Search Results (204)

Nymphs of the ambar mutant of Orius laevigatus (Fieber) are orange-colored instead of the yellowish color of the wild-type individuals. Since there were no previous studies of the pigments of this species, we searched for differences in pigments of the pteridine family between both strains. Fluorescent compounds from nymph extracts were separated by cellulose thin-layer chromatography (TLC) and by size exclusion chromatography, followed by LC/MS/MS. The present study has allowed the identification for the first time in O. laevigatus of erythropterin, leucopterin, 7-methylxanthopterin, xanthopterin, isoxanthopterin, pterin, and biopterin. The quantification was performed by fluorometry after elution of the pteridines previously separated by TLC. The results showed that the orange color in the ambar nymphs was due to the accumulation of the orange pigment erythropterin. Additionally, mutant nymphs exhibited significantly elevated levels of pterin and reduced levels of leucopterin. The possibility that these differences were due to differences in xanthine dehydrogenase (XDH) activity was tested; the results indicated that XDH deficiency is unlikely to be responsible for the mutant phenotype. Considering that the ambar mutation is recessive, the mutant phenotype should, most likely, be due to a disruption in downstream metabolic steps involved in erythropterin processing. Full article

Plants are continuously exposed to multiple environmental stressors throughout their lifecycle. Understanding their integrated physiological, biochemical, and anatomical responses under combined stress conditions is crucial for developing effective approaches to improve stress tolerance and maintain crop productivity. This study aimed to investigate the physiological, biochemical, and anatomical changes in photomorphogenic Micro-Tom plants exposed to high irradiance and water deficit—an abiotic stress combination that commonly co-occurs in natural environments but remains poorly understood in light-sensitive genotypes. We hypothesized that the high pigment 1 (hp1) mutant, due to its enhanced light responsiveness, would display improved stress acclimation compared to the wild-type when exposed to combined stress factors. This study was conducted in a controlled plant growth chamber, using a randomized block design with five replicates. Two Micro-Tom genotypes (wt and hp1) were exposed to control (soil at field capacity (FC) + 450 μmol m⁻² s⁻¹ PPFD) and combined stress (40% FC + 1800 μmol m⁻² s⁻¹ PPFD) conditions. Despite the higher concentration of chloroplast pigments in hp1, its photosynthetic performance under combined stress was not significantly improved, and its defense mechanisms did not effectively mitigate the stress impacts. Anatomically, wt exhibited greater structural adjustment, observed by adaptations in the spongy parenchyma and mesophyll. Overall, the wt genotype showed stronger defense mechanisms, while hp1 was more susceptible to combined abiotic stress. Full article

The desert locust (Schistocerca gregaria) represents one of the most destructive agricultural pests globally, renowned for its ability to form massive swarms that can devastate crops and threaten food security across vast regions. Despite the widespread application of the CRISPR/Cas9 gene-editing system in several insect orders, its utilization in locusts, particularly in the desert locust, has remained relatively unexplored. We established a CRISPR/Cas9-mediated gene-editing workflow for the desert locust using gene encoding for neuropeptide corazonin (Crz) as a target. We also analyzed the phenotypic and physiological characteristics of the mutant using paraffin sectioning, HE staining, and chitin staining techniques. Our findings revealed that while Crz knockout desert locusts were viable and maintained normal fertility, they exhibited striking phenotypic alterations, including albinism and a significant reduction in cuticle thickness. These observations not only highlight the functional role of Crz in pigmentation and cuticle development but also underscore the potential of CRISPR/Cas9 as a powerful tool for dissecting gene function in locusts. Furthermore, the successful application of CRISPR/Cas9 in desert locusts also paves the way for similar genetic studies in other non-model insects, expanding the scope of functional genomics in entomology. Full article

