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Grafting and biodynamic nanosilica-induced physiological and transcriptomic modulation of chilli (Capsicum annuum L.) under drought stress.

Chilli (Capsicum annuum L.) is an economically important vegetable crop cultivated worldwide. Increasing drought stress associated with climate change has severely reduced chilli productivity. Although grafting and silicon-based nanomaterials have each been investigated independently as drought mitigation strategies in Solanaceae crops, this study represents, to our knowledge, the first investigation of their combined physiological, yield, and genome-wide transcriptomic effects in chilli under experimentally validated drought stress. Biodynamic nanosilica (BNS) is an &#x3b1;-quartz nanoparticle preparation (20-200 nm) derived from the biodynamic agricultural preparation BD501 through a vortex-triturating process, and distinct from chemically synthesised nanosilica in preparation method and surface bioavailability, applied as a foliar spray at 50 mg L-1. Five treatments were established: well-watered (WW), drought (D), grafting + BNS + drought (G+B+D), grafting + drought (G+D), and BNS + drought (B+D), each with three independent biological replicates. Under moderate-to-severe drought conditions (DSI 62-64%; VWC ~12% v/v at 14 days), the combined G+B+D treatment significantly improved plant height (3.05-fold over D), leaf relative water content (83% vs 49% in D), net photosynthetic rate (2.0-fold over D), water-use efficiency (+40%), and antioxidant enzyme activities (SOD: 3.1-fold; CAT: 2.8-fold over D), while reducing lipid peroxidation by 76%. Root architecture was also substantially enhanced, with a 4.1-fold increase in root length and a 3.1-fold increase in root surface area relative to D. Fruit yield increased by 79% relative to drought-stressed non-grafted plants. Transcriptomic analysis using Illumina NovaSeq 6000 identified 1,051 DEGs (431 upregulated, 620 downregulated; FDR < 0.05, |log2FC| > 1). Integrated transcriptomic-phenotypic concordance analysis revealed enrichment of MAPK signalling, ABA-mediated regulation (including ABA binding and (+)-ABA 8'-hydroxylase activity), and phenylpropanoid biosynthesis as the enriched pathways. Protein-protein interaction network analysis further revealed coordinated regulation of redox homeostasis, drought-responsive hormone signalling, and water transport gene modules in the combined treatment. These findings demonstrate that integrating grafting with biodynamic nanosilica is a promising strategy to enhance drought resilience and productivity in chilli, offering a sustainable approach for vegetable production under drought.

Capsicum

Plastoglobules of leaf chloroplasts of two cultivars of Capsicum annuum.

The chloroplasts of the leaves of Capsicum annuum cv. Purple Leaf Cherry are characterized by the presence of large electron translucent plastoglobules which comprise a considerable proportion of the plastid volume. The leaf chloroplasts of Purple Leaf Yellow cultivar contain, in addition, large starch granules which are frequently in intimate association with these plastoglobules. There appears to be a controlled expulsion of plastoglobules into the vacuole by the non-degenerating chloroplasts of both cultivars, although the function of this secretion is not known. The chloroplasts of older leaves of Purple Leaf Yellow also contain plastoglobules with a nonhomogenous fine structure. Some have granular segments, membrane-like bands, or myelin-like bands, the latter suggesting the presence of glyco- and/or phospholipids in such plastoglobules.

Chloroplasts

Molecular mechanisms and agronomic strategies for thermotolerance in chili pepper (Capsicum annuum&#xa0;L.).

Heat stress is a primary environmental constraint on chili pepper (Capsicum annuum L.) productivity and fruit quality across global agricultural systems. Elevated temperatures adversely affect plant growth, flowering, fruit set, pod morphology, physiology, and metabolism. The severity of these impacts varies with the duration and intensity of heat exposure and with the stage of plant development. Reproductive development is especially vulnerable, resulting in compromised fruit formation and a decline in productivity. Heat stress disturbs photosynthesis, cellular structure integrity, and membrane stability through oxidative stress. Chili plants counter heat stress through complex molecular networks that encompass activation of antioxidant systems, heat shock protein (HSP) synthesis, osmolyte biosynthesis, and stress-responsive transcription factor expression. Recent advancements in genomics, transcriptomics, and metabolomics have elucidated fundamental regulatory pathways governing thermotolerance, highlighting the importance of heat-responsive genes, including CaHSPs, CaWRKYs, and CaNACs. This comprehensive review integrates contemporary understanding of physiological, biochemical, and molecular heat stress responses in chili pepper. Additionally, it evaluates potential agronomic and breeding strategies to strengthen crop adaptation to escalating global temperatures.

