RESUMEN
Passion fruit (Passiflora edulis) possesses a complex aroma and is widely grown in tropical and subtropical areas. Here, we conducted the de novo assembly, annotation, and comparison of PPF (P. edulis Sims) and YPF (P. edulis f. flavicarpa) reference genomes using PacBio, Illumina, and Hi-C technologies. Notably, we discovered evidence of recent whole-genome duplication events in P. edulis genomes. Comparative analysis revealed 7.6â¼8.1 million single nucleotide polymorphisms, 1 million insertions/deletions, and over 142â Mb presence/absence variations among different P. edulis genomes. During the ripening of yellow passion fruit, metabolites related to flavor, aroma, and color were substantially accumulated or changed. Through joint analysis of genomic variations, differentially expressed genes, and accumulated metabolites, we explored candidate genes associated with flavor, aroma, and color distinctions. Flavonoid biosynthesis pathways, anthocyanin biosynthesis pathways, and related metabolites are pivotal factors affecting the coloration of passion fruit, and terpenoid metabolites accumulated more in PPF. Finally, by heterologous expression in yeast (Saccharomyces cerevisiae), we functionally characterized 12 terpene synthases. Our findings revealed that certain TPS homologs in both YPF and PPF varieties produce identical terpene products, while others yield distinct compounds or even lose their functionality. These discoveries revealed the genetic and metabolic basis of unique characteristics in aroma and flavor between the 2 passion fruit varieties. This study provides resources for better understanding the genome architecture and accelerating genetic improvement of passion fruits.
Asunto(s)
Frutas , Passiflora , Frutas/genética , Odorantes , Passiflora/genética , Passiflora/metabolismo , Multiómica , Terpenos/metabolismoRESUMEN
Plants experience numerous biotic stresses throughout their lifespan, such as pathogens and pests, which can substantially affect crop production. In response, plants have evolved various metabolites that help them withstand these stresses. Here, we show that two specialized metabolites in the herbaceous perennial Belamcanda chinensis, tectorigenin and its glycoside tectoridin, have diverse defensive effects against phytopathogenic microorganisms and antifeeding effects against insect pest. We further functionally characterized a 7-O-uridine diphosphate glycosyltransferase Bc7OUGT, which catalyses a novel reversible glycosylation of tectorigenin and tectoridin. To elucidate the catalytic mechanisms of Bc7OUGT, we solved its crystal structure in complex with UDP and UDP/tectorigenin respectively. Structural analysis revealed the Bc7OUGT possesses a narrow but novel substrate-binding pocket made up by plentiful aromatic residues. Further structure-guided mutagenesis of these residues increased both glycosylation and deglycosylation activities. The catalytic reversibility of Bc7OUGT was also successfully applied in an one-pot aglycon exchange reaction. Our findings demonstrated the promising biopesticide activity of tectorigenin and its glycosides, and the characterization and mechanistic study of Bc7OUGT could facilitate the design of novel reversible UGTs to produce valuable glycosides with health benefits for both plants and humans.
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Glicosiltransferasas , Isoflavonas , Humanos , Glicosiltransferasas/genética , Isoflavonas/química , Glicosilación , Plantas/metabolismo , Uridina Difosfato , GlicósidosRESUMEN
BACKGROUND: Mammalian intestinal epithelium constantly undergoes rapid self-renewal and regeneration sustained by intestinal stem cells (ISCs) within crypts. Inducible nitric oxide synthase (iNOS) is an important regulator in tissue homeostasis and inflammation. However, the functions of iNOS on ISCs have not been clarified. Here, we aimed to investigate the expression pattern of inducible nitric oxide synthase (iNOS) within crypts and explore its function in the homeostatic maintenance of the ISC niche. METHODS: Expression of iNOS was determined by tissue staining and qPCR. iNOS-/- and Lgr5 transgenic mice were used to explore the influence of iNOS ablation on ISC proliferation and differentiation. Enteroids were cultured to study the effect of iNOS on ISCs in vitro. Ileum samples from wild-type and iNOS-/- mice were collected for RNA-Seq to explore the molecular mechanisms by which iNOS regulates ISCs. RESULTS: iNOS was physiologically expressed in Paneth cells. Knockout of iNOS led to apparent morphological changes in the intestine, including a decrease in the small intestine length and in the heights of both villi and crypts. Knockout of iNOS decreased the number of Ki67+ or BrdU+ proliferative cells in crypts. Loss of iNOS increased the number of Olfm4+ ISCs but inhibited the differentiation and migration of Lgr5+ ISCs in vivo. iNOS depletion also inhibited enteroid formation and the budding efficiency of crypts in vitro. Moreover, iNOS deficiency altered gluconeogenesis and the adaptive immune response in the ileum transcriptome. CONCLUSION: Paneth cell-derived iNOS is required to maintain a healthy ISC niche, and Knockout of iNOS hinders ISC function in mice. Therefore, iNOS represents a potential target for the development of new drugs and other therapeutic interventions for intestinal disorders.
