Ecosystem stability ensures the sustainable supply of ecosystem services. Our current understanding of ecosystem stability is mainly based on results from the initial stage after disturbance or experimental treatments, which reveal short-term responses. There is a big knowledge gap regarding whether the long-term responses would be consistent with such initial responses. We examined how grazing and mowing affect the temporal stability of aboveground biomass in a temperate semi-arid steppe and further clarified the differences in treatment impacts and primary drivers of stability between the initial four years and the subsequent five years. Both grazing and mowing significantly decreased the community stability across the nine years, and such impacts showed high stage-dependency. Grazing reduced community stability in the first four years but not in the following five. Mowing initially did not affect community stability but reduced it over the next five years. Community stability was driven by species asynchrony in the early stage, and later solely regulated by the dominant species stability. When Stipa krylovii and Cleistogenes squarrosa become dominant in the community following prolonged grazing, or when mowing leads to a notable decline in community stability over time, enclosure measures should be adopted to promote ecological recovery. Additionally, long-term enclosure may also lead to a decline in biomass, thereby affecting the maintenance of community functions. We recommend implementing a rational rotation of enclosure, grazing, and mowing in grassland management based on different stages, as a scientifically sound grassland management system is key to achieving sustainable utilization of grassland resources.

Cotton (Gossypium spp.), a globally important cash crop, is increasingly threatened by abiotic stresses that significantly affect yield and fiber quality. In this study, data on 3,016 abiotic stress-related quantitative trait loci (QTLs) described in 31 published papers were integrated through meta-QTL analysis, a total of 34 MQTLs were identified. Nine major MQTLs with numerous initial QTLs, high R² values, narrow confidence intervals (CIs), and close colocalizations were successfully detected. Combined with the transcriptome data, the candidate gene GhPCMP-E17 was identified. Through virus-induced gene silencing (VIGS) technology, the role of GhPCMP-E17 in the response to abiotic stress was clarified. Compared with the TRV:00 plants, the GhPCMP-E17-silenced plants presented more severe wilting and yellowing under drought and salt stress conditions. Silencing GhPCMP-E17 weakens the function of antioxidant enzymes, thereby increasing the accumulation of reactive oxygen species. These results indicate that downregulation of GhPCMP-E17 gene expression enhances the sensitivity of cotton plants to drought and salt stress. This research provides excellent genetic resources for adaptive abiotic crop breeding in upland cotton.

To dissect chromatin accessibility profiles in pig skeletal muscle at single-nucleus resolution, we performed single-nucleus ATAC-seq on longissimus dorsi and psoas major muscles of Large White pigs. After quality control, 26,225 nuclei were classified into seven major cell types, including myofibers, muscle stem cells, and immune cells, using snRNA-seq-based label transfer. We identified 158,438 accessible chromatin regions, with cell-type-specific differentially accessible regions (DAR) enriching for specific functions. Myonuclei subtypes (type I, IIA, IIB) showed distinct accessibility patterns, with SIX1 and MAF transcription factor motifs enriched in fast myofibers (type II). Comparative analysis between muscles revealed that myofiber composition drove chromatin differences, with psoas major featuring more type I myofibers. Cross-breed analysis (Rongchang vs. Large White) identified breed-specific DARs in myonuclei, linking MEF2-mediated regulation to myofiber hypertrophy. Pseudo-temporal analysis of myogenesis showed dynamic accessibility changes in key myogenic genes (e.g., MYF5, MYH1). This study unveils cell-type-resolved chromatin landscapes underlying myofiber specification, tissue heterogeneity, and breed-specific muscle development in pigs.

Waterlogging stress (WS) significantly threatens summer maize production. Although supplemental nitrogen (N) fertilization is a common remediation strategy, the differences in N regulation pathways across developmental stages are still unclear. The maize hybrid ‘Zhongkeyu 505’ was used as the experimental material in a 3-year field localization experiment investigating the responses and N-mediated recovery pathway in maize at the jointing stage (V6) and blister stage (R2) to WS. Over three experimental years, compared to control, WS significantly decreased grain yield at V6 and R2 by an average of 27.6 and 23.0%, respectively. Structural equation modeling showed that WS decreased grain number at V6 by inhibiting leaf photosynthetic capacity, and reduced grain capacity at R2 by limiting N allocation to grains. However, post-waterlogging nitrogen application (WF) effectively mitigated these losses, increasing grain number by 19.1% at V6 and improving grain sink capacity by 23.4% at R2. The recovery at V6 was driven by enhanced photosynthetic capacity with average increases of 23.6% in total chlorophyll content, 36.3% in phosphoenolpyruvate carboxylase activity, and 24.8% in Rubisco activity. Differently, the recovery at R2 was due to improve N utilization, where N allocation in grains increased to 73.1%, and nitrate reductase and glutamine synthetase activities in grains increased by an average of 31.7 and 35.6%, respectively. Transcriptomic analysis further confirmed upregulation of protein-processing genes (e.g. hsp18a and hsp18c), facilitating N allocation and utilization. In conclusion, the N-mediated recovery pathway varied across maize development stages, with the photosynthetic capacity and grain number restored at V6 and N allocation to grains and sink capacity was increased at R2. This study will provide significant theoretical and practical value for enhancing tolerance in maize to WS at V6 and R2.

Plastic film mulching (PM) is widely used to improve soil temperature and moisture in rainfed agriculture, but the combined effects of PM proportions and nitrogen (N) fertilization on spring maize under different hydrological conditions remain unclear. We conducted a two-year field experiment (2021 and 2022) in the Loess Plateau and improved the AquaCrop model by incorporating stage-specific temperature-increase compensation coefficients for full and wide PM treatments. Results showed that PM increased soil temperature by 2.11–3.59°C and water content by 14.3–21.2% during the first 90 days after sowing, requiring compensatory temperature increases during the sowing-tasseling stage. The modification reduced normalized root-mean-square error values of canopy cover and biomass from 14.2 and 7.6% to 8.1 and 6.2%, respectively. Using 40 years of hydrological simulation, we identified the optimal PM–N combinations: FN180 (100% PM+180 kg N ha⁻¹) for dry years to maximize precipitation use efficiency (+35.3%), and WN225 for normal and wet years to balance yield and nitrogen fertilizer agronomic use efficiency (>14 kg kg⁻¹). Our results demonstrate that tailoring PM proportions and N rates to hydrological conditions enhances water–nutrient synergy, providing a climate-resilient management approach for sustainable maize production in semi-arid agroecosystems.

