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Keywords = developmental stage–dependent

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18 pages, 5576 KB  
Article
Genome-Wide Characterization of the WOX Gene Family in Polygonatum cyrtonema and Its Applications for Regeneration
by Yan Shi, Binjie Huang, Cong Li, Jinping Si and Donghong Chen
Plants 2026, 15(11), 1745; https://doi.org/10.3390/plants15111745 - 4 Jun 2026
Abstract
Polygonatum cyrtonema is a medicinally and edible perennial herb, yet functional studies in this species are constrained by limited knowledge of regeneration-associated developmental regulators. Here, we performed a genome-wide characterization of the WUSCHEL-related homeobox (WOX) gene family in P. cyrtonema. Eleven PcWOX [...] Read more.
Polygonatum cyrtonema is a medicinally and edible perennial herb, yet functional studies in this species are constrained by limited knowledge of regeneration-associated developmental regulators. Here, we performed a genome-wide characterization of the WUSCHEL-related homeobox (WOX) gene family in P. cyrtonema. Eleven PcWOX genes were identified and classified into the ancient, intermediate, and modern/WUS clades. Comparative phylogenetic, syntenic, structural, and chromosomal analyses indicated that the PcWOX family retains a conserved evolutionary framework but also exhibits clear lineage-specific diversification. Tissue- and stage-specific expression profiling, promoter cis-element analysis, and subcellular localization further supported functional differentiation among PcWOX members, particularly between PcWOX5 and PcWUS. Weighted gene co-expression network analysis identified 33 co-expression modules, including six key modules strongly associated with PcWOX abundance patterns, and prioritized 49 candidate transcription factors (TFs) to construct PcWOX-centered regulatory networks. These TFs showed marked tissue- and stage-dependent heterogeneity. Heterologous assays in Nicotiana benthamiana showed that both PcWUS and PcWOX5 enhanced regeneration competence but produced distinct developmental outputs. These findings support PcWUS and PcWOX5 as promising candidate morphogenic regulators and provide a useful framework for future mechanistic studies, homologous validation, and regeneration improvement in P. cyrtonema. Full article
(This article belongs to the Section Plant Development and Morphogenesis)
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18 pages, 10236 KB  
Article
Heat-Stress Memory Modulates Antioxidant Metabolism and Increases Senecionine Biosynthesis Across Developmental Stages of Senecio madagascariensis
by Tamara Heck, Gustavo Maia Souza, Douglas Antônio Posso, Roque Mauricio Palacios Zuñiga and Luis Avila
Plants 2026, 15(11), 1730; https://doi.org/10.3390/plants15111730 - 3 Jun 2026
Abstract
Temperature stress strongly affects plant metabolism, and recurrent heat exposure can modify physiological responses depending on developmental stage. This study examined the biochemical and physiological adjustments of Senecio madagascariensis subjected to single (naïve) or repeated (primed) heat stress at 40 °C during vegetative [...] Read more.
Temperature stress strongly affects plant metabolism, and recurrent heat exposure can modify physiological responses depending on developmental stage. This study examined the biochemical and physiological adjustments of Senecio madagascariensis subjected to single (naïve) or repeated (primed) heat stress at 40 °C during vegetative and reproductive stages. Sampling was conducted after the second heat stress and after the subsequent recovery period. Principal component analyses (PCAs) revealed marked stage-specific contrasts. In the vegetative stage, PCA1 and PCA2 explained 75.7% of total variance, clearly separating treatments: naïve plants were associated with elevated proline, soluble sugars, phenolics, glycine betaine, hydrogen peroxide (H2O2) and lipid peroxidation, whereas primed plants were linked to enhanced superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities and reduced oxidative markers. Under stress, naïve plants showed substantial increases in soluble sugars (+198%) and proline (+66.9%), whereas primed plants exhibited attenuated oxidative responses and reduce phenolic accumulation. After recovery, primed plants exhibited markedly reduced H2O2 levels (−57.5%) and lipid peroxidation, alongside higher SOD activity. In the reproductive stage, PCA indicated more subtle priming effects, with overlapping clusters among treatments. Primed plants accumulated the highest soluble sugar levels under stress (+276.5%), while naïve plants showed higher proline and glycine betaine levels. Following recovery, osmolyte levels were similar among groups. Senecionine remained unchanged during the vegetative stage but increased in both naïve (+21.4%) and primed (+19.1%) plants during the reproductive stage after recovery. Oxidative markers revealed contrasting patterns, with primed reproductive plants showed the lowest superoxide under stress but the highest H2O2 and lipid peroxidation at both time points. Overall, the findings demonstrate that heat-stress responses in S. madagascariensis are developmentally regulated, with stronger priming effects during vegetative growth and phenology-dependent metabolic adjustments during reproduction. All results are directly supported by the measured biochemical and physiological data. Full article
(This article belongs to the Special Issue Plant Biology and Sustainable Weed Management)
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22 pages, 1054 KB  
Review
Transcriptional Heterogeneity of Oligodendrocytes: Molecular Basis of Diversity Across Development, Brain Regions, and Neurological Diseases
by Shingo Miyata, Shoko Shimizu and Yugo Ishino
Neurol. Int. 2026, 18(6), 108; https://doi.org/10.3390/neurolint18060108 - 2 Jun 2026
Viewed by 74
Abstract
Oligodendrocytes (OLs) are specialized glial cells essential for the formation and maintenance of the myelin sheath within the central nervous system (CNS). Historically, OLs were considered a functionally homogeneous population. However, the advent and widespread application of single-cell and single-nucleus RNA sequencing (scRNA-seq/snRNA-seq) [...] Read more.
