Search Results (243)

Methyl farnesoate (MF) is a sesquiterpenoid hormone that controls a variety of physiological processes in crustaceans, including morphogenesis, development, reproduction, and molting. MF action is mediated by a transcriptional signaling cascade consisting of Methoprene-tolerant (Met), Steroid receptor coactivator (Src), Krüppel homolog 1 (Kr-h1), and Ecdysone response gene 93 (E93) transcription factors (TFs), and transcriptional co-regulators CREB-binding protein (CBP) and C-terminal-binding protein (CtBP). Phylogenetic and sequence analyses revealed that these genes were highly conserved across pancrustacean species. Met and Src were characterized as basic helix-loop-helix, Period (Per)-Aryl Hydrocarbon Nuclear Translocator (ARNT)-Single-minded (Sim) protein (bHLH-PAS) TFs; Kr-h1 was characterized as a C₂H₂ zinc finger TF with seven zinc finger motifs; E93 was characterized as a helix-turn-helix, pipsqueak (HTH_Psq) TF. CBP was identified by several zinc finger-binding regions with Transcription Adaptor Zinc Finger 1 and 2, Really Interesting New Gene, Plant homeodomain, and Z-type zinc finger domains; the Kinase-inducible Domain Interacting-transcription factor docking site; the Bromodomain-acetylated lysine recognition and binding site; the histone acetyltransferase domain; and a C-terminal CREB-binding region containing a nuclear receptor co-activator-binding domain. CtBP had a dehydrogenase domain with arginine-glutamate-histidine catalytic triad. 81 Met contigs, 45 Src contigs, 136 Kr-h1 contigs, 66 E93 contigs, 60 CBP contigs, and 172 CtBP contigs were identified across pancrustacean taxa, including decapod crustaceans. Bioinformatic identification and annotation of these TFs and co-regulators in brachyuran Y-organ (YO) transcriptomes suggests that MF signaling influences YO ecdysteroidogenesis; functional tests in the YO are needed to establish causality. Full article

The basic/helix-loop-helix (bHLH) transcription factors are crucial regulators of plant development and stress responses. In this study, we conducted a genome-wide analysis of the bHLH family in Rosa roxburghii, an economically important fruit crop. A total of 89 non-redundant RrbHLHs were identified and unevenly distributed across the seven chromosomes. Phylogenetic analysis classified them into 23 subfamilies and 7 Arabidopsis subfamilies were absent, indicating lineage-specific evolutionary trajectories. Conserved motif and gene structure analyses showed that members within the same subfamily generally shared similar architectures, yet subfamily-specific variations were evident, suggesting potential functional diversification. Notably, key residues involved in DNA-binding and dimerization were highly conserved within the bHLH domain. Promoter analysis identified multiple cis-acting elements related to hormone response, stress adaptation, and tissue-specific regulation, hinting at broad regulatory roles. Expression profiling across fruit developmental stages and in response to GA₃ treatment revealed dynamic expression patterns. Furthermore, 21 duplicated gene pairs (17 segmental and 4 tandem duplicated pairs) were identified, with most evolving under purifying selection. Detailed analysis of these pairs revealed that segmental duplication, coupled with structural variations such as exon indels, dissolution/joining, and exonization/pseudoexonization, substantially contributed to their functional divergence during evolution. Our results provide a basis for understanding the evolution and potential functions of the RrbHLHs. Full article

Genome editing of late myogenic regulators provides a way to dissect the mechanisms through which transcriptional programs and growth-related signaling pathways shape muscle gene expression programs in farmed fish. This study disrupted myogenic regulatory factor 4 (MRF4) in Nile tilapia using CRISPR/Cas9 to examine downstream transcriptional changes in fast skeletal muscle across the trunk, belly, and head regions. Adult F0 crispants carried a frameshift mutation that truncated the basic helix–loop–helix domain and showed an approximate 80–85% reduction in MRF4 mRNA across the trunk, belly, and head muscles. The expression of 23 genes representing myogenic regulatory factors, MEF2 paralogs, structural and contractile components, non-myotomal regulators, cell adhesion and fusion-related transcripts, and growth-related genes within the GH–IGF–MSTN axis was quantified and compared between wild-type and MRF4-crispants. Expressions of major structural genes remained unchanged despite MRF4 depletion, whereas MyoG and MyoD were upregulated together with MEF2B and MEF2D, indicating strong transcriptional compensation. Twist1, ID1, PLAU, CDH15, CHRNG, NCAM1, MYMK, GHR, and FGF6 were also significantly elevated, while IGF1 was reduced, and MSTN remained stable. Together, these results show that MRF4 loss is associated with coordinated transcriptional changes in regulatory and growth-related pathways, while major fast-muscle structural and contractile transcript levels remain stable, thereby highlighting candidate transcriptional targets for future studies that will evaluate links to muscle phenotype and growth performance in Nile tilapia. Full article

