Important Metabolites Research Articles

Cryo-EM and photocrosslinking studies reveal the mechanism of mitochondrial protein import by the TIM complex.

Mitochondria are essential organelles that provide cells with energy through oxidative phosphorylation, as well as with many important metabolites. Mitochondria import nearly all their ∼1,000-2,000 constituent proteins from the cytosol across their double-membrane envelope. Genetic and biochemical studies have shown that the conserved protein translocase, termed the TIM23 complex, mediates import of presequence-containing precursor proteins (preproteins) into the mitochondrial matrix and inner membrane. However, the mechanism by TIM23 forms a translocation path in the membrane and enables the import process remained unclear. We have determined the cryo-EM structure of the core TIM23 complex (heterotrimeric Tim17-Tim23-Tim44) from Saccharomyces cerevisiae. Contrary to the prevailing model, Tim23 and Tim17 themselves do not form a water-filled channel, but instead have separate, lipid-exposed concave cavities that face in opposite directions. Our structural and biochemical analyses show that surprisingly, the cavity of Tim17, not Tim23, forms the protein translocation path whereas Tim23 is likely to play a structural role. The results further suggest that, during translocation of substrate polypeptides, the nonessential subunit Mgr2 seals the lateral opening of the Tim17 cavity to facilitate the translocation process. We propose a new model for the TIM23-mediated protein import and sorting mechanism, a central pathway in mitochondrial biogenesis.

Integrated transcriptome and metabolome analysis reveals that flavonoids function in wheat resistance to powdery mildew.

Powdery mildew is a fungal disease devastating to wheat, causing significant quality and yield loss. Flavonoids are important secondary plant metabolites that confer resistance to biotic and abiotic stress. However, whether they play a role in powdery mildew resistance in wheat has yet to be explored. In the present study, we combined transcriptome and metabolome analyses to compare differentially expressed genes (DEGs) and differentially accumulated flavonoids identified in plants with and without powdery mildew inoculation. Transcriptome analysis identified 4,329 DEGs in susceptible wheat cv. Jimai229, and 8,493 in resistant cv. HHG46. The DEGs were functionally enriched using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes, revealing the flavonoid synthesis pathway as the most significant in both cultivars. This was consistent with the upregulation of flavonoid synthesis pathway genes observed by quantitative PCR. Metabolome analysis indicated flavone and flavonol biosynthesis pathways as the most significantly enriched following powdery mildew inoculation. An accumulation of total flavonoids content was also found to be induced by powdery mildew infection. Exogenous flavonoids treatment of inoculated plants led to less severe infection, with fewer and smaller powdery mildew spots on the wheat leaves. This reduction is speculated to be regulated through malondialdehyde content and the activities of peroxidase and catalase. Our study provides a fundamental theory for further exploration of the potential of flavonoids as biological prevention and control agents against powdery mildew in wheat.

Micropropagation and GC–MS analysis of bioactive compounds in bulbs and callus of white squill

Medicinal herbs are the main source of bioactive compounds used in the medical industry. White squill (Urginea maritima) is an important medicinal and ornamental plant cultivated in the Mediterranean region. This study reports an efficient protocol for in vitro propagation of Urginea and investigates important bioactive compounds present in the bulbs and in vitro–produced callus. The least number of days for callus induction and shoot regeneration was achieved with Murashige and Skoog (MS) basal media supplemented with 1.0 mg L−1 1-naphthaleneacetic acid (NAA) plus 0.1 mg L−1 6-benzylaminopurine (BAP) and 1.0 mg L−1 NAA plus 0.4 mg L−1 BAP, respectively, while the highest number of shoots and fresh weight were obtained at medium supplemented with 1.0 mg L−1 NAA plus 0.5 mg L−1 BAP. Gas chromatography-mass spectrometry (GC–MS) analysis of Urginea bulb methanol extract showed the existence of important secondary metabolites, such as palmitic acid (C16H32O2), 9-hexadecenoic acid (C16H30O2), phthalic acid 2-ethylhexyl propyl ester (C19H28O4), tetradecanoic acid (C14H28O2), undecanoic acid (C11H22O2), and oleic acid (C18H34O2), in addition to other important compounds, such as 13-heptadecyn-1-ol, 9,12-octadecadienoic acid, 1-monolinoleoylglycerol trimethylsilyl ether, 2-methyl-1-hexadecanol, and octadecanoic acid. Callus methanol extracts showed a reduction in the percentages of most phyto-components compared to bulb extract except for oleic acid, 3-(octadecyloxy) propyl ester and 3-hydroxydodecanoic acid; on the other hand, some important compounds were detected only in callus extract possessing anti-cancer, antiviral, and anti-inflammatory effects, such as farnesol (C15H26O), 7-methyl-Z-tetradecen-1-ol acetate (C17H32O2), ethyl iso-allocholate (C26H44O5), 4-trifluoroacetoxypentadecane (C17H31F3O2), and 2-hydroxyhexadecanoic acid (C16H32O3).