Oligodendrocytes (OLs) are specialized glial cells essential for the formation and maintenance of the myelin sheath within the central nervous system (CNS). Historically, OLs were considered a functionally homogeneous population. However, the advent and widespread application of single-cell and single-nucleus RNA sequencing (scRNA-seq/snRNA-seq) technologies since 2015 have revealed substantial transcriptional heterogeneity, varying according to developmental stage, anatomical region, and disease state. In this review, we synthesized current advances in the understanding of OL heterogeneity. Nine OL cell classes have been identified in the mouse somatosensory cortex and hippocampal CA1 region, later expanding to 13 distinct subpopulations across ten CNS regions. Furthermore, we characterized disease-associated oligodendrocytes (DAOs)/disease-associated oligodendrocyte lineages (DOLs), identified in various neurological diseases, including multiple sclerosis (MS), Alzheimer’s disease (AD), and spinal cord injury, focusing on their molecular markers, spatial distribution, and pathophysiological roles. We summarized key transcriptional regulatory networks underlying DAO induction, including the signal transducer and activator of transcription (STAT)/interferon regulatory factor (IRF) family, the Yin Yang 1 (YY1)/nuclear factor kappa B (NF-κB) axis, and the SOX9/SOX10 regulatory system. The utility of region-specific brain analyses using spatial transcriptomics (ST) in conjunction with these approaches was also discussed. Finally, we compiled the implications of patient stratification according to white matter glial response patterns derived from large-scale snRNA-seq analyses of patients with progressive MS. Our synthesis shows that oligodendrocytes consist of multiple distinct subtypes that vary across development, brain regions, and disease conditions. In pathological states, they adopt specific disease-associated programs that reflect context-dependent responses and may influence disease progression and repair. This work provides a framework for understanding how oligodendrocyte diversity contributes to neurological disease and may support the development of targeted remyelination therapies. Full article
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17 pages, 17189 KB  
Article
Spirulina-Derived Carbon Dots Promote Context-Dependent Effects on Rice Metabolism, Yield, and Grain Quality Under Non-Stress and Heat Stress Conditions
by Luana Vanessa Peretti Minello, Shaiane Lessa dos Santos, Luana Bueno Longaray, Natan da Silva Fagundes, Sidnei Deuner, Aline Nunes, Eva Regina Oliveira, Marcelo Maraschin and Raul Antonio Sperotto
Plants 2026, 15(11), 1657; https://doi.org/10.3390/plants15111657 - 28 May 2026
Viewed by 251
Abstract
Carbon dots (CDs) have emerged as promising nanobioinputs capable of modulating plant metabolism and stress responses. However, their effectiveness under field conditions remains poorly understood. This study evaluated the effects of Spirulina-derived carbon dots (S-CDs) on rice metabolic, agronomic, and grain-quality responses under [...] Read more.