Photosynthesis serves as the primary source of nutrients synthesized in higher plants, and enhancing photosynthetic efficiency can significantly improve crop yield and fruit quality. Leaf color mutants, which are ideal materials for studying chloroplast development and photosynthesis mechanisms, have been extensively investigated in field crops. However, the study on their application in watermelon remains limited. In this study, we identified a yellow-green phenotype mutant, PKH352, from an EMS mutagenesis watermelon mutant library. The chlorophyll content and maximal photochemical efficiency in PKH352 was significantly reduced. Genetic analysis showed that the mutated trait was controlled by a single nuclear gene, which named as Clygp (Citrullus lanatus yellow-green plant). Through MutMap and linkage analysis in an F₂ population of 440 plants, we identified a single nucleotide polymorphism (SNP) mutation in ClG42_04g0106300, which encoded a signal recognition particle 54 kDa protein, as the causal variant for the yellow-green phenotype. Further validation using a CRISPR/Cas9-mediated system confirmed that knockout of ClG42_04g0106300 results in the yellow-green phenotype in watermelon. In addition, comparative transcriptomic analysis revealed that the ClG42_04g0106300 mutations greatly affected the expression of key genes associated with chloroplast development and photosynthesis, providing strong evidence that it plays a critical role in these biological pathways. Taken together, these findings provide insights into the molecular mechanisms underlying chloroplast development and photosynthetic efficiency, offering a theoretical basis for breeding watermelon varieties with high photosynthetic efficiency.

Cucumber (Cucumis sativus L.) is a major vegetable crop worldwide, and its yield and quality are closely linked to flower development. AGAMOUS-LIKE 6 (AGL6), a member of the ancient MADS-box transcription factor family, plays a crucial role in flower development. However, the specific functions of its homolog in cucumber remain poorly understood. In this study, we demonstrate that CsAGL6 is predominantly expressed in flowers, with high expression levels observed in all floral organ primordia during the early stages of floral development. The petals of Csagl6 mutants exhibit a greener color compared to wild-type plants, along with a significant increase in total chlorophyll content. Additionally, the mutants show abnormal petal morphology, including changes in size and shape, as well as enlarged sepals resembling leaves occasionally. Molecular analysis reveals that the A-class gene CAULIFLOWER (CAL) and the E-class gene SEPALLATA 4 (SEP4) are significantly downregulated in the mutants, while the chlorophyll synthesis gene Early Light-Induced Protein 1 (ELIP1) and several stress-related genes in the chloroplasts are dramatically upregulated. Our findings provide novel insights into the functional role of CsAGL6 in regulating sepal and petal development, and offer a potential avenue for understanding the genetic control of flower pigmentation and organ morphology in Cucumis species.

Flavonols and flavanones are important bioactive compounds with multiple pharmacological activities and health benefits. Transcriptional activation of flavonol and flavanone biosynthesis has been studied extensively, while little is known about the negative regulators. CRISPR/Cas9 gene-editing technology, with the advantage of precise genetic modification, is a desirable tool for breeding biofortified materials and exploring potential molecular mechanisms. In this study, a transcriptional repressor, SlMYB32, was characterized in tomato fruit. Phenotype and metabolome analysis confirmed that knockout of SlMYB32 resulted in increased accumulation of flavonols and flavanones, especially about 1 mg g^-1 FW of quercetin 3-O-rutinoside (rutin). Transcriptome analysis indicated that expression of key genes SlPAL6, Sl4CL3 and Sl4CL4 as well as five candidate SlUGTs were significantly up-regulated in slmyb32 mutants. Dual-luciferase and EMSA assay indicated SlMYB32 could bind to and repress promoter activities of SlPAL6 and Sl4CL3. Expression of 27 transcription factors belonging to twelve families was significantly changed in slmyb32 mutants, among which two SlMYBs, two SlNACs, two SlAP2s and one SlWRKY were clustered with known flavonoid regulators. Our results provide new insights into improving bioactive compounds in fruit and understanding negative regulatory mechanisms in flavonol and flavanone biosynthesis.

The bean bug, Riptortus pedestris, is a major pest of soybeans in East Asian countries. Male-released aggregation pheromones attract both adults and nymphs, offering potential for eco-friendly pest control. However, the molecular mechanisms underlying the detection of the aggregation pheromones remain unclear. In the present study, functional analysis using the Xenopus oocyte expression system demonstrated that two ORs (OR23h and OR109d) were responsible for sensing aggregation pheromones, with the primary component (E)-2-hexenyl (E)-2-hexenoate (E2HE2H) being shared by the two ORs. Further quantitative PCR (qPCR) profiling indicated that OR109d was expressed only in male antennae, while OR23h was expressed in both sexes at similar levels. RNA interference assays demonstrated that dsOR23h-treatment significantly reduced the Electroantennographic (EAG) response of (E)-2-hexenyl (Z)-3-hexenoate (E2HZ3H) in both sexes. Furthermore, simultaneous RNAi knockdown of the two ORs significantly reduced the male EAG response to E2HE2H and abolished male attraction to this compound. These results were consistent with the sex expression profile, demonstrating the sex and functional differentiation between the two ORs. Taken together, this study characterizes the ORs responsible for chemical perception and the associated aggregation behaviors driven by these pheromones. Thus, this study enhances our understanding of olfactory signaling in a hemipteran insect and contributes to the knowledge required for improved pest management.

Nitrogen (N) leaching is a major pathway of N loss in subtropical crop production systems, contributing to groundwater pollution and thus posing serious threats to human health. However, the characteristics of annual N leaching in subtropical open-field vegetable systems and the effectiveness of integrative N fertilization management practices in reducing N leaching remain poorly understood. In this study, two plot-based field experiments were conducted with open-field Chinese cabbage-pepper rotation system in subtropical southwest China to quantify annual N leaching and evaluate the effectiveness of integrated N fertilization management practices. Experiment 1 compared five N fertilizer application rates using conventional urea, while Experiment 2 compared different N sources including conventional urea, organic fertilizer, nitrification inhibitor-based fertilizer, and controlled-release urea which were all applied at the optimized N rate. Results showed that the annual N leaching under farmers’ N practice (FNP) was 251 kg N ha⁻¹, with contributions of 55, 31, and 14% from the pepper season, Chinese cabbage season, and fallow period, respectively. Total N leaching increased exponentially with N rate. The seasonal N leaching factor was 32% for pepper and 17% for Chinese cabbage in the FNP treatment, respectively. Compared to FNP, optimizing N rate based on crop requirement and soil supply significantly reduced N leaching by 68% and gray water footprint by 66−75%, while improving N use efficiency (NUE) from 35% to 54%. In Experiment 2, mixing organic and inorganic fertilizers, applying nitrification inhibitor, and using controlled-release urea further reduced annual N leaching by 27, 54, and 25%, respectively, compared to conventional urea. These practices also improved crop yields by 2−11% and NUE by 10−13%, and lowered gray water footprint by 28−58%. In summary, integrative N stewardship practices, particularly use of nitrification inhibitors under optimized N rates, effectively reduced N leaching while achieving high NUE and vegetable yields, providing a promising strategy for sustainable subtropical vegetable production.