Effects of Monascus pigments on fat metabolism in Caenorhabditis elegans

Monascus spp. are traditional eatable filamentous fungi and have been utilized in China for more than one thousand years. Their fermented products, named hongqu, are a kind of traditional fermentation food and traditional Chinese medicine. Monascus pigments, the important polyketide secondary metabolites of Monascus have been widely used in food and medicine fields. It has been proved that Monascus pigments have a series of biological activities, such as anti-obesity activity, anticancer effects, antibacterial and antioxidant effects. The effects of Monascus pigments on nabolism and catabolism of lipid in Caenorhabditis elegans were studied. It was found that Monascus pigments could significantly reduce fat accumulation in the body of C. elegans wild-type (N2), reduce the contents of TG, MDA, CAT and ROS, and increase the content of SOD. At the same time, Monascus pigments can extend the maximum life span of nematodes and the tolerance time under emergency state, and Monascus pigments can reduce the accumulation of fat by improving the movement level of nematodes. The results of quantitative PCR showed that Monascus pigments inhibited the expression level of fat-5, fat-6, fat-7, mdt-15, pod-2, acs-2 and kat-1, and increased the expression level of nhr49, atgl-1 and ech-1 in wild-type (N2) C. elegans. Monascus pigments have the activity of reducing fat accumulation obviously, and can develop new functional products for its lipid-lowering characteristics and open up the market of functional food of monascus in our country.

Identification of gene markers to predict proanthocyanidins biosynthesis in cell culture of Croton lechleri

Croton lechleri Müll. Arg. is a traditional medicinal tree growing in South America, which produces a deep red sap known as Dragon’s Blood Sap (DBS) used for its many therapeutic benefits. DBS bioactivity is mainly due to the high content of proanthocyanidins (PAs), important plant secondary metabolites involved in plant defence, but also utilised as antioxidant for human health. Since the process to directly obtain DBS from the C. lechleri tree is expensive, time-consuming and strongly dependent on the environmental conditions, the use of plant cell cultures for DBS production seems to represent a suitable alternative. The aptitude of plant cell cultures to synthesize specific secondary metabolites can be elicited by different types of abiotic stresses. In this study, the C. lechleri main genes putatively involved in the PAs biosynthetic pathway were identified and their expression was evaluated in both calli in solid medium and cell culture suspensions to identify the most suitable predictive gene markers for PAs biosynthesis. C. lechleri cell cultures were grown and elicited by N-starvation to induce the production of PAs and were then monitored by assessing the expression of previously identified gene markers, by quantifying the content of PAs and measuring few phenotypical parameters. The results show a new perspective on how PA gene transcript abundance could be used to predict the biosynthesis of useful secondary metabolites in plant cell cultures and to speed up the screening of putative elicitors.

Phase-and disorder-specific differences in peripheral metabolites of the kynurenine pathway in major depression, bipolar affective disorder and schizophrenia