The basic helix-loop-helix (bHLH) transcription factor ‘Atoh8’ is involved in the regulation of several developmental processes and pathologies. It regulates organogenesis, reprogramming, stem cell fate determination, and cancer development. However, the mechanisms underlying these observations remain unclear. Unlike many tissue-specific bHLH factors, Atoh8 is ubiquitously expressed during development as well as in adult tissues. In this study, we explored whether Atoh8 modulates basic cellular functions, which may reveal a common mechanism that could explain the diverse observations reported in the literature. Our findings demonstrate that the loss of Atoh8 impairs autophagy. In both primary myoblasts and mouse embryonic stem cells lacking Atoh8, we observed differential expression of LC3B-II, TFEB, and accumulation of p62, indicating impairment of autophagy. Furthermore, mass spectrometric analysis performed on C2C12 and Atoh8 overexpressing C2C12 myoblasts revealed significant alterations in the expression of proteins associated with mitochondrial and lysosomal functions. Finally, Cut&Tag sequencing performed in Atoh8 overexpressing C2C12 cells revealed that Atoh8 binds to multiple genes involved in autophagosome assembly. Overall, this study underscores that Atoh8 is a critical regulator of macroautophagy, and its reduction disrupts the autophagic process, whereas its overexpression results in increased autophagic flux. Full article

The basic helix–loop–helix (bHLH) transcription factors play a crucial role in plant development and stress resistance. Elucidating the structure and function of bHLH family members related to rubber tree powdery mildew (Erysiphe quercicola) is essential for breeding disease-resistant rubber tree varieties. In the rubber tree (Hevea brasiliensis Muell. Arg.) variety CATAS73397, 204 HbbHLH transcription factors were systematically identified at the genome level and classified into 15 subfamilies through evolutionary analysis. The expansion of this family was primarily driven by whole-genome duplication (WGD). Based on RNA-seq data from leaves infected with powdery mildew, 11 HbbHLH genes responsive to infection were identified. Phylogenetic analysis examined the evolutionary relationships between rubber tree bHLH genes and disease-resistant bHLH genes from other plants. Promoter analysis of the 11 differentially expressed genes revealed abundant cis-elements associated with light responses, hormones, and transcription factor binding. Quantitative Real-time polymerase chain reaction validation indicated that HbbHLH87 and HbbHLH162-2 were significantly downregulated during infection, whereas HbbHLH25 was significantly upregulated. These three genes exhibited strong responses to methyl jasmonate (MeJA) and salicylic acid (SA) treatments, suggesting their involvement in jasmonic acid and SA signal transduction pathways during the immune response. This study provides important insights into the molecular mechanisms underlying disease resistance in rubber trees and identifies potential targets for breeding disease-resistant varieties. Full article

Basic helix-loop-helix (bHLH) transcription factors represent one of the largest transcriptional regulator families in cereal crops such as rice, maize, and wheat. They play crucial and diverse roles in regulating key agronomic traits and essential physiological processes. This review provides a systematic synthesis of the functionally characterized bHLH genes across the three major cereals, offering a comparative perspective on their roles in growth, development, and stress responses. We comprehensively summarize their documented functions, highlighting specific regulators such as TaPGS1 for grain size, rice ILI subfamily for leaf angle, OsbHLH004 for seed dormancy and maize “Ms23-Ms32-bHLH122-bHLH51” cascade for the anther development. Their conserved and species-specific functions in iron homeostasis (e.g., IRO2) and in responses to drought, cold, salinity, and pathogens are also detailed. Additionally, we discuss the underlying molecular mechanisms, including specific binding to E-box/G-box cis-elements, protein dimerization, and integration with hormone signaling pathways. By integrating the current knowledge, this review serves as a consolidated and up-to-date reference that highlights the strategic potential of bHLH transcription factors in molecular breeding programs for improving yield, quality, and stress tolerance in cereals. Full article