Tomato (Solanum lycopersicum L.) is an important vegetable crop worldwide. Throughout its growth cycle, tomato is susceptible to various abiotic stresses. Among these stresses, salt stress is one of the most detrimental abiotic factors to plant growth and development. In this study, we identified a WRKY transcription factor, SlWRKY42, which is induced by salt stress. We then characterized the function of SlWRKY42 in transgenic materials under salt stress and found that SlWRKY42 positively regulates salt tolerance in tomato. Transcriptome sequencing analysis revealed that genes involved in proline biosynthesis were significantly enriched in SlWRKY42-overexpressing (SlWRKY42 OE) plants. The proline biosynthesis genes (SlP5CS1 and SlP5CS2) and proline contents were significantly upregulated in SlWRKY42 OE lines. We discovered that the promoter of the proline biosynthesis gene SlP5CS1 contains a W-box element. Further yeast one-hybrid (Y1H), luciferase, electrophoretic mobility shift assay (EMSA), and chromatin immunoprecipitation quantitative polymerase chain reaction (ChIP-qPCR) assays verified that SlWRKY42 could specifically bind to the W-box element in the promoter of SlP5CS1 and activate its expression, thereby promoting proline biosynthesis. In summary, SlWRKY42 enhances salt tolerance in tomato by regulating the expression of SlP5CS1, thereby elucidating the molecular mechanism by which the SlWRKY42 transcription factor controls salt tolerance in tomato.

Soil organic carbon (SOC) plays a crucial role as a nutrient trigger and directly impacts soil health and agricultural productivity. In China, the Well-facilitated Farmland Construction (WFC) project is a comprehensive agricultural management strategy, changing the soil environment and then influencing the SOC dynamics. However, the long-term trajectory of SOC under the implementation of the WFC project remains unclear. To address this knowledge gap, this study focused on farmland in southeastern China that completed the WFC project in 2022. A total of 202 topsoil samples (0-20 cm) were collected from the regional paddy soil in 2023. Using digital soil mapping (DSM) and the CENTURY model, we delineated key soil properties and simulated the spatio-temporal changes of SOC density (SOCD). The results revealed that the SOCD ranged from 1.23 to 6.35 kg m^-2, with an average value of 3.68 kg m^-2 in 2023. Soil pH, clay, and sand content were primary factors influencing SOCD distribution. According to CENTURY model simulations, SOCD exhibited a declining trend from 2010 to 2021, while it was projected to increase from 2022 to 2030 following the WFC implementation, which could be attributed to enhancements in irrigation and straw incorporation. Besides, the scenario without WFC results shows that SOCD would decline from 2022 to 2030, underscoring the project’s effectiveness in preventing SOC loss for paddy soil. The spatial patterns of SOCD in 2021 and 2030 were similar, and the low-value areas showed faster increase rates than the areas with high SOCD levels, indicating that the specific field plots with lower SOCD levels could sequester more carbon with improved soil management. In conclusion, the WFC project can potentially increase SOC sequestration in the paddy soil and grain yield, ensuring food security and addressing climate change.

The integration of green manure (GM) crops into traditional cropping systems has regained attention for its potential to improve soil organic carbon (SOC) content in an environmentally sustainable way. However, the effects of carbon (C) input from different GM species on the SOC accumulation and recalcitrant C fractions across soil profile remain inadequately understood. This three-year North China Plain study assessed SOC changes and C fractions of easily oxidizable organic carbon (EOC) and recalcitrant organic carbon (ROC) in fallow, rye, rapeseed, and vetch systems, with δ¹³C analysis for GM-derived C fraction and microbial C-decomposition functional genes. Our results show that SOC was significantly increased by GMs. Rapeseed was the only species that improved SOC at 20-40 cm, the rapeseed-derived C contributed 2.48% of the SOC. Rye enhanced EOC and ROC at topsoil, rapeseed increased ROC at 20-60 cm, and vetch increased EOC at 40-60 cm. At the topsoil, the abundances of cellulose- and pectin-decomposition gene were increased in vetch and decreased in rye. At 20-40 cm, the pectin- and lignin-decomposition genes were markedly improved by rapeseed, while at 40-60 cm, the chitin-decomposition gene was increased in vetch, indicating the microbial promoting effects by deep roots of vetch and rapeseed. Our results suggest GM species influence SOC deposition depth and the recalcitrance of SOC decomposition, thereby affecting the distribution of SOC accumulation through microbial-driven C decomposition activities.

Intercropping is a promising cultivation strategy that enhances the sustainable use of water and land resources while contributing to national food and oil security. To improve the yield stability of soybeans in intercropping systems, there is an urgent need to develop a scientific and efficient framework for evaluating shade tolerance. In this study, we propose an integrated shade tolerance assessment method based on high-throughput phenotyping, multienvironment trials, and machine learning (ML) approaches. Utilizing multivariate analysis, we evaluated 202 soybean accessions and partitioned their performance under intercropping into two distinct capacities, namely, shade tolerance during the cogrowth stage and recovery ability during the independent growth stage, each of which was classified into five levels from weak to strong. Preliminary trait selection was performed through correlation analysis and broad-sense heritability estimation, followed by the application of six ML models to identify the key shade tolerance traits across different growth stages. The robustness and generalizability of the selected traits were validated in three environments—a field pot, an open field, and a greenhouse—using soybean varieties with known shade tolerance levels. The results revealed that three traits—the side canopy area (SCA), top canopy area at stage 3 (TCA3), and top-view mixed entropy (TME)—were strongly associated with shade-tolerant varieties. These traits presented two distinguishing features: significantly higher values under shaded conditions and greater increases during the recovery phase. The prediction models constructed with these three traits achieved strong performance, with coefficients of determination of R⊃2;=0.776 for shade tolerance and R⊃2;=0.959 for recovery ability. In summary, this study demonstrates the potential for integrating high-throughput phenotyping with ML to efficiently identify the key indicators of shade tolerance. By measuring only three indicators—SCA, TCA3, and TME—soybean shade tolerance at the seedling stage, recovery ability during later growth, and overall shade tolerance across the full growth period can be rapidly and accurately evaluated. This method offers a powerful and practical tool for implementing shade tolerance evaluations, gene discovery, and targeted breeding of soybean cultivars that are suitable for intercropping systems.