Objectives Kynurenine, kynurenic and quinolinic acid are important metabolites in tryptophan metabolism. Due to an involvement in glutamatergic neurotransmission and immune response, previous studies have investigated this pathway in mental disorders such as major depressive disorder (MDD), bipolar disorder (BD) or schizophrenia (SCZ). Tryptophan and kynurenine have been shown to be decreased across disorders, hinting at the missing link how inflammation causes neurotoxicity and psychiatric symptoms. The main aim of our study was to investigate if individual catabolites could serve as diagnostic biomarkers for MDD, BD and SCZ. Methods We measured plasma levels of tryptophan, kynurenine, kynurenic acid, quinolinic acid and ratio of quinolinic acid/kynurenic acid using mass spectrometry in n = 175 participants with acute episodes and after remission, compared with controls. Results Decreased levels of all tryptophan catabolites were found in the whole patient group, driven by the difference between BD and HC. Manic and mixed phase BD individuals displayed significantly lower kynurenine and kynurenic acid levels. We could not find significant differences between disorders. Upon reaching remission, changes in catabolite levels partially normalised. Conclusions Our data suggests an involvement of the kynurenine pathway in mental disorders, especially BD but disqualifying those metabolites as biomarkers for differential diagnosis.

Molecular characterization of a feruloyl-CoA 6′-hydroxylase involved in coumarin biosynthesis in Clematis terniflora DC

Coumarin is an important secondary metabolite that affects plant physiology. It is a lactone of cis-o-hydroxycinnamic acid and widely exists in medicinal plants. Clematis terniflora DC. is a plant belonging to Ranunculaceae and is rich in variety of coumarins. Feruloyl-CoA 6′-hydroxylase has been reported as a key enzyme in the formation of coumarin basic skeleton only in some common plants, however, its evidence in other species is still lacking especially for the biosynthesis of coumarins in C. terniflora. In the present study, we identified a feruloyl-CoA 6′-hydroxylase CtF6′H in C. terniflora, and functional characterization indicated that CtF6′H could hydroxylate feruloyl-CoA to 6-hydroxyferuloyl-CoA. Furthermore, the expression level of CtF6′H was differed among different tissues in C. terniflora, while under UV-B radiation, the level of CtF6′H was increased in the leaves. Biochemical characteristics and subcellular location showed that CtF6′H was mainly present in the cytosol. The crystal structure of CtF6′H was simulated by homology modeling to predict the potential residues affecting enzyme activity. This study provides the additional evidence of feruloyl-CoA 6′-hydroxylase in different plant species and enriches our understanding of biosynthetic mechanism of coumarin in C. terniflora.

A novel higher polyhydroxybutyrate producer Halomonas halmophila 18H with unique cell factory attributes

For cost-competitive biosynthesis of polyhydroxybutyrate (PHB), the screening of efficient producers and characterization of their genomic potential is fundamental. In this study, 94 newly isolated halophilic strains from Turkish salterns were screened for their polyhydroxyalkanoates (PHAs) biosynthesis capabilities through fermentation. Halomonas halmophila 18H was found to be the highest PHB producer, yielding 63.72 % of its biomass as PHB. The PHB produced by this strain was physically and chemically characterized using various techniques. Its genome was also sequenced and found to be large (6,713,657 bp) and have a GC content of 59.9 %. Halomonas halmophila 18H was also found to have several copies of PHB biosynthesis genes, as well as 20 % more protein-coding genes and 1075 singletons compared to other high PHB producers. These unique genomic features make it a promising cell factory for the simultaneous production of PHAs and other biotechnologically important secondary metabolites.

Uncovering a miltiradiene biosynthetic gene cluster in the Lamiaceae reveals a dynamic evolutionary trajectory

The spatial organization of genes within plant genomes can drive evolution of specialized metabolic pathways. Terpenoids are important specialized metabolites in plants with diverse adaptive functions that enable environmental interactions. Here, we report the genome assemblies of Prunella vulgaris, Plectranthus barbatus, and Leonotis leonurus. We investigate the origin and subsequent evolution of a diterpenoid biosynthetic gene cluster (BGC) together with other seven species within the Lamiaceae (mint) family. Based on core genes found in the BGCs of all species examined across the Lamiaceae, we predict a simplified version of this cluster evolved in an early Lamiaceae ancestor. The current composition of the extant BGCs highlights the dynamic nature of its evolution. We elucidate the terpene backbones generated by the Callicarpa americana BGC enzymes, including miltiradiene and the terpene (+)-kaurene, and show oxidization activities of BGC cytochrome P450s. Our work reveals the fluid nature of BGC assembly and the importance of genome structure in contributing to the origin of metabolites.