Carbon dots (CDs) have emerged as promising nanobioinputs capable of modulating plant metabolism and stress responses. However, their effectiveness under field conditions remains poorly understood. This study evaluated the effects of Spirulina-derived carbon dots (S-CDs) on rice metabolic, agronomic, and grain-quality responses under non-stress and heat stress conditions. Two independent field experiments were conducted under ambient and heat stress conditions, the latter imposed using temporary greenhouse structures during the reproductive stage. S-CDs were applied by foliar spraying (0.2 mg mL−1) at key developmental stages. Their effects were assessed through metabolic, agronomic, and grain quality analyses. Under non-stress conditions, daily average temperatures ranged from 20 to 27 °C, while daily maxima ranged from 24 to 38 °C. Heat stress increased daily average temperature by 3.9 °C and daily maximum temperature by 12.5 °C, with temperature peaks frequently exceeding 45 °C for several hours. Under non-stress conditions, S-CDs induced modest changes in antioxidant, carbon, and nitrogen metabolism, but without consistent improvements in grain yield or yield components. Under heat stress, however, S-CD application reduced spikelet sterility and increased grain yield despite limited changes in the metabolic variables evaluated. Grain quality and nutritional composition also responded differently depending on the environmental condition, indicating context-dependent effects. S-CDs showed limited agronomic relevance under favorable field conditions, but contributed to yield stability under heat stress. These findings support the potential of S-CDs as complementary nanobiostimulants to improve rice resilience under climate-related stress conditions. Full article
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23 pages, 6449 KB  
Review
Vimentin’s Journey from “Background Scaffold” to Multi-Scale Regulator of Neuronal Growth and Function: Historical, Conceptual and Epistemic Perspectives
by Blen Amare Gebreselase and Alexander A. Minin
Int. J. Mol. Sci. 2026, 27(11), 4869; https://doi.org/10.3390/ijms27114869 - 28 May 2026
Viewed by 195
Abstract
Neurons achieve their highly polarized architecture by coordinating cytoskeletal systems across space and time, enabling axons to extend over remarkable distances and dendrites to elaborate complex arbours. Early neuroanatomists described intracellular “neurofibrils,” yet these structures remained poorly understood until electron microscopy resolved them [...] Read more.
Neurons achieve their highly polarized architecture by coordinating cytoskeletal systems across space and time, enabling axons to extend over remarkable distances and dendrites to elaborate complex arbours. Early neuroanatomists described intracellular “neurofibrils,” yet these structures remained poorly understood until electron microscopy resolved them into three distinct polymer systems: microtubules, actin filaments, and intermediate filaments. Although this framework clarified neuronal ultrastructure, it simultaneously established a conceptual hierarchy in which microtubules and actin were regarded as the principal drivers of neurite growth, while intermediate filaments were relegated to a passive, supportive role. Unlike prior reviews that document vimentin dynamics primarily from a cell-biological standpoint, this review integrates historical, conceptual, and epistemological perspectives to examine both how and why that hierarchy arose and how it has been dismantled. This review traces how that hierarchy arose and why it has been increasingly reconsidered in favour of intermediate filaments, focusing on vimentin as a case study. Evidence from live cell imaging, molecular manipulation, and genetic models shows that vimentin is dynamically regulated rather than static. Vimentin networks remodel continuously, exchange subunits with soluble pools, and move in coordination with microtubules. Most recently, sparse single-filament labelling combined with correlative volume electron microscopy has demonstrated that individual vimentin filaments remain motile even within dense perinuclear networks previously assumed to be static, a finding that fundamentally redefines what filament density implies about cytoskeletal organization. In neural and neural precursor cells, vimentin expression is developmentally regulated and is prominent during early differentiation stages associated with neurite initiation giving way to neurofilaments in mature neurons. Functional studies further link vimentin to neurite formation and extension, cytoskeletal coordination, organelle positioning, and cellular stress responses. Philosophical analysis reveals that these empirical advances were inseparable from shifts in imaging technology and conceptual framing, and that epistemic risks including model dependency and confirmation bias can be mitigated through methodological pluralism and explicit model disclosure. Taken together, these findings support a revised understanding of intermediate filaments as active, context-dependent contributors to neuronal development and plasticity, and illustrate the value of integrating biological evidence with historical and philosophical reflection. Full article
(This article belongs to the Special Issue Recent Prospects in Neurons)
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20 pages, 2250 KB  
Review
Hermaphroditism in Cannabis sativa L.: Impacts, Inducers, and Industry Implications
by Chaylen Douglas Richards, Byeong-Ryeol Ryu, Gyeong-Ju Gim and Sang-Hyuck Park
Plants 2026, 15(11), 1643; https://doi.org/10.3390/plants15111643 - 27 May 2026
Viewed by 273
Abstract
Cannabis sativa L. is a predominantly dioecious species, but sex expression is highly plastic and can be modified by genetic, hormonal, developmental, and environmental factors. This plasticity has major implications for commercial production because hermaphroditic expression in female plants can cause unintended pollination, [...] Read more.