The Type A protein phosphatase 2C (PP2C-A) gene family is vital for regulating the ABA signaling pathway and plant stress responses. In this research, 14 SlPP2C-A genes were identified in the tomato genome, distributed across six chromosomes. Most SlPP2C-A genes contain cis-acting elements associated with growth, development, light, hormones, and stress responses. Collinearity analysis revealed high homology between the tomato and Arabidopsis PP2C-A gene families. Tissue-specific expression analysis indicated that SlPP2C7 is highly expressed in flowers, leaves, and mature fruits, and is significantly induced by saline-alkali stress. Gene-edited SlPP2C7 knock-out mutants subjected to saline-alkali stress confirmed that SlPP2C7 negatively regulates saline-alkali tolerance in tomato. Combined transcriptomic and metabolomic analyses showed that under saline-alkali stress, metabolic pathways such as flavonoid biosynthesis, isoflavonoid biosynthesis, flavone and flavonol biosynthesis, phenylpropanoid biosynthesis, and phenylalanine metabolism were significantly enriched. These outcomes imply that SlPP2C7 may enhance tolerance to saline-alkali stress through modulating flavonoid biosynthesis pathways. This research reveals comprehension of the physiological and molecular mechanism responsible for saline-alkali stress tolerance mediated by SlPP2C7 in tomato.

Floral scent is an important ornamental shape of garden plants. Monoterpenes in terpenoids are the main components of lily floral scents. 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) plays a role in the second enzymatic reaction of methylerythritol phosphate (MEP) pathway, which is responsible for monoterpene synthesis. However, the function of DXR gene in the floral monoterpene synthesis pathway of Lilium 'Sorbonne' remains unclear. In this study, the Lilium oriental 'Sorbonne' was used as the experimental material, and the differentially expressed LiDXR gene was selected using the early transcriptomic data. It was found that it had a high consistency with the rater rate process from synthesis to termination of floral substances in the flowering stage of lily. Therefore, the LiDXR gene was cloned and bioinformatics analyzed. A total of 472 amino acids are encoded. The expression of LiDXR gene was the highest at the Sorbonne half opening stage, and the expression of LiDXR gene in petals was significantly higher than that in other flower organs. The results of subcellular localization showed that LiDXR protein was localized in chloroplasts of leaf epidermal cells. A virus-induced gene silencing (VIGS) assay showed that silencing LiDXR can reduce monoterpene levels by down-regulating TPS gene expression downstream of the MEP pathway. Meanwhile, the results of HS-SPME-GC-MS showed that the total volatile terpene content of lily decreased significantly after silenced. The results of overexpressed plants A. thaliana and petunia showed that the transgenic plants had stronger growth potential and advanced flowering time. The GC-MS results of transgenic petunias showed that the content of volatile total terpenes in transgenic strains was 78% higher than that of wild type. Overexpression of LiDXR gene would affect the expression level of MEP pathway genes, and then affect the synthesis of terpenes including monoterpenes downstream of MEP pathway. The purpose of this study was to analyze the function of LiDXR gene and provide theoretical basis for floral breeding of lily and ornamental plants.

Chloroplasts are important organs for photosynthesis, which is essential for increasing the yields of pak choi. In this study, we evaluated a delayed chloroplast development mutant ‘M136’ during self-crossing of the pak choi inbred ‘136’. The newborn true leaves of ‘M136’ were yellow and gradually green with maturation. Chloroplast development, pigment contents and photosynthesis parameters were impaired and gradually recovered with growth in ‘M136’, and chlorophyll fluorescence parameters were also impaired in ‘M136’. Based on genetic analysis and bulk segregant analysis (BSA)-seq, the mutant phenotype was controlled by a single recessive gene, identified as BraA06g011520.3.5C (BrECB2), which encoded a DYW-type pentatricopeptide repeat (PPR) protein. In ‘M136’, a T-to-C single nucleotide polymorphism (SNP) in the 4th PPR motif of BrECB2 caused a Threonine-to-Isoleucine amino acid substitution. BrECB2 was mainly expressed in young leaves. The chloroplast RNA editing efficiency of ‘M136’ was affected and fully recovered after the leaf turned green, and the editing efficiency was partially restored in complementary lines. The plastid-encoded RNA polymerase activity was not affected in ‘M136’. Functional complementation analyses revealed that the transient overexpression of BrECB2 partially rescued the mutant phenotype and the RNA editing efficiency of ‘M136’. In summary, this study indicate that BrECB2 is involved in early chloroplast development and RNA editing, providing a theoretical basis for understanding the regulatory network involved in chloroplast development in pak choi.

African swine fever (ASF), caused by African swine fever virus (ASFV), is a highly contagious disease that has spread globally, posing a significant threat to swine production and international trade. As rapid diagnosis is crucial for controlling ASF, its major capsid protein, p72, has become a key target for diagnostic and vaccine development. In this study, we generated five monoclonal antibodies (mAbs) against the p72 protein by immunizing mice with inactivated virus. Using phage display technology, we identified the epitope for one mAb as a novel linear B-cell epitope within amino acids 130-152 of the p72 protein. Structural and homology analyses revealed that this epitope is highly conserved across diverse ASFV genotypes and is exposed on the surface of the p72 trimer. Importantly, the epitope showed strong reactivity with sera from ASFV-positive swine. These findings offer a foundation for creating improved serological diagnostics and designing epitope-based vaccines against ASFV.

Thirty-six walnut cultivars were analyzed for apparent, nutritional, processing and protein properties. Systematic cluster analysis (SCA) was applied to classify 36 walnut cultivars, while multivariate linear regression (MLR) analysis was used to develop a model for evaluating walnut protein solubility. The walnut cultivars were classified into three distinct clusters. Wen 185 and Xinguang had protein purity of 64.42, 70.57, and solubility of 27.04, 30.04%, respectively. Wen 185 and Xinguang were identified as the more suitable cultivars for extracting and processing soluble proteins. The MLR model revealed critical factors influencing protein solubility, such as arginine (Arg), glutamic acid (Glu), threonine (Thr), lysine (Lys), histidine (His), and crude fat. Glu (r=-0.64) and Arg (r=-0.57) showed a significant negative correlation with solubility. With a R² of 0.832 between predicted and experimental values, the model was validated. This study has improved the efficiency of walnut protein during the processing and pointed out the direction for the processing and utilization of different cultivars of walnuts.

Geminiviruses mainly infect economically important dicot plants and cause serious damages in agriculture. Here we report that the dicot plant Nicotiana benthamiana microtubule-associated E3 ligase (MEL) plays a dual role in regulating geminivirus infection in N. benthamiana. On the one hand, NbMEL functioned as a defense factor to mediate resistance against geminiviruses with single-stranded, circular DNA genomes by promoting the degradation of plant immune negative regulator. On the other hand, NbMEL could specifically recognize geminivirus-encoded V2 protein, a viral gene silencing suppressor and effector, for polyubiquitination and degradation to suppress geminivirus infection. These findings provide a fundamental basis for utilizing MEL to generate crop for broad-spectrum resistance in dicot plants.

