High-quality reference genomes at the population scale are fundamental for advancing pan-genomic research. However, high-quality genome assembly at the population scale is costly and time-consuming. To overcome these limitations, we developed Reference-Assisted Genome Assembly (RAGA), a hybrid computational tool that combines and reference-based assembly approaches. RAGA efficiently employs existing reference genomes from the same or closely related species in combination with PacBio HiFi reads to produce high-quality alternative long sequences. These sequences can be integrated with assemblies to improve assembly quality across population-scale datasets. The performance of RAGA across various plant genomes demonstrated its ability to reduce the number of contigs, decrease gaps, and correct genome assembly errors. The implementation of RAGA (available at https://github.com/wzxie/RAGA) significantly streamlines population-scale genome assembly workflows, providing a robust foundation for comprehensive pan-genomic investigations. This tool represents a substantial advancement in making large-scale genomic studies more accessible and efficient.

Clubroot is a devastating soil-borne disease that parasitizes cruciferous crops, posing a severe threat to rapeseed production. To date, no clubroot-resistant (CR) genes have been successfully cloned in cabbage (). This study aimed to identify CR genes and elucidate the molecular mechanisms underlying clubroot resistance in . A BC mapping population was developed from a cross between CR cabbage W12 and clubroot-susceptible cabbage Z5. A major CR locus, , was identified on chromosome C07 by Bulk Segregant Analysis. Subsequently, the was fine-mapped to a 170.2-kb interval using linkage analysis. Two candidate genes, and , exhibiting sequence variations between the parents were induced upon infection. Overexpression of (CR cabbage W12) in and rapeseed significantly reduced the disease index compared to the wild type (WT) after inoculation. In contrast, plants overexpressing (the susceptible cabbage Z5), , and exhibited symptoms comparable to those of WT, indicating that is a CR gene. RNA-seq analysis revealed that may mediate resistance to by modulating pathways related to reactive oxygen species, cell wall metabolism and modification, as well as secondary metabolite synthesis. In addition, long noncoding RNAs were found to play a significant role in regulating gene expression associated with interaction. This study broadens the repertoire of CR genes, offering a solid foundation for breeding CR cruciferous crops. Additionally, it provides novel insights into resistance mechanisms in response to infection in .

EIN3 binding F-box (EBF) proteins have been reported to play important roles in ethylene signaling pathway by mediating the ubiquitin-dependent degradation of EIN3-Like (EIL) proteins, but little is known about their roles in postharvest disease resistance. Here, we showed that SlEBF3 confers resistance against by ubiquitin-mediated degradation of SlBBX20. Overexpression of enhanced resistance to and increased the expression levels of genes related to PR (pathogenesis-related) and JA (jasmonic acid) in tomato, while knockdown of does not affect tomato resistance to . Further study demonstrated that SlEBF3 interacts with SlBBX20 the interaction between SlEBF3 and SlBBX20 promotes SlBBX20 degradation via the 26S proteasome, which confers enhanced resistance to through the JA signaling pathway mediated by the SlBBX20-SlMYC2-SlMED25 module. Meanwhile, SlEBF3 extends fruit shelf life by remodeling cell wall composition and promoting cuticular accumulation. Additionally, SlEBF3 is involved in carotenoid metabolism regulation by interacting with SlBBX20, SlRIN, SlFUL1, and SlTAGL1, which is independent of the degradation of EIL proteins. Overall, this study revealed the molecular mechanism by which responds to JA signaling to regulate resistance, enriched the roles of in the regulatory network of carotenoids metabolism, and provided new insights into the extension of fruit shelf life.

Hydrogen sulfide (HS), a gasotransmitter molecule, plays critical roles in stomatal closure and cellular bioenergetics. Alternative splicing (AS) is a key regulatory mechanism during plant development and stress responses; however, the interplay between HS signaling and AS in drought tolerance remains unexplored in Chinese cabbage. In this study, we found that the mitochondrial inner membrane enzyme succinate dehydrogenase (SDH) responds to HS signaling during stomatal closure. Silencing of impaired the effects of HS on stomatal closure, SDH activity, and ATP production. RNA-Seq analysis revealed that HS modulates the AS of , resulting in transcript variants with differential expression. Overexpression of and in enhanced drought resistance, whereas had no significant effect. HS enhanced SDH activity and ATP production, promoted stomatal closure, and reduced excess reactive oxygen species (ROS) in OE- and OE-C lines but not in OE-. Furthermore, biotin-switch assays demonstrated that HS induced persulfidation of BrSDH1-1A and BrSDH1-1C, with no effect on variant BrSDH1-1B. These findings reveal a novel regulatory mechanism by which HS modulates splicing to mediate stomatal closure and improve drought tolerance, offering valuable molecular insights for enhancing stress resilience in horticultural crops.