Cannabis sativa L. is a predominantly dioecious species, but sex expression is highly plastic and can be modified by genetic, hormonal, developmental, and environmental factors. This plasticity has major implications for commercial production because hermaphroditic expression in female plants can cause unintended pollination, seed formation, reduced floral quality, and losses in cannabinoid yield. This review summarizes current understanding of sex determination and sex-expression instability in C. sativa, with emphasis on hermaphroditism and its agronomic significance. We examine the genomic architecture of sex determination, the roles of ethylene and gibberellin signaling, and the effects of exogenous chemical treatments used to alter sexual phenotype. Particular attention is given to silver-based ethylene inhibitors, especially silver thiosulfate, which remain the most effective tools for induced masculinization and feminized seed production. We also assess the role of environmental stressors in sex instability and review current approaches for early detection, including visual inspection, Raman spectroscopy, and sex-linked molecular markers. Overall, the available evidence supports a multilayered and context-dependent model in which genotype, treatment regime, developmental stage, and environmental conditions interact to shape sexual phenotype. Improved understanding of these processes will be essential for reducing hermaphroditic risk, improving breeding strategies, and supporting stable, high-value cannabis production. Full article
(This article belongs to the Special Issue Medicinal Cannabis: Phytochemistry and Biotechnological Advances)
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22 pages, 2047 KB  
Review
The Role of the Rhizosphere, Endophytes, and the Influence of Plant-Growth-Promoting Bacteria: Take the Cannabis Microbiome as an Example
by Piotr Stanisław Wiszpolski and Mariusz Jerzy Stolarski
Int. J. Mol. Sci. 2026, 27(11), 4802; https://doi.org/10.3390/ijms27114802 - 26 May 2026
Viewed by 166
Abstract
Cannabis sativa L. is a multipurpose crop of increasing agricultural and medical relevance, whose productivity and phytocannabinoid profile are influenced not only by genotype and environmental factors but also by the composition of its microbiota. This review synthesizes current knowledge (2020–2026) on the [...] Read more.
Cannabis sativa L. is a multipurpose crop of increasing agricultural and medical relevance, whose productivity and phytocannabinoid profile are influenced not only by genotype and environmental factors but also by the composition of its microbiota. This review synthesizes current knowledge (2020–2026) on the rhizosphere and endophytic microbiota of hemp, with particular emphasis on plant growth-promoting bacteria (PGPB) and their mechanisms of action. Molecular studies indicate that hemp-associated bacterial communities are dominated by Proteobacteria, Actinobacteriota, Firmicutes and Bacteroidota, with genotype-, tissue- and developmental-stage-dependent variation. PGPB influence plant performance through direct mechanisms, including biological nitrogen fixation, phosphate solubilization, siderophore production and phytohormone synthesis (indole-3-acetic acid (IAA), gibberellins, cytokinins, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase), as well as indirect mechanisms such as antibiosis, enzyme-mediated pathogen inhibition and induction of systemic tolerance to abiotic stress. Experimental studies demonstrate that inoculation with selected strains or consortia can enhance biomass accumulation, improve germination and root architecture, increase resistance to Fusarium oxysporum and modulate cannabinoid and terpene profiles. Importantly, plant responses are cultivar-specific, highlighting the need for genotype-tailored microbial formulations. Full article
(This article belongs to the Section Molecular Plant Sciences)
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15 pages, 1610 KB  
Article
EtG6PI Is Implicated in Host Cell Invasion and Maduramycin Resistance in Eimeria tenella
by Fanghe Zhao, Yu Yu, Ke Xiao, Qiping Zhao, Shunhai Zhu, Jinwen Wang, Jiayu Bai, Wenqi Han, Shanbo Wu, Hui Dong and Hongyu Han
Microorganisms 2026, 14(6), 1204; https://doi.org/10.3390/microorganisms14061204 - 26 May 2026
Viewed by 145
Abstract
Chicken coccidiosis, caused by obligate intracellular protozoa of the genus Eimeria, inflicts substantial economic losses on the global poultry industry. The extensive use of anticoccidial drugs has led to widespread resistance, underscoring the need for molecular markers associated with this resistance. Our [...] Read more.
