Search Results (154)

Colored wheat lines, which feature elevated anthocyanin content and associated traits, represent valuable genetic resources for enhancing the plant’s nutritional and aesthetic properties. This genome-wide association study (GWAS) utilized a set of radiation-induced mutant lines to identify genetic loci linked to agricultural and biochemical traits. The GWAS models Fixed and Random Model Circulating Probability Unification, and the Bayesian-information and Linkage-Disequilibrium Iteratively Nested Keyway were employed to increase the reliability of marker–trait associations (MTAs). In total, 35 significant MTAs were identified, and seven single-nucleotide polymorphisms (SNPs) were commonly detected by both models. To explore candidate genes, a ± 1.5-Mb window around each significant SNP was analyzed according to the estimated linkage disequilibrium decay, revealing 635 genes. Among these, several genes were annotated as transcription factors and enzymes associated with flavonoid biosynthesis and modification, including MYB, WD-repeat proteins, and UDP-glycosyltransferases. Expression profiling and RT-qPCR further supported the functional relevance of selected SNP–gene pairs, particularly for anthocyanin accumulation and seed color variation. In summary, the integration of GWAS, gene annotation, and expression data could provide valuable insights into the genetic basis of complex traits in wheat, providing data for future molecular studies and marker-assisted breeding of colored wheat mutant cultivars. Full article

UDP-glycosyltransferases (UGTs) play essential roles in various biological processes, such as phytohormone homeostasis, abiotic stress adaptation, and secondary metabolite biosynthesis. Populus euphratica is a model species for investigating stress adaptation; however, the PeUGT gene family has yet to be systematically characterized. Here, we identified 134 UGT genes in P. euphratica. Phylogenetic analysis classified these genes into 16 major groups (A–P), and UGT genes within the same groups showed similar structural characteristics. Tandem duplication events were identified as the predominant mechanism driving the expansion of the PeUGT family. Cis-acting element analysis revealed an enrichment of motifs associated with developmental regulation, light response, phytohormone signaling, and abiotic stress in the promoters of PeUGT genes. Expression profiling demonstrated spatiotemporal regulation of the PeUGT genes under drought stress. Among them, PeUGT110 was significantly induced by PEG treatment in the leaf, root, and stem tissues of P. euphratica. Overexpression of PeUGT110 enhanced drought tolerance in transgenic Arabidopsis. Furthermore, the PeUGT110-OE lines exhibited reduced malonaldehyde accumulation, elevated proline content, higher superoxide dismutase activity, and upregulated expression of stress-related genes under drought stress. The results demonstrated that PeUGT110 plays a critical role in plant drought resistance. These findings establish a foundation for elucidating the function of PeUGT genes. Full article

Daptomycin (DAP) is a therapeutic option for vancomycin-resistant Enterococcus faecium (VRE) infections, but DAP resistance may occur during treatment. Previously, we identified a mutation within the E. faecium lafB gene that induces hypersusceptibility to DAP. The lafB gene encodes a glycosyltransferase involved in lipoteichoic acid anchor synthesis, which makes it a promising target for enhancing DAP efficacy. In this study, we characterized E. faecium LafB protein (EfLafB) biophysical properties, used AlphaFold3 to predict LafB in silico three-dimensional structure, and determined lafB gene mutation’s role in virulence, comparing E. faecium HBSJRP18 (DAP-hypersusceptible) and a lafB revertant, HBSJRP18_2.7, and analyzing bacterial growth kinetics, biofilm formation ability, and virulence in a Galleria mellonella model. After gene cloning and expressing and purifying EfLafB, circular dichroism and SEC-MALS assays revealed its monomeric nature under in vitro conditions, with approximately a 40 kDa molecular mass and a melting temperature of 50 °C. In silico prediction indicated that LafB is an αβ-type protein with two domains conforming to the GT-4 family glycosyltransferases. These results are further supported by the highly conserved amino acids (E257, D91, R184, and K185), likely involved in UDP-Glc binding. The studied lafB gene mutation resulted in a significant decrease in bacterial growth and virulence in the invertebrate model. Full article

