as key sensitive indicators characterizing the degree of leaf injury. Elucidating the response patterns of these parameters offers a scientific basis for developing stress-resilient cultivation strategies for soybean.

【Objective】 The objective of this study is to investigate pathogen species and dominant pathogenic fungi of rice bakanae disease in Heilongjiang Province, and to provide a basis for precise prevention and control of rice bakanae disease. 【Method】 In 2023, rice bakanae disease samples were collected from 8 cities and 15 counties (districts) in Heilongjiang Province. A total of 172 single-spore strains were isolated and purified from the samples using tissue separation method and dilution plating method. Morphological identification, combined with multilocus phylogenetic analysis using ITS, RPB2, TEF1-α, LSU, and TUB2, and validation via Koch’s postulates, was used to complete the identification of the pathogen causing rice bakanae disease. 【Result】 172 strains of the pathogenic fungi causing rice bakanae disease were classified into 31 morphotypes based on their morphological characteristics. Multilocus phylogenetic analysis identified 15 genotypes, and the isolated pathogenic fungi included four species complexes: FFSC, FNSC, FOSC, and FIESC. Among them, the dominant pathogenic fungus was Fusarium fujikuroi in the FFSC. A total of 148 strains were obtained, accounting for 86.05% of the total number of isolated strains, and comprising 4 genotypes and 18 morphotypes. Nine strains of Fusarium madaense were also obtained from this species complex, accounting for 5.23% of the total strains, with 1 genotype and 3 morphotypes. In addition, 1 strain of Fusarium subglutinans was identified, representing 0.58% of the total, with 1 genotype and 1 morphotype. In the FIESC, 2 strains of Fusarium wereldwijsianum were obtained, accounting for 1.16% of the total strains, with 2 genotypes and 2 morphotypes; 7 strains of Fusarium ipomoeae were also identified, representing 4.07% of the total, with 5 genotypes and 5 morphotypes. From each of the FNSC and FOSC, 4 strains of Fusarium arbusti and 1 strain of Fusarium cugenangense were obtained, accounting for 2.33% and 0.58% of the total strains, respectively, with 1 genotype and 1 morphotype for each species. All 7 Fusarium species were pathogenic, but with varying degrees of pathogenicity. The dominant pathogenic fungus, F. fujikuroi, caused rice bud rot and etiolation; F. arbusti induced only rice bud rot; F. ipomoeae, F. subglutinans and F. wereldwijsianum inhibited rice growth; F. madaense and F. cugenangense resulted in rice etiolation. 【Conclusion】 The pathogens causing rice bakanae disease in Heilongjiang Province include 7 species: F. fujikuroi, F. madaense, F. ipomoeae, F. arbusti, F. wereldwijsianum, F. subglutinans, and F. cugenangense. Obvious differences exist in pathogenic types and geographical distribution depending on the pathogen species. Of these, F. fujikuroi and F. arbusti exhibited the strongest pathogenicity; F. fujikuroi was detected in all regions of Heilongjiang Province and served as the dominant pathogenic fungus.

