Premature bolting under elevated temperatures is a major constraint on lettuce production because it shortens the vegetative harvest period and reduces leaf quality. Bolting, floral transition, and flowering are closely connected but distinguishable processes that are regulated by overlapping genetic and environmental factors. Recent advances in lettuce genome assemblies, super-pangenomes, population resources, multi-omics analyses, and genome editing have enabled a more integrated understanding of this complex trait. This review summarizes the genomic architecture and natural variation associated with reproductive timing, the integration of temperature with photoperiodic and stress-responsive pathways, and the hormonal and metabolic changes that support inflorescence stem elongation. Particular attention is given to phytochrome-associated variation, heat-responsive transcription factors, LsSOC1, auxin and gibberellin signaling, and carbohydrate remodeling. We further discuss how these findings can inform genome editing, marker-assisted and genomic selection, and cultivation strategies. Together, current evidence indicates that heat-induced bolting arises from coordinated environmental signal integration, reproductive commitment, and hormone-dependent stem growth rather than from a single linear thermosensory pathway.
Several plant-derived metabolites that enhance plant resilience in response to biotic and abiotic stresses have been developed as senolytics after their apoptotic effects were discovered. Several ginsenosides have also been reported to exert apoptotic effects. We therefore hypothesized that ginsenosides from wild P. ginseng, which survives for long periods in harsh environments, may act as senolytics for skin longevity. The phytochemical characteristics of wild P. ginseng were investigated by LC–MS/MS-based comparison of 38 ginsenosides between cultivated and wild P. ginseng. Wild P. ginseng had a 2.7-fold higher total ginsenoside content in the underground parts. Intermediate ginsenoside metabolites, particularly ginsenoside C-Mc1, were especially abundant and characteristic of wild P. ginseng. To isolate trace amounts of ginsenoside C-Mc1, adventitious roots were mass-cultured from tissue segments of wild P. ginseng. The cultured roots were subjected to a four-step column chromatography procedure (HP-20, silica gel twice, and reversed-phase) to obtain both a crude saponin fraction and purified ginsenoside C-Mc1. These were then combined to prepare a C-Mc1-fortified ginsenoside preparation derived from wild ginseng adventitious roots, designated wild ginseng ginsenosides (WGGs), thereby increasing the level of the naturally occurring characteristic metabolite C-Mc1. The senolytic activity of WGGs was evaluated in senescence-induced fibroblasts. While WGGs maintained apoptotic cell levels in normal fibroblasts at 8–9
The periderm protects storage roots from water loss and pathogen invasion, but the developmental timing of periderm initiation and suberin deposition has not been systematically characterized in Panax ginseng (P. ginseng), a perennial medicinal crop cultivated for several years. In this study, periderm development in first- and second-year P. ginseng roots was examined using histological staining and Fluorol Yellow fluorescence microscopy to visualize suberin deposition. Periderm initiation was detected at approximately 3 weeks after germination (WAG), before rupture of the primary cortex at 4 WAG. Suberin deposition began during early periderm differentiation and progressively increased as the storage root expanded. Quantitative analysis showed a continuous increase in phellem cell layers, indicating sustained cork cambium activity during secondary growth and after dormancy release. These results demonstrate that early periderm formation and suberin deposition establish a continuous protective barrier before primary tissue rupture, while progressive phellem expansion reinforces root protection during ginseng development. These findings provide an anatomical framework for future studies on periderm formation, root surface integrity, and their potential relevance to root quality in ginseng cultivation.
Doubled haploid (DH) technology rapidly generates completely homozygous lines and is widely used in modern crop breeding. In Brassica crops such as Pak–choi (Brassica rapa ssp. chinensis), traditional DH methods, such as microspore culture, remain highly genotype–dependent and technically challenging due to whole–genome triplication, gene redundancy, and low regeneration efficiency. Consequently, the development of in planta genome–editing strategies is vital for overcoming these limitations and providing a practical alternative to conventional tissue culture–based approaches. Here, we review CRISPR/Cas9–mediated haploid induction in plants, focusing on three pathways: centromere–mediated genome elimination by modifying the centromere–specific histone CENH3, fertilization–related maternal haploid induction via the gynoecium–expressed patatin–related phospholipase pPLAIIγ, and male gametophyte–dependent haploid induction controlled by DMP genes. We compare these pathways regarding their molecular mechanisms, developmental timing, gene essentiality, and suitability for genome editing, with particular emphasis on Brassica species. Special attention is given to haploid inducer gene homologs in Pak–choi and to the challenges imposed by ancestral genome triplication in Brassica rapa. Finally, we discuss how to use CRISPR/Cas9–based haploid induction systems in Brassica breeding programs, highlighting their value for accelerating doubled haploid production and functional genomics in genetically complex crop species.