The effects of sustained and diel-cycling hypoxia on high-latitude fish Phoxinus lagowskii

High-latitude fish are subjected to sustained and diel-cycling hypoxia. Oxygen deficiency could pose a serious threat to fish, but little information is available regarding the response mechanisms employed by high-latitude fish to sustained and diel-cycling hypoxia. In this study, a combination of transcriptomics and metabolomics were used to examine the molecular response mechanisms actioned by sustained and diel-cycling hypoxia in the high-latitude fish, Phoxinus lagowskii. P. lagowskii was divided into normoxic control (6.0–7.0 mg/L dissolved oxygen), sustained (1.5 mg/L dissolved oxygen), and diel-cycling hypoxic treatment (6.0–7.0 mg/L between 07:00–21:00, and 3.0–4.0 mg/L between 21:00–07:00) tanks for 28 days. Differentially expressed genes (DEGs) and significantly different metabolites (DMs) related to digestive proteases, lipid metabolism, estrogen signaling pathway, steroid hormone biosynthesis, glutathione metabolism, and tryptophan metabolism were identified from comparative metabolomic and transcriptomic data expression profiles within the liver. The current study found that P. lagowskii had significantly different responses between sustained and diel-cycling hypoxia. P. lagowskii faced with sustained hypoxia may enhance their tolerance capacity through phospholipid and glutathione metabolism. Our data provide new insights into the high latitude fish coping with changes in hypoxia and warrants further investigation into these potentially important genes and metabolites.

Microfluidic Mixer for In Situ Ammonia Analysis of Single Cells in Mass Spectrometry.

Mass spectrometry (MS) is a powerful tool for exploring single-cell heterogeneity. However, due to the ultralow absolute content of most substances in a single cell, existing methods can only analyze high-content substances; conventional methods are incompetent for quantitative analysis of important trace-amount small-molecule metabolites such as ammonia and sulfide. Herein, a method integrating single-cell extraction, online derivatization, and MS for multifunctional and more accurate MS analysis is reported. For application, ammonia content in a single cell was analyzed, where the cellular heterogeneity in ammonia metabolism was revealed. First, the extraction room of a microfluidic probe was covered on the target single cell, and the extraction fluid was allowed to flow through a single cell and dissolve cellular ammonia. Then, the ammonia was mixed and reacted with the pretreatment reagent input from another inlet to achieve the derivatization and signal amplification, enhancing the analysis sensitivity on MS. Finally, the sample was sent to MS, and the ammonia content was successfully quantitatively evaluated by analyzing its derivative urotropine, demonstrating the potential of this method to advance single-cell mass spectrometry analysis to higher sensitivity.

Systematic Identification of Suitable Reference Genes for Quantitative Real-Time PCR Analysis in Melissa officinalis L.

Melissa officinalis L. is well known for its lemon-scented aroma and various pharmacological properties. Despite these valuable properties, the genes involved in the biosynthetic pathways in M. officinalis are not yet well-explored when compared to other members of the mint family. For that, gene expression studies using quantitative real-time PCR (qRT-PCR) are an excellent tool. Although qRT-PCR can provide accurate results, its accuracy is highly reliant on the expression and stability of the reference gene used for normalization. Hence, selecting a suitable experiment-specific reference gene is very crucial to obtain accurate results. However, to date, there are no reports for experiment-specific reference genes in M. officinalis. Therefore, in the current study, ten commonly used reference genes were assessed for their suitability as optimal reference genes in M. officinalis under various abiotic stress conditions and different plant organs. The candidate genes were ranked based on BestKeeper, comparative ΔCt, geNorm, NormFinder, and RefFinder. Based on the results, we recommend the combination of EF-1α and GAPDH as the best reference genes to normalize gene expression studies in M. officinalis. On the contrary, HLH71 was identified as the least-performing gene. Thereafter, the reliability of the optimal gene combination was assessed by evaluating the relative gene expression of the phenylalanine ammonia lyase (PAL) gene under two elicitor treatments (gibberellic acid and jasmonic acid). PAL is a crucial gene involved directly or indirectly in the production of various economically important secondary metabolites in plants. Suitable reference genes for each experimental condition are also discussed. The findings of the current study form a basis for current and future gene expression studies in M. officinalis and other related species.

Integration of transcriptomics and metabolomics reveals a novel gene signature guided by FN1 associated with immune response in oral squamous cell carcinoma tumorigenesis.

