Protein phosphatase type-2Cs (PP2Cs) are widely involved in regulating plant growth and development, cell division, and, importantly, responses to abiotic stresses through reversible protein phosphorylation. We investigated the PP2C gene family in alfalfa through genome-wide identification and expression profiling analysis. Overall, 104 MsPP2C members identified in the alfalfa genome were classified into 13 subfamilies. Phylogenetic relationships, chromosomal distributions, duplication events, gene structures, and conserved motifs of MsPP2Cs were systematically analyzed. Additionally, transcriptomic and real-time quantitative PCR analyses revealed 14 MsPP2C genes were significantly differentially expressed in alfalfa under alkali stress. Among these, MsPP2C8, MsPP2C38, MsPP2C60, MsPP2C62, MsPP2C102,and MsPP2C103 (subfamily A) were rapidly and markedly upregulated in response to alkali stress. KEGG enrichment analysis revealed these genes were involved in plant hormone signal transduction and MAPK signaling pathway. MsPP2C38 had a core regulatory role in a predicted protein-interaction network, and interacted strongly with the proteins WRKY40 and DREB2A. Subcellular localization assays indicated MsPP2C8, MsPP2C38, MsPP2C62, and MsPP2C103 to be localized in the nucleus. These findings improve our understanding of the PP2C family, and clarify the critical regulatory roles of subfamily A members in mediating salt–alkali stress responses and tolerance in alfalfa.

Sweet osmanthus (Osmanthus fragrans), an evergreen shrub valued for its ornamental and aromatic qualities, is widely used in landscaping and the perfume industry. However, its sensitivity to cold stress limits its cultivation to specific climatic regions, constraining both its horticultural uses and the full exploitation of its genetic resources. In this research, we conducted an identification and examination of the members of RING-type E3 ubiquitin ligase found in sweet osmanthus, discovering a total of 705 members and categorizing them into seven unique RING subtypes. We screened 96 candidate cold stress-responsive genes, of which members of the RING-HC subfamily accounted for the greatest proportion (45/96), suggesting their more significant functional roles under cold stress. Expression profiling and quantitative reverse qRT-PCR analyses revealed that the RING-HC gene OfRNF182-like, was the member whose expression was most strongly induced under cold stress. Using biochemical assays and functional analysis based on transient transformation in sweet osmanthus callus, we demonstrated that OfRNF182-like overexpression reduced cold tolerance, as evidenced by attenuated antioxidant capacity, aggravated oxidative stress, excessive ROS accumulation and OfCBF1, OfDREB2A, OfPOD expression down regulation. Conversely, silencing of OfRNF182-like expression in callus tissue enhanced antioxidant activity and upregulated expression of cold-responsive genes. Interestingly, we found the OfRNF182-like can interact with the cold stress-positive regulator protein OfPP2-B10-like, promoting its degradation under cold stress. Our findings revealed that OfRNF182-like was an important gene that mediated cold tolerance through ROS homeostasis, a previously unrecognized regulatory mechanism, thus identifying it as a potential gene target for breeding cold-tolerant crops.

Bud dormancy is a key adaptive strategy in perennial plants, enabling them to survive in adverse environmental conditions. However, it generates challenges in crop cultivation, especially in fruit crops like apple, where synchronized bud break is crucial for consistent growth and yield. The synthetic cytokinin 6-benzylaminopurine (6-BA) promotes dormancy release, but its molecular and metabolic mechanisms remain poorly understood. This study investigates dormancy release in Xianheng 01 apple rootstock nursery plants through integrated transcriptomic, metabolomics, and hormonal analyses. Dormant buds were treated with 6-BA, followed by morphological, biochemical, and molecular profiling conducted over 30 days. 6-BA treatment increased plant height and leaf emergence by increasing the level of endogenous hormones such as cytokinins (DHZR/IPA) and decrease in level of abscisic acid (ABA). Transcriptomics analysis identified 7,009 differentially expressed genes (DEGs) in response to 6-BA treatment. The cytokinin-responsive gene A-ARR8 exhibited a distinct expression pattern, remained upregulated at 1, 3 and 6 d post-treatment but downregulated at 11 d. In contrast ABA related genes SnRK2a/b, PP2C and ABF3 were consistently downregulated throughout the treatment period. Metabolomics analysis identified 2,053 metabolites, showing early-phase dominance of phenylpropanoids and flavonoids, followed by a shift towards ABC transporter-mediated nutrient mobilization. Conjoint analysis highlighted coordinated activation of secondary metabolite biosynthesis and cytokinin signaling. These results demonstrate that 6-BA induces dormancy release through cytokinin-ABA antagonism and phased metabolic reprogramming from stress protection to growth promotion. Our findings provide a comprehensive framework for optimizing dormancy management in apple cultivation and highlight 6-BA as an effective agrochemical for enhancing temperate fruit production.

2026年5月，大西洋邮轮暴发安第斯病毒疫情，多国出现关联输入病例，引发全球公共卫生关注。中国农科院上海兽医研究所联合多家高校研究团队指出，农业与野生动物养殖环境是汉坦病毒传播最被忽视的核心热点，呼吁采用One Health策略破解相关公共卫生难题。

汉坦病毒为经啮齿动物排泄物传播的人兽共患病毒，可引发高致死率的汉坦病毒肺综合征及肾综合征出血热。中国是全球汉坦病毒防控主战场，承担全球70%-90%的肾综合征出血热病例负担。目前我国虽发病率已显著下降，但农民群体始终占总病例的大多数，且近十年无明显变化，病例高峰与春秋农耕周期高度重合。同时，国外文献显示野生动物养殖业成为新的高风险领域，相关调查表明养殖户血清阳性率较高，养殖环境本身是独立风险因素。

研究团队强调汉坦病毒并非区域性公共卫生问题，其跨地域传播风险已在本次邮轮疫情中得到显现。据此，研究团队建议亚太地区三大防控优先方向：将汉坦病毒纳入兽医继续教育和农场生物安全指南；建立野生动物养殖的专项监管体系；依托中国成熟的监测网络和疫苗技术，引领区域职业风险防控工作。

此次全球对汉坦病毒的关注，为补齐农业领域公共卫生短板提供了关键窗口期，唯有打通人、动物与环境健康的防控壁垒，才能从根源上消除这一隐藏但危害严重的健康风险。

Leaf color is one of the most important agronomic traits of Chinese cabbage(Brassica rapa L. ssp. Pekinensis). However, the molecular mechanism underlying leaf color development in Chinese cabbage remains largely unexplored. In this study, a juvenile leaves yellowing (jly) mutant was screened from an EMS-mutagenized mutant library of Chinese cabbage A03. The jly mutant displayed reduced chlorophyll and starch content, impaired photosynthetic efficiency, accumulated reactive oxygen species (ROS) in yellowed juvenile leaves, and abnormal chloroplast development. Using MutMap and KASP technologies, the gene BrRNE was identified as regulating the juvenile leaves yellowing phenotype in Chinese cabbage. BrRNE was preferentially expressed in leaves, and BrRNE was localized in chloroplasts. VIGS validation confirmed a significant association between the BrRNE gene and the leaves yellowing phenotype. Mutation of the BrRNE induced alternative splicing variants. Western blot analysis demonstrated that the BrRNE mutation impaired the expression of photosynthesis-related proteins in the jly mutant. Transcriptome analysis revealed significant activation of the retrograde signaling in the jly mutant, accompanied by marked upregulation of the chloroplast quality control system. Expression of genes involved in chlorophyll metabolism, sugar metabolism, and photosynthesis-related pathways was suppressed. Additionally, miRNAs involved in regulating the expression of genes in the chlorophyll metabolism and sugar metabolism pathways within the forward signal pathway were identified, including bra-miR824-p3, mtr-miR171b, bra-miR390-3p, and ath-miR158a-3p. This study identifies the gene BrRNE as a regulator of the juvenile leaves yellowing phenotype and constructs a comprehensive miRNA-mRNA co-regulation network for plant leaf color development, viewing the process of leaves color development from the perspective of intracellular signaling exchange.