Chicken coccidiosis, caused by obligate intracellular protozoa of the genus Eimeria, inflicts substantial economic losses on the global poultry industry. The extensive use of anticoccidial drugs has led to widespread resistance, underscoring the need for molecular markers associated with this resistance. Our previous RNA-seq analysis revealed differential expressions of glucose-6-phosphate isomerase (EtG6PI) between drug-sensitive (DS) and maduramycin-resistant (MRR) strains of Eimeria tenella. In this study, we examined EtG6PI expression across developmental stages using qPCR and Western blotting, finding that both transcription and translation peaked in second-generation merozoites. Furthermore, EtG6PI expression was significantly upregulated in MRR strains in a dose-dependent manner and was also elevated in field isolates exhibiting maduramycin resistance. Indirect immunofluorescence localized EtG6PI to the parasite surface, cytoplasm, and parasitophorous vacuole membrane (PVM), with signal intensity increasing during intracellular development. In addition, anti-rEtG6PI polyclonal antibodies significantly inhibited sporozoite invasion of host cells in vitro. These results indicate that EtG6PI plays a role in host cell invasion, a process essential for parasite proliferation, and is associated with maduramycin resistance in E. tenella, supporting its potential as a biomarker for resistance detection in field settings. Full article
(This article belongs to the Section Molecular Microbiology and Immunology)
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27 pages, 699 KB  
Review
When Fertilization Is Not Enough: Maternal-Zygotic Transition as a Determinant of Embryo Competence in IVF
by Charalampos Voros, Fotios Chatzinikolaou, Georgios Papadimas, Ali Can Gunes, Aristotelis-Marios Koulakmanidis, Ioannis Papapanagiotou, Athanasios Karpouzos, Diamantis Athanasiou, Kyriakos Bananis, Antonia Athanasiou, Aikaterini Athanasiou, Charalampos Tsimpoukelis, Maria Anastasia Daskalaki, Christina-Maria Trakatelli, Nikolaos Thomakos, Panagiotis Antsaklis, Dimitrios Loutradis and Georgios Daskalakis
Int. J. Mol. Sci. 2026, 27(11), 4787; https://doi.org/10.3390/ijms27114787 - 26 May 2026
Viewed by 119
Abstract
A significant concern with IVF is that many embryos fail to develop despite proper fertilization. This gap indicates that factors outside sperm-oocyte fusion influence developmental competency. The maternal-zygotic transition (MZT) is a crucial developmental phase during which control shifts from maternally inherited transcripts [...] Read more.
A significant concern with IVF is that many embryos fail to develop despite proper fertilization. This gap indicates that factors outside sperm-oocyte fusion influence developmental competency. The maternal-zygotic transition (MZT) is a crucial developmental phase during which control shifts from maternally inherited transcripts to activation of the embryonic genome. During the early stages following fertilization, the embryo depends only on maternal mRNA and proteins amassed throughout oogenesis. For successful development, these transcripts must be expeditiously removed with the concurrent genome activation. Any disruption, whether due to inadequate maternal mRNA degradation, aberrant translational control, or delayed genome activation, has been associated with premature developmental stoppage and diminished blastocyst formation. Principal regulators, such as BTG4, CPEB1, DAZL, and components of the translational machinery, govern this modification and seem to be affected by the quality of the oocyte and the age of the mother. Increasing evidence suggests that disruption of MZT may account for instances of suboptimal embryo development that conventional assessment techniques cannot elucidate. MZT offers a biological framework for assessing embryo competency beyond simple appearance. If scientists had a deeper understanding of this process, they might identify molecular markers and enhance the selection of embryos in IVF. Full article
(This article belongs to the Collection Advances in Cell and Molecular Biology)
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17 pages, 5783 KB  
Article
Effect of the Entomopathogenic Fungus Beauveria bassiana (Balsamo) Vuillemin (Strain PPRI5339) on Nymphs of the Calliptamus italicus (Orthoptera: Acrididae) Under Laboratory Conditions
by Spyridon Mantzoukas, Constantina Stamou, Aimilia Lachlou, Vasilis Kotsantis, Ioannis Lagogiannis, Stergios Bitivanos, Dimitris Servis and Panagiotis A. Eliopoulos
Insects 2026, 17(6), 545; https://doi.org/10.3390/insects17060545 - 23 May 2026
Viewed by 305
Abstract
The entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) is widely used as a biological control agent, yet its efficacy against orthopteran pests remains poorly characterized. In this study, we evaluated the susceptibility of second-, third-, and fourth-instar nymphs of the Italian locust, Calliptamus italics [...] Read more.
The entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) is widely used as a biological control agent, yet its efficacy against orthopteran pests remains poorly characterized. In this study, we evaluated the susceptibility of second-, third-, and fourth-instar nymphs of the Italian locust, Calliptamus italics (Orthoptera: Acrididae), to B. bassiana strain PPRI5339 under controlled laboratory conditions. Nymphs were exposed to six conidial concentrations (1 × 103 to 1 × 108 conidia/mL) via treated food, with mortality recorded over 10 days. The fungus caused significant, dose- and time-dependent mortality across all instars. Second-instar nymphs exhibited the highest susceptibility, with an LC50 of 5.8 × 105 conidia/mL and an LT50 of 3.1 days at the highest concentration. Susceptibility decreased with advancing developmental stage; fourth-instar nymphs required higher concentrations and longer exposure to achieve comparable mortality (LC50 = 2.3 × 106 conidia/mL; LT50 = 4.5 days). Cox proportional hazards analysis confirmed that the mortality hazard was significantly lower for third (HR = 0.66) and fourth (HR = 0.51) instars compared to second instars (HR = 1.00). These results demonstrate that B. bassiana is pathogenic to C. italicus, with pronounced stage-dependent susceptibility. Full article
(This article belongs to the Special Issue Insect Pathogens as Biocontrol Agents Against Pests)
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24 pages, 602 KB  
Review
Integrating Envirotyping and Phenomics for AI-Enabled Multi-Environment Genomic Prediction in Crop Breeding
by Xiongwei Liang, Shaopeng Yu, Yongfu Ju, Yingning Wang and Dawei Yin
Agronomy 2026, 16(10), 1019; https://doi.org/10.3390/agronomy16101019 - 21 May 2026
Viewed by 387
Abstract
Genomic prediction is now routine in crop improvement, but its main bottleneck has shifted from marker density to environmental complexity. Breeders rarely need predictions for one fixed environment; they need to rank genotypes across target populations of environments that differ in weather, soils, [...] Read more.
Genomic prediction is now routine in crop improvement, but its main bottleneck has shifted from marker density to environmental complexity. Breeders rarely need predictions for one fixed environment; they need to rank genotypes across target populations of environments that differ in weather, soils, management, and stress timing. This makes genotype-by-environment interaction a primary breeding problem rather than a secondary statistical nuisance. This review examines how genomic, environmental, and phenomic information can be integrated to improve multi-environment prediction in crop breeding pipelines. The review is narrative rather than PRISMA-style, but the literature search and selection logic were structured and explicitly defined. Peer-reviewed English-language studies were identified through structured searches of Web of Science Core Collection and Scopus, supplemented by backward citation screening, with emphasis on literature published from January 2023 to March 2026. Four conclusions emerge. First, environmental information is most useful when it is developmentally aligned, biologically interpretable, and matched to the target population of environments. Second, strong structured statistical baselines remain highly competitive, especially in moderate-sized or highly unbalanced datasets, whereas gains from more flexible machine-learning and deep-learning approaches are most evident in large, sparse, heterogeneous, and multimodal settings. Third, phenomic markers often improve prediction for complex traits, especially yield, because they capture realized crop responses not fully represented by markers alone. Fourth, practical value depends less on isolated gains in predictive accuracy than on evaluation under realistic deployment scenarios, including untested genotype and untested environment settings. Progress therefore requires transparent reporting, benchmark design, stage-aware envirotyping, multimodal integration, uncertainty reporting, and cost-aware deployment. Full article
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16 pages, 8200 KB  
Article
Potential of RNAi Targeting Juvenile Hormone Acid Methyltransferase (JHAMT) for Controlling Dendroctonus valens LeConte (Coleoptera: Scolytidae)
by Qin Cao, Yue Sun, Dejun Kong, Jinbin Han, Jianrong Wei and Jigang Li
Forests 2026, 17(5), 628; https://doi.org/10.3390/f17050628 - 21 May 2026
Viewed by 148
Abstract
Dendroctonus valens LeConte represents a major invasive pest species in China. Both larvae and adults primarily feed on the phloem of the tree trunk base and roots, disrupting nutrient transport and leading to host tree mortality, which poses a severe threat to forest [...] Read more.