Oral squamous cell carcinomas (OSCCs) are primary head and neck malignant tumours with a high incidence and mortality. However, the molecular mechanisms involved in OSCC tumorigenesis are not fully understood. OSCC and paired para-carcinoma samples were collected and used to perform multi-omics study. Transcriptomic analysis was used to reveal significant alterations in inflammatory and immune processes in OSCC. Ingenuity Pathway Analysis (IPA) combined with the LASSO Cox algorithm was used to identify and optimize a crucial gene signature. Metabolomics analysis was performed to identify the important metabolites which linked to the crucial gene signature. The public data TCGA-HNSCC cohort was used to perform the multiple bioinformatic analysis. These findings identified a FN1-mediated crucial network that was composed of immune-relevant genes (FN1, ACP5, CCL5, COL1A1, THBS1, BCAT1, PLAU, IGF2BP3, TNF, CSF2, CXCL1 and CXCL5) associated with immune infiltration and influences the tumour microenvironment, which may contribute to OSCC tumorigenesis and progression. Moreover, we integrated the relevant genes with altered metabolites identified by metabolic profiling and identified 7 crucial metabolites (Glu-Glu-Lys, Ser-Ala, Ser-Ala, N-(octadecanoyl) sphing-4-enine-1-phosphocholine, N-methylnicotinamide, pyrrhoxanthinol and xanthine) as potential downstream targets of the FN1-associated gene signature in OSCC. Importantly, FN1 expression is positively correlated with immune infiltration levels in HNSCC, which was confirmed at the single-cell level. Overall, these results revealed the differential genetic and metabolic patterns associated with OSCC tumorigenesis and identified an essential molecular network that plays an oncogenic role in OSCC by affecting amino acid and purine metabolism. These genes and metabolites might, therefore, serve as predictive biomarkers of survival outcomes and potential targets for therapeutic intervention in OSCC.

Beyond metabolic waste: lysine lactylation and its potential roles in cancer progression and cell fate determination.

Lactate is an important metabolite derived from glycolysis under physiological and pathological conditions. The Warburg effect reveals the vital role of lactate in cancer progression. Numerous studies have reported crucial roles for lactate in cancer progression and cell fate determination. Lactylation, a novel posttranslational modification (PTM), has provided a new opportunity to investigate metabolic epigenetic regulation, and studies of this process have been initiated in a wide range of cancer cells, cancer-associated immune cells, and embryonic stem cells. Lactylation is a novel and interesting mechanism of lactate metabolism linked to metabolic rewiring and epigenetic remodeling. It is a potential and hopeful target for cancer therapy. Here, we summarize the discovery of lactylation, the mechanisms of site modification, and progress in research on nonhistone lactylation. We focus on the potential roles of lactylation in cancer progression and cell fate determination and the possible therapeutic strategies for targeting lysine lactylation. Finally, we suggest some future research topics on lactylation to inspire some interesting ideas.

Energy metabolism disorder dictates chronic hypoxia damage in heart defect with tetralogy of fallot.

Tetralogy of Fallot (TOF) belongs to cyanotic heart damage, which is the most common in clinic. In the chronic myocardial hypoxia injury related to TOF, the potential molecular mechanism of cardiac energy metabolism remains unclear. In our study, microarray transcriptome analysis and metabonomics methods were used to explore the energy metabolism pathway during chronic hypoxia injury. The gene expression omnibus (GEO) dataset GSE132176 was obtained for analyzing the metabolic pathways. The clinical samples (right atrial tissues) of atrial septal defect (ASD) and TOF were analyzed by metabonomics. Next, we screened important pathways and important differential metabolites related to energy metabolism to explore the pathogenesis of TOF. Gene set enrichment analysis (GSEA) indicated that fructose 6-phosphate metabolic process, triglyceride metabolic process, and et al. were significantly enriched. Gene set variation analysis (GSVA) results showed that significant difference of ASD group and TOF group existed in terpenoid metabolic process and positive regulation of triglyceride metabolic process. Pathways with significant enrichment (impact > 0.1) in TOF were caffeine metabolism (impact = 0.69), sphingolipid metabolism (impact = 0.46), glycerophospholipid metabolism (impact = 0.26), tryptophan metabolism (impact = 0.24), galactose metabolism (impact = 0.11). Pathways with significant enrichment (impact > 0.1) in ASD are caffeine metabolism (impact = 0.69), riboflavin metabolism (impact = 0.5), alanine, aspartate and glutamate metabolism (impact = 0.35), histidine metabolism (impact = 0.34) and et al. Disturbed energy metabolism occurs in patients with TOF or ASD, and further investigation was needed to further clarify mechanism.