【Objective】 Soil-borne disease has become a prevalent and intractable issue in current agricultural production, while the existing chemical control agents often lead to pesticide residues and phytotoxicity to crops. In contrast, botanical fungicides possess distinct advantages, including broad-spectrum inhibitory activity, environmental friendliness, as well as easy metabolism and degradation in the ecosystem. This study aimed to target two typical soil-borne pathogens (Phytophthora capsici and Fusarium oxysporum f. sp. niveum), determine the inhibitory activities of extracts from 75 tested plants, and to screen out plant resources with high activity, thereby laying a foundation for the development of novel botanical fungicides. 【Method】 The growth rate method was employed to determine the inhibitory activities of ethanol extracts of 75 kinds of plants against P. capsici and F. oxysporum f. sp. niveum. The pot efficacy of the screened high-activity extracts against pepper phytophthora disease and watermelon fusarium wilt was determined by means of the root irrigation and the root injury method. 【Result】 The mycelial inhibition rates of five ethanol extracts of Magnolia officinalis, Notopterygium incisum, Syringa pinnatifolia, Platycladus orientalis and Ilex chinensis against P. capsici were over 60% at the concentration of 1 mg·mL^-1. The inhibition rates of 11 other ethanol extracts, including Rubia cordifolia and Lindera aggregata, ranged from 30% to 60%, while the remaining 59 extracts showed no significant inhibitory activity. Among them, the ethanol extract of M. officinalis had the highest toxicity against P. capsici, with EC₅₀ of 0.023 mg·mL^-1. The mycelial inhibition rates of three ethanol extracts of M. officinalis, Ligusticum chuanxiong and Curcuma longa against F. oxysporum f. sp. niveum were over 60% at the concentration of 1 mg·mL^-1. The inhibition rates of 12 other ethanol extracts, such as Alpinia officinarum and S. pinnatifolia, were between 30% and 60%, and the remaining 60 extracts displayed no significant inhibitory activity. Among them, the ethanol extract of C. longa had the highest toxicity against F. oxysporum f. sp. niveum, with EC₅₀ of 0.606 mg·mL^-1. Pot efficacy results showed that the ethanol extract of M. officinalis exhibited high protective and curative efficacy against pepper phytophthora disease at the concentration of 8 mg·mL^-1, with efficacy rates of 64.74% and 63.25%, respectively. The ethanol extract of S. pinnatifolia showed high curative efficacy against pepper phytophthora disease at the concentration of 8 mg·mL^-1, with efficacy rate of 63.14%. However, the efficacy of both extracts was lower than the control fungicide shenqinmycin (protective efficacy: 81.05%; curative efficacy: 73.72%). The ethanol extract of C. longa showed high protective efficacy against watermelon fusarium wilt at the concentration of 8 mg·mL^-1, with efficacy rate of 69.94%, which was comparable to the control fungicide carbendazim (66.65%). When applied at 2 mg·mL^-1, this extract demonstrated a high curative efficacy against watermelon fusarium wilt, with efficacy rate of 78.08%, which was better than the control fungicide carbendazim (62.42%). In contrast, the efficacy of A. officinarum ethanol extract against watermelon fusarium wilt was less than 50% at all tested concentrations. 【Conclusion】 Ethanol extracts from S. pinnatifolia, C. longa, and M. officinalis exhibit potent inhibitory activities against soil-borne pathogens, which provides a scientific basis for the subsequent development of new botanical fungicides.

【Objective】 To address the issues of excessive nitrogen application and water scarcity in the irrigation area of northern Henan, the aim of this study was to explore the regulating effect of annual water and nitrogen optimization (reducing water and nitrogen by 25%) on crop growth and nitrogen loss in the wheat-maize rotation system, so as to provide a theoretical basis and technical support for the sustainable agricultural practices in the wheat-maize rotation system of this region. 【Method】 A five-year (2018-2023) field experiment was conducted in Huaxian County, Anyang City of Henan province, and three water and nitrogen management treatments were set up: local traditional water and nitrogen treatment (TP, annual nitrogen application of 552.0 kg·hm^-2 combined with irrigation of 270.0 mm), nitrogen factor optimization treatment (OF, annual nitrogen application of 414.0 kg·hm^-2 combined with irrigation of 270.0 mm), and water and nitrogen factor optimization treatment (OWF, annual nitrogen application of 414.0 kg·hm^-2 combined with irrigation of 202.5 mm). The effects of annual water and nitrogen optimization on the yield, biomass, nitrogen accumulation, nitrogen harvest index (NHI), nitrogen partial factor productivity (NPFP), and nitrogen uptake and physiological efficiency (NUPE), soil moisture and nitrate nitrogen content, nitrogen leaching and runoff, and annual nitrogen balance of the wheat-maize rotation system were studied. 