牛病毒性腹泻（BVD）是由牛病毒性腹泻病毒（BVDV）引起的一种急性、高传染性疾病，对全球养牛业构成严重威胁。由于持续性感染及新型病毒株的不断出现，开发高效的抗病毒药物成为控制BVDV感染的迫切需求。牛干扰素（BoIFNs）是一类具有抗病毒和免疫调节功能的多活性糖蛋白。其中BoIFNα主要发挥抗病毒作用，BoIFNγ发挥免疫调节作用，为实现BoIFNα与BoIFNγ的协同效应，本研究通过原核表达系统将二者进行串联融合表达，利用镍柱亲和层析纯化重组融合蛋白（rBoIFNα/γ）。体外分别通过MTT法、细胞病变抑制法、qPCR等方法对rBoIFNα/γ的细胞毒性、抗病毒活性以及免疫调节活性进行测定。同时，为进一步提升rBoIFNα/γ的体内稳定性和递送效率，采用离子凝胶技术将rBoIFNα/γ封装于壳聚糖纳米颗粒中，构建了负载rBoIFNα/γ的壳聚糖纳米颗粒（CS-rBoIFNα/γ NPs），并利用动态光散射、傅里叶变换红外光谱等对纳米颗粒表征进行分析。最后，通过小鼠体内实验评价CS-rBoIFNα/γ NPs对BVDV的防治效果。结果表明rBoIFNα/γ在细胞水平上安全可控，最高无毒浓度达62.5 μg·mL^-1，抗病毒活性可达34,667 U·mg^-1，显著高于单体牛干扰素。同时rBoIFNα/γ能够高效激活JAK-STAT信号通路。成功制备CS-rBoIFNα/γ NPs，该纳米颗粒呈球形，平均直径为170 nm，粒径分布均匀，聚集趋势低，傅里叶变换红外光谱分析证实了壳聚糖特征峰的存在。细胞毒性实验表明，CS-rBoIFNα/γ NPs对MDBK细胞的最大无毒浓度达1 mg·mL^-1。在最优质量比（rBoIFNα/γ : CS = 1:5）条件下，封装率达76.3%，载药率达67.2%。体内实验表明，rBoIFNα/γ能促进T淋巴细胞的增殖和分化，且CS-rBoIFNα/γ NPs组表现出更优的效果。在BVDV感染的小鼠模型中，与rBoIFNα/γ组相比，CS-rBoIFNα/γ NPs组表现出更优的疗效，有效缓解了白细胞和淋巴细胞的急剧下降；显著降低脾脏、十二指肠、血液和粪便中的病毒载量；明显下调血清中炎症因子IL-6和TNF-α的水平。组织病理学分析进一步证实，CS-rBoIFNα/γ NPs组小鼠仅见轻微的肠绒毛水肿和极少量淋巴细胞坏死，而BVDV组和rBoIFNα/γ组则出现明显的淋巴细胞点状坏死。综上所述，CS-rBoIFNα/γ NPs显著提升了抗BVDV感染的疗效，为牛病毒性腹泻的治疗提供了具有前景的纳米蛋白药物策略。

Fall armyworm (FAW), Spodopterafrugiperda, is one of the most destructive pests of maize worldwide. Since invading Africa in 2016 and China in 2019, it has caused substantial crop losses, especially across Sub-Saharan Africa. To identify practical, smallholder-friendly pest management tactics, the FAO-China South-South Cooperation FAW Project conducted on-farm trials at two sites in Kenya (Bungoma and Embu) and two in Ghana (Kpong and Pampaso). Baseline surveys revealed infestation rates of 73.3–93.7%, with larval densities of 95–532 larvae per 100 plants. Six treatments were compared against an untreated control: neem oil, Bacillusthuringiensis (Bt), emamectin benzoate and chlorantraniliprole (Kenya) or lambda-cyhalothrin (Ghana), each applied either as a knapsack-spray or as sand granules dropped into the maize whorl. Leaf damage levels (LDL) and number of living larvae (NLL) per plant were recorded weekly. After one season, emamectin benzoate both sprayed and sand-applied consistently provided the lowest leaf damage level ((1.06±0.02)–(1.35±0.07)) compared with the control ((3.21±0.16)–(4.60±0.16)) in Kpone and Pampaso of Ghana. In Kenya, emamectin benzoate also resulted in lower leaf damage levels ((1.21±0.03)–(3.17±0.09)) compared with the control ((4.48±0.05)–(6.30±0.18)) at Embu and Bungoma. Bt sand granules were highly effective in Ghana ((1.20±0.04)–(1.29±0.07)) but performed less well in Kenya ((3.98±0.08)–(6.97±0.16)). Neem oil provided moderate suppression with leaf damage levels ((1.31±0.06)–(2.04±0.12)) in Ghana and ((3.13±0.08)–(3.58±0.16)) in Kenya, whereas lambda-cyhalothrin failed completely. In parallel, we screened five Ghanaian and eleven Kenyan local maize varieties for natural resistance; The Omankwa variety showed significantly lower leaf damage level ((1.91±0.12)–(2.73±0.21)) than the Lake (hybrid) variety ((2.78±0.31)–(3.93±0.17)) in Ghana; In Kenya, Pioneer (2.52±0.12), SC 73 Tumbo (2.58±0.14) and WH508 (2.44±0.11) showed significantly lower leaf damage levels compared with DK777 (5.44±0.08), W301 (5.47±0.07), Tembo (Seedco) (5.71±0.07), and Sungura (Seedco) (3.76±0.07). These trials offer smallholder farmers direct, comparative evidence to support cost-effective and locally adapted FAW management decisions.

Optimizing heading date and key agronomic traits is critical for wheat adaptation to diverse cropping systems. Genome-wide association studies (GWAS) have identified numerous loci for these traits. However, the stability and transferability of these associations across diverse genetic backgrounds remain limited. Here, we conducted GWAS for six agronomic traits using 461 Korean wheat accessions and validated major loci in a globally diverse panel of 206 accessions from 48 countries. We identified 22 marker–trait associations (MTAs), including six major-effect loci with phenotypic variance explained (PVE)≥10%. Five of these were converted to Kompetitive Allele-Specific PCR (KASP) markers, and all five showed consistent allelic effects in the global validation panel, confirming their transferability across distinct genetic backgrounds. Three validated loci were linked to functionally characterized genes: GIGANTEA on chromosome 3B for heading date, the Rht8-linked region on 2D for spike compactness, and the B1 interval on 5A for awn inhibition. The GIGANTEA-linked allele advanced heading by 5.3–7.7 days without reducing kernels per spike. The Rht8-linked locus reduced spike length while maintaining spikelet number. The B1-linked locus shortened awns without yield penalties. Two additional validated loci for heading date (2B) and culm length (5A) were associated with previously uncharacterized genes. These results demonstrate that major loci identified through regional GWAS can maintain stable effects across globally diverse genetic backgrounds and provide transferable KASP markers for wheat breeding worldwide. This highlights the value of integrating region-specific discovery with cross-population validation.

Yellow stem borer (YSB; Scirpophagaincertulas) is a major constraint on rice production in Asia. To dissect the genetic basis of YSB resistance, we developed 265 F₉ recombinant inbred lines (RILs) from the resistant parent A232 and the susceptible parent G46B and performed whole-genome resequencing for bin mapping and linkage analysis, generating 3,327 bins. Resistance was evaluated as dead heart index in two field trials under natural infestation. Composite interval mapping consistently detected a reproducible QTL, qTir04, on chromosome 4, with a peak LOD of 15.14 and 21.8% phenotypic variance explained. Local mapping with nine InDel markers resolved qTir04 into two linked sub-loci, qTir04.1 (~300 kb) and qTir04.2 (~1.14 Mb). Validation in a BC₃F₂-derived NIL population confirmed the effect of qTir04, prioritized qTir04.1 as the primary and most reproducible resistance interval, and indicated that the effect of qTir04.2 was conditional or less stable under our validation conditions. Transcriptome profiling and qRT-PCR in NILs, together with parental fixed SNPs/InDels and structural variants, identified five genes within qTir04.1 showing genotype- and/or infestation-associated expression divergence and prioritized receptor-like kinases and subtilisin-like proteases as leading candidates. These results provide a genetically validated QTL interval and closely linked markers for future marker-assisted breeding, and establish a foundation for functional validation and cloning of the prioritized qTir04.1 candidate genes.