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Células de Paneth , Nicho de Células Madre , Animales , Ratones , Homeostasis , Mucosa Intestinal/metabolismo , Intestinos , Mamíferos/metabolismo , Ratones Noqueados , Ratones Transgénicos , Óxido Nítrico Sintasa de Tipo II/metabolismo , Células de Paneth/metabolismo , Receptores Acoplados a Proteínas G/metabolismoRESUMEN
Response surface methodology was used to optimize the medium composition to improve the production of the heat-tolerant ß-glucosidase from Bacillus sp. ZJ1308. Three significant factors were found to be corn cob, beef extract, and MnSO4 ·H2 O. The final medium compositions optimized were corn cob (51.8 g/L), beef extract (23.8 g/L), salicin (0.5 g/L), MnSO4 ·H2 O (0.363 g/L), MgSO4 ·7H2 O (0.4 g/L), and NaCl (5 g/L). Under the optimal conditions, the activity of ß-glucosidase was up to 4.71 U/mL. ß-Glucosidase was purified to homogeneity with a recovery rate of 5% and a specific activity of 110.47 U/mg. The optimal pH and temperature were 7.0 and 60 °C, respectively. ß-Glucosidase was stable within a pH range of 6.0-8.0 and showed an extremely high thermostability at 80 and 90 °C, retaining 56% and 38% of its maximal activity, respectively. Ni(2+) and Ba(2+) heavily inhibited the ß-glucosidase activity. The purified ß-glucosidase showed a high substrate specificity. The kinetic parameters revealed that it had a high catalytic efficiency toward the substrate p-nitrophenyl-ß-d-glucopyranoside (Kcat /Km = 700). It also showed a high catalytic activity toward the natural substrate salicin. This study provides a new insight into the future development and use of ß-glucosidase from Bacillus sp. ZJ1308.
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Bacillus/metabolismo , Biotecnología/métodos , Calor , beta-Glucosidasa/biosíntesis , beta-Glucosidasa/genética , Bacillus/aislamiento & purificación , Microbiología del Suelo , Especificidad por Sustrato , beta-Glucosidasa/aislamiento & purificaciónRESUMEN
Glycosylation plays a very important role in plant secondary metabolic modifications. Neodiosmin, identified as diosmetin-7-O-neohesperidoside, not only acts to mitigate bitterness and enhance the flavor of food but also serves as a pivotal metabolite that reinforces plant immunity. Investigating its biosynthetic pathway in plants is crucial for optimizing fruit quality and fortifying plant immune responses. In this study, through analysis of transcriptomic data from Astilbe chinensis, we identified two novel uridine diphosphate (UDP)-glycosyltransferases (UGTs): Ach14791 (AcUGT73C18), responsible for flavonoid 7-O-glycosylation and Ach15849 (AcUGT79B37), involved in flavonoid-7-O-glucoside-2â³-O-rhamnosylation. By delving into enzymatic properties and catalytic promiscuity, we developed a biosynthesis route of neodiosmin by establishing a one-pot enzyme-catalyzed cascade reaction. Simultaneously, lonicerin and rhoifolin were also successfully synthesized using the same one-pot dual-enzyme catalytic reaction. Taken together, our findings not only identified two novel UGTs involved in neodiosmin biosynthesis but also provided important biocatalytic components for the microorganism-based biosynthesis of flavonoid-7-O-disaccharide compounds.
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Flavonoides , Glicosiltransferasas , Glicosiltransferasas/genética , Glicosiltransferasas/metabolismo , Glicosilación , Flavonoides/metabolismo , Uridina Difosfato/metabolismo , Glicósidos , FilogeniaRESUMEN
Radiation exposure often leads to serious health problems in humans. The intestinal epithelium is sensitive to radiation damage, and radiation causes destruction of the intestinal epithelial barrier, which leads to radiation enteritis (RE), the loss of fluids, and the translocation of intestinal bacteria and toxins; radiation can even threaten survival. In this study, we aimed to explore the influence of IVIg on the integrity of the intestinal epithelial barrier after RE. Using a RE mouse model, we investigated the protective effects of intravenous immunoglobulin (IVIg) on the epithelial junctions of RE mice and validated these findings with intestinal organoids cultured in vitro. In addition, transmission electron microscopy (TEM), western blotting (WB) and immunostaining were used to further investigate changes in intestinal epithelial ferroptosis and related signaling pathways. When RE occurs, the intestinal epithelial barrier is severely damaged. IVIg treatment significantly ameliorated this damage to epithelial tight junctions both in vivo and in vitro. Notably, IVIg alleviated RE by inhibiting intestinal epithelial ferroptosis in RE mice. Mechanistically, IVIg promoted activation of the mTOR pathway and inhibited ferroptosis in the intestinal epithelium of mice. Rapamycin, which is a potent inhibitor of the mTOR protein, significantly abolished the protective effect of IVIg against radiation-induced damage to intestinal epithelial tight junctions. Overall, IVIg can prevent RE-induced damage to the intestinal epithelial barrier and inhibit ferroptosis by activating the mTOR pathway; this study provides a new treatment strategy for patients with RE caused by radiotherapy or accidental nuclear exposure.