【Result】 Compared with TP and OF treatments, OWF treatment had no significant effect on the annual average yield, aboveground biomass, grain nitrogen accumulation, and aboveground nitrogen accumulation of wheat and maize, but significantly increased NPFP by 31.4% and 0.9%, NUPE by 28.5% and 0.1%, respectively. There was a significant positive correlation among yield, biomass, grain nitrogen accumulation, and aboveground nitrogen accumulation. The annual optimization of water and nitrogen management significantly reduced soil moisture content and NO₃^--N content at different growth stages. Compared with TP and OF treatments, OWF treatment resulted in a significant reduction in soil moisture content by 6.6% and 9.6%, respectively, and a significant decrease in NO₃^--N content by 46.1% and 37.9%, respectively. The wheat season and maize season accounted for 2.9% and 97.1% of the annual average nitrogen leaching loss, respectively. The annual average nitrogen leaching losses under TP, OF, and OWF treatments were 52.9, 45.8, and 39.6 kg·hm^-2, respectively. Compared with TP and OF treatments, OWF treatment significantly reduced the annual average NO₃^--N leaching loss by 25.2% and 13.5%, respectively, and decreased NO₃^--N surface runoff by 32.0% and 18.1%, respectively. The annual average nitrogen surplus in the irrigation area of northern Henan was 60.6 kg·hm^-2. Compared with TP treatment, the nitrogen surplus under OWF treatment was reduced by 81.9%, while compared with OF treatment, it was increased by 20.8%. 【Conclusion】 Annual water and nitrogen optimization could maintain stable crop yields in the wheat-maize rotation system while enhancing the efficient utilization of water and nitrogen resources. This approach significantly reduced soil nitrate nitrogen (NO₃^--N) content across the 0-100 cm soil layers, mitigated NO₃^--N leaching, promoted soil nitrogen balance, and reduced the risk of non-point source pollution in the irrigation area of northern Henan. These results demonstrated that water and nitrogen optimization served as a green and efficient management measure for the wheat-maize rotation system in this region.

【Objective】 Based on the long-term fertilization station in fluvo-aquic soil region, this study investigated how bacterial community structure, ecological network, and key species in response to fertilization. 【Method】 Based on the long-term experiment started from 1990, soil was sampled after wheat maturity in 2023, and high-throughput sequencing and ecological network analysis were used to examine dynamics in soil organic carbon (SOC) and other nutrients, enzyme activity, bacterial community composition, ecological network and stochastic process under four treatments: no fertilization (CK), mineral nitrogen (N), phosphorus (P), and potassium (K) (NPK), NPK+straw (NPKS), and NPK+manure (NPKM). 【Result】 The combined application of organic and inorganic fertilizers significantly increased SOC and other soil nutrients, as well as soil enzyme activity. Compared with CK, SOC content under NPKS and NPKM increased by 52.1% and 81.9%, respectively, and particulate carbon increased by 60.6% and 137.4%, respectively, and easily oxidized organic carbon increased by 45.3% and 63.4%, respectively; additionally, β-N-acetylglucosaminidase activity increased by 7.2% and 12.6%, respectively, while alkaline phosphatase activity increased by 166.4% and 216.2% (P<0.05), respectively. Notably, β-1,4-glucosidase activity was the highest under NPKS (63.82 μmol·g^-1·d^-1). In terms of bacterial diversity, α-diversity significantly decreased under NPKS compared with CK, with reductions of 5.4%, 5.2%, and 2.6% in the Ace, Chao1, and Shannon indices, respectively (P<0.05). Fertilization treatment alerted bacterial community structure, while NPKS and NPKM exhibited similar compositions. Compared with CK, NPKS significantly reduced the relative abundance of Chloroflexi, Gemmatimonadota, and Methylomirabilota, while NPKM significantly increased the relative abundance of Bacteroidota (P<0.05). Redundancy analysis identified ammonium nitrogen (NH₄⁺-N), nitrate nitrogen (NO₃^--N), and organic carbon (SOC) as the primary environmental factors shaping microbial community structure. Network analysis showed, compared with CK, NPKM increased the complexity, stability of the bacterial community network and the proportion of positive correlations between species. Furthermore, both NPKS and NPKM significantly enhanced the relative abundances of eight keystone taxa, including members of Actinobacteriota (order Microtrichales), Chloroflexi (order Thermomicrobiales), Bacteroidota (orders Chitinophagales and Cytophagales), Myxococcota (uncultured order bacteriap25), and Proteobacteria (order Burkholderiales) (P<0.05). These keystone taxa were closely associated with soil carbon, nitrogen, phosphorus, and other material cycling, as well as plant growth promotion. Partial least squares path modeling suggested that fertilization did not directly impact key species but exerted an indirect influence by significantly affecting soil pH, regulating community composition, and increasing soil nutrient availability. 【Conclusion】 Long-term combined application of organic and inorganic fertilizers enhanced soil nutrient content and extracellular enzyme activity, regulated microbial community composition and structure, affected bacterial network complexity and stability, and increased the relative abundance of key species involved in soil nutrient cycling and material transformation. These findings provided valuable insights into the interactions between soil properties and microbial communities under long-term fertilization, contributing to a deeper understanding of bacterial community dynamics and key species in agricultural ecosystems.