Compound agricultural drought and heat events (CADHEs) pose an escalating threat to global food security, yet current assessments often overlook the distinct roles of stress duration and severity in driving yield loss. To address this gap, we developed a process-based classification framework incorporating antecedent cumulative effects to decouple CADHEs into duration-dominant and severity-dominant types, and to assess their spatiotemporal dynamics and impacts on maize yield in the Huang-Huai-Hai (HHH) Plain. The results showed that drought duration-dominant events (CADHE_dd) occurred more frequently than heat duration-dominant events (CADHE_hd) and concurrent events (CADHE_co),with occurrences concentrated in the V0–V6 stages across central-southern Hebei, most of Henan, and northern Shandong. Furthermore, compared with heat severity-dominant events (CADHE_hs), drought severity-dominant events (CADHE_ds) were more frequent, persisted longer, and exhibited higher severity and intensity. Critically, maize yield responses to compound events exhibited strong phenological dependence and clear asymmetry between duration-dominant and severity-dominant stress. Duration-dominant CADHEs, especially CADHE_dd, acted as chronic constraints during the vegetative-to-reproductive transition, and were governed by a cumulative deficit mechanism, whereby yield loss was most sensitive to prolonged exposure, as evidenced by a decline rate of 2.63% d^-1 (P<0.01). In contrast, severity-dominant CADHEs, particularly CADHE_hs, functioned as acute stressors during the reproductive phase, acting as severity-dependent amplifiers with a significantly steeper marginal sensitivity (-6.39%, P<0.01) than CADHE_ds (-4.88%, P<0.01). The R1–R3 stage emerged as the most vulnerable window, where the average yield loss rate exceeded 15% under heat severity-dominant conditions (P<0.01). These findings indicate that while drought imposes widespread cumulative penalties, heat severity acts as a critical tipping point for yield collapse. Sustainable agricultural risk management should therefore shift from static protocols to driver-specific strategies, prioritizing buffering measures against chronic drought and avoidance strategies against acute heat stress.

High-vigor seeds are fundamental to ensuring high-quality seedling establishment in rice (Oryzasativa L.) and play a key role in reducing cultivation costs and improving yield. To elucidate the genetic basis of rice seed vigor, we performed a genome-wide association study (GWAS) on vigor-related traits using 237 accessions from the 3K Rice Core Collection, identifying 16 quantitative trait loci (QTL). Among these, four QTLs identified in the present study—qOsGR5 (germination rate), qOsGP5 (germination potential), qOsGI5 (germination index), and qOsSS5 (seed storability)—were found to co-localize at a single locus on chromosome 5. Given its pleiotropic association with these vigor-related traits, this locus was designated qOsSV5 (rice seed vigor on chromosome 5). Candidate gene analysis preliminarily identified the bHLH transcription factor Os05g0541400 as the corresponding gene for qOsSV5. Knockout of OsSV5 significantly reduced seed vigor and led to a marked increase in reactive oxygen species (ROS) accumulation in seeds. Domestication analysis indicated that OsSV5 has undergone strong artificial selection in japonica rice. Haplotype analysis revealed three haplotypes of OsSV5, among which the dominant OsSV5^Hap3 was associated with high seed vigor. This favorable haplotype has been widely utilized in rice breeding programs in southern China. Collectively, our findings demonstrate that OsSV5 is a key regulator of rice seed vigor and that its dominant haplotype has been extensively applied in breeding practice.

碳青霉烯类、黏菌素和替加环素是临床治疗多重耐药革兰氏阴性菌感染的最后防线抗生素。肺炎克雷伯菌是临床上重要的条件致病菌，也是耐药基因水平传播的重要储存库。一旦菌株同时对这三类抗生素产生耐药性，将严重威胁公共卫生安全。

研究方法：在常规监测工作中，从江苏某养鸡场随机采集了40份新鲜粪便样本。通过PCR检测鉴定出3株同时携带bla_NDM-7与mcr-8.2基因的肺炎克雷伯菌，其中1株还同时携带tmexCD1-toprJ1基因簇。采用微量肉汤稀释法测定菌株的抗菌药物敏感性；通过接合试验验证三类耐药基因的接合转移能力；开展全基因组测序，分析菌株的基因组特征；同时，从NCBI数据库下载相关菌株进行比较基因组分析，以阐明菌株系统发育关系及三类耐药基因的分布特征。

研究结果：3株菌株均为多重耐药菌，对所有受试的8种抗菌药物均表现出耐药。3株菌株中的bla_NDM-7基因均可成功接合转移至受体菌，而mcr-8.2与tmexCD1-toprJ1基因不具备转移能力。基因组分析显示，3株菌株中的bla_NDM-7基因均位于IncX3型可接合质粒上，mcr-8.2基因位于IncR/IncFII型融合质粒上，tmexCD1-toprJ1基因则位于IncFIB(Mar)型质粒上。mcr-8.2阳性质粒与tmexCD1-toprJ1阳性质粒可能与其他质粒发生重组，进而促进其在不同宿主菌间传播。3株菌株均属于ST1型肺炎克雷伯菌，将其与NCBI数据库中的156株ST1型肺炎克雷伯菌共同开展系统发育分析，发现ST1型菌株地域与宿主分布范围广泛，本研究的3株菌株与数据库中3株同时携带bla_NDM-1、mcr-8.2及tmexCD1-toprJ1的菌株亲缘关系较近，表明克隆传播与质粒水平转移共同推动三类耐药基因的汇聚与扩散。此外，将我们的菌株与数据库中638株bla_NDM-7阳性、65株mcr-8.2阳性及310株tmexCD1-toprJ1阳性肺炎克雷伯菌进行分析，发现其序列分型、地理来源及宿主均呈现高度多样性，总体而言，tmexCD1-toprJ1、 bla_NDM-1/bla_NDM-5和mcr-1/mcr-8的共存组合最为常见。

创新性：该研究报道了禽源ST1型肺炎克雷伯菌中bla_NDM-7、mcr-8.2及tmexCD1-toprJ1三种耐药基因共存的现象。并发现mcr-8.2阳性质粒与tmexCD1-toprJ1阳性质粒具备与其他质粒重组形成新型融合质粒的潜能，从而加速耐药基因在不同宿主菌中扩散。

Weak seedling vigor of machine-transplanted rice during the recovery stage often limits basal-tillering nitrogen (N) uptake and yield, particularly under the split urea application with the increasing N. In this experiment, the effect of tandem long-mat seedlings (TLMS) transplanted with seedling fertilizer (SF) on yield and N use efficiency (NUE) was studied. Three-season field experiments at two sites consisting of TSF (transplanting with 7.0 kg ha^-1 SF) and T (transplanting without SF) based on five different dosages of basal-tillering N were conducted to comprehensively study the effect of SF in field. The results show that SF released rapidly after transplanting and significantly increased dry matter accumulation, N uptake and their rates during tillering stage. Consequently, TSF showed enhanced growth with early emerging tillers, significantly higher proportion of effective tillers and panicle numbers, and 7.8% higher yield than T. Consistently, basal tillering and total NUE (13.8%) of TSF was significantly higher than that of T. Notably, in some growing seasons, even with a 37.5 kg ha^-1 reduction in basal-tillering N, TSF still achieved a comparable dry weight, N uptake and yield to that of T, supporting the quantitative significance of seedling fertilizer in TLMS. Further quantification through regression analysis of yield and dosage verified that 7.0 kg ha^-1 N supplied via SF was equally effective as 24.1 kg/ha basal-tillering N, in terms of yield response. Overall, TLMS transplanting with SF is an effective strategy to enhance the early growth vigor, improve yield and NUE, and reduce basa-tillering N input in machine transplanted rice. This study successfully integrates the soilless nursery establishment with SF within mechanical rice-transplanting system and quantitatively demonstrating its contribution to post-transplantation performance of rice seedlings.

