Lithospermum erythrorhizon is a medicinal plant valued for its ability to produce shikonin and its derivatives, a compound with important pharmacological properties. While R2R3-MYB transcription factors are known regulators of plant metabolism, no genome-wide analysis had been conducted in L. erythrorhizon. In this study, we present the comprehensive identification and evolutionary study of R2R3-MYBs in L. erythrorhizon, characterizing 176 LeMYB genes. Structural and conserved motif analyses revealed that the R2 and R3 domains of LeMYBs are relatively conserved, while sequence diversity has progressively emerged during evolution. Phylogenetic analysis classified the LeMYBs into 31 subfamilies, expanded mainly through whole-genome duplication, transposed duplication, and dispersed duplication. Expression profiling identified that certain LeMYBs exhibit root/leaf/flower-specific expression, with LeMYB1 predominantly expressed in roots, showing a 63-fold increase in transcript levels compared to leaves and significant upregulation under shikonin-inducing conditions. Subcellular localization confirmed that LeMYB1 is a nucleus-localized protein. Finally, yeast one-hybrid (Y1H) and dual-luciferase reporter assays collectively demonstrate that LeMYB1 binds to the promoters of key shikonin biosynthetic enzyme genes (LeHMGR2, LeGPPS1, LePGT1, LeGHQ3’’H2, and LeSAT1) and activates their transcription, thereby promoting shikonin biosynthesis. This study provides the first genome-wide evolutionary analysis of R2R3-MYBs in L. erythrorhizon and reveals that LeMYB1 coordinately regulates multiple shikonin biosynthetic genes, laying the foundation for elucidating the regulatory mechanisms of shikonin biosynthesis and for mining potential transcription factors for genetic improvement.

Arnebia tschimganica is a vulnerable species within the Boraginaceae (Boraginales), which has long been taxonomically debated due to inconsistent molecular and morphological characteristics. Shikonin and its derivatives, which are found in the roots of Boraginaceae species, possess significant pharmacological and industrial potential; however, the regulatory mechanisms underlying their biosynthesis are not yet fully comprehended. The lack of reference genomes for Arnebia species has hindered further research in these fields. Here, this study sequenced and assembled the chromosome-level genome of A. tschimganica, revealing that Boraginales is sister to Lamiales within the lamiids and suggesting that the taxonomic status of A. tschimganica should be regressed from Arnebia to Lithospermum. Arnebia tschimganica has undergone a recent whole-genome duplication that is shared with other Boraginaceae species, and this event has driven the evolution of shikonin biosynthesis. Multi-omics analysis revealed significant differences in shikonin production between A. tschimganica and Lithospermum erythrorhizon, attributing reduced shikonin productions in A. tschimganica to low transcript levels of key biosynthetic genes postdivergence. Furthermore, AtsDSH1, the enzyme responsible for catalyzing the hydroxylation of deoxyshikonin to shikonin in A. tschimganica, was identified and functionally characterized. Two ERF transcription factors were identified as conserved regulators of the dehydroshikonin hydroxylase gene DSH1, potentially regulating shikonin biosynthesis. These findings provide a chromosome-level genomic perspective to clarify the taxonomy of this controversial swing species and advance valuable insights for shikonin biosynthesis regulation.

Methicillin-resistant Staphylococcus aureus (MRSA) infections pose a major threat to public health due to multi-drug resistance, necessitating the urgent need for novel therapeutic approaches. Synergistic combinations of plant-derived bioactive compounds with conventional antibiotics offer a promising therapeutic alternative. Leveraging the structural diversity and inherent anti-MRSA activity of naphthoquinones from Arnebia euchroma, this study aimed to screen their derivatives for synergistic effects with clinical antibiotics. We investigated the synergy between 5,8-dihydroxy-1,4-naphthoquinone (DHNQ), the most potent derivative among 35 screened shikonin compounds, and eight clinical antibiotics against MRSA. DHNQ exhibited synergistic activity with ursolic acid (UA), Cefazolin (CZ) and gentamicin (GM) against MRSA (FICI≤0.5). Remarkably, the DHNQ-GM combination reduced the MIC of DHNQ by 40-fold (from 8 to 0.2 μg/mL) and that of GM from 64 to 28.8 μg/mL. In vitro, this combination achieved over 80% growth inhibition of MRSA, with negligible cytotoxicity toward HaCaT human keratinocytes and L929 murine fibroblasts, and hemolysis rates below 5%. In a murine model of MRSA-infected wounds, DHNQ-GM combination therapy accelerated wound healing, reduced bacterial loads by 4 log10 CFU, and alleviated systemic pathology. Mechanistically, the combination disrupted MRSA biofilm integrity, as directly visualized by electron microscopy, which revealed characteristic cell shrinkage and membrane fragmentation, accompanied by extensive leakage of intracellular contents. Furthermore, the treatment suppressed arginine and pyrimidine biosynthesis, as indicated by downregulation of key enzymes (ArcC, ArgF, CarB, and PyrF), impaired energy metabolism, evidenced by reduced ATP levels and diminished glucose uptake, and perturbed enzymatic activity, reflected in decreased lactate dehydrogenase (LDH) and increased alkaline phosphatase (AKP) activities. Collectively, these findings identify DHNQ-GM as a promising synergistic regimen against MRSA, combining potent antibacterial efficacy with a favorable safety profile and multi-target mechanisms, thereby offering a potential therapeutic alternative to combat antibiotic resistance.