The expansion of wheat cultivation has intensified the challenges of traditional disease detection, highlighting the need for efficient intelligent monitoring. Automated disease recognition is thus critical for smart agriculture, yet existing methods often exhibit delayed detection and high false-positive rates in field applications. This study introduces a novel recognition approach based on an enhanced RegNet architecture with a dual-attention mechanism. The model first expands the pooling layer’s input units and output dimensions to improve complex-feature processing and spatial detail capture. Second, a deformable attention mechanism refines feature discrimination. Third, a graph attention network facilitates node-level interaction within graph-structured data. Experimental results demonstrate an overall classification accuracy of 97.72
Understanding how plants maintain whole-plant water balance under heterogeneous soil moisture, such as partial root-zone drying (PRD), is crucial for improving crop drought resilience. This study investigated the physiological and molecular mechanisms underlying PRD adaptation in foxtail millet (Setaria italica L.), using a split-root hydroponic system with well-watered (WW), uniform drought (WD), and PRD (WM, with one compartment in osmotic stress and one well-watered) treatments. PRD plants maintained shoot biomass closer to WW levels than WD plants, demonstrating effective adaptation. This was achieved through a spatially coordinated strategy: PRD significantly increased whole-plant root exudation compared to WD concurrent with a reduction in leaf transpiration. Physiological and transcriptomic analyses revealed asymmetric resource allocation and gene expression. Roots in the drying compartment (WM-D) accumulated abscisic acid (ABA) and exhibited upregulated expression of ABA biosynthetic genes (NCEDs), whereas roots in the WM-W compartment showed significant accumulation of soluble sugars (fructose, glucose, sucrose) and upregulated expression of sugar transporter (SiSUT3), starch catabolism (SiBAM2) genes, and multiple aquaporins. Compartment-specific sugar profiling showed that fructose levels in WM-W roots were 47–56
Sweetpotato (Ipomoea batatas [L.] Lam) is known for its high and stable yields, robust adaptability, and rich nutritional value, making it an ideal food source acknowledged by the World Health Organization. But soil salinization and the global temperature change have limited their ability to grow. Universal stress proteins (USP) play a key role in organismal responses to adverse environments, yet research on them in the field of sweetpotato is extremely limited. The IbUSP32 gene was successfully separated from sweetpotato in this study. The IbUSP32 was shown to be expressed in different tissues of sweetpotato by the qRT-PCR analysis, with comparatively high expression in the roots of the plant. When stresses such as salt, drought, high temperature, low temperature, ABA, and CdSO4 were present, the gene was upregulated. Under mannitol and salt stress, the wild type of Arabidopsis showed notably lower germination rate, root length, and fresh weight compared to the overexpression lines of IbUSP32. Heterologous overexpression of IbUSP32 upregulated the mRNA levels of antioxidant enzyme genes AtSOD, AtCAT, and AtAPX, and increased antioxidant enzyme activity under soil drought and salt stress. It also reduced the levels of malondialdehyde, superoxide anion (O2·−), and hydrogen peroxide (H2O2), and accumulated osmoregulatory substances, proline, and soluble sugars. The results showed that IbUSP32 increased stress tolerance by activating the osmotic regulation pathway and the antioxidant defense pathway. In addition, overexpression of IbUSP32 significantly upregulated CBL‑interacting protein kinase gene AtCIPK6 expression in Arabidopsis, indicating the two genes have possible functional association. In summary, this study identifies IbUSP32 as a candidate gene for improving stress tolerance in sweetpotato.