Reproductive performance is a key determinant of efficiency and sustainability in sheep production, yet the molecular basis of prolificacy remains incompletely understood. Here, we constructed an integrative framework for sheep ovarian biology by combining multi-tissue transcriptomics, differential expression analysis, cross-species comparative transcriptomics, and single-cell transcriptomics. We first profiled ovary-specific expression across tissues and identified 3,483 ovary-specific genes, defining a reproductive program enriched for key regulators such as FOXL2 and BMPR1B. We then compared high- and low-prolificacy groups across different FecB genotypes. When comparing BB-high-prolificacy with ++-low-prolificacy sheep, genes involved in the ovulation cycle, including BMPR1B, IGF1R, and GDF9, were significantly enriched, indicating a close association with the follicular regulatory axis. In contrast, when comparing ++-high-prolificacy with ++-low-prolificacy sheep, the BMP signaling pathway, including ACVR2B, BMPR1A, and BMP4, was enriched, suggesting that prolificacy in sheep lacking the FecB mutation may be governed by a distinct molecular mechanism. Cross-species analysis further identified conserved reproductive genes, including LHX9 and WNT4, supporting an evolutionarily conserved ovarian program. At single-cell resolution, BMPR1B was also prominently expressed in eGCs 3 granulosa cells. Finally, integrative analyses prioritized IGF1R as a candidate regulator in the FecB-assoc

Wheat is one of the most important food crops in the world, and grain number per spike (GNS) is one of the most important factors affecting wheat grain yield. Therefore, identifying genes controlling GNS is important for wheat production. However, wheat is an allohexaploid species and has a highly complex genome, and the isolation of wheat genes is challenging. In this study, we identified a candidate gene, TraesCS4B02G047100 (TaRLK-4B), associated with GNS using both quantitative trait locus (QTL) mapping and genome-wide association study (GWAS) based on RNA-Seq. We obtained three homozygous mutant lines, bb-1, bb-2 and bb-3, using the CRISPR/Cas9 gene editing system. Compared with Fielder (wild type, WT), the three mutant lines exhibited significant reductions in GNS, total spikelets per spike (TSS) and spike length (SL). These results indicated that TaRLK-4B positively regulates GNS and its related traits TSS and SL. The TaRLK-4B gene contains two exons and one intron and belongs to the largest subfamily of LRR-RLKs. We performed RNA-Seq analysis using spikes from the WT and bb-2 mutant line at the tillering stage. A total of 1,193 differentially expressed genes (DEGs) were identified, including previously cloned TaSPL17 homologous genes in the three subgenomes and their orthologous gene OsIPA1 (OsSPL14) in rice. Combining RNA-Seq data of TaRLK-4B and DAP-Seq data of the TaSPL17-7D, we identified 136 overlapping genes which are likely downstream targets of TaSPL17-7D. Therefore, we hypothesized that TaRLK-4B represents a novel putative component of the TaSPL17-centered regulatory network. In addition, haplotype analysis revealed that Haplotype 2 (Hap2) is a favorable haplotype for increasing GNS, but the thousand-grain weight (TGW) was not significantly different between Haplotype 1 (Hap1) and Hap2.

The rapid development of precision livestock farming highlights the need for accurate and complete high-throughput phenotypic data. However, in conventional milking systems, anomalies and missing records are common, leading to reduced reliability of downstream analyses of milk yield, such as quantification of milk losses and derivation of resilience indicator traits. This study proposed a data-cleaning framework for high-throughput session milk yield records by integrating anomaly detection and imputation of missing data. The Isolation Forest-based strategy using residuals with daily milk yield as a mediating variable was developed for anomaly detection. For missing-data imputation, we proposed a hybrid strategy combining linear weighted moving average and nearest-neighbor methods. Simulated datasets were constructed for stepwise validations, including five control methods for anomaly detection and four for imputation. In simulations, the proposed strategy outperformed all control methods. Compared to baseline strategies, the F1 score for anomaly detection improved by 0.08, and the R² for imputation improved by 0.09. Furthermore, we applied the proposed strategy to real records from 28,745 Chinese Holstein cows. After data cleaning, the average R² of lactation curve fitting improved by 0.10, and root mean squared error (RMSE) decreased by 0.57 kg (12.64%). Using the complete dataset, cluster analysis revealed four patterns of daily variability in session milk yields across lactations. Both hierarchical clustering and subspace clustering identified persistent high-variability groups (2.1-12.2% of lactations). Notably, over 20% of cows changed the variation groups across parities, demonstrating inconsistent inter-parity stability. Overall, this study provides a reliable tool for enhancing the accuracy and completeness of milk yield records to be used for daily herd management and breeding purposes.

Symbiotic nitrogen fixation (SNF) is an essential nitrogen (N) source for peanut growth and development, significantly contributing to sustainable and efficient peanut cultivation. However, the molecular mechanisms underlying efficient nodulation and N fixation in peanuts are unclear. This study involved reciprocal grafting using the high-nodulation peanut (XH11) and the non-nodulation peanut (DH9), which are derived from different parental lines and share no common parents. Physiological, transcriptomic, and metabolomic data of peanut roots were examined on days 25 and 35 after grafting. The results demonstrated that grafted combinations with XH11 as the rootstock (XH/XH and DH/XH) formed nodules after 35 days, with XH/XH exhibiting a higher nodule number, leghemoglobin, ureide, and allantoic acid contents  than DH/XH. The scion genotype also played a regulatory role: the non-nodulating scion DH9 limited nodule formation even when grafted onto a nodulating rootstock (DH/XH). Nodular combinations exhibited greater root length, root surface area, root volume, and root dry weight  than non-nodular combinations (DH/DH and XH/DH). Weighted co-expression network analysis of root transcriptome data identified 40 hub genes, including the LHY protein (Late Elongated Hypocotyl, a core component of the circadian clock), serine/threonine kinases, and ethylene-responsive transcription factors, and their module‑specific expression patterns may partially account for the difference in nodule number between XH/XH and DH/XH. Integrated transcriptomic and metabolomic analyses demonstrated significant enrichment of DAMs/DEGs in the glutathione metabolism pathway in nodular peanut roots. Downregulation of GPX, GSR, and ODC genes, coupled with upregulation of GST genes and increased GSSG and spermidine levels, synergistically promoted nodulation, glutamate accumulation, and N assimilation efficiency. This study systematically clarifies the molecular regulatory network governing root symbiosis in peanut-grafted systems, providing a reference for the genetic enhancement of high-efficiency N-fixing varieties in legume crops.

Plant synthetic biology offers transformative potential for sustainable biotechnology, precision agriculture, and recombinant protein production. However, the field faces persistent challenges, including lengthy development timelines, gene silencing, limited transformation efficiency, and regulatory hurdles surrounding stable genetic modifications. To overcome these bottlenecks, virus-based transient expression systems have emerged as powerful solutions, enabling rapid, high-level, and modular gene expression in plants without the need for stable integration. Agrobacterium-mediated delivery of deconstructed viral vectors has revolutionized the plant design-build-test-learn (DBTL) cycle. These platforms facilitate efficient expression of synthetic circuits, multigene metabolic pathways, and therapeutic proteins in model hosts such as Nicotiana species and edible species such as lettuce. When integrated with controlled environment agriculture (CEA) systems, transient expression offers a closed-loop, reproducible environment, supporting regulatory-grade biomanufacturing. Applications include rapid prototyping of genetic circuits, genome editing through CRISPR/Cas systems, and production of complex biologics such as monoclonal antibodies and vaccines. Case studies such as ZMapp and Medicago's COVID-19 VLP vaccine illustrate the clinical relevance of this approach. Emerging innovations, including synthetic viral chassis, standardized part libraries, and AI-driven construct optimization, promise to further enhance transient expression as a foundational tool in plant synthetic biology. Collectively, these advancements are shaping a future in which plants can serve as programmable, scalable biofactories for health and industrial applications.