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Enteritis , Ferroptosis , Exposición a la Radiación , Humanos , Ratones , Animales , Inmunoglobulinas Intravenosas/farmacología , Inmunoglobulinas Intravenosas/uso terapéutico , Intestinos , Mucosa Intestinal , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
Intestinal stem cells (ISCs) are pivotal for the maintenance and regeneration of the intestinal epithelium. Berberine (BBR) exhibits diverse biological activities, but it remains unclear whether BBR can modulate ISCs' function. Therefore, we investigated the effects of BBR on ISCs in healthy and radiation-injured mice and explored the potential underlying mechanisms involved. The results showed that BBR significantly increased the length of the small intestines, the height of the villi, and the depth and density of the crypts, promoted the proliferation of cryptal epithelial cells and increased the number of OLFM4+ ISCs and goblet cells. Crypts from the BBR-treated mice were more capable of growing into enteroids than those from untreated mice. BBR alleviated WAI-induced intestinal injury. BBR suppressed the apoptosis of crypt epithelial cells, increased the quantity of goblet cells, and increased the quantity of OLFM4+ ISCs and tdTomato+ progenies of ISCs after 8 Gy WAI-induced injury. Mechanistically, BBR treatment caused a significant increase in the quantity of p-S6, p-STAT3 and p-ERK1/2 positive cryptal epithelial cells under physiological conditions and after WAI-induced injury. In conclusion, BBR is capable of enhancing the function of ISCs either physiologically or after radiation-induced injury, indicating that BBR has potential value in the treatment of radiation-induced intestinal injury.
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Berberina , Mucosa Intestinal , Ratones Endogámicos C57BL , Células Madre , Animales , Berberina/farmacología , Berberina/uso terapéutico , Células Madre/efectos de los fármacos , Ratones , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/efectos de la radiación , Mucosa Intestinal/patología , Masculino , Apoptosis/efectos de los fármacos , Apoptosis/efectos de la radiación , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Traumatismos Experimentales por Radiación/tratamiento farmacológico , Traumatismos Experimentales por Radiación/patología , Células Caliciformes/efectos de los fármacos , Células Caliciformes/efectos de la radiación , Células Caliciformes/patología , Traumatismos por Radiación/tratamiento farmacológico , Traumatismos por Radiación/patología , Factor de Transcripción STAT3/metabolismo , Intestino Delgado/efectos de los fármacos , Intestino Delgado/efectos de la radiación , Intestino Delgado/patología , Intestino Delgado/lesiones , Intestinos/efectos de los fármacos , Intestinos/efectos de la radiaciónRESUMEN
Mitochondria are critical for cellular energetic metabolism, intracellular signaling orchestration and programmed death regulation. Therefore, mitochondrial dysfunction is associated with various pathogeneses. The maintenance of mitochondrial homeostasis and functional recovery after injury are coordinated by mitochondrial biogenesis, dynamics and autophagy, which are collectively referred to as mitochondrial quality control. There is increasing evidence that mitochondria are important targets for melatonin to exert protective effects under pathological conditions. Melatonin, an evolutionarily conserved tryptophan metabolite, can be synthesized, transported and metabolized in mitochondria. In this review, we summarize the important role of melatonin in the damaged mitochondria elimination and mitochondrial energy supply recovery by regulating mitochondrial quality control, which may provide new strategies for clinical treatment of mitochondria-related diseases.
RESUMEN
A fluorescence enhancement phenomenon in the europium (Eu)-Ofloxacin (OF)-Sodium Dodecyl Benzene Sulfonate (SDBS) fluorescence system was observed when Gd(3+) was added. The fluorescence intensity of the systems was measured (lambda (ex)/lambda (em) = 280/612 nm) at pH 7.8. Under optimum conditions, a linear relationship between the enhanced fluorescence intensity and the Eu(3+) concentration in the range of 5.0 x 10(-10) approximately 2.0 x 10(-7) mol x L(-1) was observed. The detection limit of Eu(3+) was 1.46 x 10(-10) mol x L(-1) (S/N = 3). This method was used for the determination of trace amounts of europium in synthetic rare earth samples with satisfactory results. In addition, the interaction mechanism is also studied.