目的：近年来，因Ⅱ相鞭毛基因被耐药基因或噬菌体序列替代而产生的沙门菌单相变异株不断出现并流行。碳青霉烯耐药株的出现对公共卫生安全构成严重威胁。本研究旨在解析一株NDM-13阳性印第安那沙门菌单相变异体的基因组特征、耐药谱、遗传演化特点及产生机制。

方法：采用微量肉汤稀释法或琼脂稀释法检测菌株对14种抗菌药物的敏感性；同时利用传统玻片凝集法进行血清学分型，并结合Salmonella In Silico Typing Resource（SISTR）对基因组进行血清型预测。随后，应用多位点序列分型（MLST）和核心基因组MLST（cgMLST）解析其系统发育特征；通过比较基因组学揭示前噬菌体整合与Ⅱ相鞭毛基因缺失的机制；并对所携带质粒的耐药基因组成和结构进行系统解析，结合接合实验评估其水平传播潜力。

结果：血清学鉴定结合MLST和cgMLST分析结果证实，该菌株为印第安那沙门菌的单相变异体。比较基因组学进一步揭示，其Ⅱ相鞭毛基因簇fljAB及邻近七个基因被来源于S. Mbanada的一个8.9 kb截短前噬菌体（ΔΦSM）取代。ΦSM介导的不精确切除导致了fljAB基因簇及周边序列丢失，从而驱动了单相变异体的形成。耐药性分析表明，该菌株携带一条IncHI2质粒，包含两个多重耐药区（MRRs）。除常见的bla_TEM-1B、bla_CTX-M-65、strA、strB和sul2外，质粒中还存在一段长达51.3 kb的额外MRR，插入于dam基因，携带aph(3’)-IIa、aadA22、bla_NDM-13、mph(A)和erm(B)，赋予菌株对卡那霉素、链霉素、碳青霉烯类、阿奇霉素和林可霉素的耐药性。但由于IS26介导的转座导致接合区缺失，该质粒在实验条件下未能实现接合转移。

结论：本研究阐明了前噬菌体不精确切除介导NDM-13阳性印地安纳单相变异体产生的分子机制，并揭示了其携带的复杂耐药质粒结构。结果表明，前噬菌体与可移动遗传元件的协同作用可以推动耐药性沙门菌单相变异体产生与演化。

创新性：本研究首次证实前噬菌体不精确切除在NDM-13阳性印第安纳单相变异体形成中的关键作用，并揭示了其独特的耐药质粒进化特征。这不仅拓展了对单相型沙门菌分子进化机制的认识，也为基于全基因组测序的耐药性监测与食源性病原菌预警提供了新的理论依据和实践思路。

Microorganisms carrying cbbL, pmoA and coxL genes play crucial roles in regulating soil-atmosphere exchanges of carbon trace gases (CO₂, CH_4, and CO). However, the geographical distribution patterns of these functional genes in agricultural ecosystems and their environmental drivers remain poorly understood. Here, we surveyed agricultural soils across four climate zones (tropical, subtropical, warm temperate, and mid-temperate) in eastern China to quantify the abundances of CO₂-assimilating bacteria (cbbL gene), methanotrophs (pmoA gene), and CO-oxidizing bacteria (coxL gene). We found significant ecosystem-specific patterns: the cbbL gene was more abundant in upland soils (averaging 9.46×10⁹ copies g^-1) than in paddy soils (6.44×10⁹ copies g^-1). In contrast, methanotrophs abundance was 1 to 3 orders of magnitude higher in paddy (averaging 1.17×10⁸ copies g^-1) than in upland (5.78×10⁶ copies g^-1) soils. The coxL gene maintained similar abundance levels across both soil types (averaging 6.12×10⁸ vs. 5.91×10⁸ copies g^-1). Structural equation models revealed that spatial factors primarily shaped cbbL and pmoA in uplands, whereas total bacterial abundance was the dominant predictor for all three genes in paddy soils. These results highlight distinct ecological controls on microbial functional groups and provide a predictive framework for how land use and climate change may regulate microbial mediation of carbon gas fluxes across a continental-scale transect in eastern China.

African swine fever (ASF) represents a highly contagious and fatal condition affecting both domestic and wild pigs, necessitating mandatory reporting status as designated by the World Organisation for Animal Health (WOAH). Currently, the primary strategy for preventing and controlling ASF revolves around early detection and stringent culling practices. However, the swift dissemination of ASF in both newly affected and previously impacted countries and regions underscores the absence of efficient measures to effectively curb the disease. To address this threat, a diverse array of methodologies is being employed globally in the pursuit of developing vaccines to combat ASF. In this context, we delve into the advancements achieved in ASF vaccine research over the past decade, encompassing the challenges and prospects associated with attenuated vaccines, subunit/live vector vaccines, and more. A profound comprehension of the virus's genetic diversity, pathogenic mechanisms, as well as the strengths and weaknesses of vaccine-induced immune protection, will pave the way for the development of novel vaccines in the future.

Loquat (Eriobotrya japonica.) is an evergreen fruit tree native to China, with a flowering period that typically occurs in winter (October to January), making it vulnerable to low-temperature stress during critical reproductive stages. However, the molecular mechanisms underlying cold tolerance in loquat remain largely unclear. In this study, transcriptome data from multiple loquat cultivars were analyzed using Weighted Gene Co-expression Network Analysis (WGCNA), identifying two gene modules (brown and turquoise module) highly associated with cold treatment. Among the cold-responsive candidates, the Rab5 family GTPase EjRabF2b was consistently upregulated under low-temperature conditions. Functional validation revealed that overexpression of EjRabF2b in Arabidopsis thaliana and tomato significantly enhanced cold tolerance, while its silencing in loquat compromised stress resistance. Mechanistically, EjRabF2b contributed to maintaining cell membrane integrity and enhancing antioxidant enzyme activity. Promoter analysis and interaction assays further confirmed that the C2H2-type transcription factor EjZAT10 directly binds to the EjRabF2b promoter and activates its transcription under cold stress. Collectively, this study uncovers a regulatory module composed of EjZAT10 and EjRabF2b that participates in loquat cold adaptation through vesicle-mediated antioxidant defense and membrane protection, offering a theoretical foundation and potential targets for the molecular breeding of cold-tolerant cultivars.

Aspergillus species are ubiquitous fungi that produce mycotoxins (secondary metabolites) known as sterigmatocystin, aflatoxin, ochratoxin A, and cyclopiazonic acid in many different kinds of foods, which leads to serious contamination in agricultural products thereby endangering human health. With the rapid advancement of molecular biology technology, extensive studies on Aspergillus fungi have been conducted on growth and development, mycotoxin biosynthesis, and their interactions with environment. Here, we summarized a series of functional genes of the main Aspergillus fungi relative to toxins occurrence in foods, which revealed the signal transduction mechanisms of their involvement in growth and development, toxin production, and response to environmental changes, anticipating providing theoretical guidance on developing control and prevention technologies for mycotoxin contamination in agricultural products to ensure food safety.