Aquaporins (AQPs) are integral membrane channel proteins that facilitate water transport and contribute significantly to plant adaptation under drought stress. However, the evolutionary origins and mechanisms of functional diversity of this gene family remain to be elucidated. A comprehensive genome-wide analysis was therefore performed on 104 representative species spanning the green plant lineage, from algae to angiosperms. This study used two datasets: Taxon I (algae to eudicots) and Taxon II (angiosperms including drought-tolerant and drought-sensitive plants). By systematically optimizing the gene structure, codon preferences, motifs, and cis-elements of these two datasets, the molecular mechanisms of AQP genes in plant adaptation evolution and drought-tolerance evolution were revealed. The results of phylogenetic analysis indicate that the AQP gene family is divided into five main subfamilies: PIPs, NIPs, TIPs, SIPs, and XIPs. Through in-depth analysis of the evolution characteristics of each subfamily, it was found that the emergence and loss of different subclusters are related to the ecological adaptation needs of specific species. By systematically analyzing the evolutionary history of the members of PIPs and TIPs subfamilies and subclusters, and combining their gene expression patterns, it was confirmed that PIP2, TIP1, and TIP4 subcluster members exhibit more significant expression response characteristics under drought stress. This study is the first to analyze the evolutionary patterns and drought-tolerance mechanisms of the AQP gene family at a multidimensional scale, providing important molecular targets for crop drought resistance breeding.

Fusarium wilt caused by () is one of the most destructive diseases in global banana production. The response of root system to infection through gene expression in multiple cell types is crucial for understanding the disease resistance mechanism in banana. Here, we reported a single-cell transcriptional landscape of banana root tips in response to f. sp. tropical race 4 ( TR4) infection. We characterized 10 major cell types from 19 cell clusters. We explored differentiation trajectories of meristematic cells, root cap cells, and pericycle cells through pseudotime analysis, and identified pericycle cell as the dominant root cell type under TR4 infection. Moreover, we identified 11 co-expression regulatory networks, of which eight were significantly associated with TR4 response. Specifically, was co-expressed with two Zn -dependent genes ( and ) in M5 module, which was associated with pericycle cell type and responded to TR4 infection. Further analysis demonstrated that MaKAN4 protein could interact with the promoters of and to promote their expression levels, highlighting a crucial role of in banana disease resistance by regulating the Zn -dependent / module. These findings provide a comprehensive view of cell fate determination in banana root tips and highlight novel insights into the regulatory mechanisms of major cell types in response to TR4 infection, laying a useful foundation for developing disease-resistant banana cultivars.

Centromeres are essential for centromere-specific histone H3 (CENH3) recruitment and kinetochore assembly, ensuring accurate chromosome segregation and maintaining genome stability in plants. Although extensively studied in model species, the structural organization of centromeres in nonmodel plants, such as fruit trees, remains poorly explored. Our previous study revealed that jujube centromeres lack the typical tandem repeat (TR)-rich structure, complicating their precise identification. In this study, we updated the genome assembly of jujube ( Mill. 'Dongzao') to a haplotype-resolved T2T version, enabling accurate mapping and comparison of centromeres between haplotypes using CENH3 ChIP-seq. These centromeres, ranging from 0.75 to 1.40 Mb, are largely conserved between haplotypes, except for a localized inversion on chromosome 10. Unlike the TR-rich centromeres found in many plant species, jujube centromeres are predominantly composed of -type long-terminal repeat retrotransposons (LTR-RTs). Among these, we identified a centromere-enriched LTR family, centromeric retrotransposons of jujube (CRJ), which is particularly abundant in terminal LTRs compared to the internal transposon regions. Comparative analysis across plant species revealed that centromeric retrotransposons primarily fall into three subfamilies-, , and -highlighting strong subfamily specificity. Notably, early insertions of CRJ-derived LTR segments contributed to the formation of TR-like structures, suggesting a mechanistic link between transposable elements and the evolution of centromeric tandem repeats. This work provides the first in-depth characterization of a TE-dominated centromere architecture in a fruit tree, offering new insights into the diversity and evolution of plant centromeres.