Dendroctonus valens LeConte represents a major invasive pest species in China. Both larvae and adults primarily feed on the phloem of the tree trunk base and roots, disrupting nutrient transport and leading to host tree mortality, which poses a severe threat to forest ecosystems and the forestry economy. Juvenile hormone acid methyltransferase (JHAMT) is a key enzyme in insect juvenile hormone (JH) biosynthesis. In this study, we identified a JHAMT-encoding gene, DvJHAMT, in D. valens via bioinformatic analysis. RT-qPCR analysis revealed that DvJHAMT is predominantly expressed during the egg and larval stages. In the fourth-instar larvae, the highest expression levels were observed in the head and epidermis, suggesting a central regulatory role during this critical developmental period. To investigate its function via RNA interference (RNAi), a nanomaterial, star polycation (SPc), was employed for the transdermal delivery of dsRNA into the fourth-instar larvae. The results demonstrated that DvJHAMT knockdown significantly downregulated mRNA levels, resulting in marked decreases in larval survival, pupation, and eclosion rates. Notably, treatment with 0.7 µg dsDvJHAMT-SPc resulted in a 96.67% mortality rate and a reduced pupation rate of 41.67% at 34 days post-treatment. Furthermore, RNAi led to developmental deformities and significant weight loss in larvae. ELISA assays confirmed that DvJHAMT silencing led to reduced JHAMT enzyme activity and JH III titers in a dose-dependent manner. In conclusion, our findings demonstrate that DvJHAMT plays a vital role in JH biosynthesis and that its suppression exhibits potent lethal effects, suggesting that DvJHAMT is a promising candidate for RNAi-based management of D. valens. Full article
(This article belongs to the Special Issue Advances in Wood Borer Control and Management)
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21 pages, 2619 KB  
Article
Stage-Specific Expression of Lens-Associated Structural Genes During Early Embryogenesis in European Seabass (Dicentrarchus labrax)
by Andreas Tsipourlianos, Nikolaos Veliotis, Rafael Angelakopoulos, Themistoklis Giannoulis and Katerina A. Moutou
Genes 2026, 17(5), 590; https://doi.org/10.3390/genes17050590 - 21 May 2026
Viewed by 243
Abstract
Background/Objectives: Lens development is an essential component of visual-system development during fish embryogenesis, yet its transcriptional timing remains poorly characterized in European seabass (Dicentrarchus labrax). This study aimed to provide a stage-resolved transcriptomic characterization of lens-associated gene expression in D. labrax [...] Read more.
Background/Objectives: Lens development is an essential component of visual-system development during fish embryogenesis, yet its transcriptional timing remains poorly characterized in European seabass (Dicentrarchus labrax). This study aimed to provide a stage-resolved transcriptomic characterization of lens-associated gene expression in D. labrax embryos and to relate these patterns to classical embryological stages. Methods: Publicly available RNA-seq data from embryos at the mid-gastrula, late somitogenesis, and hatching stages were analyzed. A targeted lens-associated gene set was defined using Gene Ontology annotations, with emphasis on genes assigned to the structural constituent of the eye lens category. Expression patterns were examined using normalized counts, variance-stabilized data, principal component analysis, and pairwise differential expression analysis. Results: Lens-associated genes displayed clear stage-dependent expression dynamics. Principal component analysis separated samples primarily by developmental stage, with the first two components explaining 89.3% of the total variance. The strongest biological shift occurred between mid-gastrula and late somitogenesis, when transcripts encoding β-crystallins and lens-fiber architecture components increased markedly. Among the most pronounced changes were the induction of crybb1l3 and cryba4, along with increased expression of membrane and cytoskeletal genes, such as the lim2 paralogs and bfsp1. By hatching, this structural-gene expression pattern was partly maintained, while specific crystallin-related loci, including crybg1a, showed further stage-associated increases. Conclusions: These findings define stage-specific patterns of lens-associated gene expression in D. labrax embryos and indicate that lens-associated structural gene expression is most pronounced during late somitogenesis among the stages analyzed. This work provides a useful reference for future studies of visual development in European seabass and for aquaculture-oriented investigations of early sensory ontogeny. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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11 pages, 268 KB  
Protocol
Sleep in Autism Across the Lifespan: A Protocol for a Cross-Sectional Survey with Nationwide Dissemination in Spain
by María Luisa Sánchez de Ocaña-Moreno, Ana María García-Muñoz, Isabel María Timón, Guillermo Benito Ruiz, Marta Plaza Sanz, Ruth Vidriales Fernández, Elena Martínez-Cayuelas, Laura Gisbert-Gustemps, Jorge Lugo-Marín, Gonzalo Pin-Arboledas, Isabel Mengual-Luna, Juana Mulero-Cánovas, Pilar Zafrilla, Begoña Cerdá, Beatriz Rodríguez-Morilla and Pura Ballester-Navarro
Healthcare 2026, 14(10), 1398; https://doi.org/10.3390/healthcare14101398 - 20 May 2026
Viewed by 275
Abstract
Background: Autism spectrum disorder (ASD) is consistently associated with a high prevalence of sleep disturbances across the lifespan, with reported rates ranging from 60% to 86% depending on age and clinical characteristics. Although this issue has been widely described in the international literature, [...] Read more.