Visual behaviour in zebrafish, often measured by the optokinetic reflex (OKR), serves as a valuable model for studying aspects of human neurological and ocular diseases and for conducting therapeutic or toxicology assays. Traditional methods for OKR analysis often rely on binarization techniques (threshold-based conversion of images to black and white) or costly software, which limits their utility in low-contrast settings or hypopigmented disease models. Here, we present a novel deep learning pipeline for OKR analysis, using ResNet-50 within the DeepLabCut framework in a Python Version 3.10 environment. Our approach employs object tracking to enable robust eye movement quantification, regardless of variations in contrast or pigmentation. OKR responses were elicited in both wild-type and slc45a2 (albino) mutant zebrafish larvae at 5 days post-fertilisation, using a mini-LED arena with a rotating visual stimulus. Eye movements were recorded and analysed using both conventional software and our deep learning approach. We demonstrate that the deep learning model achieves comparable accuracy to traditional methods, with the added benefits of applicability in diverse lighting conditions and in hypopigmented larvae. Statistical analyses, including Bland–Altman tests, confirmed the reliability of the deep learning model. While this study focuses on 5-day-old zebrafish larvae under controlled conditions, the pipeline is adaptable across developmental stages, pigmentation types, and behavioural assays. With appropriate adjustments to experimental parameters, it could be applied to broader behavioural studies, including social interactions and predator–prey dynamics in ocular and neurological disease models. Full article

The chloroplast signal recognition particle (cpSRP) components cpSRP43 and cpSRP54 not only form a complex with light-harvesting chlorophyll (Chl)-binding proteins to direct them to the thylakoid membrane, but also serve other functions. cpSRP43 independently acts as a chaperone for some tetrapyrrole biosynthesis (TBS) enzymes, while cpSRP54 participates in the co-translational targeting of plastid-encoded proteins. However, it remains unclear to what extent the two cpSRP components are coregulated—despite their distinct functions—and whether both participate in genomes-uncoupled (GUN)-mediated retrograde signaling. Here, we demonstrate that cpSRP43 and cpSRP54 accumulation is strongly interdependently controlled: overexpression of one protein increases the level of the other, while a deficiency in one of the two proteins leads to a simultaneous decrease in the other component. Disruption of this balance, e.g., by combining the overexpression of one component with a knockout of the other, results in severe chlorosis, stunted growth, and reduced levels of Chl and tetrapyrrole intermediates. Moreover, cpSRP43 deficiency exacerbates the pale-green phenotype of gun4 and gun5 mutants, highlighting a synergistic impact on TBS; however, cpSRP43 overexpression fails to rescue these defects. Remarkably, loss of cpSRP43 does not affect the expression of nuclear-encoded photosynthetic genes under intrinsic plastid stress, clearly demonstrating that cpSRP43 is not involved in plastid-to-nucleus retrograde signaling. Overall, our findings underscore that the fine-tuned expression of cpSRP43 and cpSRP54 is crucial for proper chloroplast function and pigment biosynthesis, while cpSRP43 alone does not participate in the retrograde signaling pathway. Full article

Leaf colour is a valuable morphological phenotype for studying plant metabolism and physiology. To elucidate the mutation mechanism of leaf colour variation in maize, we compared the ethyl methylsulfonate (EMS)-induced maize mutant zmpgl, which has light green leaves, with the wild-type maize line B73. At the seedling stage, the zmpgl mutant presented distinct light green leaf colouration. Comprehensive analyses revealed that both the photosynthetic parameters and pigment contents of the mutant seedlings were significantly lower than those of the wild-type seedlings. Transmission electron microscopy of the mutant leaves revealed alterations in the chloroplast structure, which consequently impaired the photosynthetic efficiency and accumulation of organic matter. Through integrated transcriptomic and metabolomic profiling, we identified differentially expressed genes (DEGs) and differentially abundant metabolites associated with the zmpgl phenotype. These molecular components were associated with pathways related to plant metabolism, chloroplast structure-associated hormone signalling, and redox homeostasis. Further investigation revealed a significant differential expression of genes involved in several critical biological processes, including tetrapyrrole synthesis, lipid metabolism (related to leaf photosynthesis), amino acid metabolism (associated with chlorophyll synthesis and the light response), and abscisic acid (ABA) biosynthesis. These processes are crucial for plant photosynthesis, respiration, and catalytic functions. This study not only provides a valuable resource for further investigation of plant photosynthetic systems but also establishes a foundational framework for the comprehensive functional characterisation of genes involved in the leaf colour change in the zmpgl mutant. These findings contribute to our understanding of the molecular basis of leaf colour variation and its impact on photosynthetic performance in maize. Full article