Bee pollination enhances crop productivity and food security globally. However, its impact on pollen performance within pistil tissues and the underlying regulatory mechanisms remain unclear. In this study, artificial self-pollination yielded the highest pollen deposition on stigmas (119879.33 ± 43037.92 grains), followed by bee pollination (95464.60 ± 3985.01 grains). Conversely, bee pollination achieved the highest seed set rate (55.21% ± 1.84%), significantly exceeding the artificial self-pollination rate (7.27% ± 1.87%). A positive correlation was observed between pollen load on the stigmatic pollination band and seed set rate. Bee pollination delivers ample high-quality pollen to the stigmas of oil tree peony, enhancing seed production. Moreover, a trend high correlation was observed between pollen deposition on the stigmatic pollination band and seed set rate. Fluorescence microscopy and endogenous hormone analyses revealed that bee pollination stimulated a rapid increase in ZR, IAA, and GA levels in the pistil tissues, promoting pollen germination and pollen tube growth. Transcriptome analysis identified , a key candidate gene involved in pollen development, in the pistil tissues after bee pollination. This gene exhibits high homology with genes found in other crops. The gene localizes to the cell membrane, validating earlier predictions, and exhibits strong transcriptional activity. Silencing disrupts pollen development in 'Fengdan' manifesting as structural defects in pollen walls and significantly reduces pollen viability. In conclusion, bees enhance fertilization in oil tree peony by delivering high-quality pollen that promotes germination and pollen tube growth. Crucially, we identified , a membrane-localized key gene regulating pollen development. This study establishes a crucial foundation for deciphering the molecular mechanisms by which bee pollination and phytohormone signaling mediate pollen development.

Cytokinins play crucial roles in regulating the flower bud differentiation in fruit trees. However, the molecular mechanisms by which cytokinins promote flowering in plants are largely unknown. The litchi ( Sonn.) is a typical subtropical fruit tree that suffers from severe alternate fruiting due to unstable flowering. Here, we developed a novel alternate-fruiting management, which can ensure 100% flowering of the on-year trees, while the off-year trees hardly flower at all. The abundance of two types of cytokinins (tZR, iPR) and expression in the leaves of on-year trees were continuously increased throughout the period of floral bud physiological differentiation. In contrast, these corresponding indicators in off-year trees were maintained at a significantly lower level. Exogenous application of 40 mg/kg 6-BA significantly promoted flowering and increased expression level in the leaves of the off-year trees. , encoding a pivotal rate-limiting enzyme in cytokinin biosynthesis, was identified as the key gene determining the differences in cytokinin levels between on-year trees and off-year trees. Interestingly, we discovered that both and are directly activated by LcARR11, a type-B cytokinin response factor, as demonstrated through both and assays. Furthermore, ectopic expression of in resulted in elevated expression and cytokinin content, alongside increased expression and earlier flowering. Collectively, our findings suggest that cytokinin-mediated promotion of flowering in litchi might be orchestrated by LcARR11, which could promote floral bud physiological differentiation by activating both and .

Apple replant disease (ARD) poses a major threat to global orchard productivity, yet its biological causes remain poorly understood. Although microbial dysbiosis in replant soils has been recognized as a major contributing factor, the specific pathogenic agents involved and the efficacy of synthetic microbial communities in mitigating ARD remain unclear. In this study, we integrated physiological, transcriptomic, metabolomic, and microbiome analyses to investigate the effects of replant soils on the growth of rootstock M26. Absolute quantification amplicon sequencing of 16S rRNA and ITS regions revealed a marked decline in rhizospheric microbial diversity in replant soils compared to fallow controls, accompanied by an enrichment of fungal genera such as , , and . Pathogenicity assays and seedling colonization experiments verified strong pathogenicity for five isolates- sp., , , , and -implicating them as potential causal agents of ARD. High-throughput culturing and confrontation assays were used to isolate and screen candidate microbial antagonists. A synthetic microbiota (SynMs) composed of 12 bacterial strains and sp. was developed. Inoculation with SynMs significantly inproved plant height by 133% ( < 0.05) and total root length by 186% ( < 0.01), and effectively suppressed pathogen proliferation of the five pathogenic isolates in replant soils. Collectively, these findings identify key fungal pathogens underlying ARD and propose a sustainable microbiota-based strategy for its effective mitigation, offering both mechanistic insights and practical solutions for microbiome-informed orchard management.