Background: Autism spectrum disorder (ASD) is consistently associated with a high prevalence of sleep disturbances across the lifespan, with reported rates ranging from 60% to 86% depending on age and clinical characteristics. Although this issue has been widely described in the international literature, Spain currently lacks large-scale data to estimate the prevalence of sleep disturbances or to examine their relationship with factors such as age, intellectual disability, and co-occurring conditions. This study aims to estimate the prevalence and severity of sleep disturbances in individuals with autism spectrum disorder in Spain and to examine their associations with developmental stage, intellectual disability, affective symptoms, and contextual factors. Methods: This is a cross-sectional observational survey with nationwide dissemination approved by the Ethics Committee of the Universidad Católica San Antonio de Murcia. Data will be collected through an online survey (SurveyMonkey) including validated instruments: the Children’s Sleep Habits Questionnaire–Autism (CSHQ-Autism) and the Sleep Disturbance Scale for Children (SDSC) for pediatric participants; the Pittsburgh Sleep Quality Index (PSQI) for adolescents and adults without intellectual disability; and the Diagnostic Assessment for the Severely Handicapped–II (DASH-II) for adults with intellectual disability. Anxiety and depressive symptoms will be assessed using the Child Behavior Checklist (CBCL) in children and adolescents and the Hospital Anxiety and Depression Scale (HADS) and DASH-II. Statistical analyses will be conducted using SPSS v22 by applying parametric or non-parametric tests according to data distribution. Conclusions: This study represents one of the first survey protocols with nationwide dissemination designed to assess sleep disturbances in individuals with ASD in Spain. The resulting findings are expected to help identify vulnerability profiles, inform public health strategies, and support the development of multidisciplinary interventions aimed at improving sleep and, consequently, the quality of life of individuals with autism and their families. Full article
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Article
Mature-Stage Eisenia fetida Proteins Suppress Macrophage Inflammation via NF-κB and MAPK Pathways
by Hind Althagafi, Hussam A. Althagafi, Fahad Alharthi, Abdullah A. A. Alghamdi, Abdullah M. Almotayri, Ibrahim Jafri, Leena S. Alqahtani, Atif Abdulwahab A. Oyouni, Abdulaziz Albogami and Deyala M. Naguib
Int. J. Mol. Sci. 2026, 27(10), 4568; https://doi.org/10.3390/ijms27104568 - 19 May 2026
Viewed by 251
Abstract
Earthworm-derived bioactive compounds are emerging as promising pharmaceutical agents; however, the immunomodulatory effects of Eisenia fetida proteins at different developmental stages remain unclear. This study evaluated, for the first time, the stage-dependent immunomodulatory activity of E. fetida protein extracts in RAW 264.7 macrophages. [...] Read more.
Earthworm-derived bioactive compounds are emerging as promising pharmaceutical agents; however, the immunomodulatory effects of Eisenia fetida proteins at different developmental stages remain unclear. This study evaluated, for the first time, the stage-dependent immunomodulatory activity of E. fetida protein extracts in RAW 264.7 macrophages. Soluble proteins isolated from juvenile, mature, and senescent worms were lyophilized and tested for their effects on cell viability, phagocytic activity, nitric oxide (NO), reactive oxygen species (ROS), and inflammatory gene expression. Amino acid profiling and Western blot analysis were additionally performed to investigate biochemical composition and signaling mechanisms. Mature-stage extracts exhibited the highest protein yield, minimal cytotoxicity, enhanced macrophage phagocytosis, and significant suppression of LPS-induced NO, ROS, and proinflammatory cytokines. In contrast, juvenile-stage extracts showed moderate immunomodulatory activity, whereas senescent-stage extracts induced oxidative stress and inflammatory responses. Western blot analysis demonstrated that mature-stage proteins strongly inhibited phosphorylation of NF-κB and MAPK signaling proteins, including p65, IκBα, p38, ERK1/2, and JNK, while senescent-stage extracts maintained elevated pathway activation. Amino acid analysis further revealed enriched immunologically relevant amino acids in mature-stage extracts. These findings demonstrate that developmental stage strongly influences the biological activity of E. fetida proteins and highlight mature-stage extracts as promising natural immunomodulatory agents. Full article
(This article belongs to the Special Issue Advances in Bioactivity and Molecular Mechanisms of Natural Products)
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