Fusarium verticillioides is a globally prevalent phytopathogenic fungus responsible for multiple diseases in maize and a major producer of the mycotoxin fumonisin B1 (FB1), a highly toxic fungal secondary metabolite (FSM). The histone code, which includes reversible modifications such as acetylation and methylation, plays a critical role in regulating chromatin structure and gene expression. In fungi, di- and tri-methylation of histone H3 at lysine 9 (H3K9me2/3) serves as a key epigenetic mark associated with heterochromatin formation and transcriptional repression. In this study, we identified and characterized a putative heterochromatin protein 1 (HP1) family member in F. verticillioides, designated FvHP1, based on conserved domain architecture and phylogenetic analyses. FvHP1 retains essential residues required for H3K9me2/3 recognition, supporting its functional conservation within the HP1 protein family. Phenotypic analysis of the ΔFvHP1 mutant revealed impaired vegetative growth, reduced conidiation and virulence, and altered FB1 mycotoxin production. Additionally, the accumulation of red pigment in the mutant was linked to the deregulation of secondary metabolism, specifically the overproduction of fusarubin-type naphthoquinones, such as 8-O-methylnectriafurone. These results support the role of FvHP1 in facultative heterochromatin-mediated repression of sub-telomeric biosynthetic gene clusters, including the pigment-associated PGL1 cluster. Our findings provide new insights into the epigenetic regulation of fungal pathogenicity and metabolite production, as well as the first evidence of a functional HP1 homolog in F. verticillioides. Full article

Chrysanthemum indicum L. is characterized by a high concentration of flavonoid compounds, which exert multifaceted influences on the organoleptic properties, chromatic stability, and therapeutic efficacy of capitulum-derived extracts. These components exhibit diverse biological activities, including heat-clearing, antibacterial, and hepatoprotective properties. A novel white C. indicum variant lacking linarin was recently identified, but its metabolic and transcriptional differences from traditional yellow varieties remain unclear. This study compared flavonoid metabolism in white mutant (BHYJ) and yellow (HJ06) varieties through integrated metabolomic and transcriptomic analyses. Metabolomics identified 491 flavonoids, revealing distinct accumulation patterns: BHYJ accumulated dihydroflavones/chalcones (eriodictyol, hesperetin-8-C-glucoside-3′-O-glucoside, naringenin chalcone), while HJ06 showed higher flavones/flavonols (linarin, rhoiflolin, vitexin, rutin, nicotiflorin). Transcriptomics identified 43 differentially expressed enzyme genes, with key regulators FNSII, F3′H, and F3H showing expression patterns correlating with metabolite profiles. Integrated analysis revealed metabolic divergence at the naringenin node: BHYJ produced less naringenin than HJ06 and preferentially channeled it toward eriodictyol synthesis rather than linarin production. This metabolic shift explains the reduced linarin accumulation in BHYJ. Experimental validation confirmed the coordinated expression patterns of key enzymes. These findings provide foundational insights into transcriptional regulation of flavonoid divergence in pigmented C. indicum varieties, establishing a framework for elucidating enzymatic control of flavonoid biosynthesis in capitulum development. Full article

Fusarium acuminatum is a major pathogenic fungus causing root rot in alfalfa (Medicago sativa). DNA polymerase theta is known to play a crucial role in repairing DNA double-strand breaks. However, its biological function in F. acuminatum remains unknown. In this study, the POLQ gene was deleted by homologous recombination using Agrobacterium tumefaciens-mediated transformation. Compared to the wild type (with the POLQ gene), the mutants (without the POLQ gene) showed significant phenotypic changes: they produced brown-yellow pigments instead of pink, slowed mycelial growth, and exhibited changes in macroconidia size and shape. The virulence of the mutants was greatly reduced, inducing only mild symptoms in alfalfa. In addition, FITC-WGA staining showed impaired spore germination and hyphal growth. These results suggest that POLQ is a key gene regulating growth and development of F. acuminatum, indicating that DNA repair may play an essential role in the pathogenicity of the pathogen in alfalfa. The POLQ gene could thus be a promising target for limiting F. acuminatum infections in alfalfa. Full article