UDP-glycosyltransferases (UGTs) are widely distributed enzymes in living organisms that catalyze the transfer of glycosyl groups from donor molecules to acceptor molecules’ glycoside ligands. These enzymes are pivotal for detoxifying and eliminating both endogenous and exogenous toxic substances in insects. In this study, bioinformatics methods were used to analyze the UGT gene superfamily in the fall armyworm (Spodoptera frugiperda), resulting in the identification of 48 UGT genes located across 10 chromosomes, including 23 tandem duplication pairs. The predicted SfUGT proteins mainly exhibit α-helical secondary structures. Intron numbers varied significantly, with high diversity observed in amino acid sequences. Phylogenetic analysis grouped UGT genes from three insect species into three distinct subfamilies, revealing a closer evolutionary relationship between S. frugiperda and Spodoptera litura, supported by a greater number of orthologous genes. Expression profiling showed that SfUGT16 and SfUGT21 are highly expressed in the first and fourth larval instars, respectively; SfUGT16 is predominantly expressed in the Malpighian tubules and midgut, implying roles in digestion, metabolism, and detoxification. Meanwhile, SfUGT21, SfUGT30, and SfUGT48 exhibited elevated expression in the hemolymph, suggesting functions in metabolism and transport, whereas SfUGT40 showed high expression in both the midgut and hemolymph, indicating involvement in detoxification and metabolic processes. These findings provide a foundation for further exploration of the biological functions of the UGT gene family. Full article

Uridine diphosphate glycosyltransferase (UGT) is a core protein for glycosylation of plant natural products and other small molecules. Although many studies on functional identification of UGTs are now available, analysis of UGTs in Platycodon grandiflorus is still relatively scarce. We identified 107 PgUGTs genome-wide from P. grandiflorus and investigated their phylogenetic relationships, chromosomal localisation, collinearity, cis-regulatory elements, motifs, domains, and gene structures. PgUGT29 and PgUGT72 were two putative glycosyltransferases for platycodins biosynthesis in P. grandiflorus according to our previous study and bioinfornatical analyses. In vitro enzyme activity showed that PgUGT29 can catalyse the glycosylation of the C3 position of Platycodin D (PD) to generate Platycodin D3 (PD3), while candidate enzyme PgUGT72 does not function as a glycosyltransferase. Molecular docking indicated that T145, D392, Q393, and N396 may be the crucial residues for PgUGT29 to catalyse the generation of PD3 from UDP-Glc and PD. In this study, we identified and cloned PgUGT29, elucidated its catalytic function in converting PD to PD3, and predicted key residues critical for its enzymatic activity. These findings provide a theoretical foundation and technical framework for future targeted metabolic engineering and directional regulation of medicinal components in Platycodon grandiflorus. Full article

Dihydroflavonol 4-reductase (DFR) genes contribute greatly to anthocyanin biosynthesis in plants. Up to now, however, research on the DFR gene family and the key anthocyanin-related DFR members in blueberries (Vaccinium corymbosum) has been limited. In this study, we performed a genome-wide identification of the blueberry DFR gene family, identifying 36 VcDFR genes categorized into five subfamilies. Gene expression analysis showed that three Subfamily III members (VcDFR11/29/34) and four Subfamily V members (VcDFR4/7/30/33) are highly expressed in blueberry fruits, particularly at late ripening stages. Transient overexpression analysis in apple fruits verified the contributions of VcDFR11 and VcDFR30 to anthocyanin biosynthesis, with VcDFR11 showing better promoting effects. Blueberry fruit-based transient overexpression further confirmed the promoting effects of VcDFR11 on anthocyanin accumulation and the expression of anthocyanin-related structural genes (especially its downstream anthocyanindin synthase (ANS) and UDP-glucose: flavonoid 3-O-glycosyltransferase (UFGT) genes). The VcDFR11 promoter contains binding sites for both bHLH and MYB transcription factors (TFs). Consistently, yeast one-hybrid and dual-luciferase assays confirmed that anthocyanin-related VcMYB-1 and VcbHLHs can bind to and activate the VcDFR11 promoter. Furthermore, co-overexpressing VcMYB-1/VcbHLHs with VcDFR11 led to much higher anthocyanin accumulation than overexpressing VcDFR11 alone, indicating that these TFs positively regulate anthocyanin biosynthesis by upregulating VcDFR11. In summary, our study characterized the blueberry DFR gene family and demonstrated the role of VcDFR11 in anthocyanin biosynthesis. Full article