Phytoene synthase (PSY) is the primary rate-limiting enzyme in the carotenoid biosynthesis pathway and plays a crucial role in regulating carbon flux toward carotenoid production. The Orange (OR) protein functions as a molecular chaperone for PSY, increasing its activity and facilitating carotenoid accumulation. Notably, the mutation of OR to OR^His further amplifies this effect. However, the interaction dynamics between OR/OR^His and multiple PSY copies in plants remain unclear, particularly whether these proteins interact uniformly with all PSY variants or exhibit differential effects. We here cloned three BoaPSY homologs from Chinese kale and demonstrated that all identified BoaPSYs contribute to carotenoid accumulation in engineered bacterial systems and Chinese kale calli. Among them, BoaPSY1 exerted the most pronounced effect, followed by BoaPSY2 and BoaPSY3. Additionally, BoaOR/OR^His directly interacted with BoaPSYs, and the overexpression of BoaOR/OR^His–BoaPSYs complexes further enhanced carotenoid accumulation in Chinese kale calli, with the calli overexpressing BoaOR^His-BoaPSY1/2 showing the highest carotenoid content. Binding energy analyses and α-galactosidase activity assays revealed that BoaOR^His has the strongest binding affinity for BoaPSY2. Overexpression of the BoaOR^His–BoaPSY2 complex in Chinese kale plants increased carotenoid and chlorophyll levels but reduced their degradation during postharvest storage. Our findings advance the understanding of carotenoid metabolic regulation and propose a novel strategy for improving both preharvest quality and postharvest shelf life of Chinese kale, thereby increasing its economic value and market potential.

Orphan genes (OGs) are usually species specific and lack recognizable domains, posing challenges for their functional analysis. Flowering time is a key agronomic trait affecting the yield and adaptability of Chinese cabbage (Brassica rapa). However, the molecular mechanism by which Chinese cabbage OGs regulate flowering remains unclear. Here, we verified that OG BOLTING RESISTANCE 2 (BR2) is a key inhibitory factor for flowering in Chinese cabbage. The BR2 loss-of-function mutant ‘br2-M’, exhibited a significantly earlier flowering time. In contrast, the BR2 overexpression line ‘BR2OE’ delayed flowering. Further analysis showed that the expression of BR2 was directly repressed by vernalization treatment. To clarify its mechanism, we used yeast two-hybrid library screening and protein interaction experiments and found that BR2 specifically bound to the MADS-box domain of flowering integrator BrSOC1c. Molecular mechanism studies showed that BrSOC1c directly bound to the promoter of downstream flowering gene BrLFYb and activated its transcription, whereas BR2 significantly inhibited its transcriptional activation ability by interacting with BrSOC1c. Vernalization treatment relieved the inhibitory effect of BR2 on the BrSOC1c–BrLFYb pathway, promoting flowering. In conclusion, this study identified BR2 as a negative regulatory factor induced by vernalization that delays the flowering process by directly inhibiting the activity of BrSOC1c. These results uncover the molecular mechanism underlying vernalization-mediated flowering regulation in Chinese cabbage.

Sporopollenin, a core structural component of spore and pollen walls in land plants, is critical for pollen formation, yet its metabolic mechanism in tomato (Solanum lycopersicum) remains unclear. In this study, an anther-specific gene SlFAR3 encoding a fatty acyl-CoA reductase was identified, which participates in sporopollenin biosynthesis by regulating fatty alcohol metabolism. Phenotypic analysis revealed that slfar3 mutants exhibited abnormal anther and pollen development, ultimately leading to pollen-free male sterility. Cytological evidence demonstrated defective Ubisch body formation and significantly reduced accumulation of sporopollenin precursors in slfar3 anthers. In vitro enzymatic assays confirmed that SlFAR3 specifically catalyzes the reduction of palmitoyl-ACP (C16:0-ACP) to hexadecanol (C16:0-OH). Mass spectrometry analysis of mutant anthers showed a marked decrease in cutin monomers, altered composition of epicuticular waxes, and distinct changes in soluble fatty acid and alcohol profiles. Integrated transcriptomic and metabolomic analyses revealed significant enrichment of differentially expressed genes and metabolites associated with the phenylpropanoid biosynthesis pathway in slfar3, suggesting that SlFAR3-derived hexadecanol contributes to sporopollenin precursor formation via phenylpropanoid metabolism. This study systematically elucidates the molecular mechanisms by which SlFAR3 regulates sporopollenin synthesis to ensure pollen development in tomato, integrating insights from molecular, cellular, and metabolic perspectives. These findings advance the understanding of plant male gametophyte development and provide a foundation for engineering tomato male sterile lines.

To enhance the commercial value of cucumbers and reduce the burden of manual grading, this study proposed an automated grading method based on computer vision and deep learning. The method included both equipment and algorithm design. In terms of equipment, a compact grader suitable for rod-shaped agricultural products was developed to provide hardware support for intelligent grading. On the algorithmic side, the study proposed an innovative approach that uses morphological key points to obtain shape parameters of rod-shaped agricultural products, leading to the development of the CukeposeNet model. The method predicts cucumber key point coordinates in an end-to-end manner. By combining geometric computation and threshold-based decisions, six morphological parameters were detected and used for multiple indicators grading. Experimental results showed that CukeposeNet achieved a key point detection accuracy of 98.5% and a processing speed of 50 FPS, outperforming six other key point detection algorithms. The correlation coefficients (R) for length-related parameters (length, neck length, arch height) were all above 0.90, and the mean absolute errors (MAE) for diameter-related parameters (stalk, middle, stem) were all below 0.15 cm, indicating good overall accuracy. The model also demonstrated strong generalization performance in parameter detection of other rod-shaped crops such as bitter melon, green pepper, and luffa; the R reached 0.985. In an online multiple indicators grading experiment, the grader achieved a grading accuracy of 87%，which is 20% higher than manual grading and operated at twice the manual efficiency, thereby meeting production requirements. This study confirmed the feasibility of morphological parameter detection based on key points and provided a practical solution for the online external quality detection and grading of cucumbers and similar agricultural products.

Fruit neck affects the appearance and commercial quality of melon. In the present study, we investigated the genetic basis of fruit neck in melon by utilizing a segregating population derived from M1-15 (without fruit neck, ssp. agrestis, cultivar) and PI 614174 (with fruit neck, ssp. agrestis, wild type). The results indicated that a single-dominant gene controls melon fruit neck trait (designated as CmFn, fruit neck). Bulk segregant analysis (BSA-seq) and genetic mapping narrowed down the CmFn locus into a 116.693-kb region on chromosome 2, harbors 11 predicted genes. Sequence variation analysis in the promoter and coding regions combined with gene expression analysis between parental lines indicated MELO3C015395.2 (late embryogenesis abundant protein), MELO3C015398.2 and MELO3C015399.2 (both encoding cysteine synthase-like) may represent the most promising candidate genes for CmFn locus. Our finding provided target genes for further melon fruit development research.

Cold stress negatively affects melon growth and development. Previous studies have shown that exogenous trehalose (Tre) alleviates the cold damage in melon seedlings; however, the specific mechanism is not fully clarified. Here, Tre treatment increased the indole-3-acetic (IAA) content and upregulated an early response factor of IAA (small auxin up RNA gene, CmSAUR1) gene expression. Inhibition of IAA signal transport or silencing of CmSAUR1 decreased the activities of superoxide dismutase, ascorbate peroxidase and glutathione reductase. This led to increased susceptibility to cold damage in melon seedlings and diminished the beneficial impact of Tre. Through the analysis of transcriptome and promoter of CmSAUR1, the upstream transcription factor CmTCP9/12/25 of CmSAUR1 was excavated. Protein–DNA interaction experiments further verified that CmTCP9/12/25 could transcriptionally regulate CmSAUR1 expression. Yeast two-hybrid, luciferase complementation imaging and bimolecular fluorescence complementation experiments further confirmed that CmTCP9/12 interacted with CmTCP25 to form protein complexes. Silencing CmTCP9/12/25 genes significantly aggravated the cold damage of melon seedlings and weakened the efficacy of Tre. All of them downregulated the expression of CmSAUR1. In conclusion, application of Tre induced the upregulation of CmTCP9/12/25, and CmTCP9/12/25 could bind to the CmSAUR1 promoter and activate its expression. These eventually increased the cold tolerance of melon seedlings.