The precise manipulation of genome sequences through gene targeting (GT) is beneficial; however, the low efficiency of homology-directed repair (HDR) in seed plants has made GT difficult to achieve. Generation of double-strand breaks (DSBs) at the target DNA site of interest represents a promising approach to facilitate HDR-mediated GT in organisms. Despite recent advances, GT remains a significant challenge in seed plants. To address these challenges, we propose that the efficiency of CRISPR/Cas9-mediated GT could be enhanced by the exclusive selection of plants that exhibit high levels of HDR activity. To test this hypothesis, a surrogate screening system was developed, which consists of a nonfunctional split-selection marker gene. In this system, DSBs generated by CRISPR/Cas9 at the linker sequence of the tandem repeat will be repaired via single-strand annealing (SSA), a subtype of HDR, resulting in the achievement of antibiotic resistance in plants. This approach allows for a 2- to 23-fold increase in precise and heritable GT efficiency in Arabidopsis and rice. The results indicate that screening with SSA-mediated surrogate system can enrich cells and plants with high HDR activity as well as DSB activity, thus facilitating the establishment of highly efficient GTs at target loci in these plants.

Plant-parasitic root-knot nematodes ( species) are highly polyphagous parasites that alter cellular identity of terminally differentiated root cells to induce the formation of giant cells and knot-like structures known as galls, whose ontogeny remains largely unknown. In this study, we generated single-nucleus RNA-seq data of galls and neighboring root tissues at two distinct stages of infection of tomato () plants. Analysis of 35 393 high-quality nuclei resulted in the identification of three stele-associated cell clusters that captured young and more differentiated giant cells, where 772 genes were preferentially expressed. Giant cell-specific expression patterns of a set of these genes were validated using promoter activity assays. We used pseudotime analysis to trace how gene activity changes as giant cells develop. Developmental trajectory analysis revealed a gradual activation of more complex gene regulatory networks as young giant cells adopt specific fates and become more differentiated. Functional assays using gene silencing confirmed the functional importance of giant cell-expressed genes in mediating plant susceptibility to . Cell type-specific gene expression analysis revealed that xylem, phloem, stele, endodermal, and protophloem cells undergo extensive transcriptome reprograming, which facilitates coordinated cellular responses to nematode infection, including immune signaling, structural support, and metabolic adjustments. Together, our analyses represent the first single-nucleus transcriptomic map of nematode-induced giant cells and provide novel insights into the molecular events leading to the formation of a new plant organ and feeding cells orchestrated by an animal parasite.

BAK1 was initially identified as a coreceptor of BRI1 in regulating the brassinosteroid-triggered signaling pathway in . Over the past two decades, increasing pieces of evidence have demonstrated that BAK1 and its close paralogs form receptor-coreceptor complexes with distinct ligand-binding receptors. Through ligand-induced heterodimerization with receptor-like protein kinases or receptor-like proteins, BAK1 thereby regulates a variety of physiological events such as plant development, immunity, and stress responses. Thus, BAK1 plays a central role in directly governing the trade-offs of multiple signaling pathways. Deciphering the molecular mechanisms underlying how BAK1 coordinates plant growth and defense, with specific emphasis on its coreceptor functions, holds significant potential for future advancements in crop breeding. This review seeks to explore the latest insights into how BAK1 impacts the intricate equilibrium between plant development and immunity, as well as its roles in coordinating the activation of pattern-triggered immunity and effector-triggered immunity or programmed cell death. Furthermore, it offers significant perspectives on why BAK1 has been chosen as a shared component in determining plant growth and defense mechanisms across model plants to horticultural crops.