Fungal infections of crops pose a threat to global agriculture. Fungi of the genus Fusarium cause widespread diseases in cereal crops. Fusarium graminearum reduces yields and produces harmful mycotoxins such as deoxynivalenol (DON). Plants mitigate DON toxicity through glucose conjugation mediated by UDP-glycosyltransferases (UGTs), forming deoxynivalenol-3-O-glucoside (DON-3-Glc). Few such UGTs have been identified, predominantly from Fusarium-susceptible crops. Given that the presence of this activity in diverse plants and across broader UGT subfamilies and groups was underexplored, we screened a library of 380 recombinant plant UGTs and identified and characterized eight novel enzymes glycosylating DON in vitro. Among these, ZjUGT from Ziziphus jujuba stood out with the highest activity, showing an apparent k_cat of 0.93 s⁻¹ and k_cat/K_m of 2450 M⁻¹ s⁻¹. Interestingly, four enzymes produced primarily a novel, still uncharacterized glucoside. Furthermore, we evaluated the in vivo resistance provided by these UGTs when expressed in a DON-sensitive yeast strain. At least six of the novel UGTs conferred some level of resistance, allowing growth at concentrations of up to 120 mg/L of DON. This study contributes to potential strategies to enhance DON resistance in cereal crops in the future. Full article

Flower color is one of the most ornamental values of Meconopsis wilsonii, but very limited studies have been reported on its flower color formation. The UDP-glycosyltransferase (UGT) gene family plays a crucial role in plant flower color formation. In this study, the full-length transcriptome data of M. wilsonii was used to identify MwUGTs, focusing on protein physicochemical properties’ subcellular localization, and phylogenetic relationships. In addition, sequence analysis, expression pattern analysis, subcellular localization, and functional validation of MwUGT2 were also performed. A total of 26 MwUGTs were identified in full-length transcriptome and clustered into eight subgroups. Phylogenetic analysis and KEGG database annotation showed that MwUGT2 is associated with anthocyanin synthesis and accumulation. Subsequently, based on the expression of MwUGT2 during flower development and in different tissues, it was preliminarily determined that MwUGT2 plays a role in the flower bud stage. Subcellular localization assays suggested that MwUGT2 is present in the nucleus and cytoplasm. Overexpression in Nicotiana tabacum showed that MwUGT2 significantly increased the content of Cyanidin-3-O-glucoside and resulted in dark pink flowers in transgenic plants. In summary, our findings suggest that MwUGT2 plays a crucial role in the biosynthesis of anthocyanin and will also contribute to understanding the mechanisms of flower color formation in M. wilsonii. Full article

As the main active ingredient in Anoectochilus roxburghii, kinsenoside has important health and medical effects including hepatoprotective, anti-oxidant, and bacteriostasis, among others. In recent years, with the limited application of high-throughput technology to A. roxburghii, there has been no research on the key regulatory genes involved in the synthesis of kinsenoside. Therefore, we examined three species of A. roxburghii that are widely planted in mainland China and Taiwan Province, A. roxburghii cultivar ‘Jian ye’, Anoectochilus formosanus, and Anoectochilus burmannicus, determining the content of kinsenoside, performing transcriptomic and metabolomic sequencing, identifying UDP glycosyltransferases, and screening for UDP glycosyltransferases that may be involved in kinsenoside synthesis. The results showed that among the three species of A. roxburghii, the content of kinsenoside in A. roxburghii cv. ‘Jian ye’ was the highest. Transcriptome and metabolome data showed that A. roxburghii cv. ‘Jian ye’ and the two other species of A. roxburghii have 3702 and 5369 differentially expressed genes and 69 and 120 differentially accumulated metabolites, respectively. Meanwhile, differentially expressed genes and differentially accumulated metabolites are enriched in the glucose metabolism and hormone pathways. We also treated the A. roxburghii samples with exogenous auxin and characterized the related genes. In A. roxburghii, we identified 73 members of the UDP glycosyltransferase family. Through phylogenetic tree, transcriptome data expression profile, and qPCR analyses, we screened for members that may be involved in the synthesis of kinsenoside. In summary, the results of this study provide insights for breeding high-kinsenoside-content and high-intron varieties of A. roxburghii. Full article