Medullary bone is a labile calcium store for eggshell deposition. The dysfunctional mineralization function of medullary bone in aged laying hens leads to decreased eggshell mineralization and increased egg breakage. Understanding the mechanisms underlying age-related decline in medullary bone mineralization function is critical for developing novel strategies to improve eggshell quality in aged laying hens. Hy-Line Brown laying hens were sampled at different ages (48, 61, 74, and 87 wk of age; n=12 at each age). Analyses were conducted to determine the effect of aging on eggshell mineralization, medullary bone remodeling, and physiological and molecular biological parameters for bone marrow microenvironment senescence. The results showed that: (1) compared with 48- and 61-wk-old la`ying hens, 74- and 87-wk-old laying hens had decreased (P<0.05) eggshell quality; (2) micro computed tomography scans illustrated a progressive decrease (P<0.05) with age in femoral mineralization; (3) compared with 48-wk-old laying hens, 61-, 74- and 87-wk-old laying hens had decreased (P<0.05) osteoblastic function, characterized by decreased serum alkaline phosphatase levels and decreased protein expression of osteogenic markers (runt-related transcription factor 2, osteopontin and alkaline phosphatase) of femoral medullary bone; (4) compared with 48- and 74-wk-old laying hens, 87-wk-old laying hens had increased (P<0.05) protein expression of osteoclastic marker (tartrate resistant acid phosphatase) of femoral medullary bone; (5) compared with other ages, 87-wk-old laying hens had increased (P<0.05) mRNA expression of senescence-associated secretory phenotype in the femoral marrow microenvironment; (6) compared with 48- and 61-wk-old laying hens, 74- and 87-wk-old laying hens had increased (P<0.05) protein expression of senescence biomarkers (gamma H2A.X, galactosidase beta 1 and tumor suppressor protein 53) in the femoral marrow microenvironment; (7) compared with 48-wk-old laying hens, 61-, 74- and 87-wk-old laying hens had increased (P<0.05) femoral adipose deposition. Age-related decline in mineralization function of medullary bone was accompanied by an uncoupling of medullary bone osteoblastic and osteoclastic functions, which may link to a senescent femoral marrow microenvironment in aged laying hens. Methods are needed to monitor and manage the bone marrow microenvironment in the later laying periods.

The effect of adding hydroxycinnamic acids (caffeic acid, sinapic acid, p-coumaric acid and chlorogenic acid) in Cabernet Sauvignon dry red wine before and after fermentation was investigated, taking into account the color, anthocyanins and other polyphenols in the wine samples. The copigmentation effect of malvidin-3-O-glucoside and sinapic acid was further explored in model solution and through theoretical calculations. The results indicated that the addition of hydroxycinnamic acids significantly enhanced the wine's color with sinapic acid (before the fermentation) showing the most pronounced color protection effect. Furthermore, the content of total phenols and total anthocyanins increased by 36% and 28%, respectively. Thermodynamic analysis revealed that the interaction between sinapic acid and malvidin-3-O-glucoside was spontaneous and exothermic. Theoretical studies identified hydrogen bonding and dispersion forces as the main contributors to binding, with the carboxyl group of sinapic acid playing a critical role, while the anthocyanin backbone also influenced the interaction.

Cropland parcels are the basic unit for agricultural production, and their size and shape may change due to human activities, e.g. land consolidation. Remote sensing has been increasingly used for mapping cropland parcel, yet detecting changes in cropland parcels by wall-to-wall mapping is time-consuming. This paper proposes a new algorithm to identify whether and where cropland parcel changes have been undertaken without generating complete parcel maps. We use the number of edge pixels derived from remote sensing imagery as a proxy indicator for cropland parcel changes. First, we apply a Sobel operator to delineate the total edge pixels of parcels from dual-time images. Second, we apply the connected-components labeling to remove pseudo-edges arising from non-cropland built structures and transmission towers. We then perform topological optimization, including morphological dilation and skeleton extraction, to eliminate redundant edge pixels for parcel structure. Finally, we detect whether parcel changes have been undertaken by counting and comparing the number of edge pixels derived from dual-time images. We applied this innovative framework in five regions in East Asia where land consolidation has significantly changed cropland parcels. Our method demonstrated robust detection results, with stable accuracy, precision, recall, and F1-score, all exceeding 0.85. Screening redundant edge pixels reduced noise and permitted efficient detection of changes in cropland parcels. Our method extends the traditional detection of semantic change to structural change and can quickly detect cropland parcel changes with high accuracy. This capability offers the potential to identify hotspot areas of cropland changes on a larger scale without the need to produce full cropland maps, which is particularly useful for monitoring land consolidation programs.

A novel variegated tea cultivar exhibiting a stable variegated phenotype was recently identified, demonstrating significantly elevated amino acid content concomitantly with reduced polyphenolic compound levels compared to conventional green-leaf varieties. Nevertheless, the underlying mechanism remains unclear. Here, variegated leaves and normal leaves of ‘Huangshanzhong’ tea plant were used to perform pigment content analysis and comparative transcriptome analysis. The chlorophyll content in variegated leaves significantly decreased compared to normal leaves, while the ratio of Chl a to Chl b was enhanced. Multiple genes (CsrpiA, CsGAPDH, CsgpmI, CsPK and CsOGDH) involved in sugar metabolism exhibited downregulated expression in variegated leaves. Key genes involved in the photosynthetic pathway were down-regulated in variegated leaves, such as light-harvesting protein complex chlorophyll a/b binding proteins (CsLhca1, CsLhca4, CsLhcb1 and CsLhcb3) and photosystem II complex proteins (CspsbP and CspsbW). Meanwhile, genes involved in chlorophyll degradation metabolism (CsSGR, CsCLH1) were upregulated in variegated leaves. Compared to the wild type, transgenic plants of CsCLH1 and CsCLH2 exhibited no significant changes in chlorophyll content. Enzyme activity assays showed that CsCLH1 could degrade chlorophyll in vitro. Subcellular localization results revealed that CsCLH1 and CsCLH2 were localized in the cytoplasm and nucleus. These findings suggest that impaired photosynthetic system function, suppressed carbohydrate synthesis, and accelerated degradation of photosynthetic pigments collectively contribute to the variegated phenotype in tea leaves. This study advances our understanding of mechanisms underlying plant leaf variegation.