Extracellular vesicles (EVs) are crucial mediators of intercellular communication in various biological systems, including plants, where they facilitate the sharing of proteins, lipids, and nucleic acids. While plant-derived EVs (PDEVs) have documented intrinsic functions in stress and host–pathogen responses in model species, methods for isolating and characterizing these vesicles from agriculturally important crops, such as cucumber (Cucumis sativus) apoplast, remain unoptimized. This methodological study evaluates and compares current protocols to establish an efficient and standardized technique for PDEV isolation from apoplastic washing fluid (AWF). We directly compared three key isolation methods PEG precipitation, ultracentrifugation, and size exclusion chromatography (SEC) assessing their performance based on cost, scalability, purity, and vesicle preservation/integrity. Transmission electron microscopy (TEM) and immunoblot analyses revealed the morphology and proteome profile of the isolated EVs, validating vesicle integrity and marker enrichment obtained from each method. Results confirm that while ultracentrifugation serves as the gold standard for isolation, it risks compromising vesicle integrity. Conversely, PEG precipitation offers a low-cost, scalable method, despite co-precipitating soluble proteins. We demonstrate the value of using SEC as a critical secondary purification step to enhance the purity of the isolated EVs. In conclusion, TEM and immunoblot characterization together supported that SEC-based purification yields the most morphologically intact and biochemically enriched EVs. We further demonstrate the cross-species applicability of AtTET8 and AtPEN1 antibodies in cucumber and validate CsTET8 as an EV-associated marker, underscoring the importance of standardized workflows for crop PDEV research. Establishing a reliable, optimized protocol is an essential first step to enable rigorous functional studies of crop EVs and to unlock their potential applications in both biomedicine and sustainable agriculture.
Oil palm, a tropical woody crop, faces challenges in tissue culture regeneration due to the genotype backgrounds and low regeneration efficiency. Somatic embryogenesis (SE) is a key process in plant regeneration and propagation. However, identifying plant materials capable of forming embryogenic callus is challenging and cannot be reliably determined through visual or macroscopic observation alone. In this study, we established histological markers to assess tissue culture materials based on their microstructural and cytological characteristics. Paraffin sectioning was performed on various oil palm explants (zygotic embryos, shoot tips, young leaves, young root tips, young female flowers, young male flowers, root primordia, and buds), as well as on embryogenic/non-embryogenic calli, and somatic embryos for histological analysis. Results revealed that meristematic stem cells showed explant-specific localization. In zygotic embryos, these cells were localized in the cotyledon vascular bundles, whereas in the shoot tips, they were concentrated in the apical meristem. Callus induction sites varied depending on the type of explant, such as the hypocotyl/cotyledon incisions in zygotic embryos, and pericycle cells in roots. Embryogenic callus exhibited a regular shaped, dense cytoplasm, a high nucleus-to-cytoplasm ratio, abundant inclusions, and significant accumulation of starch grains and proteins, which distinguished them from the irregular, vacuole-rich non-embryogenic callus. Somatic embryos developed from surface or internal cells of embryogenic callus via globular, heart-shaped, torpedo, and mature embryo stage. These findings provide reliable histological markers for distinguishing embryogenic potential in oil palm tissues, offering a valuable tool to improve tissue culture regeneration and accelerate clonal propagation.
Electrical stimulation represents an emerging physical biotechnology for enhancing plant in vitro culture and micropropagation, offering potential solutions to persistent challenges including genotype-dependent recalcitrance and suboptimal metabolite yields that limit conventional chemically-based approaches. This review critically synthesizes current knowledge on the applications and mechanisms of electrical stimulation modalities including direct current (DC), alternating current (AC), and pulsed electric fields (PEF), across diverse tissue culture contexts. We examine documented effects on shoot and root organogenesis, somatic embryogenesis, protoplast transformation, and secondary metabolite production, where treatments have achieved 2–5-fold enhancements in regeneration efficiency and significant increases in valuable compounds such as taxanes, camptothecin, and phenolics. The mechanisms proposed to underlie these responses include transient membrane permeabilization under pulsed electric field conditions, electrically induced changes in ion flux and Ca²⁺ signaling, possible alterations in hormone distribution, and ROS-associated stress signaling. However, except for electroporation under appropriate pulse conditions, several of these pathways remain incompletely resolved and may be context dependent. Despite reproducible benefits demonstrated over four decades, the field remains transitional between laboratory proof-of-concept and practical implementation, hindered by critical methodological inconsistencies including lack of standardized electrical parameters, inadequate reporting of electrode configurations, and insufficient mechanistic understanding of how exogenous fields interface with endogenous bioelectric networks. Species- and genotype-specific variability further complicates protocol optimization, while scale-up to commercial bioreactors presents unresolved engineering challenges. Future progress depends on developing international reporting guidelines, conducting rigorous multi-laboratory validation studies, and employing multi-omics approaches to elucidate signaling pathways, ultimately establishing electrical stimulation as a standardized tool for plant biotechnology applications.