【Objective】 This study aimed to investigate the characteristics of soil fungal diversity, functional groups, and community assembly in tea garden soils subjected to varying long-term nitrogen applications. Additionally, it sought to determine the optimal nitrogen fertilizer application rate for tea gardens, thereby providing a scientific foundation for the sustainable and rational use of nitrogen in tea cultivation. 【Method】 The field experiment (small cement pond) was set up in the base of the Tea Research Institute, Fujian Academy of Agricultural Sciences, (beginning in 2011), and four nitrogen levels were applied: N0 (0), N1 (112.5 kg N·hm^-2), N2 (225 kg N·hm^-2), N3 (450 kg N·hm^-2), and each treatment was repeated four times. ITS high-throughput sequencing was used to analyze the effects of long-term nitrogen applications on the soil fungal diversity, functional groups, and community assembly. 【Result】 Compared with N0, long-term nitrogen increased the yield of spring and autumn tea by 137.79%-430.20% and 33.43%-67.49%, respectively, but there was no significant difference between N2 and N3 treatments. The soil fungal diversity tended to initially increase and then decrease with increasing nitrogen addition in two seasons. Compared with N0, N3 significantly increased the Ace, Chao1 and Shannon of fungal. Results of non-metric multidimensional scaling analysis (NMDS) and permutational multivariate analysis of variance (PERMANOVA) showed that long-term nitrogen application drastically changed the community structures of soil fungi in tea plantations. The functional prediction with FUNGuild showed that long-term nitrogen application significantly changed the functions of soil fungi during the spring period, but the changes were not significant during the autumn. Compared with the N0 treatment, N2 and N3 treatments decreased the relative abundance of plant pathogens and soil saprotrophs fungi (especially during the spring tea), and N2 treatment increased the relative abundances of two beneficial fungi, namely ectomycorrhizal fungi and arbuscular mycorrhizal fungi. Redundancy analysis (RDA) showed that the soil pH, nitrate nitrogen, ammonium nitrogen and available potassium were the main factors for determining fungal community structure and functional groups. Compared with N0 treatment, the application of nitrogen fertilizer increased the number of edges, average degree, average clustering coefficient, and network density of the fungal network, and so enhanced the stability of the fungal community and improved its resistance to disturbances. In the process of community succession, stochastic processes dominated the construction of the tea garden soil fungal community under long-term nitrogen application, and the deterministic processes were enhanced in the community assembly under the low and medium nitrogen treatments. 【Conclusion】 From the perspective of yield, community diversity, enrichment of enrichment of beneficial fungi, and network stability, the applying nitrogen fertilizer of 225 kg N·hm^-2 was a reasonable amount for tea plant.