Capsicum

Engineering cold stress resilience in capsicum annuum through functional genomics and precision breeding.

This review synthesizes the molecular mechanisms of cold tolerance in pepper, integrating multi-omics data,genome editing, and precision breeding strategies to accelerate the development of cold-resilient cultivars. Cold stress is a significant environmental factor that affects the growth, productivity, and fruit quality of Capsicum annuum by impairing membrane integrity photosynthesis and cellular redox homeostasis. Although pepper has several endogenous cold-responsive regulators such as CaNAC035 and CabHLH035, along with antioxidant defense systems, its cold tolerance remains limited due to low transcriptional activation of key regulators, functional redundancy among cold-responsive genes, and the polygenicity of cold tolerance. These complexities, combined with low genetic diversity and linkage drag, have hindered the improvement of cold-resistant cultivars through conventional breeding. This review brings together the recent progress in understanding the molecular mechanisms of cold stress perception, signal transduction, transcriptional regulation, metabolic reprogramming, and phytohormone interactions in pepper. Precision Breeding 2.0 is a new innovation that combines the integration of multi-omics-based target identification with next-generation genome-editing techniques, allowing precise and multiplex engineering of complex and interconnected regulatory networks instead of single genes. We cover new approaches such as engineering the DREB/CBF pathway, allele-specific editing and targeted disruption of negative regulators to enhance the pathway(s) involved in cold response. Moreover, we propose a roadmap for integration of transcriptomics, proteomics, metabolomics, high-throughput phenomics, and speed breeding to accelerate the identification, validation, and deployment of superior alleles to boost cold tolerance. This review provides a foundation for developing climate-resilient pepper cultivars by connecting functional genomics with precision genome engineering approaches to maintain productivity under variable environmental conditions.

Capsicum

Occurrence of lanosterol and lanostenol in seeds of red pepper (Capsicum annuum).

Cycloartenol and cycloartanol are the major components of the 4,4-dimethylsterol fraction separated from the seeds of red pepper (Capsicum annuum, Solanaceae). Lanosterol and lanostenol, whose occurrence in higher plants has hitherto been known as extremely rare, are identified in the fraction by gas liquid chromatography, combined gas liquid chromatography-mass spectrometry, and isolation as their epoxides.

Chromatography, Gas

Cercaricidal activity of some essential oils of plants from Brazil.

Extended screening tests on the cercaricidal activity of various plants from northeast Brazil have shown that the extracts of the leaves and fruits of Piper marginatum, Protium heptaphyllum, and Capsicum annuum show a remarkable effect on the cercaria of Schistosoma mansoni. In the case of the oils of Piper marginatum and Capsicum annuum, 90--96% of the cercaria of Schistosoma mansoni were killed within 15 min. According to the NMR spectra, the active principles of the extracts seem to be water-soluble unsaturated compounds from the oils or their hydrolysis products. Natural substances provide an attractive alternative for preventive actions against schistosomiasis, as the perturbation of the ecological equilibrium of natural waters can be avoided in this manner.

Animals

Systematic identification pepper CaE2F transcription factor reveals the role of CaDPb in drought stress response.

The EARLY 2 FACTOR (E2F) transcription factor (TF) family plays a pivotal role in regulating plant development and adaptations to environmental stresses. However, the physiological function of E2Fs in pepper (Capsicum annuum L.) are not well elucidated. In this work, we conduct a comprehensive genome-wide annotation of the E2F family within the Zunla-1 pepper genome and further explore the biological roles of CaDPb in response to drought stress. Through systematic bioinformatics analysis, we identify a total of nine CaE2F genes within the Zunla-1 genome, categorizing them into three distinct subgroups. Additionally, we discover multiple cis-regulatory elements in the CaE2F promoter regions associated with responses to plant hormones and drought stress. Public RNA-seq datasets reveal distinct expression profiles of CaE2F genes across various pepper tissues and their responses to environmental stimuli and plant hormones. Subsequently, the CaDPb gene is further functionally verified in drought response. Our findings indicate that TRV2:CaDPb silenced pepper plants are more sensitivity to drought. Furthermore, we show that CaDPb participates in the regulation of reactive oxygen species (ROS) production, the expression of drought-responsive genes, and the modulation of stomatal aperture. Taken together, our findings provide a comprehensive characterization of E2F genes in pepper and offer insights into the biological function of CaDPb in pepper drought stress response.

Capsicum

Genome-wide identification of CHY zinc finger and RING finger (CHYR) genes in pepper and functional characterization of CaCHYR5 in response to Phytophthora capsici infection.