Epigenetic modifications, such as DNA methylation, histone modifications, chromatin remodeling, and RNA-associated silencing, play critical roles in regulating gene expression without altering the DNA sequence. In horticultural crops, these mechanisms control key biological processes, including fruit development and ripening, flowering time, stress adaptation, and phenotypic plasticity. Driven by high-throughput sequencing and multi-omics technologies, researchers have begun to uncover the dynamic landscape of plant epigenomes. Notably, the Encyclopedia of DNA Elements (ENCODE) project was developed to systematically map functional elements within the genome. Inspired by this initiative, similar strategies have been increasingly applied to plants to identify regulatory elements, chromatin states, and transcriptional networks. This review integrates recent findings on epigenetic regulation in model and horticultural species, emphasizing the role of epigenomic tools and ENCODE-like approaches in annotating cis-regulatory elements, epigenetic markers, and long non-coding RNAs (lncRNAs). We discuss how epigenetic modifications mediate developmental transitions and responses to environmental cues. Finally, we propose a framework for integrating ENCODE-derived insights with precision breeding to improve yield, quality, and stress resilience in horticultural crops. These advancements offer exciting opportunities for translating epigenomic knowledge into practical crop improvement strategies.

Tea plant [ (L.) O. Kuntze] is a globally important crop but is severely threatened by infestations, which impact yield and safety. However, the response of tea plants to aphid feeding remains largely unexplored. This study investigates the feeding behavior of on different cultivars and identifies 'Huangjinya' and 'Qiancha 1' as susceptible and resistant cultivars, respectively. Transcriptome analysis revealed that was significantly upregulated in response to infestation. biochemical assays demonstrated that encodes a flavonoid 7-glycosyltransferase that catalyzes the conversion of flavonoids and UDP-glucose into flavonoid 7--glucosides. , silencing significantly reduced flavonoid glycoside accumulation. To further clarify the role of in tea plant resistance to , we used tobacco and tea flowers to evaluate aphid feeding and reproduction under chemical treatment, gene silencing, and gene overexpression conditions. Statistical analysis showed that, compared with flavonoids, the application of flavonoid 7--glycosides significantly reduced reproductive capacity. Furthermore, compared with the control, overexpression of significantly reduced the reproductive ability of aphids, while its silencing increased reproductive rates. Overall, our findings demonstrate that mediates flavonoid glycosylation and enhances insect resistance in tea plants by increasing flavonoid glycoside levels, offering new insights into the role of flavonoid glycosides in the insect resistance of .

The domestication of ornamental plants is primarily driven by aesthetic values and usually involves frequent hybridization events. spp., a globally famous woody flower, exemplifies the complex origins and extensive phenotypic variation. Here, based on the whole genome resequencing 220 germplasms, we developed Camellia21K, a high-density SNP array enabling cost-effective genome-wide genotyping. We demonstrated that Camellia21K accurately resolves 69 cultivars with complex hybridization histories. For molecular identification of closely related varieties, we developed a set of fingerprinting SNPs to support variety discrimination. To dissect the genomic basis of ornamental traits, we performed a genome-wide association study (GWAS) analysis of five leaf shape traits using the Camellia21K array and screened 31 SNP loci significantly associated with the traits. Further, by analyzing the genotypes of the SNP loci and the haplotypes of the surrounding segments, we identified potential genes regulating leaf tip length, thus demonstrating the versatility of the array. To enhance breeding efficiency, we evaluated and optimized four genomic selection (GS) models for leaf trait prediction. We found that the number of SNPs and model selection significantly affected prediction performance, with optimal predictive accuracy (PC) from 0.362 to 0.542, which was positively correlated with heritability. Finally, we integrated fixed-effects SNPs from GWAS and found significant enhancement of PC (24.7%-64.7%), indicating that the combination of GWAS and GS is indispensable for precision breeding applications. We demonstrated that Camellia21K is effective in discriminating the origin of varieties, in genetic analysis of traits and in genomic prediction, and thus informative for crop breeding.

UDP-dependent glycosyltransferases (UGTs) play a critical role in producing glycosylated metabolites that mediate plant-environment interactions. Recent studies have examined the role of genes across various plant genomes. However, the evolutionary history and functional divergence of the pan-gene family in the genus have not yet been explored. This study integrated data from 61 tomatoes and 9 representative genomes, ranging from algae to angiosperms, to identify 12 073 genes. The phylogeny of the UGT gene family reveals a clear evolutionary trajectory from unicellular algae to ferns, mosses, gymnosperms, and angiosperms. The study identified a significant number of tomato-specific genes and explored the expansions of and subfamilies. The entire genes (10 769) in tomato were classified into 118 orthologous gene groups, including 58 core, 31 softcore, 10 dispensable, 19 private orthologous gene groups, and the core groups contained 7811 genes, representing 72.53% of the total genes. Analysis of gene family expansion revealed that whole-genome triplication and tandem duplication events play significant roles in the expansion of the gene family. Selection pressure analysis revealed that the genes experienced purifying selection in the genus . Additionally, expression profiles of some genes in different tissues demonstrated expression divergence of multicopy genes across different subfamilies due to the increase in gene dosage. Subcellular localization prediction revealed that most genes are localized in the chloroplast. These findings provide critical insights into the evolution and function of the genes in tomato, laying a foundation for further exploration in adaptive evolution.