Postharvest anthracnose is a critical disease affecting the quality of papaya fruit and the development of the papaya industry. However, the molecular mechanisms influencing this disease remain unclear. Since major latex proteins (MLPs) are pivotal components in the defensive responses of many plants, it is worthwhile to explore the functions and regulatory mechanisms of MLPs in papaya anthracnose resistance. In this paper, we performed a comprehensive genome-wide analysis of the papaya MLP gene family, identifying a total of 15 CpMLPs and providing information on their gene structures, protein motifs, phylogenetic relationships, and stress-related cis-regulatory elements. Based on bioinformatics and previous findings, CpMLP14 was selected for in-depth functional investigation. CpMLP14 was inhibited by the infection of Colletotrichum brevisporum, and CpMLP14 protein localized in nucleus and cytoplasm. Through transient overexpression of CpMLP14 in papaya, and heterologous expression CpMLP14 in tomato, as well as inoculation and phenotypic analysis, it was found that CpMLP14 negatively regulates papaya resistance to C. brevisporum. Further results demonstrated that CpMLP14 can inhibit the accumulation of reactive oxygen species (ROS) as well as the expression of pathogenesis related 1 (PR1) and PR4 genes. Additionally, our results revealed that CpMLP14 was regulated by the transcription factor CpWRKY50, which has been proven to be a key transcription factor positively regulating papaya's resistance to anthracnose. CpWRKY50 could directly bind to the W-box motif in the promoter of CpMLP14 and inhibit its expression. Collectively, these results provide a systematic understanding of the papaya MLP gene family, establish CpMLP14 as a direct target of CpWRKY50, and highlight the critical role of the CpWRKY50-CpMLP14 module in modulating papaya resistance against anthracnose. These results lay a foundation for future genetic improvement of disease resistance in papaya.

Broccoli sprouts, valued for their high nutritional content and richness in sulforaphane (SF), exhibit quality traits that are significantly modulated by light quality. However, the mechanisms underlying light regulation of glucosinolate (GSLs) biosynthesis in sprouts remain elusive. This study employed broccoli 'Xianglv No. 3' sprouts as experimental material, comparing sprouts grown in darkness (control) with those under five distinct LED light spectra. We assessed sprout growth, quantified GSLs and SF levels, measured total antioxidant capacity, and evaluated cytotoxicity against cancer cells. Integrated metabolomic and transcriptomic analyses were subsequently conducted. Results demonstrated that blue light significantly enhanced GSLs biosynthesis and SF accumulation in broccoli sprouts compared to the dark control, concurrently boosting antioxidant capacity and reducing cancer cell proliferation. Metabolomic profiling revealed significant alterations in 19 GSL metabolites under blue light, with 6 up-regulated and 13 down-regulated. Aliphatic GSLs exhibited the most pronounced changes, while SF increased markedly. Transcriptome analysis identified 9,128 differentially expressed genes (DEGs) in response to blue light. The results suggested that blue light regulated GSLs biosynthesis, particularly AGS, potentially by inhibiting carbon chain elongation and secondary modifications in their side chains. We propose a regulatory model of the HY5-MYB28-CYP83A1 signaling cascade. In this model, blue light induces the up-regulation of the light-responsive transcription factor HY5. HY5 subsequently activates the expression of its downstream target, MYB28, which in turn upregulates CYP83A1, ultimately driving enhanced GSL and SF production. This study elucidates the molecular basis for light quality regulation of broccoli sprout quality and provides valuable insights for advancing efficient cultivation and quality enhancement strategies.

The intensification of agriculture exacerbates continuous cropping obstacles, threatening global food security. Chemical fumigation effectively suppresses soil-borne diseases, however, it also reduces microbial diversity, disrupts ecological networks, and impairs essential ecosystem functions. Although organic amendments exhibit great potential in restoring soil health, a comprehensive understanding of post-fumigation microbial recovery and functional reconstruction strategies still remains elusive. This review introduces the “ecological vacuum” concept and proposes a “targeted reconstruction” framework based on fumigation-amendment synergy. We systematically summarize the antimicrobial mechanisms of chemical fumigants and their adverse effects on microbial diversity, network interactions, resistance and resilience and key ecosystem services. Building on this, we further elucidate how organic amendments promote beneficial microbes colonization, guide functional recovery and stabilize soil micro-ecosystem following fumigation. It was demonstrated that compost, straw and wormcast could provide resources and habitats for beneficial microbes colonization and community assembly. Seaweed extracts and bio-organic fertilizers promote nutrient cycling, pathogen suppression, and plant growth through enzymes, antibiotics, and signaling molecules. Humic acid and biochar improve soil structure, alleviate acidification and strengthen ecosystem stability. These insights advance sustainable soil management strategies that combine fumigation with precision amendments to enhance soil health and agricultural productivity.

Wheat, a staple food crop essential to global food security, has seen remarkable advances in genomics over the past decade, with Chinese scientists making prominent contributions alongside international collaborations.  This review focuses on the achievements of Chinese researchers in wheat genomics, covering cutting-edge basic research, applied technologies, and intelligent production management, integrating global progress.  In basic research, breakthroughs in genome assembly, functional genomics, and genetic variation analysis have clarified the genetic basis of key agronomic traits.  In applied research, high-efficiency genotyping tools and cloned genes related to yield, quality, and stress resistance have promoted the translation of genomics knowledge into practical breeding guidance.  Future research will focus on precise genome editing, multi-omics integration, and intelligent breeding.  Artificial intelligence and precision agriculture have improved production efficiency and resource utilization, enhanced the efficiency of water and fertilizer use, and reduced environmental impacts and labor costs.  All of these are expected to provide strong technical support for global and Chinese food security and sustainable wheat production.

Based on a comprehensive dataset from 348 field experiments in China (2000-2023), this study developed and validated a season-specific intelligent fertilizer recommendation system for radish, based on the Nutrient Expert (NE) framework. The Quantitative Evaluation of the Fertility of Tropical Soils (QUEFTS) model was used to quantify distinct nutrient requirements for spring and autumn radishes, revealing that autumn radishes need substantially higher nutrients, notably approximately double the phosphorus, for equivalent yields. The analysis established a robust quadratic relationship between yield response and agronomic efficiency for N, P, and K in each season, forming the core of the radish NE system. Multi-location field trials demonstrated that, compared to farmers’ practice (FP) and soil testing (ST), the NE significantly reduced total application rates of fertilizer over 30% (especially P₂O₅ by >40%) and total fertilizer cost, while achieving the highest marketable root yields. NE also notably enhanced recovery efficiency (RE) of N, P, and K by 52.75%, 151.30%, and 87.34%, respectively; enhanced agronomic efficiency (AE) of N, P, and K by 76.88%, 291.75%, and 120.34%, respectively; enhanced partial factor productivity (PFP) of N, P, and K by 31.28%, 98.96%, and 31.57%, respectively; and brought apparent N (14.86 kg N ha⁻¹) and P (6.44 kg P ha⁻¹) balances close to zero, thereby reducing the environmental effects of reactive N and P. This study presents a data-driven, adaptive decision-support tool that enables radish farmers to optimize productivity, economic returns, and environmental sustainability, aligning with Chinese agricultural goal of reducing fertilizer input while increasing efficiency.