Anthocyanins play a crucial role in plant growth, development, reproduction, and stress response. Additionally, anthocyanins enhance the quality of fruits and vegetables due to their antioxidant properties. While numerous previous research has been conducted on anthocyanins, limited information exists regarding their composition and the role of the anthocyanin pathway gene DFR (dihydroflavonol 4-reductase) in chili pepper leaves. In this study, we used a purple leaf pepper cultivar H18 with anthocyanins on the leaves decreasing as they grow and develop. Targeted anthocyanin metabolite assays revealed that the contents of delphinidin, malvidin, and petunidin derivatives followed the same trend as the overall anthocyanin content, with delphinidin derivatives being the predominant component of H18 pepper leaves. Transcriptome sequencing was performed on H18 leaves at four different stages. The results showed that DEGs at various stages were primarily associated with biological processes and flavonoid metabolic pathways. Through evolutionary tree and expression analysis, three candidate genes involved in DFR function were identified. Substrate catalysis assays of CaDFRs demonstrated that only CaDFR1 was active, catalyzing DHQ, DHM, and DHK. VIGS-mediated silencing of CaDFR1 resullted in a significant decrease reduction in anthocyanin levels in H18 pepper leaves and stems and along with a decreased reduction in the expression levels of other candidate functional genes in the anthocyanin metabolic pathway. This study identifies the key anthocyanin components in the leaves of H18 peppers and validates the function of CaDFR1, providing a theoretical foundation for modifying anthocyanin content in pepper plants through molecular breeding.

Triticeae represents one of the most significant sources of cereal crops in Poaceae, including wheat, barley, and rye. Global annual production reaches 900 million tons, constituting 30% of total grain production. The utilization of wild relatives is crucial for enhancing crop resilience. Sea barley (Hordeum marinum Huds), a wild relative species of wheat and barley, demonstrates exceptional salt/waterlogging tolerance and other valuable traits. Moreover, it exhibits partial cross-compatibility with common wheat. Sea barley has emerged as an essential donor of elite genes for crop breeding, with potential applications both as a de novo domesticated crop and as forage cultivated in saline-alkali soils and waterlogged areas. This review synthesizes current knowledge regarding sea barley, emphasizing its origin, evolution, genome characteristics, genetic transformation, mechanisms of stress tolerance, fungal resistance, and cross-compatibility with wheat. Additionally, we identify key knowledge gaps and future research directions to enhance its utilization for crop breeding and novel crop development, aiming to transform sea barley from an underutilized wild grass into a genetic resource for climate-smart agriculture.

Soil salinization severely impairs mungbean (Vigna radiata (L.) Wilczek) seedling uniformity and productivity. In this study, genome-wide association study (GWAS) was conducted using a natural population of 374 mungbean accessions and 4,875,143 SNPs. By evaluating the population under two independent environments and applying two statistical models, we identified a significant SNP (Chr01_26769549) associated with relative germination traits under salt stress. Based on this locus, a Kompetitive Allele-Specific PCR (KASP) marker was successfully developed for marker assisted selection. Integrated haplotype and expression analyses confirmed polygalacturonase gene VrPG1 as a key candidate gene regulating salt tolerance during seed germination. Two haplotypes of VrPG1 (Hap1/Hap2) were identified, with a mutation in the Hap1 promoter region enhancing its transcriptional activity. Overexpression of VrPG1 in Arabidopsis thaliana significantly increased germination rates under salt stress by promoting endosperm cell wall softening. Salt-tolerant mungbean varieties exhibit higher polygalacturonase activity and earlier loosening of thin-walled cell walls during the germination period, which promotes seed imbibition and radicle emergence. Collectively, these findings demonstrate that VrPG1 enhances salt tolerance during germination through cell wall remodeling. This study provides novel genetic targets and efficient marker-assisted selection tools for breeding salt-tolerant mungbean. This study provides novel genetic targets and efficient marker-assisted selection tools for breeding salt-tolerant mungbean varieties.

The rapid and accurate identification of functional major blast-resistance genes represents a crucial and essential step in rice blast-resistance breeding. This study focused on five major blast-resistance genes at the Piz/Pi9 locus: Piz, Pi2, Pi9, Pizt, and Pigm. Molecular markers were developed for each gene, and one Pyricularia oryzae differential strain containing only the avirulence gene Avr-Pizt was identified through screening. This screening utilized a set of rice monogenic lines containing 24 major blast-resistance genes and a landrace Gumei 4 containing the major blast-resistance gene Pigm. Analysis of 193 rice varieties from Heilongjiang province using the molecular markers identified 42 varieties containing Pizt and 54 ones containing Piz. Subsequently, using the differential strain of Avr-Pizt, 29 varieties, including Longgeng 31, Longgeng 3013, and Longgeng 1614, were confirmed to contain functional Pizt. This research establishes an approach that combines molecular markers and P. oryzae differential strains for efficient and precise identification of functional major blast-resistance genes in rice.

Low phosphorus (LP) stress induces tissue-specific anthocyanin biosynthesis and sugar accumulation in plants. However, the relationship between sugar levels and phosphate (Pi) availability in regulating anthocyanin remains unclear. This study investigated the spatiotemporal patterns of sugar accumulation and anthocyanin biosynthesis in maize seedlings, and conducted experiments modifying sugar status to examine the significance of sugar accumulation for LP-induced anthocyanin biosynthesis. The results demonstrated that, under LP conditions, anthocyanin biosynthesis and sucrose accumulation were spatially and temporally coupled, with leaf sheaths exhibiting the lowest Pi content and highest sucrose and anthocyanin levels. Artificially increasing endogenous sucrose through cold-girdling promoted anthocyanin biosynthesis, whereas reducing sucrose via leaf-shading inhibited it. Analysis revealed a significant positive correlation between sucrose and anthocyanin levels. In vitro incubation of leaves and sheaths with different sugars further confirmed that sucrose accumulation was indispensable for LP-induced anthocyanin biosynthesis. Therefore, the temporal and spatial patterns of anthocyanin biosynthesis under LP are determined by both tissue Pi levels and sucrose accumulation, and anthocyanin distribution can be modulated by altering Pi and sucrose patterns. Transcriptome analysis of LP-treated leaf sheaths, with or without sucrose accumulation, suggested that PHR1 may mediate the interaction between sugar and LP signaling pathways in regulating anthocyanin biosynthesis. These insights elucidate the mechanisms governing tissue-specific anthocyanin biosynthesis under LP conditions, while providing potential targets for improving phosphorus use efficiency via anthocyanin regulation.

Cotton leaves are fundamental components for cotton growth and serve vital roles in photosynthesis and transpiration. The completion of point cloud data on cotton leaf morphology is critically important for examining the interaction between morphological parameters and the environment. Previous methods have shown effective performance in capturing objects with regular shapes and continuous surfaces, particularly for industrially produced 3D-modeled objects. However, these techniques demonstrate limitations in processing plants with diverse morphological structures. This study proposes PCompNet (a segmentation and improved completion network) for cotton leaf point cloud completion, reconstructing complete geometries from whole plants with diverse shapes and discontinuous surfaces through morphological part segmentation technique with deep hierarchical point-set feature learning. Additionally, a unified loss function was implemented to effectively penalize the average distance discrepancy between patch centers and their nearest neighbors in PF-Net, preventing the generated missing point clouds of cotton leaves from excessive concentration. The experimental results demonstrated that PCompNet achieved substantial reductions in Chamfer distance (CD) on the Cotton3D dataset compared to PMP-Net, GRNet, SnowfakeNet, FoldingNet, and PF-Net, with reductions of 95.46, 98.45, 97.46, 100.00, and 84.93%, respectively. Moreover, PCompNet accurately completed missing regions at different scales while maintaining the geometry of the input point cloud. Even with 75% of data missing, the CD value remained at 0.115. These results demonstrate the effectiveness and robustness of PCompNet in completing point cloud data for cotton leaves, indicating its potential for applications in cotton growth and environmental studies.