CHY zinc finger and RING finger (CHYR) proteins play crucial roles in the growth and development, as well as stress response. To date, no systematic or comprehensive analysis of the CHYR gene family has been performed in pepper (Capsicum annuum L.). In this study, we identified 8 CaCHYR genes (CaCHYR1-CaCHYR8), which were classified into 3 groups based on phylogenetic relationships. CaCHYR members within the same group exhibited similar distributions of conserved motifs and exon-intron structures. Chromosomal localization analysis showed that 8 CaCHYR genes were unevenly distributed on 6 chromosomes. Segmental duplication, rather than tandem duplication, was found to be the major contributor to the expansion of this gene family. CaCHYR genes feature a variety of cis-elements involved in developmental processes, phytohormone responses, and stress adaptation. Expression analysis based on RNA-seq data revealed that CaCHYR genes exhibited distinct spatial expression patterns across different tissues and in response to Phytophthora capsici infection (PCI), and quantitative real-time PCR (qRT-PCR) further confirmed that three of them (CaCHYR2, CaCHYR3, and CaCHYR5) exhibited altered expression under PCI. Furthermore, transient overexpression of CaCHYR5 in pepper leaves increased susceptibility to PCI, suggesting its potential negative regulatory role in pepper defense against P. capsici. Collectively, these findings reveal the expression patterns and regulatory functions of pepper CHYR genes in growth and development, laying a groundwork for breeding pepper cultivars tolerant to PCI.

Phytophthora capsici infection (PCI)

A Novel Approach to Engineering Tomato Spotted Wilt Virus Infectious Clones by Disarming Key Nodes in Antiviral Defenses.

Tomato spotted wilt virus (TSWV) is an economically devastating pathogen that rapidly overcomes genetic resistance in major crops. Reverse genetic systems are crucial for investigating plant-virus interactions and resistance-breaking mechanisms, and developing these tools for segmented ambisense RNA viruses remains a crucial challenge. Current TSWV clones rely on extensively modified Asian isolates requiring co-delivery of multiple replication helpers and viral silencing suppressors. Streamlining these systems for regionally significant strains with minimal genetic alterations is essential. Here, we developed the first infectious clone of a U.S. TSWV isolate (PA01). Three binary plasmids contain cDNAs for the antigenomic L and S segments, as well as the genomic M segment, with enhanced GFP replacing NSs on the S segment. Co-delivery of the cucumovirus 2b alone or in combination with tombusvirus P19 or begomovirus AL2 achieved a high proportion of systemically infected Nicotiana benthamiana and Capsicum annuum plants. In N. tabacum, co-delivering the Caenorhabditis elegans cell death suppressor CED-9 or using NahG transgenic plants produced 30 to 33% systemically infected plants. Co-delivery of 2b boosted infection levels in NahG plants to 62%. These data indicate that in addition to the antiviral RNA-silencing machinery, additional host defense pathways influence TSWV rescue and systemic infection from cDNA. [Formula: see text] Copyright &#xa9; 2026 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Tospovirus

Incidence of aflatoxin-potential contamination in Spanish sausages.

525 strains of moulds were isolated from 8 samples of red pepper (Capsicum annuum) and from 3 samples of black pepper (Piper nigrum). These spices were commonly added to the various kinds of spanish sausages, specially to the spanish "chorizo". Of these 525 isolated moulds, 100 belonged to the Aspergillus flavus group. 69 were isolated from red pepper and 31 from black pepper. Of these strains, 28 strains isolated from red pepper were found positive in producing aflatoxin B1 while only 8 of the strains isolated from black pepper produced aflatoxin B1. 16 of the strains isolated from red pepper also produced aflatoxin G1. After experimental contamination by these aflatoxicogenic strains of moulds of various kinds of spanish sausages, and after drying them under similar storage conditions used in the sausage industry, the presence of aflatoxin were detected only on the skin but not in the meat of the sausage.

Aflatoxins

Evolution and domestication-trait associations of ultra-long centromere haplotypes in pepper plants.

Centromeric and pericentromeric regions of most eukaryotic genomes are highly repetitive and strongly recombination-suppressed, confounding efforts to resolve genetic variation, population structure and phenotypic associations. Pepper (Capsicum annuum) centromeres are nearly devoid of satellite repeats, facilitating assembly and population-level comparison of centromeric regions. Here we integrate 9 near-complete genome assemblies, CENH3 ChIP-seq profiles from 26 diverse accessions, and resequencing and phenotypic data from ~400 cultivated and wild accessions to investigate population-level diversity and phenotypic relevance of pepper peri/centromeric regions. Functional centromere positions are largely fixed on 8 of 12 chromosomes, whereas the remaining 4 carry distinct centromeric epialleles shaped mainly by centromere repositioning and pericentromeric inversions. Pepper centromeres are embedded within ultra-long centromere-spanning haplotype (cenhap) blocks, ranging from 29.8 to 112.9&#x2009;Mb and collectively covering 23.96% of the genome; each block contains only 1-4 major haplotypes. Some cenhaps may act as supergene-like units and are strongly associated with fruit traits, probably because recombination-suppressed intervals harbour multiple fruit-related genes, including OFP and F-box genes. F2 segregation assays further reveal transmission distortion of chromosomes carrying alternative cenhaps. Together, these findings highlight peri/centromeric regions as underrecognized reservoirs of agronomically important variation.