UDP-glycosyltransferases (UGTs) contribute to catalyzing the glycosylation of numerous functional natural products and novel derivatives with improved bioactivities. UDP-glucose sterol glucosyltransferase (SGT) is normally involved in the synthesis of sterol glycosides in a variety of organisms. SGT was derived from Salinispora tropica CNB-440 and heterologously expressed in Escherichia coli BL21 (DE3). Novel 12-O-glucosylginsenoside Rh2 was identified using HPLC, high-resolution MS (HR-MS), and NMR analysis. The cell viability assay was performed on 12-O-glucosylginsenoside-treated AGS stomach cancer, HeLa cervical cancer, U87MG glioma, and B16F10 melanoma cell lines. Protein structure modeling, molecular docking, and dynamics simulations were performed using AutoDock 4.2 and GROMACS 2020.1 software. The SGT gene is comprised of 1284 nucleotides and codes for 427 amino acids. The 12-O-glucosylginsenoside Rh2 may be a potential anticancer agent due to its potent viability inhibition of cancer cells. Structural analysis showed critical perspectives into the intermolecular interactions, stability, and binding energetics of the enzyme–ligand complex, with outcomes complementing the experimental data, thereby deepening our understanding of the structural basis of SGT-mediated glycosylation and its functional implications. This report presents a novel ginsenoside, 12-O-glucosylginsenoside Rh2, utilizing reshuffled SGT derived from S. tropica, and provides a promising candidate for anticancer drug research and development. Full article

L-ascorbic acid (vitamin C, AA) is widely present in plants, but humans lack the ability to synthesize it independently. As a potent reducing agent, AA is susceptible to oxidation, making the enhancement of its stability crucial. 2-O-β-D-glucopyranosyl-L-ascorbic acid (AA-2βG) is a stable natural derivative of AA with glycosylation, initially discovered in the fruits of Lycium barbarum. Understanding the biosynthesis of AA-2βG is crucial for enhancing its production in L. barbarum. While the established biosynthesis pathway of AA constitutes the upstream of AA-2βG biosynthesis, the conclusive step of β-glycosylation remains unclear. We identified a L. barbarum cultivar by UPLC, ZN01, with a high content of AA-2βG, and compared its leaves, immature fruits, and mature fruits to a normal AA-2βG content L. barbarum cultivar for metabolomic and transcriptomic analysis. The RNA-seq and RT-qPCR analysis revealed that the expression levels of genes involved in the AA biosynthesis pathway did not consistently correlate with AA-2βG content, suggesting that the final glycosylation step may be a key determinant of AA-2βG accumulation. Subsequently, utilizing phylogenetic and co-expression analysis, we identified ten UDP-glycosyltransferases (UGTs) and three β-glucosidases (BGLUs) which may be involved in the crucial step of the conversion from AA to AA-2βG, and the UGTs’ activities were predicted through molecular docking. Lastly, we speculated that the presence of the glycosylation process of AA might have a crucial role in maintaining AA homeostasis in L. barbarum, and deliberated on potential correlations between AA, carotenoids, and anthocyanins. Our integrated multi-omics analysis provides valuable insights into AA-2βG biosynthesis in L. barbarum, identifying thirteen candidate genes and highlighting the complex interplay between AA, carotenoids, and anthocyanins. These findings have implications for improving AA-2βG content in L. barbarum. Full article

Mutagenesis breeding, combined with the application of corresponding herbicides to develop herbicide-resistant rice germplasm, provides great promise for the management of weeds and weedy rice. In this study, a topramezone-resistant rice mutant, TZR1, was developed from the indica rice line Chuangyu 9H (CY9H) through radiation mutagenesis and topramezone selection. Dose–response curves revealed that the resistance index of TZR1 to topramezone was 1.94-fold compared to that of CY9H. The resistance mechanism of TZR1 was not due to target-site resistance. This resistance could be reversed by a specific inhibitor of glutathione S-transferase (GST). The activity of antioxidant enzymes was analyzed. SNPs and Indels were detected using whole-genome resequencing; differentially expressed genes were identified through RNA sequencing. Then, they underwent Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. Key candidate genes associated with topramezone resistance were validated via a real-time quantitative PCR assay. Five GST genes, two UDP-glycosyltransferase genes, and three ATP-binding cassette transporter genes were identified as potential contributors to topramezone detoxification in TZR1. Overall, these findings suggest that GST enzymes possibly play an important role in TZR1 resistance to topramezone. This study will provide valuable information for the scientific application of 4-hydroxyphenylpyruvate dioxygenase inhibitors in paddy fields in future. Full article