【Objective】 To explore the elite germplasm resources of citron and lemon, this study conducted genetic diversity analysis and comprehensive evaluation based on resequencing and phenotypic data. The research aims to elucidate the population genetic structure of citron and lemon germplasm resources, providing foundational materials for lemon germplasm innovation. 【Method】 In November 2023, 13 fruit traits and 43 volatile functional components in the peel of 158 citron and lemon germplasm resources were measured. In June 2023, the contents of 4 flavonoids in immature fruits were analyzed. Genetic diversity was assessed using the Shannon-Wiener index and coefficient of variation. Combined with correlation analysis, cluster analysis, principal component analysis and other methods, the citron, lemon germplasm resources were comprehensively evaluated. SNP data were used to construct the phylogenetic tree and analyze population genetic structure. Specific accessions were screened based on phenotypic data. Genome-wide association analysis (GWAS) was performed to identify significant variation sites associated with alloocimene. Candidate genes within 10 kb of these sites were annotated and analyzed. 【Result】 The fruit traits of 158 citron, lemon germplasm resources displayed rich genetic diversity and extensive variation, The coefficient of variation of the 13 phenotypic traits (single fruit weight FW, single fruit peel weight PW, fruit transverse diameter FD, fruit longitudinal diameter FL, fruit shape index FSI, peel thickness PT, number of segments SNPF, seed number per fruit SNF, single seed weight SW, soluble solids content TSS, ascorbic acid content AA, titratable acid content TA, solid acid ratio TSS /TA) ranged from 14.81%-143.47%, and the genetic diversity index ranged from 1.21 to 2.01.The genetic diversity index of 4 flavonoids (rutin, naringin, hesperidin and neohesperidin) in fruits ranged from 0.75 to 1.89, and the genetic diversity index of 43 volatile compounds in peel ranged from 0.25 to 1.93. The principal component analysis on 60 quantitative traits of citron, lemon germplasm resources was carried out, the top 5 principal components explained 66.55% of cumulative variance. A comprehensive evaluation was conducted based on the results of principal component analysis, and comprehensive evaluation score ranged from -0.09 to 2.14, specific germplasms such as Volkamer (Citrus volkameriana Pasquale), Fantastico Bergamot (Citrus bergamia Risso), Castagnaro Bergamot (Citrus bergamia Risso), Amber Lemon (Citrus limon (L.) Burm.f.) and Verna Lemon (Citrus limon (L.) Burm.f.) were identified. Germplasm resources were divided into 7 sub-groups, with high consistency between phenotypic clustering and phylogenetic analysis, except Lime (Citrus aurantifolia Swingle) and Citrus limonia sub-groups showed slight divergence. The elite germplasm clustered predominantly in sub-group 3, indicating shared genetic background. The population genetic structure analysis was consistent with the phylogeny analysis results except that the hybrid varieties (e.g., sweet lemon) formed a distinct sub-group. Significant SNP loci associated with alloocimene content were identified. Two cytochrome P450 family genes (C71D7_SOLCH, C71DB_LOTJA) were proposed as potential regulators of alloocimene metabolism. 【Conclusion】 Citron and lemon germplasm exhibit extensive genetic diversity in fruit quality traits. Phenotypic clustering analysis was similar to phylogenetic tree analysis and population genetic structure analysis, it showed that gene introgression from mandarin and pomelo likely drove sub-group differentiation. Citron and lemon germplasm resources with higher scores in the comprehensive evaluation were more concentrated in the same sub-group in phylogenetic tree, indicated similar genetic background were presented in these elite and specific germplasms. The candidate genes of C71D7_SOLCH and C71DB_LOTJA may be involved in the metabolism of alloocimene, a volatile substance in the peel.