Pinellia ternata is a high-value medicinal plant increasingly produced under controlled cultivation systems to address rising market demand. However, hydroponic substrates may act as reservoirs for microbial colonization, potentially affecting plant health and raw material quality. In this study, we characterized culturable fungal contaminants (molds and yeasts) associated with sponge substrates used for nutrient film technique (NFT) hydroponic cultivation of P. ternata. Fungal communities were investigated using a multi-media isolation approach across seven agar formulations, followed by macroscopic and microscopic phenotyping and molecular identification using 18 S (SSU) and/or 28 S (LSU) rDNA sequencing. After 7 days of culture, diverse colony morphotypes were recovered, and a non-redundant panel of 20 purified isolates was assembled. Among these, 12 isolates yielded interpretable sequences with high similarity to reference accessions (query coverage 98–100
This study aimed to identify, collect, and conserve semi-oriental tobacco genetic resources. In 2022, systematic surveys were conducted across ten provinces in Türkiye (Hatay, Adıyaman, Diyarbakır, Bingöl, Batman, Mardin, Bitlis, Muş, Trabzon, and Burdur) where semi-oriental tobacco populations are distributed. Seed material representing 126 genotypes exhibiting substantial phenotypic variation was collected. Genetic diversity was assessed using nine simple sequence repeat (SSR) markers and by morphological, agronomic, and chemical traits through two field trials conducted at the Tokat and Samsun locations. Dendrogram analysis based on SSR data revealed that the genotypes were clustered into eight major groups, with 82
Immune checkpoint inhibitors (ICIs) have demonstrated substantial therapeutic efficacy across multiple carcinomas by restoring T-cell–mediated antitumor immune responses. In particular, monoclonal antibodies targeting the programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1) axis effectively overcome tumor immune evasion mechanisms in several cancers. However, conventional mammalian cell-based antibody production platforms are limited in large-scale manufacturing due to high cost and complex processes. Plant-based expression systems have therefore emerged as attractive platforms for therapeutic protein production, owing to lower production cost and the absence of zoonotic infection risks associated with animal cells or bacteria. In this study, a pembrolizumab-based anti-PD-1 monoclonal antibody (mAb) was transiently expressed in Nicotiana benthamiana using an Agrobacterium-mediated protein expression system, and its purification and functional properties were evaluated. Transient expression was achieved by incorporating an endoplasmic reticulum retention signal (KDEL) into pEAQ-HT vector, followed by transformation into Agrobacterium tumefaciens and agroinfiltration of plant leaves. Antibody mRNA expression was confirmed by reverse transcription–polymerase chain reaction (RT-PCR), while protein expression was verified by Western blot analysis. Antibody size and purity were assessed using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and size-exclusion high-performance liquid chromatography (SEC-HPLC) analysis. Furthermore, protein-based enzyme-linked immunosorbent assay (ELISA) demonstrated that the plant-derived anti-PD-1 antibody specifically bound recombinant PD-1 protein. Collectively, these findings demonstrate the feasibility of a plant-based transient expression system for the production and purification of structurally intact, antigen-binding immune checkpoint antibodies.