The root parasitic weed Phelipanche aegyptiaca (Pers.) Pomel poses a serious threat to solanaceous crops, leading to yield losses of up to 80% in tomato (Solanum lycopersicum L.). Strigolactones (SLs), derived from the carotenoid metabolic pathway, serve as key host-recognition signals for root-parasitic plants. This study investigated the molecular mechanisms of host resistance, focusing on the suppression of SL biosynthesis through altered carotenoid metabolism in the high-pigment tomato mutants hp-1 and og^c. Both pot experiment and in vitro seed germination assays demonstrated that the mutants exhibited reduced susceptibility to P. aegyptiaca and triggered lower germination rates in broomrape seeds compared to the wild-type cultivar AC. Quantitative RT-PCR analysis revealed a significant downregulation of SL biosynthesis genes (SlD27, SlCCD7, SlCCD8, SlMAX1, SlP450, SlDI4) in hp-1 at various parasitic stages post-inoculation, with a more pronounced suppression observed in hp-1 than in og^c. Notably, the extent of downregulation correlated with the enhanced resistance phenotype in hp-1. These findings highlight a synergistic resistance mechanism involving the coordinated regulation of carotenoid metabolism and SL biosynthesis, providing new insights into the molecular defense network underlying tomato-broomrape interactions. Full article

Floral coloration in Freesia hybrida is predominantly attributed to anthocyanins, with glycosylation playing a critical role in their stability and diversity. This study investigates the molecular mechanisms underlying color variation between F. hybrida cultivars, focusing on anthocyanin 5-O-glucosyltransferases (An5GTs). HPLC analysis revealed that ‘Pink Passion’ petals accumulate 3,5-O-diglucosidic anthocyanins, absent in ‘Red River^®’ and ‘Ambiance’. RNA-seq identified seven candidate Fh5GT genes, with phylogenetic and subcellular localization analyses confirming their classification as cytosolic glycosyltransferases. Expression profiling highlighted elevated transcript levels of Fh5GT1, Fh5GT3, and Fh5GT7 in ‘Pink Passion’, correlating with its di-glucosidic anthocyanin accumulation. In vitro enzymatic assays demonstrated that Fh5GT3 and Fh5GT7 preferentially glucosylate 3-O-monoglucosidic anthocyanins to form stable 3,5-O-diglucosides, with minimal activity on anthocyanidins to generate 5-O-glucosidic anthocyanins. Heterologous expression of Fh5GT3 and Fh5GT7 in Arabidopsis complemented anthocyanin deficiency in 5gt mutants, restoring pigmentation. These findings elucidate the potential role of 5GTs in modulating floral color diversity through anthocyanin modification, providing insights for targeted breeding strategies to enhance ornamental traits in horticultural species. Full article

Chloroplast biogenesis is a crucial biological process in plants. Endoribonuclease E (RNase E) functions in the RNA metabolism of chloroplast and plays a vital role for chloroplast development in Arabidopsis. However, despite sharing 44.7% of its amino acid sequence identity with Arabidopsis RNase E, the biological function of rice OsRNE (Oryza sativa RNase E) remains unknown. Here, we identified a white leaf and lethal 1 (wll1) mutant that displayed white leaves and died at the seedling stage. The causal gene OsRNE was isolated by MutMap+ method. CRISPR/Cas9-mediated knockout of OsRNE resulted in white leaves and seedling lethality, confirming OsRNE as the causal gene for the wll1 phenotype. The albino phenotype of osrne mutant was associated with decreased chlorophyll content and abnormal thylakoid morphology in the chloroplast. The absence of OsRNE led to a significant reduction in the Rubisco large subunit (RbcL), and the 23S and 16S chloroplast rRNAs were nearly undetectable in the osrne mutant. OsRNE transcripts were highly expressed in green tissues, and the protein was localized to chloroplasts, indicating its essential role in photosynthetic organs. Furthermore, transcriptome analysis showed that most of the genes associated with photosynthesis and carbohydrate metabolism pathways in the osrne mutant were significantly down-regulated compared with those in WT. Chlorophyll- and other pigment-related genes were also differentially expressed in the osrne mutant. Our findings demonstrated that OsRNE plays an important role in chloroplast development and chlorophyll biosynthesis in rice. Full article