The basic Helix-Loop-Helix (bHLH) transcription factor family is crucial for plant growth, development, and stress response regulation. Despite previous studies on the bHLH gene family in Lagerstroemia indica, many bHLH genes remain unidentified, hindering further research on LibHLHs. Here, we identified 150 LibHLHs from the genome of L. indica and categorized them into 12 subfamilies (comprising 25 subgroups) showing conservation within subgroups. Cis-acting element analysis suggests roles in plant development, and responses to light, hormones, and stress. Examination of gene expression patterns highlighted the potential involvement of specific genes, such as LibHLH25 in subgroup IIIf, LibHLH68, LibHLH106, and LibHLH142 in subgroup IIIb, and LibHLH112 in subgroup VIIa, in anthocyanin biosynthesis in leaves of L. indica. This investigation enhances our comprehension of the complexity of the bHLH gene family and highlights the potential roles of LibHLHs in anthocyanin biosynthesis in L. indica, offering valuable insights for future genetic breeding endeavors. Full article

Background/Objectives: The basic helix–loop–helix (bHLH) transcription factor family regulates plant development, metabolism, and stress responses. Yet, its genome-wide composition remains unexplored in Lilium bakerianum var. rubrum (LBVR), an ornamental lily valued for its floral traits. This study aimed to identify, classify, and profile the bHLH family in LBVR using full-length transcriptomic resources. Methods: PacBio HiFi full-length transcriptome sequencing was combined with Illumina RNA-seq for accurate structural annotation and expression quantification. Candidate bHLHs were identified by iTAK and HMMER-Pfam, and their physicochemical properties, secondary structures, motifs, and phylogenetic positions were examined. Expression patterns were analyzed across four floral stages (bud, initial bloom, full bloom, and late bloom). Results: A total of 113 high-confidence bHLH genes were identified, with ~90% successfully annotated. The proteins displayed variation in molecular weight, isoelectric point, structural features, and motif composition. Phylogenetic analysis placed them into 13 clades consistent with Arabidopsis subfamilies, revealing lineage-specific expansions and contractions. Expression profiling showed that 95 genes were active in at least one stage, with two transcriptional waves: a strong bud-to-initial-bloom activation and a secondary wave spanning anthesis. Seventeen genes were expressed exclusively at the bud stage, suggesting roles in early floral-organ initiation and pigmentation. Conclusions: This work provides the first genome-wide characterization of bHLHs in LBVR. The integrated sequencing approach generated a robust catalogue and developmental expression map, offering candidates for functional studies and resources for breeding in lilies. Full article

Rice (Oryza sativa), a crucial global staple, grapples with environmental stress and resource constraints, necessitating sustainable farming. This review explores the transformative role of transcription factors (TFs) in revolutionizing rice agriculture and their potential impact on global food security. It underscores TFs’ pivotal role in gene expression, particularly in responding to environmental stimuli, presenting a promising avenue for enhancing rice resilience. Delving into key TF families in rice, it highlights their multifaceted roles in abiotic stress responses, defense mechanisms, yield improvement, nutrient uptake, seed development, photosynthesis, and flowering regulation. Specific TFs, including DREB (Dehydration-Responsive Element-Binding), WRKY, NAC, MYB (Myeloblastosis), AP2/ERF (APETALA2/Ethylene Responsive Factor), and bHLH (basic Helix–Loop–Helix), are examined for their contributions to stress resilience, defense mechanisms, and yield enhancement. Concrete examples from cutting-edge research illustrate the tangible benefits of harnessing these molecular regulators. However, manipulating TFs presents challenges, necessitating innovative approaches such as predictive models, collaborative field testing, and transparent communication to navigate intricate regulatory networks and regulatory hurdles. Ultimately, a promising future emerges where manipulating rice TFs leads to the development of resilient, high-yielding, and nutritious varieties. Embracing research advancements and addressing existing challenges is imperative to unlock the full potential of these concealed regulators, ensuring sustainable food security for a growing global population. Full article