Sweetpotato (Ipomoea batatas) is a genetically complex allohexaploid and a major source of calories and micronutrients in many stress-prone regions; however, it has historically lagged behind model crops in terms of genomic and molecular resources. This review synthesizes recent advances in sweetpotato proteomics, encompassing genome-enabled proteogenomics, stress physiology, storage-root development, metabolism, and postharvest biology. The advent of haplotype-resolved reference genomes, improved genome annotations, and optimized protein extraction protocols has enabled high-coverage LC–MS/MS–based workflows, including tandem mass tag–based quantification and data-independent acquisition. Together, these advances have transformed static protein catalogs into dynamic, systems-level network maps. Key discoveries include the proteomic “lignin–starch switch” underlying storage-root formation, the dual roles of sporamin and β-amylase in both storage and stress responses, and conserved stress-signaling modules that mediate cross-tolerance to heat, drought, cold, and pathogen or nematode attack. Despite these advances, several challenges remain. Matrix effects in phenolic- and starch-rich tissues, limited datasets on post-translational modifications, and the lack of single-cell and subcellular proteomic resolution collectively constrain functional interpretation and the translation of proteomic insights into breeding applications. Future priorities include the proteogenomic refinement of gene models, signaling studies centered on post-translational modifications, organelle- and tissue-resolved proteomics, and the integration of proteomics with multi-omics datasets and quantitative genetics. Such efforts will facilitate the identification of proteomic biomarkers for precision breeding in sweetpotato.
Plant productivity is severely constrained by diverse pathogens, among which oomycetes represent some of the most destructive threats to global agriculture. These filamentous microorganisms cause devastating diseases, including potato late blight and downy mildew, leading to significant yield losses in major crops. Successful infection relies on the formation of haustoria through which oomycetes deliver numerous effector proteins that manipulate host cellular processes and suppress both pattern-triggered and effector-triggered immunity. To date, three major classes of oomycete effectors, including RXLR, Crinkler, and CHXC, along with a putative class YxSL [RK], have been identified in oomycetes. These effector molecules, along with the recently identified apoplastic effectors, play key roles in governing compatible and incompatible interactions and establishing disease in the host plant. Plants perceive these effectors by deploying multilayered immune strategies including plasma-membrane localized pattern-recognition receptors (PRRs) and intracellular NLR receptors that induce redox- and hormone-regulated defense pathways, and dynamic remodeling of transcriptional and metabolic networks. Understanding these effectors and how they manipulate host defense is a prerequisite for the generation of disease-resistant plants. In this review, we discuss the recent progress in the oomycete effectors, their secretion system, and their targets in the plant cells. By integrating pathogen strategies with host immune responses, we highlight how effector-mediated manipulation of plant signaling provides new opportunities for breeding and engineering broad-spectrum and durable resistance against oomycete pathogens.
The in vitro propagation of Rosa hybrida ‘White Beauty’ is often hindered by physiological disorders such as shoot-tip necrosis and leaf chlorosis, primarily attributed to ethylene accumulation in sealed culture vessels. This study aimed to evaluate the effectiveness of three ethylene inhibitors—silver nitrate, aminoethoxyvinylglycine (AVG), and silver nanoparticles (AgNPs)—in alleviating these disorders and improving plant quality. Treatment with 3.0 mg L−1 AgNO3 and 2.0 mg L−1 AgNPs significantly reduced shoot-tip necrosis and leaf chlorosis, while enhancing shoot growth and soil plant analysis development (SPAD) values. While AVG treatments reduced necrosis, their impact on plant vigor was limited. The role of ethylene in shoot-tip necrosis and leaf chlorosis was confirmed through co-application with 1-aminocyclopropane-1-carboxylic acid (ACC), an ethylene precursor. AgNPs effectively reversed ACC-induced shoot-tip necrosis and leaf chlorosis. Additionally, treatment with 0.9 and 1.8 mg L−1 indole-3-acetic acid (IAA) significantly improved the rooting percentage, root number, and post-acclimatization growth, indicating enhanced root system architecture and transplant performance. These results demonstrate that the use of AgNO3 or AgNPs with IAA resulted in the optimal, high-quality micropropagation of Rosa hybrida ‘White Beauty.’ This strategy holds promise for commercial-scale production and warrants further investigation into the mechanisms underlying hormonal signaling and the field performance of regenerated plantlets.