Accurate estimation of the winter wheat yield is critical for agricultural management and food security. Crop models are essential tools for simulating crop growth and predicting yield. However, uncertainties owing to variability in crop model parameters cannot be ignored. We integrated the enhanced WOrld FOod STudies (E-WOFOST) and PROSAIL models to compare different data assimilation schemes for yield prediction across 23 field sites in Henan Province. A comparison between data assimilation schemes was conducted based on three aspects: (i) remote-sensing crop data, (ii) soil moisture (SM), and (iii) assimilation algorithms. These aspects were crosscombined to form 14 assimilation schemes. The assimilation results of different remote-sensing crop data showed that vegetation indices (VIs) outperformed Sentinel-2 (S2) reflectance, while the latter generally surpassed the use of satellite products and measured values. Reanalysis of products incorporating deep-layer SM achieved better results than using only surface-layer measured SM. Among the five VIs, the chlorophyll index red edge (CI_red-edge) demonstrated the best performance (R²=0.70; RMSE=732.71 kg ha^-1; nRMSE=15.46%). Relative to the Genetic Algorithm and Shuffled Complex Evolution-University of Arizona (SCE-UA), Particle Swarm Optimization (PSO) reduced the RMSE/nRMSE by 13.25 ha^-1/0.28% and 201.40 ha^-1/4.25%, respectively. Among the 14 assimilation schemes, the highest accuracy was obtained by assimilating ERA5-Land SM, constraining E-WOFOST with CI_red-edge, and optimizing via PSO (R²=0.71, RMSE=719.46 kg ha^-1, and nRMSE=15.18%). Finally, application of the best scheme demonstrated strong interannual performance in Henan (2018–2020 and 2024: R²=0.50–0.63; RMSE=719.76–771.67 kg ha^-1) and transferred effectively to Shandong (2025: R²=0.63; RMSE=951.11 kg ha^-1), demonstrating its robust applicability across different years and regions.

Soybean grain yield was largely determined by flower differentiation and effective pod set. In maize-soybean strip intercropping, changes in light conditions affect organ development and nutrient allocation, affecting flower and pod set—though the exact physiological mechanisms remain unclear. To address this, a two-year field experiment was conducted from 2021 to 2022. This study aimed to analyze the regulatory effects of strip intercropping versus monocropping on soybean flower pod formation and abscission, and yield components, examining both temporal dynamics and spatial distribution. The experiment employed a split-plot design with two factors: the main plots consisted of four soybean cultivars (Gongqiudou 8 (GQ8), Guixia 7 (GX7), Nanxiadou 25 (ND25), and Nannong 99-6 (NN996)), and the subplots comprised three planting patterns (sole soybean (SS), maize-soybean strip intercropping (MSI), and maize-soybean relay strip intercropping (MSR). Results showed that compared with MSI, MSR extended the flowering and pod-setting periods by 4–7 days and improved branch development (reflected in increased branch node number and length). This enhanced architecture provided more potential flowering sites on branches, ultimately resulting in a distinct bimodal flowering curve. By optimizing the spatial distribution of reproductive organs towards the middle and lower canopies, MSR promotes flower and pod formation on branches and directs assimilate allocation to these pods, thereby improving yield potential. Compared with MSI, MSR increased the number of effective pods on branches at the bottom space by 40.2%, 86.6%, and 102.0% for varieties GX7, ND25, and NN996, respectively. We conclude that MSR combined with lately branching soybean variety mitigates the yield penalty commonly seen in intercropping systems by optimizing the spatiotemporal distribution of reproductive structures, where early shading is compensated by prolonged flowering and a strategic shift in assimilate allocation to branch pods, thereby stabilizing yield through efficient pod retention and set.

In acidic soils, phosphorus (P) from water-soluble phosphate fertilizers is readily immobilized into less available forms, thereby limiting soil P availability and crop P uptake. To address this constraint, two “in situ” value-added mono-ammonium phosphate (MAP) fertilizers (VP₁ and VP₂) were evaluated, and their effects on seedling growth of wheat and rapeseed and the underlying mechanisms were studied by soil incubation and pot culture experiments. The results showed that VP treatments maintained significantly higher P availability, reduced Fe-bound P fixation in acidic soil, and increased soil available Fe and Zn concentrations compared with MAP. Moreover, VP treatments were accompanied by higher acid phosphatase activity (7.6–50.0%), upregulated phoC expression (2.5–3.7-fold), enrichment of Stenotrophomonas, and enhanced mineralization of NaHCO₃-extractable organic P at 5 days. VP application significantly expanded the spatial distribution of available P in the fertisphere, thereby enhancing early-stage P uptake. Consequently, under VP treatments, wheat and rapeseed exhibited higher biomass, P concentration, and P uptake compared with MAP, even with a 50% reduction in P input. These findings suggest that VPs improved soil P availability and crop performance in acidic soil through three coordinated processes involving reduced chemical P fixation, enhanced organic P mineralization, and optimized spatial distribution. These integrated effects highlight the potential of VPs to improve P management in acidic soil systems.

Field crops are frequently exposed to fluctuating light (FL), which limits photosynthetic carbon assimilation and crop productivity. Mechanisms of soybean adaptation to FL remain largely unclear. This study investigated two soybean cultivars (ND12 and GX7) with contrasting responses to FL using integrated phenotypic, physiological, anatomical, transcriptomic and proteomic analyses. Under FL conditions, both cultivars showed consistent phenotypic and physiological changes: reduced specific leaf weight and leaf thickness, retarded vein development, a significant positive correlation between dynamic FL intensity and net photosynthetic rate, and asynchronous responses between stomatal movement and photosynthetic induction. Notably, ND12 enhanced CO₂ diffusion by optimizing mesophyll structure and exhibited demand-driven stomatal regulation tightly coordinated with photosynthetic activity. In contrast, GX7 showed greater structural sensitivity and adopted a preparatory stomatal opening pattern, maintaining partial aperture under low light to mitigate transient CO₂ limitation during light transitions. At the molecular level, both cultivars shared a conserved regulatory framework centered on the jasmonic acid (JA) signaling pathway and α-linolenic acid metabolism. Differential expression of JA biosynthetic and blue light-responsive genes contributed to cultivar divergence, with ND12 maintaining relatively stable JA-related gene expression, whereas GX7 displayed sustained downregulation and additional blue light-responsive differentially expressed genes. These findings clarify the coordinated physiological and molecular basis of soybean adaptation to FL and provide potential targets for breeding cultivars tolerant to FL environments.

High-density planting is a pivotal strategy for safeguarding food security, yet it is frequently accompanied by a pronounced escalation in lodging risk. This study aimed to elucidate how a spatially heterogeneous configuration mitigates lodging and stabilizes yield in wheat. Field experiments, together with machine-learning approaches and path modeling, were conducted to identify the key constraints underlying lodging resistance under contrasting planting configurations. Relative to the high-density homogeneous distribution (HD), the high-density heterogeneous distribution (HD-h) effectively alleviated within-canopy shading, thereby stabilizing the assimilate carbon flux delivered to the stem. This sustained carbon supply, in turn, enhanced the activity and transcriptional expression of key enzymes/genes involved in cellulose biosynthesis, promoting the development of mechanical tissues and increasing stem mechanical load-bearing capacity. Machine-learning further revealed a density-dependent shift in the primary limiting factors for lodging resistance: constraints transitioned from fluctuations in canopy light environment under lower resource pressure to a mechanically oriented bottleneck under high density, dominated by carbon metabolism and structural construction. By stabilizing source-derived carbon flux, HD-h coordinated the maintenance of high lodging resistance with concurrent support for spike number per unit area and individual-spike traits, thereby relaxing the conventional trade-off between yield and structural stability under dense planting. Optimizing spatial configuration improves lodging resistance and yield stability under high-density conditions by promoting assimilate conversion into structural carbon.