【Objective】 Oil content is a key indicator determining the quality and economic value of oil crops. Tea plant (Camellia sinensis (L.) O. Kuntze), as an economically significant plant, holds potential application value in oil resource development through its fruit, tea seeds. The glycerol-3-phosphate dehydrogenase gene (GPDH) is a crucial regulatory gene in plant oil biosynthesis. Cloning CsGPDH and conducting an in-depth analysis of its expression characteristics and function aims to provide a theoretical basis for elucidating the molecular mechanism by which CsGPDH regulates oil synthesis in tea plants. 【Method】 Using ‘Jincha No.18’ as plant material, the full-length sequence of CsGPDH was cloned. Bioinformatics tools were employed to analyze the structural characteristics and evolutionary relationships of its encoded amino acid sequence. Real-time quantitative PCR (qRT-PCR) was used to detect the expression pattern of CsGPDH in different tea plant tissues and during key stages of oil synthesis. Subcellular localization of the encoded protein was observed using laser scanning confocal microscopy. Agrobacterium-mediated genetic transformation was utilized to generate CsGPDH-overexpressing Arabidopsis thaliana plants. Phenotypic analysis was performed on transgenic and wild-type Arabidopsis plants, measuring indicators such as seed oil content and fatty acid composition. 【Result】 CsGPDH was successfully cloned, with a coding sequence (CDS) length of 1 128 bp. The protein encoded by CsGPDH possesses a typical glycerol-3-phosphate dehydrogenase domain and exhibits high sequence similarity with homologous proteins from other species. Subcellular localization revealed that the CsGPDH protein is localized to the plasma membrane and cytoplasm. qRT-PCR analysis showed that CsGPDH expression significantly increased during the middle and late stages of seed development, highly coinciding with the period of rapid oil accumulation. Functional validation in transgenic Arabidopsis demonstrated that overexpression of CsGPDH increased seed oil content by 20.6%-25.2% and significantly elevated the proportion of unsaturated fatty acids (C18:1, C18:2). 【Conclusion】 CsGPDH expression is regulated by the seed developmental process, showing high expression during the critical phase of oil synthesis. Overexpression of CsGPDH can significantly enhance seed oil content in plants and plays an important positive regulatory role in the plant oil biosynthesis metabolic pathway.

【Background】 In China’s drought-prone grape-growing regions, high summer temperatures and scarce precipitation often lead to prolonged water deficits, exposing grapevines to heat stress. This results in accelerated fruit ripening, elevated sugar content, reduced acidity, and compromised wine flavor and quality. Under such water-scarce conditions, balancing irrigation water conservation with raw material quality improvement has become a critical challenge. 【Objective】 This study investigated the combined effects of regulated deficit irrigation (RDI) and anti-transpirant (AT) application on fundamental quality indicators of Cabernet Sauvignon grape berries and wine quality. The objective was to identify an optimal combination strategy that simultaneously could achieve water conservation and enhance wine quality. 【Method】 The experiment was conducted at Longyu Winery in Xixia District, Yinchuan City, Ningxia in 2024, using Cabernet Sauvignon grapes as materials. From early July (berry swelling stage), regulated deficit irrigation treatments (RDI-1: 40% ETc (evapotranspiration), RDI-2: 60% ETc, and RDI-3: 80% ETc) were implemented, and ATs were sprayed twice in August. Starting from late July, grapes were harvested every two weeks to measure basic physicochemical indicators. On September 12, the grapes were harvested and used to produce Cabernet Sauvignon dry red wine. After fermentation, the physicochemical indicators and various phenolic compound indicators of the wine were measured. 【Result】 The synergistic application of regulated deficit irrigation (RDI) and ATs significantly enhanced soluble solids and total acid content in grapes while markedly suppressing reducing sugar accumulation, which contributed to reduced alcohol content in wine. The RDI-2-AT group exhibited the lowest alcohol content (11.94% vol). The RDI-1 group showed the lowest total organic acid content (5.01 g·L^-1), whereas the RDI-2-AT group demonstrated a 12.40% increase in tartaric acid content compared with the RDI-2 group. Total phenolics exhibited a gradient increase with intensified water deficit, but anti-transpirant-treated groups showed significant reductions (14.83%, 21.33%, 22.59%, and 32.45% lower than that under CK group). Specifically, the RDI-1-AT group had 26.89% lower phenolic acid monomers than the RDI-1 group. The RDI-3 group recorded the highest total monomeric phenolics (163.74 mg·L^-1), 3.54% higher than that under CK group, while the RDI-2-AT group had the lowest value (115.4 mg·L^-1), significantly below the CK group. For monomeric anthocyanins, the CK group had the lowest total value (23.38 mg·L^-1), whereas the RDI-1 group had the highest value (34.82 mg·L^-1). Both RDI-1-AT and RDI-3-AT groups showed significantly lower monomeric anthocyanin contents than their non-anti-transpirant counterparts (RDI-1 and RDI-3 groups). 【Conclusion】 The combined application of 60% ETc regulated deficit irrigation (RDI-2) and ATs (RDI-2-AT) could increase total acid and phenolic content in wine while reducing volatile acidity and alcohol content. This synergistic approach thereby enhanced the taste profile and stability, demonstrating the most significant improvement in wine quality.