Seed germination is a critical phase in the plant lifecycle of Camellia oleifera (oil tea), directly influencing seedling establishment and crop reproduction. In this study, we examined transcriptomic and physiological changes across five defined germination stages (G0–G4), from radicle dormancy to cotyledon emergence. Using RNA sequencing (RNA-seq), we assembled 169,652 unigenes and identified differentially expressed genes (DEGs) at each stage compared to G0, increasing from 1708 in G1 to 10,250 in G4. Functional enrichment analysis revealed upregulation of genes associated with cell wall organization, glucan metabolism, and Photosystem II assembly. Key genes involved in cell wall remodeling, including cellulose synthase (CESA), phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase (4CL), caffeoyl-CoA O-methyltransferase (COMT), and peroxidase (POD) showed progressive activation during germination. A Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed dynamic regulation of phenylpropanoid and flavonoid biosynthesis, photosynthesis, carbohydrate metabolism, and hormone signaling pathways. Transcription factors such as indole-3-acetic acid (IAA), ABA-responsive element binding factor (ABF), and basic helix–loop–helix (bHLH) were upregulated, suggesting hormone-mediated regulation of dormancy release and seedling development. Physiologically, cytokinin (CTK) and IAA levels peaked in G4, antioxidant enzyme activities were highest in G2, and starch content increased toward later stages. These findings provide new insights into the molecular mechanisms underlying seed germination in C. oleifera and identify candidate genes relevant to rootstock breeding and nursery propagation. Full article

Brain muscle ARNT-like1 (Bmal1) is a transcriptional factor, consisting of basic helix–loop–helix (bHLH) and PER-ARNT-SIM (PAS) domains, that plays a central role in circadian clock activity. However, the precise roles of the BMAL1-PAS domain, a circadian rhythm-regulating structure, remain unexplored in monocytes. Here, we highlight the BMAL1-PAS domain as a key structure in monocyte pleiotropic functions by using human monocytic cell line THP-1. THP-1 cells lacking the BMAL1-PAS-B domain (THP-1#207) abrogated the circadian expression of core clock genes. THP-1#207 cells exhibited less proliferation, glycolysis and oxidative phosphorylation activity, and LPS-induced IL-1β production, but exhibited more production of LPS-induced IL-10 than THP-1 cells. A quantitative proteomics analysis revealed significant expression changes in ~10% metabolic enzymes in THP-1#207 cells compared to THP-1 cells, including reduction in a rate-limiting enzyme hexokinase2 (HK2) in the glycolytic pathway. Importantly, treatment of THP-1 with 2-deoxy-D-glucose (2-DG), an HK2 inhibitor, largely recapitulated the phenotypes of THP-1#207 cells. These findings suggest that the BMAL1-PAS-B domain is an important structure for the regulation of proliferation, cellular energetics, and inflammatory response in THP-1 cells, at least in part, via the control of glycolytic activity. Thus, the BMAL1-PAS-B domain may become a promising pharmacological target to control inflammation. Full article

Background: Pinus massoniana is a significant lipid-producing tree species in China and a susceptible host for both the pine wood nematode and its insect vector, Monochamus alternatus. The basic helix–loop–helix (bHLH) family of transcription factors play a crucial role in responding to both biotic and abiotic stresses. However, the role of bHLH in terpene-induced defense in P. massoniana remains poorly studied. Results: Transcriptome sequencing using DNA Nanoball Sequencing (DNBSEQ) and PacBio Sequel platforms was performed, revealing differences in gene expression in P. massoniana branch under the simulated feeding treatment of methyl jasmonate (MeJA) spraying. Fifteen bHLH genes were cloned and analyzed, among which eight highly upregulated PmbHLH genes showed similar temporal expression after MeJA treatment and M. alternatus adult feeding. Five highly upregulated bHLH genes with nuclear localization were highly expressed in P. massoniana after M. alternatus feeding and interacted with the promoter of the terpene synthase gene Pm TPS (−)-α-pinene, confirming their involvement in the defense response of P. massoniana against the M. alternatus adult feeding. Conclusions: Our results unveil the temporal changes and the regulation of the induced defense system in P. massoniana mediated by both MeJA signaling and M. alternatus feeding treatment. The potential application for transgenic experiments and the breeding of resistant species in the future were discussed. Full article