Conventional agricultural practices heavily rely on chemical inputs, posing risks to environmental and food safety. Seed biopriming with beneficial microorganisms offers a sustainable alternative to improve crop performance and resilience. Previously, we showed that the mycovirus-mediated hypovirulent strain DT-8 of Sclerotinia sclerotiorum functioned as a plant vaccine that significantly increased wheat and rapeseed yield. In this study, we utilized a laboratory-formulated plant vaccine based on strain DT-8 as a seed biopriming agent for rice. Through systematic optimization of seed priming parameters with plant vaccine, we defined that treatment at 24°C for 12 hours with a 3-day fermented broth (50 mL kg^-1 seed; mycelial fresh weight: 50 g L^-1) achieved more than 80% colonization efficiency without compromising germination. Microscopy confirmed root colonization by the plant vaccine, showing both intercellular and intracellular hyphal growth along the root axis without appressorium formation. Under both greenhouse and field conditions, plant vaccine-primed rice plants exhibited substantially enhanced growth traits, including plant height, chlorophyll content, dry weight, tiller number and yield. Furthermore, plant vaccine priming markedly reduced the incidence of rice false smut caused by Ustilaginoidea virens, under both artificial inoculation and natural field conditions. Transcriptome analysis further revealed the upregulation of growth-related genes. Concurrently, a significant enrichment of defense response terms and the diterpenoid biosynthesis pathway was observed, indicating the activation of key antimicrobial defenses. Our finding demonstrated that mycovirus-mediated plant vaccine serves as an effective seed priming agent, promoting rice growth and triggering systemic resistance.

Soil organic matter (SOM) and bacterial communities are key regulators of soil fertility, yet the synergistic regulatory effect remains poorly elucidated. In this study, 180 black soil samples spanning a wide and continuous fertility gradient were collected through large-scale field sampling in Northeast China. Metabonomics and high-throughput sequencing were employed to characterize the SOM molecular composition and bacterial community structure. Multidimensional analysis identified a response threshold (integrated fertility index, IFI of around 0.8) for both bacterial communities and SOM molecular variables to soil IFI, where the key driver of soil fertility shifted from SOM molecules to bacterial communities. Based on the response threshold, all the samples were divided into two groups: the low-fertility group with IFI<0.8 and the high-fertility group with IFI>0.8. Specifically, in the low-fertility group (IFI<0.8), IFI was only positively correlated with SOM molecular composition, which explained 52.81% of the total variation in IFI. While in the high-fertility group (IFI>0.8), IFI was only positively correlated with bacterial richness, which explained 70.77% of the total variation in IFI. These findings highlight the importance of targeted soil management strategies: prioritizing SOM monitoring in low-fertility soils and bacterial community assessment in high-fertility soils. This study provides a basis for precision agricultural practices aimed at optimizing and sustaining soil fertility in the black soil region of Northeast China.

Plant-beneficial bacteria (PBB) play a critical role in sustaining soil health and enhancing crop productivity through nutrient mobilization, pathogen suppression, and stress resilience. Yet, how environmental factors shape the biogeography and assembly of PBB in agricultural soils remains unclear. To fill this knowledge gap, we conducted a large-scale study across eastern China farmland to investigate: (i) the diversity, composition, and spatial distribution of PBB, and (ii) the key environmental drivers and mechanisms governing PBB community assembly. We found that plant growth-promoting (PGP), biocontrol (BIOC), and stress resistance (SR) functional groups accounted for 4.6–13.2%, 0.7–4.8%, and 0.1–1.3% of total bacterial community, respectively. The α-diversity of total bacteria was positively correlated with that of PBB, suggesting that overall microbial richness supports beneficial taxa. Soil pH acted as the primary driver of β-diversity for SR and BIOC, while mean annual precipitation (MAP) was the main factor influencing PGP β-diversity. Community assembly analysis revealed that species replacement contributed more to PBB β-diversity than richness differences. Mechanistically, homogeneous selection was the dominant process structuring PGP, BIOC, and SR assemblages, whereas dispersal limitation primarily governed the total bacterial community. Accordingly, all three PBB functional groups showed higher dispersal capacity and contained more generalist taxa than the total bacteria. The high abundance and broad distribution of PGP, BIOC, and SR underscored their potential ecological benefits, which are modulated by key environmental factors such as soil pH and precipitation. Our findings highlight key environmental filters for PBB assembly and identify widely distributed, generalist taxa, providing a framework for the targeted screening of regionally adapted microbial inoculants.

Understanding the spatiotemporal dynamics of soil organic C (SOC) quantity and origins, along with the potential microbial-mineral mechanisms influencing these changes, is crucial for long-term SOC preservation in agroecosystems; however, this knowledge is lacking during apple cultivation. By analyzing the temporal variations in SOC, microbial-derive C (MNC), plant-derived C (PC), iron oxides, exo-enzymes, and microbial communities across soil profiles over apple plantation establishment time-series (6, 14, 28, 35, and 42 years), this study investigated the synergistic roles of iron oxides and microbial attributes in regulating SOC cycling. Results showed that the SOC contents ranged from 5.5 to 12.3 g kg⁻¹, increased with increasing plantation ages; meanwhile, the pH values showed opposite trends with SOC during orchard development. Averaged across all soil profiles (0−60 cm), the elder orchard (i.e., 35 and 42 years) soils exhibited greater C resource levels (i.e., 45.0%−83.1% higher MNC and 91.4%−202.4% higher PC) than the younger orchard (i.e., 6 and 14 years) soils; meanwhile, long-term apple orchard cultivation (i.e., 35 and 42 years) enlarged PC but reduced MNC contribution to SOC. Notably, the above-mentioned C resources exhibited pronounced ‘surface aggregation’ patterns across soil profiles, with PC and MNC contents in the 0–20 cm layer being 42.0%–78.9% and 70.8%–302.0% higher than those in deeper layers. Besides, the contents of amorphous (Feo) and complex iron-oxides (Fep) increased with increasing plantation ages, accompanied by elevated iron-bound SOC contents and C:Fe molar ratios, but microbial biomass (evidenced by total PLFAs) and hydrolase activities firstly increased, peaking at 28 years orchards with mean values of 11.6 nmol g^–1 and 36.7, and subsequently decreased across all soil profiles. Partial least squares path model (PLS-PM) verified that increased soil acidification impedes microbial growth while promoting iron oxides accumulation. Random forest model revealed that the contribution of PC (or MNC) to SOC were mainly regulated by microbial biomass, nitrate-N, and Feo (or Fep, pH, and microbial biomass). The PLS-PM and Mantel test jointly confirmed that MNC was mainly regulated by mineral indices (i.e., iron oxides), rather than microbial attributes; meanwhile, the PC was primarily governed by mineral indices (i.e., Feo and activation index) and biotic variables (e.g., hydrolase activities). Collectively, these findings underscore the crucial role of pH in modulating mineral-microbial interactions for SOC preservation during orchard cultivation, thereby deepening our understanding of SOC dynamics within orchard ecosystems in China.

Soil organic nitrogen (SON) constitutes the dominant reservoir of soil N and governs its retention and supply. While the impact of biochar on soil N cycling has received increasing attention, due to the heterogeneous properties of SON fractions, the long-term effects of biochar on the conversion of fertilizer N into specific SON fractions remain unclear. Through a 7-year field experiment with ¹⁵N isotopic tracing, we investigated the fate of fertilizer N under combined applications of different biochar rates. Results showed that biochar amendment improved fertilizer N recovery by 20.85‒64.42% in the soil and by 2.72‒25.67% in plants relative to the NPK treatment. Fertilizer N was initially allocated mainly to acid-hydrolyzable ¹⁵N (H-¹⁵N), within amino acid ¹⁵N (AA-¹⁵N), hydrolyzable unknown ¹⁵N (HU-¹⁵N), and hydrolyzable ammonium ¹⁵N (HA-¹⁵N) being the labile fractions that were readily mineralized for plant uptake. Conversely, the non-hydrolyzable ¹⁵N (NH-¹⁵N) increased steadily over time despite its initially low proportion, contributing to enhanced soil N retention. Biochar promoted the conversion of fertilizer N into AA-¹⁵N, HA-¹⁵N, amino sugar ¹⁵N (AS-¹⁵N) and NH-¹⁵N, while suppressing its allocation to HU-¹⁵N in a dose- and time-dependent manner. Amino acid N (AAN) functioned as a transitional N pool that directly regulated the internal transformation of different SON fractions, thereby mediating the fate of residual fertilizer N. Overall, biochar simultaneously enhanced the content of labile and recalcitrant SON fractions, achieving a synergistic improvement in both the rapid supply and long-term retention capacities of soil N. This study provides a scientific basis for the long-term agricultural application of biochar and its impact on soil N cycling.