Kiwifruit bacterial canker, caused by pv. actinidiae, poses a critical threat to global kiwifruit production. Previous studies implicated jasmonic acid (JA) signaling in kiwifruit responses to this pathogen; however, the molecular mechanisms underlying JA-mediated regulation remain largely unclear. Here, we identified and characterized AcJAZ2L2, a pivotal jasmonate-signaling regulator that confers substantial resistance against pv. actinidiae. Transcriptomic profiling coupled with consensus co-expression network analysis revealed that AcJAZ2L2 expression is uniquely up-regulated in resistant kiwifruit cultivars after pathogen infection. Functional validation through genome editing with the clustered regularly interspaced short palindromic repeat-associated protein 9 nuclease and, through transgenic overexpression, confirmed the essential role of AcJAZ2L2 in resistance. Specifically, lines overexpressing AcJAZ2L2 displayed markedly reduced disease symptoms, lower pathogen colonization, and decreased stomatal density, whereas knockout lines exhibited increased susceptibility. Mechanistically, AcJAZ2L2 directly interacts with AcMYC2-like transcription factors, repressing downstream JA-responsive genes (AcVSP2L1 and AcVSP2L2) and maintaining stomatal closure to prevent pathogen entry. Promoter analysis further revealed cultivar-specific allelic divergence that drives differential AcJAZ2L2 transcriptional activation, explaining genotype-dependent resistance levels. Our findings establish a novel JAZ-MYC regulatory module that links JA signaling to stomatal immunity in kiwifruit and provide precise genetic targets for breeding cultivars with enhanced resistance to bacterial canker.

Root tips, which represent the initial stage of taproot development, serve as an ideal model for investigating plant growth and secondary metabolism. However, studies of root tips in species have been limited, restricting our understanding of cell fate transitions during early root development and the cellular heterogeneity associated with ginsenosides biosynthesis. To address this gap, we conducted single-cell RNA sequencing (scRNA-seq) and spatial metabolomics analyses on the root tips of three species: , , and . Our research reconstructed the developmental trajectory of the early endodermis and revealed epidermis-specific expression patterns of key enzyme genes involved in ginsenosides biosynthesis. We identified several novel transcription factors (TFs): (which positively regulates endodermis suberization) and / (positive regulators of ginsenosides biosynthesis), validated by dual-LUC reporter and electrophoretic mobility shift assay (EMSA). Conserved and divergent ligand-receptor interaction patterns across the three species were discovered, with the gene family exhibiting tissue- and species-specific expression. Cell-specific genes expression was confirmed by RNA hybridization. Mass spectrometry imaging (MSI) mapped ginsenosides spatial distribution, while LC-MS/MS verified species-specific biosynthesis. This study presents a single-cell transcriptional landscape of early differentiation and cell type-specific ginsenosides accumulation in the genus.

accumulates abundant α-linolenic acid (ALA) and its metabolic volatiles, which hold significant potential for applications in healthcare and agriculture. However, the genetic basis underlying their biosynthesis has not been systematically investigated. Here, we present a high-quality genome assembly for (614.46 Mb). Despite a high repetitive sequence content (70.46%), it avoided excessive expansion due to the efficient elimination of long terminal repeat retrotransposons. Phylogenomic analyses revealed that experienced two whole-genome duplication (WGD) events, with WGD-derived genes predominating in oil biosynthesis. Notably, , a WGD-duplicated fatty acid desaturase, was identified as a key seed-specific gene for ALA biosynthesis. Its regulation by the transcription factor was functionally validated through yeast one-hybrid, luciferase, β-glucuronidase, and transgenic functional assays. Furthermore, (E)-2-hexenal, the predominant ALA-derived volatile in leaves, exhibited potent antifungal activity against (minimum inhibitory concentration: 0.188 ml/l), with its biosynthesis linked to . These findings provide genomic and functional insights into ALA biosynthesis and metabolic volatiles in , supporting its potential in sustainable agriculture and bioactive compound development.