【Objective】 The meat of Lueyang black-bone chickens is delicious and nutritious, and has the value of medicine and food therapy, which has a special flavor. However, there are few studies on its flavor substances, especially the lack of systematic sensory flavor analysis. This study provided the theoretical basis for the selection of marketing time of Lueyang black-bone chickens by analyzing the body size traits in the growth and development and comparing the flavor differences of breast muscles. 【Method】 The experiment selected 30 Lueyang black-bone chickens randomly, and the body weight and body sizes at the age of 3, 6, 8, 10, 13, and 16 weeks were measured to analyze dynamic growth and development process. At 13 and 16 weeks of age, 10 chickens were randomly selected for slaughter for collecting the chest muscle and analyzing their volatile flavor substances by comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (GC×GC-TOF MS) combined with multivariate data. 【Result】 The growth rate of Lueyang black-bone chickens was faster before 13 weeks, and there was a significant difference in body weight and body size (P<0.05), while the growth rate slowed down after 13 weeks, there was no significant difference in body weight between 13 and 16 weeks (P>0.05). Body size indexes (chest width, chest depth, keel length, etc.) also tended to be stable after 13 weeks (P>0.05). The results of the meat quality analysis showed that there was no significant difference in pH, shear force and dripping loss between 13 and 16 weeks (P>0.05), indicating that the 13 weeks had reached physiological maturity. The results of flavoromics analysis showed that 642 and 660 flavor substances were identified in the chest muscle of 13 and 16 weeks Lueyang black-bone chickens respectively, among which alcohols, aldehydes, esters and hydrocarbons were the main components. Sensory analysis showed that the breast muscles of 13 weeks Lueyang black-bone chickens were significantly better than those of 16 weeks in aroma note types of sweety, green and fruity, and this difference was closely related to fatty acid composition. 43 key differential flavor substances were further screened, and Phenol, Pyrazine, 2-Cyclopenten-1-one, 3-methyl- and Trichloromethane were found to be the main volatile compounds responsible for the flavor difference between 13 and 16 weeks chest muscles. Correlation analysis showed, 1H-pyrrole,2-methyl-, 2-Cyclopenten-1-one,3-methyl-, 2-Hexene,3,5,5- trimethyl-, 2(3H)-Furanone,5-butyldihydro-, Ethanone,1-(4,5-dihydro-2-thiazolyl)-, Formamide, N, N-dibutyl-, Phenol, Pyrazine, Pyrazine, methyl- and Trichloromethane (green and fruity substances) were significantly negative correlation with the total amount of fatty acids, and 1-Heptanol,2-propyl- and Biphenyl (lipid fragrance precursor substance) were significantly positive correlation with the total amount of fatty acids (|r|>0.5，P<0.05). 【Conclusion】 The chest muscle at 13 weeks had a more obvious floral, fruity, and sweety aroma. The higher fatty acids content in the chest muscle at 16 weeks might be richer after cooking. Considering the growth rate, flavor characteristics and breeding cost, it was possible to market the Lueyang black-bone chickens at 13 weeks of age in advance.