RNA-Seq reveals the key pathways and genes involved in the light-regulated flavonoids biosynthesis in mango (Mangifera indica L.) peel.

Flavonoids are important water soluble secondary metabolites in plants, and light is one of the most essential environmental factors regulating flavonoids biosynthesis. In the previous study, we found bagging treatment significantly inhibited the accumulation of flavonols and anthocyanins but promoted the proanthocyanidins accumulation in the fruit peel of mango (Mangifera indica L.) cultivar 'Sensation', while the relevant molecular mechanism is still unknown. In this study, RNA-seq was conducted to identify the key pathways and genes involved in the light-regulated flavonoids biosynthesis in mango peel. By weighted gene co-expression network analysis (WGCNA), 16 flavonoids biosynthetic genes were crucial for different flavonoids compositions biosynthesis under bagging treatment in mango. The higher expression level of LAR (mango026327) in bagged samples might be the reason why light inhibits proanthocyanidins accumulation in mango peel. The reported MYB positively regulating anthocyanins biosynthesis in mango, MiMYB1, has also been identified by WGCNA in this study. Apart from MYB and bHLH, ERF, WRKY and bZIP were the three most important transcription factors (TFs) involved in the light-regulated flavonoids biosynthesis in mango, with both activators and repressors. Surprisingly, two HY5 transcripts, which are usually induced by light, showed higher expression level in bagged samples. Our results provide new insights of the regulatory effect of light on the flavonoids biosynthesis in mango fruit peel.

Beyond the Gut, Emerging Microbiome Areas of Research: A Focus on Early-Life Microbial Colonization.

Undoubtedly, the human body harbors trillions of microbes of different kinds performing various physiological activities, such as priming the immune system, influencing host metabolism, and improving health by providing important metabolites such as short-chain fatty acids. Although the gut is considered the "microbial organ" of our body as it hosts the most microbes, there are microbes present in various other important anatomical locations differing in numbers and type. Research has shown the presence of microbes in utero, sparking a debate on the "sterile womb" concept, and there is much scope for more work in this area. It is important to understand the early-life microbiome colonization, which has a role in the developmental origins of health and disease in later life. Moreover, seminal studies have indicated the presence of microbes beyond the gut, for example, in the adipose tissue and the liver. However, it is still unclear what is the exact source of these microbes and their exact roles in health and disease. In this review, we appraise and discuss emerging microbiome areas of research and their roles in metabolic health. Further, we review the importance of the genital microbiome in early-life microbial interactions.

Cadmium toxicity in medicinal plants: An overview of the tolerance strategies, biotechnological and omics approaches to alleviate metal stress.

Medicinal plants, an important source of herbal medicine, are gaining more demand with the growing human needs in recent times. However, these medicinal plants have been recognized as one of the possible sources of heavy metal toxicity in humans as these medicinal plants are exposed to cadmium-rich soil and water because of extensive industrial and agricultural operations. Cadmium (Cd) is an extremely hazardous metal that has a deleterious impact on plant development and productivity. These plants uptake Cd by symplastic, apoplastic, or via specialized transporters such as HMA, MTPs, NRAMP, ZIP, and ZRT-IRT-like proteins. Cd exerts its effect by producing reactive oxygen species (ROS) and interfere with a range of metabolic and physiological pathways. Studies have shown that it has detrimental effects on various plant growth stages like germination, vegetative and reproductive stages by analyzing the anatomical, morphological and biochemical changes (changes in photosynthetic machinery and membrane permeability). Also, plants respond to Cd toxicity by using various enzymatic and non-enzymatic antioxidant systems. Furthermore, the ROS generated due to the heavy metal stress alters the genes that are actively involved in signal transduction. Thus, the biosynthetic pathway of the important secondary metabolite is altered thereby affecting the synthesis of secondary metabolites either by enhancing or suppressing the metabolite production. The present review discusses the abundance of Cd and its incorporation, accumulation and translocation by plants, phytotoxic implications, and morphological, physiological, biochemical and molecular responses of medicinal plants to Cd toxicity. It explains the Cd detoxification mechanisms exhibited by the medicinal plants and further discusses the omics and biotechnological strategies such as genetic engineering and gene editing CRISPR- Cas 9 approach to ameliorate the Cd stress.