Lime application represents an established approach for ameliorating soil acidity, and understanding its effects on the interactions between aluminum (Al) and iron (Fe) oxides and soil organic carbon (SOC) fractions is essential for promoting sustainable agricultural practices that enhance carbon sequestration. This investigation examined the interactions among Al and Fe oxides and SOC fractions under long-term fertilization and liming. A long-term field experiment was implemented with five treatments: CK (no fertilizer), N (nitrogen fertilizer), NCa (N plus lime), NPK (nitrogen, phosphorus, and potassium fertilizer), and NPKCa (NPK plus lime). Soil samples were obtained from three depths: 0–10, 10–20, and 20–30 cm. The findings revealed that lime application increased SOC by 20.84% under the N treatment but decreased SOC by 9.97% under NPK, compared with CK. At the 0–10 cm depth, dissolved organic carbon (DOC) was substantially higher under NCa (410.51 mg kg^-1) and NPKCa (372.83 mg kg^-1) compared with CK. Particulate organic carbon (POC) and mineral-associated organic carbon (MAOC) demonstrated consistent enhancement under NPK and NPKCa across all soil depths compared with CK. DOC exhibited significant positive correlations with both aluminum (Ald), reactive aluminum (Alo) and aluminum (Alp), indicating a key role of organically bound and reactive Al in carbon dynamics. Compared to the CK treatment, SOC stock increased significantly by 43.49% under NPK and by 36.82% under NPKCa. Structural equation modeling demonstrated that lime application mitigated the negative effects of free Al (Ald) on carbon sequestration, while Fe oxides (Fed) contributed positively to SOC stabilization. DOC showed no significant impact on carbon sequestration rate (CSR), while easily oxidizable carbon (EOC) negatively affected CSR directly. These results highlight the crucial role of lime in improving acidic soil conditions and enhancing the stability and sequestration of soil organic carbon.

Lime application represents an established approach for ameliorating soil acidity, and understanding its effects on the interactions between aluminum (Al) and iron (Fe) oxides and soil organic carbon (SOC) fractions is essential for promoting sustainable agricultural practices that enhance carbon sequestration. This investigation examined the interactions among Al and Fe oxides and SOC fractions under long-term fertilization and liming. A long-term field experiment was implemented with five treatments: CK (no fertilizer), N (nitrogen fertilizer), NCa (N plus lime), NPK (nitrogen, phosphorus, and potassium fertilizer), and NPKCa (NPK plus lime). Soil samples were obtained from three depths: 0–10, 10–20, and 20–30 cm. The findings revealed that lime application increased SOC by 20.84% under the N treatment but decreased SOC by 9.97% under NPK, compared with CK. At the 0–10 cm depth, dissolved organic carbon (DOC) was substantially higher under NCa (410.51 mg kg^-1) and NPKCa (372.83 mg kg^-1) compared with CK. Particulate organic carbon (POC) and mineral-associated organic carbon (MAOC) demonstrated consistent enhancement under NPK and NPKCa across all soil depths compared with CK. DOC exhibited significant positive correlations with both aluminum (Ald), reactive aluminum (Alo) and aluminum (Alp), indicating a key role of organically bound and reactive Al in carbon dynamics. Compared to the CK treatment, SOC stock increased significantly by 43.49% under NPK and by 36.82% under NPKCa. Structural equation modeling demonstrated that lime application mitigated the negative effects of free Al (Ald) on carbon sequestration, while Fe oxides (Fed) contributed positively to SOC stabilization. DOC showed no significant impact on carbon sequestration rate (CSR), while easily oxidizable carbon (EOC) negatively affected CSR directly. These results highlight the crucial role of lime in improving acidic soil conditions and enhancing the stability and sequestration of soil organic carbon.

Soybeans, a crucial grain and oil crop, are valued for their high protein and oil content.  Soil salinization presents a significant abiotic stress that negatively impacts soybean growth and development, leading to reduced yield and quality.  The germination period represents a critical phase in soybean development.  This study evaluated salt tolerance in 165 soybean mutant lines during germination, resulting in the identification of five elite salt-tolerant germplasm resources.  Multi-environment Genome-wide association studies (GWASs) identified 11 significantly associated and 44 suggestive SNPs, alongside five novel QTLs linked to salt tolerance.  Analysis of candidate regions qtl5-1 and qtl5-2 identified Glyma.05G097200 and Glyma.05G240200 as promising candidate genes, exhibiting distinct expression patterns between salt-tolerant and salt-sensitive genotypes. Functional characterization in Arabidopsis demonstrated that overexpression of the soybean gene GmMACPF1 induced salt sensitivity, while the macpf1 mutant of Arabidopsis displayed enhanced salt tolerance.  Additionally, GmMACPF1 underwent selection during soybean domestication, with haplotypes Hap1 and Hap3 conferring improved salt tolerance.  These results indicate that GmMACPF1 functions as a negative regulator of salt tolerance during germination, offering novel insights into the molecular mechanisms governing soybean response to salt stress during this crucial developmental stage.

Microbial necromass carbon (MNC) serves a crucial function in the formation and stabilization of soil organic carbon (SOC). Although biochar amendment is recognized as a promising approach for enhancing SOC sequestration, its impact on MNC accumulation across the paddy soil profile remains uncertain. Through a 4-year field experiment, this study examined the effect of biochar amendment on MNC accumulation across three soil layers (0–15, 15–30, and 30–45 cm) in a paddy soil profile by combining vertical soil profiling, microbial community dynamics, and biomarker analysis. The results showed that biochar amendment reduced MNC by 10.5% (0–15 cm), 7.5% (15–30 cm), and 9.6% (30–45 cm), respectively, compared to the unamended control. In the topsoil (0–15 cm), the reduction in MNC under biochar amendment was attributed to decreases in both fungal and bacterial necromass carbon (C), whereas in the subsoil (15–45 cm), it primarily resulted from the decrease in bacterial necromass C. Biochar amendment reduced MNC content by decreasing microbial biomass and increasing nitrogen (N) acquisition enzyme activities, mainly due to a shift in the microbial community toward K-strategists and intensified microbial N limitation. This study provides novel insights into the microbially-mediated SOC dynamics in response to biochar amendment.