Acetic acid bacteria (AAB) are a group of bacteria, most of which can produce pigments. However, the mechanism of pigment production by AAB is unclear. A strain of AAB, Gluconacetobacter tumulisoli FBFS 97, which can produce a large amount of brown pigment (BP), was isolated in our previous research. In the current study, it was found that the BP yield of the FBFS 97 strain was enhanced in the presence of tyrosine, and an intermediate of melanin, L-3,4-dihydroxyphenylalanine (L-DOPA), was identified using ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The structural properties of BP were analyzed by pyrolysis gas chromatography–mass spectrometry (Py-GC-MS). All these analyses suggest that BP may be eumelanin, a type of melanin. Then, the eumelanin biosynthetic pathway was investigated in the FBFS 97 strain, and three related genes with eumelanin including pheA, yfiH, and phhB in its genome were found and knocked out, respectively. The results showed that eumelanin production increased 1.3-fold in the pheA deletion mutant compared to the wild-type FBFS 97 strain, but when either yfiH or phhB was knocked out, the eumelanin production in the mutants was the same as that in the wild-type FBFS 97 strain. Finally, a possible biosynthetic pathway for eumelanin in the FBFS 97 strain is proposed. Full article

Leaf coloration, a critical trait in ornamental foliage plant breeding, is influenced by chlorophyll, carotenoids, and flavonoids, which dictate plant aesthetic and economic value. The regulatory role of MYB transcription factors in leaf pigmentation is well recognized. However, their specific influence on Cymbidium leaf coloration remains obscure despite the genus’s global economic importance. This study utilized a novel orchid mutant with leaf variegation as the experimental material to investigate the role of CcR2R3-MYB genes. This research has successfully identified and cloned a novel MYB transcription factor, namely CcR2R3-MYB, from a leaf variegation mutant of Cymbidium. The expression level of CcR2R3-MYB was significantly higher in the mutant plants, with the protein predominantly localized in the nucleus. Phylogenetic analysis indicates that the gene is closely related to AtMYB106 and DhMYB1 and regulates leaf cell morphogenesis and color variation in Cymbidium. Overexpression of CcR2R3-MYB resulted in a yellowish-green and a reduction in photosynthetic pigment content in the Dendrobium. These findings not only lay a foundation for unraveling the mechanism by which CcR2R3-MYB regulates the development of orchid foliage art but also hold significant implications for creating new orchid germplasm and the enhancement of varietal traits. Full article

Carotenoids are crucial for photosynthesis, playing key roles in light harvesting and photoprotection. In this study, spheroidene and bacteriochlorophyll a (Bchl a) were reconstituted into the chromatophores of the carotenoidless mutant Rhodobacter sphaeroides R26.1, resulting in the preparation of high-quality LH2 complexes. Global and target analyses of transient absorption data revealed that incorporating B800 Bchl a significantly enhances excitation energy transfer (EET) efficiency from carotenoids to Bchl a. EET predominantly occurs from the carotenoid S₂ state, with additional pathways from the S₁ state observed in native LH2. Unique relaxation dynamics were identified, including the generation of the carotenoid S* state in reconstituted LH2 with both spheroidene and B800 Bchl a and the formation of the carotenoid T₁ state in reconstituted LH2. These findings underscore the critical influence of pigment composition and spatial organization on energy transfer mechanisms. They provide valuable insights into the molecular interplay that governs excitation energy transfer in photosynthetic light-harvesting systems. Full article