【Objective】 The aim of this study was to analyze the cellular heterogeneity of duck testes before and after maturation using single-cell sequencing technology, to construct a developmental map of duck testicular cells, and to further reveal the process and regulatory mechanisms of testicular maturation. 【Method】 Each three testes samples from 10 week (immature testis group, IMT) and 23 week old (mature testis group, MT) ducks were collected, and single-cell suspensions were prepared for single-cell sequencing. The quality control, standardization, dimensionality reduction, and cluster analysis on the obtained sequencing data were performed to construct testicular maps of ducks at different developmental stages. Cell subpopulations were annotated, intergroup differences were analyzed, and key regulatory genes and signaling pathways of testicular development and maturation were screened. 【Result】 More than 30 million high-quality sequencing data were obtained from the each six samples. After quality control, 54 702 valid cells were obtained (27 756 in the IMT group and 26 946 in the MT group). Through cell clustering and identification of marker genes, 21 cell sub-populations were successfully identified, including 10 germ cell sub-populations and 11 somatic cell sub- populations. During the development and maturation of duck testes, spermatogonial stem cells, spermatogonia, primary spermatocytes, and Sertoli cells type 1 and type 3 decreased significantly, while intermediate spermatocytes, secondary spermatocytes, round spermatids, elongated spermatids, and Sertoli cells type 2 and type 4 increased significantly. Differential gene analysis identified 4 495 differential genes (1 737 up-regulated and 2 758 down-regulated). Among them, genes such as Gm614, SPATC1, TSSK6, ZIC1, PHF7, Armc12 and FAM166C were significantly up-regulated in germ cells, and genes such as PKIA, SNX22, Wnt6, LAMA1, SCN4B, CDH6 and FAM110C were significantly down-regulated in Sertoli cells. Functional enrichment analysis showed that the top 20 GO terms all belonged to the cell component category. KEGG analysis indicated that signaling pathways such as protein processing in the endoplasmic reticulum and cellular senescence played key regulatory roles. Furthermore, protein-protein interaction network analysis of differentially expressed genes of these two signaling pathways showed that SEC63 regulated the protein processing process in the endoplasmic reticulum through interactions with ERLEC1, AMFR and RAD23B, etc., while CDK6 might regulate the cellular senescence signaling pathway through interactions with CHEK2, CCNA2, CCNA1 and CDC25, etc. 【Conclusion】 This study constructed the single-cell transcription map of duck testes before and after maturity for the first time, revealed the dynamic change rules of cell sub-populations during testicular development, discovered a large number of differentially expressed genes, and suggested that SEC63 and CDK6 might play important roles in the testicular maturation mechanism by regulating the endoplasmic reticulum stress response and cell cycle progression, respectively.

Under the impetus of economic and social development and the rural revitalization strategy, China’s grain output has maintained bumper harvests for many consecutive years, but it still faces challenges such as the low production efficiency and the large income gap among farmers. Given the national condition of “a large country with small-scale agriculture”, promoting the connection between small farmers and modern agriculture is of great significance, and agricultural socialized services have become a key bridge linking them. This paper focused on China's agricultural socialized services, aiming to analyze their development status and regional characteristics, draw lessons from international experience, and put forward development suggestions, so as to provide the reference for promoting agricultural modernization. In terms of the current development situation, the policy system for agricultural socialized services has been gradually improved, with increasingly clear policy goals, supporting the diversification of service subjects, the diversification of policy tools, and regional differentiation. In terms of the regional service structure, taking the socialized service situation in Henan Province as the analysis object, the agricultural socialized service market has formed a three-level market structure of "leading enterprises - agricultural machinery professional cooperatives - agricultural machinery professional households". The government took the lead in providing financial support, and market entities at different levels had complementary advantages. In terms of regional service characteristics, China has initially built a new agricultural socialized service system characterized by “government guidance, multi-stakeholder collaboration and functional complementarity”. This system took public service institutions as the basic support, cooperative economic organizations as the grassroots network, leading agricultural enterprises as the industrial hub, and social forces such as scientific research institutions and industry associations as important supplements. It combined public welfare services with commercial services and coordinates special services with comprehensive services. In terms of international experience, the United States has built a three-dimensional collaborative agricultural socialized service system adapting to large-scale agricultural production, which consisted of government public services, private enterprise services and farmer cooperative systems. Japan has established an agricultural socialized service system centered on agricultural associations, which was suitable for small-scale family operations. In the reform of their service systems, both countries have achieved upgrades in organizational structure, service content and technical empowerment. The common points of the two countries' service systems lay in connecting farmers with modern agriculture through organizational embedding, promoting agricultural modernization with technical empowerment, and innovating service content to enhance industrial competitiveness. Based on China's national conditions and international experience, the following policy suggestions were put forward: We should strengthen government guidance and institutional guarantees to consolidate the service foundation, cultivate diversified service subjects and build a collaborative service network, deepen financial and digital empowerment to improve service quality and efficiency, and improve the interest linkage mechanism to protect the rights and interests of small farmers.