Prunus mume, a traditional ornamental species native to China, is highly valued for both its captivating weeping variety and economic value. The glycosylation of metabolites, which is mediated by UDP-glycosyltransferases (UGTs), is essential for the regulation of secondary metabolic pathways in plants. Here, we systematically identified and analyzed the UGTs in P. mume. A total of 182 PmUGTs were identified using genomic data and categorized into 16 distinct subfamilies (A–P). All PmUGTs were distributed unevenly across the eight chromosomes, with clear evidence of tandem duplication. Additionally, synteny analysis revealed a close evolutionary relationship between P. mume and Prunus persica. A promoter cis-acting element analysis indicated that PmUGTs may respond to light, hormones, and external stresses. A heatmap analysis revealed that PmUGTs had specific expression patterns across different tissues, under various hormone treatments, and in different developmental stages of stem lignification. Notably, qRT-PCR verification showed significant differences in PmUGT163 expression between straight and weeping stems, underscoring its role in regulating plant architecture formation. Taken together, our study elucidates the evolutionary trajectory of PmUGTs and lays the groundwork for the further validation of the candidate genes involved in plant architectural formation. Full article

The release of herbivore-induced plant volatiles (HIPVs) has been recognized to be an important strategy for plant adaptation to herbivore attack. However, whether these induced volatiles are beneficial to insect herbivores, particularly insect larvae, is largely unknown. We used the two important highly polyphagous lepidopteran pests Spodoptera frugiperda and S. litura to evaluate the benefit on xenobiotic detoxification of larval exposure to HIPVs released by the host plant maize (Zea mays). Larval exposure of the invasive alien species S. frugiperda to maize HIPVs significantly enhanced their tolerance to all three of the well-known defensive compounds 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), chlorogenic acid, and tannic acid in maize and the two commonly used insecticides methomyl and chlorpyrifos. HIPV exposure also improved the larval tolerance of S. litura third instars to chlorogenic and tannic acids. Furthermore, larval exposure to either maize HIPVs or DIMBOA induced the activities of cytochrome P450 enzymes (P450s), glutathione-s-transferase (GST), and carboxylesterase (CarE) in the midguts and fat bodies of the two insects, while the induction was significantly higher by the two components together. In addition, the expression of four genes encoding uridine diphosphate (UDP)-glycosyltransferases (UGT33F28, UGT40L8) and P450s (CYP4d8, CYP4V2) showed similar induction patterns in S. frugiperda. Cis-3-hexen-1-ol, an important component in maize HIPVs, also showed the same functions as maize HIPVs, and its exposure increased larval xenobiotic tolerance and induced the detoxification enzymes and gene expression. Our findings demonstrate that HIPVs released by the pest-infested host plants are conductive to the xenobiotic tolerance of lepidopteran insect larvae. Hijacking the host plant HIPVs is an important strategy of the invasive alien polyphagous lepidopteran pest to counter-defend against the host plant’s chemical defense. Full article

Vibrio sp. KMM 8419 (=CB1-14) is a Gram-negative bacterium isolated from a food-net mucus sample of marine polychaete Chaetopterus cautus collected in the Sea of Japan. Here, we report the structure and biosynthetic gene cluster of the capsular polysaccharide (CPS) from strain KMM 8419. The CPS was isolated and studied by one- and two-dimensional ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy. The molecular weight of the CPS was about 254 kDa. The CPS consisted of disaccharide repeating units of D-glucose and sulfated and acetylated L-rhamnose established as →2)-α-L-Rhap3S4Ac-(1→6)-α-D-Glcp-(1→. To identify the genes responsible for CPS biosynthesis, whole-genome sequencing of KMM 8419 was carried out. Based on the genome annotations together with the Interproscan, UniProt and AntiSMASH results, a CPS-related gene cluster of 80 genes was found on chromosome 1. This cluster contained sets of genes encoding for the nucleotide sugar biosynthesis (UDP-Glc and dTDP-Rha), assembly (glycosyltransferases (GT)), transport (ABC transporter) and sulfation (PAPS biosynthesis and sulfotransferases) of the sulfated CPS. A hypothetical model for the assembly and transportation of the sulfated CPS was also proposed. In addition, this locus included genes for O-antigen biosynthesis. Further studies of biological activity, the structure–activity relationship in the new sulfated polysaccharide and its biosynthesis are necessary for the development of potent anticancer agents or drug delivery systems. Full article