Centromere

CaMYB121-CaABF2 negative feedback loop modulates CaNHX2 expression to confer salt tolerance in pepper.

Salt stress is a major abiotic factor that severely restricts pepper (Capsicum annuum) production. Although abscisic acid (ABA) is vital for salt tolerance, the transcriptional regulatory networks governing ABA-mediated salt defense remain largely unknown. Here, we uncovered a negative feedback loop between CaMYB121 and CaABF2.1/2 that modulates the expression of CaNHX2.1/2/3, thereby enhancing salt tolerance in pepper plants. RNA-seq analysis revealed that CaMYB121 displayed an expression pattern consistent with that of CaNHX2 after salt treatment. Silencing CaMYB121 markedly reduced salt tolerance and inhibited root growth. Mechanistically, CaMYB121 directly binds to the CaNHX2 promoter to activate transcription, thereby promoting salt resilience. Salt stress also robustly triggered ABA signaling genes, with CaABF2.1/2 displaying expression patterns closely mirroring those of CaMYB121. Transient silencing of CaABF2.1/2 results in phenotypes similar to those observed with CaMYB121 suppression. Notably, CaMYB121 activates CaABF2.1/2 transcription by binding to its promoters, whereas CaABF2.1/2 represses CaMYB121 expression by directly targeting its promoter, forming a self-regulating feedback loop that prevents excessive defense activation. Collectively, our findings reveal a CaMYB121-CaABF2 feedback circuit that dynamically balances growth and defense to optimize salt tolerance in pepper plants.

Salt Tolerance

Quantitative microanalysis of capsaicin, dihydrocapsaicin and nordihyrocapsaicin using mass fragmentography.

A mass fragmentographic method for the quantitative microanalysis of capsaicin, dihydrocapsaicin and nordihydrocapsaicin in the fruits of Capsicum annuum has been developed. The molecular ions at m/e 377, 379 and 365 in the mass spectra were used for monitoring the trimethylsilyl derivatives of capsaicin, dihydrocapsaicin and nordihydrocapsaicin, respectively. The ratios of the height of each molecular ion to that of an internal standard (cholestane) were linear over the range 5-60 ng. The purification of individual capsaicinoids by high-performance liquid, thin-layer and gas-liquid chromatography is also described.

Capsaicin

A chromosome-scale genome of Capsicum pubescens provides insights into candidate terpene-associated gene clusters and pan variation of terpene synthases.

A chromosome-scale genome of Capsicum pubescens and comparative pan-TPS analysis support structural characterization and gene-level prioritization of a chromosome-9 terpene-associated candidate locus in this accession. Capsicum pubescens is one of the five domesticated Capsicum species, mainly cultivated in mid- to high-elevation regions of the Americas. Despite its distinctive morphology and fruit traits, genomic resources for C. pubescens remain less developed than those for the widely cultivated C. annuum. Here, we assembled a chromosome-scale reference genome for accession HNUCP0001, spanning 3.70&#xa0;Gb with a scaffold N50 of 278.01&#xa0;Mb. Comparative genomics revealed 679 significantly expanded gene families enriched in sesquiterpenoid and triterpenoid biosynthesis. Genome-wide biosynthetic gene-cluster mining identified multiple terpene-associated candidate loci, which were subsequently prioritized using genome-derived structural criteria and Capsicum pubescens-specific expression evidence. Subsequently, we curated the terpene synthase (TPS) repertoire and, across 16 Capsicum genomes, resolved 36 TPS orthogroups with pronounced presence/absence variation, highlighting dynamic lineage-specific diversification. Together, these analyses establish HNUCP0001 as an accession-specific genomic resource and provide a comparative framework for prioritizing terpene-associated TPS genes and candidate BGCs in Capsicum. These candidate loci, together with accession-level transcriptomic and metabolomic evidence, offer testable hypotheses for future functional studies of specialized terpenoid metabolism in C. pubescens.

Alkyl and Aryl Transferases