The ferritin gene, designated BrFer1, was isolated from the cDNA library of Chinese cabbage leaves treated with sodium nitroprusside (SNP) nitric oxide donor. The BrFer1 cDNA encoded a putative polypeptide of 254 amino acids with a molecular mass of 28,183 Da. Its amino acid sequence was highly homologous to several ferritins from other cruciferous plant species like Brassica napus, B. juncea and Arabidopsis thaliana. BrFer1 gene expression was rapidly up-regulated in the leaves sprayed by SNP (0.5 mM) and gradually increased in dose-dependent. Exogenous FeCl2 distinctly increased BrFer1 expression, whereas FeCl3 did not affect the gene regulation. BrFer1 expression was early responsive to superoxide anion radical generator methyl viologen (MV), but it was hardly induced by hydrogen peroxide (H2O2). BrFer1 was inducible by FeCl2, SNP and MV, but FeCl2- and MV-triggered BrFer1 gene expression was mediated by iron-mediated signalling. Overexpression of BrFer1 in the transgenic Arabidopsis conferred tolerance to MV-triggered oxidative stress. These results suggest that NO-induced BrFer1 expression may play a role in dealing with superoxide stress in Chinese cabbage leaves.
Cotton serves as a crucial economic crop and raw material for the global textile industry. However, cotton leaf diseases such as brown spot, verticillium wilt, target spot, and fusarium wilt impair cotton yield and quality. For accurate early detection of cotton leaf diseases, we propose an integrated deep learning framework that integrates an improved ResNeXt convolutional neural network (CNN) with the efficient channel attention (ECA) module and call it RECA. Specifically, we first extracted fundamental and critical feature information using two convolutional layers paired with max-pooling layers. Second, we modified the ResNeXt network to reduce the number of computational parameters. Third, we incorporated three ECA modules into ResNeXt to enhance classification accuracy. Finally, the Softmax classifier was utilized to determine the categories of cotton leaf diseases. Experimental results show that the proposed method achieves an overall classification accuracy of 99.04
Papaya ringspot virus (PRSV) is one of the most destructive viruses of papaya, causing heavy yield losses and serious economic damage to farmers. Its impact is seen in almost all major papaya-growing regions across the world, making it a major constraint to papaya production and trade. This review presents a comprehensive overview of PRSV, beginning with its emergence, historical distribution and molecular biology, including genome structure and mechanisms of pathogenesis. The viral infection cycle, covering host invasion, replication and systemic movement is discussed alongside characteristic symptoms that facilitate field identification. Diagnostic approaches are evaluated, spanning conventional, serological and nucleic acid-based techniques, with an emphasis on advanced platforms such as next-generation sequencing. Strategies for managing PRSV are critically reviewed, including traditional control methods, breeding-based resistance and cross-protection. Special focus is placed on genetic engineering approaches, such as coat protein-mediated resistance, RNA interference and replicase gene-based resistance, which offer potential for long-term and stable control. Overall, this article brings together both classical and modern strategies for managing PRSV and aims to support future efforts to develop papaya varieties with durable resistance.
This study explored the application of tobacco plant expression system for production of dual target monoclonal antibody (mAb) proteins and computational analysis to evaluate their biochemical characteristics. The murine anti-colorectal cancer large single-chain antibody (CL) and H-13F6 human anti-Ebola virus full-size monoclonal antibody (EF) or anti-Ebola large single-chain antibody (EL) co-expressed in F1 plants through crossbreeding transgenic plants expressing each specific antibody. The binding activities of these antibodies to their respective target antigens were then examined. PCR analysis confirmed the presence of the CL gene and the heavy (HC) and light chain (LC) genes of EF in F1 plants resulting from the crossbreeding. Additionally, both CL and EL genes were identified in F1 plants produced through the crossbreeding of transgenic plants expressing each CL and EL. RT-PCR and immunoblot analyses confirmed the mRNA and protein expression of all transgenes in F1 plants. CL, EF, and EL were successfully purified from F1 plants. Indirect ELISA analysis revealed that CL × EF proteins exhibited binding activity to Ebola virus-specific antigenic protein, whereas CL × EL lost this binding activity. Conversely, both CL × EF and CL × EL proteins exhibited enhanced binding activity to the colorectal cancer-specific antigenic protein GA733. In addition, computational analyses [Molecular Dynamics (MD) and Molecular Mechanics Poisson-Boltzmann Surface Area (MM/PBSA) approach] were performed for Ebola virus (EBOV) antigen-antibody complexes and possible three combinations of Fc consistent with experimental results, supporting our assumption and interpretation.