Drought stress is one of the primary environmental factors affecting plant survival rates and productivity, and it is a key bottleneck restricting the development of the world kiwifruit industry. Therefore, studying the drought resistance-related genes and drought resistance mechanisms of kiwifruit is essential. The bHLH (basic helix-loop-helix) TF family plays a crucial role in the resistance of kiwifruit to abiotic stresses such as drought stress. In this study, we analyzed the response of the AcbHLH gene in kiwifruit under drought stress based on the kiwifruit genome database, transcriptome data, and metabolome data. One hundred eighty-seven AcbHLH genes were identified via bioinformatics and divided into eighteen subfamilies via phylogenetic analysis. The cis-acting elements of the AcbHLH gene are mainly hormone-related cis-acting elements. Under drought stress, 64 AcbHLH genes were significantly different, 5 AcbHLH genes whose expression significantly differed were randomly selected for qRT-PCR verification, and the correlation between the qRT-PCR results and the transcriptome data was high. The determination of plant hormone contents revealed that the contents of plant hormones, such as JA, changed markedly before and after drought stress. Through the combined analysis of transcriptome and metabolome data, it was speculated that AcbHLH84 and AcbHLH97 have functions similar to those of the MYC2 transcription factor and are the main downstream effectors in the JA signaling pathway; these functions could be activated and participate in the JA signaling pathway and that the activation of the JA signaling pathway would inhibit the production of reactive oxygen species. In turn, the drought resistance of kiwifruit is improved. The AcbHLH84 and AcbHLH97 genes could be candidate genes for breeding new transgenic drought-resistant kiwifruit varieties. Full article

The occurrence of anthocyanins in rice (Oryza sativa) and barley (Hordeum vulgare) varies among cultivars, with pigmented varieties (e.g., black rice and purple barley) accumulating higher concentrations due to genetic and environmental factors. The biosynthesis of anthocyanins is regulated by a complex network of structural and regulatory genes. Key enzymes in the pathway include chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), and UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT). These genes are tightly controlled by transcription factors (TFs) from the MYB, bHLH (basic helix–loop–helix), and WD40 repeat families, which form the MBW (MYB-bHLH-WD40) regulatory complex. In rice, OsMYB transcription factors such as OsMYB3, OsC1, and OsPL (Purple Leaf) interact with OsbHLH partners (e.g., OsB1, OsB2) to activate anthocyanin biosynthesis. Similarly, in barley, HvMYB genes (e.g., HvMYB10) coordinate with HvbHLH TFs to regulate pigment accumulation. Environmental cues, such as light, temperature, and nutrient availability, further modulate these TFs, influencing the production of anthocyanin. Understanding the genetic and molecular mechanisms behind the biosynthesis of anthocyanins in rice and barley provides opportunities for the development of biofortification strategies that enhance their nutritional value. Full article

Apple (Malus domestica Borkh.) is an economically important fruit. The use of nitrate by plants plays a crucial role in their growth and development, and its absorption and dispersal are controlled by nitrate transport proteins (NRTs). In this study, we investigated the potential function of MdNRT2.4 under low-nitrogen (N) stress by overexpressing it in tobacco. Compared with plants treated with a normal nitrogen level (5 mM), the MdNRT2.4 overexpression lines under low-N stress (0.25 mM) exhibited significantly greater plant height and width, as well as larger leaves and a higher leaf density, than wild-type plants, suggesting that the overexpression of MdNRT2.4 enhances the low-N tolerance of tobacco. Enhanced antioxidant enzyme activities in the MdNRT2.4 overexpression plant lines promoted the scavenging of reactive oxygen species, which reduced damage to their cell membranes. GUS staining of pMdNRT2.4::GUS-transformed Arabidopsis thaliana lines showed that MdNRT2.4 was expressed in the roots, vascular bundles, seeds in fruit pods, and young anther sites, suggesting that MdNRT2.4 mediates the transport of nitrate to these tissues, indicating that MdNRT2.4 might promote nitrate utilization in apple and improve its tolerance to low-N stress. Experiments using yeast one-hybrid and dual-luciferase assays revealed that MdbHLH3 binds to the MdNRT2.4 promoter and activates its expression. MdbHLH3 belongs to the basic helix–loop–helix (bHLH) transcription factor (TF). It is speculated that MdbHLH3 may interact with the promoter of MdNRT2.4 to regulate N metabolism in plants and enhance their low-N tolerance. This study establishes a theoretical framework for investigating the regulatory mechanisms of low-N responsive molecules in apple, while simultaneously providing valuable genetic resources for molecular breeding programs targeting low-N tolerance. Full article