TRAIL mediated apoptosis ruling and anticancer trigger by fine-tuned nano spheres of Fagonia cretica methanolic extracts as novel cancer regime

Fagonia cretica L. is a tropical plant of family Zygophyllaceae with wide range of medicinally important secondary metabolites. The low cellular uptake of the polar compounds in the extract of the plant limits its biological application. In present study efficacy of F. cretica modified bioactive nano-formulations for in vitro modulation of TRAIL mediated extrinsic apoptotic pathway as cancer therapy was investigated. F. cretica methanolic extracts were formulated at nano-scale for green synthesis of silver nanoparticles, albumin conjugation and liposomes encapsulation to enhance targeted bioactivity against cancer. Physical characterization of the synthesized nanoparticles was done by SEM, EDX and Zeta potential analyzer. In vitro cell viability assay MTT was done for MCF-7, Hep-2, HUH-7 and HCEC cell lines. Relative expression variation of the apoptotic pathway-associated genes was done by qRT-PCR. SEM revealed spherical shape of 56.62 ± 8.04, 143 ± 11.54 and 83.36 ± 38.73 nm size and zeta potential − 18.6, − 15.5 and − 18.3 mV for liposomes, silver and albumin nanoparticles. Silver nanoparticles showed highest anticancer activity in vitro than albumin and liposomes nanoparticles with IC50 0.101 ± 0.004, 0.177 ± 0.03 and 0.434 ± 0.022 mg/mL in MCF-7, Hep-2 and HUH-7 respectively. F. cretica albumin and silver nanoparticles upregulated the in vitro TRAIL, DR4, DR5 and FADD gene expression at statistically significant levels in Hep-2 cell lines. Nano-formulations of F. cretica proved therapeutically important biomolecules in vitro. The hypothesized modulation of extrinsic apoptosis pathway genes through the plant nanoparticles proved novel medicinal options for effective treatment of cancer and enhancing the bioavailability of the active plant metabolites.

QTL analysis of important agronomic traits and metabolites in foxtail millet (Setaria italica) by RIL population and widely targeted metabolome.

As a bridge between genome and phenotype, metabolome is closely related to plant growth and development. However, the research on the combination of genome, metabolome and multiple agronomic traits in foxtail millet (Setaria italica) is insufficient. Here, based on the linkage analysis of 3,452 metabolites via with high-quality genetic linkage maps, we detected a total of 1,049 metabolic quantitative trait loci (mQTLs) distributed in 11 hotspots, and 28 metabolite-related candidate genes were mined from 14 mQTLs. In addition, 136 single-environment phenotypic QTL (pQTLs) related to 63 phenotypes were identified by linkage analysis, and there were 12 hotspots on these pQTLs. We futher dissected 39 candidate genes related to agronomic traits through metabolite-phenotype correlation and gene function analysis, including Sd1 semidwarf gene, which can affect plant height by regulating GA synthesis. Combined correlation network and QTL analysis, we found that flavonoid-lignin pathway maybe closely related to plant architecture and yield in foxtail millet. For example, the correlation coefficient between apigenin 7-rutinoside and stem diameter reached 0.98, and they were co-located at 41.33-44.15 Mb of chromosome 5, further gene function analysis revealed that 5 flavonoid pathway genes, as well as Sd1, were located in this interval . Therefore, the correlation and co-localization between flavonoid-lignins and plant architecture may be due to the close linkage of their regulatory genes in millet. Besides, we also found that a combination of genomic and metabolomic for BLUP analysis can better predict plant agronomic traits than genomic or metabolomic data, independently. In conclusion, the combined analysis of mQTL and pQTL in millet have linked genetic, metabolic and agronomic traits, and is of great significance for metabolite-related molecular assisted breeding.