(R)-shikonin/(S)-alkannin and their derivatives (collectively denoted as shikonins) are abundantly distributed in Boraginaceae roots. Acetylshikonin, as a crucial derivative of (R)-shikonin, possesses substantial research significance. Previous investigations have shown that LeSAT2, specifically catalyzes the acylation of (R)-shikonin to yield various acylated (R)-shikonin derivatives, including acetylshikonin. Additionally, electrophoretic mobility shift assays (EMSA) indicated that LeWRKY24 transcription factors may be involved in the transcriptional regulation of LeSAT2. For this study, the interaction between LeWRKY24 and LeSAT2 was further verified using dual-luciferase reporter (DLR) and yeast one-hybrid (Y1H) assays. The LeWRKY24 overexpression vector was constructed and introduced into Echium plantagineum to induce transgenic hairy roots. High-performance liquid chromatography (HPLC) analysis showed that in LeWRKY24-overexpressing hairy root lines, a significant increase was observed in the total pigment content, and in particular, the content of acetylshikonin was dramatically upregulated by as much as 200-fold. Transcriptome data revealed that genes associated with shikonin biosynthesis in the overexpression lines were significantly upregulated, activating the metabolic pathway, which explains the molecular mechanism underlying this regulatory effect. Our results also confirm that LeWRKY24 possesses cross-species regulatory universality in acylshikonin biosynthesis, laying a foundation for its widespread application in the metabolic engineering of Boraginaceae plants.

Invasive fungal infections (IFIs), predominantly caused by Candida albicans, are a significant threat to immunocompromised individuals. The emergence of drug-resistant strains has intensified the need for novel antifungal agents. Natural naphthoquinones, including 5,8-dihydroxy-1,4-naphthoquinone (DHNQ, also as PNP-02), have broad-spectrum antimicrobial properties, but their antifungal potential against C. albicans remains underexplored. This study evaluates the antifungal activity of DHNQ derivatives and elucidates their mechanisms of action. The antifungal properties of these compounds were evaluated using the Kirby-Bauer disk diffusion method, broth microdilution assays, and phenotypic screening. DHNQ was identified as the most effective compound, and further investigation focused on its effects on C. albicans growth, biofilm formation, hyphal development, and underlying mechanisms, including oxidative stress induction and mitochondrial dysfunction. In a murine candidiasis model, DHNQ significantly reduced the fungal burden in both the kidneys and the skin, with a minimum inhibitory concentration (MIC) ranging from 2 to 8 μg/mL, exceeding the activity of fluconazole against clinical isolates of fluconazole-resistant C. albicans strains by over 32 times. Mechanistic investigations revealed that DHNQ exerts its antifungal effects through a multi-pronged approach: inhibiting glycolysis, disrupting biofilm and hyphal formation, and inducing oxidative stress-mediated mitochondrial dysfunction. Notably, DHNQ exhibited low cytotoxicity in vitro and no observable toxicity in vivo. These properties make it a promising lead molecule for future optimization and development of treatments for C. albicans infections, pending crucial evaluations of its selectivity and safety in host environments.IMPORTANCEIn summary, this study demonstrated that DHNQ exhibits potent and broad-spectrum antifungal activity, showing significant efficacy against C. albicans both in vitro and in vivo. Unlike conventional antifungals, DHNQ disrupted the virulence of C. albicans by inhibiting glycolysis, suppressing biofilm formation, and inducing oxidative stress-mediated mitochondrial dysfunction. These findings not only highlight the promising potential of DHNQ as a treatment for C. albicans infections but also provide critical insights that may facilitate the development of new antifungal agents.

Given the increasing concern regarding antibacterial resistance, the antimicrobial properties of naphthoquinones have recently attracted significant attention. While 1,4-naphthoquinone and its derivatives have been extensively studied, the antibacterial properties of 5,8-dihydroxy-1,4-naphthoquinone derivatives remain relatively unexplored. This study presents a comprehensive in vitro and in vivo analysis of the antibacterial activity of 35 naturally sourced and chemically synthesized derivatives of 5,8-dihydroxy-1,4-naphthoquinone. Kirby-Bauer antibiotic testing identified three compounds with activity against methicillin-resistant Staphylococcus aureus (MRSA), with one compound (PNP-02) demonstrating activity comparable to vancomycin in minimum inhibitory concentration, minimum bactericidal concentration (MBC), and time-kill assays. Microscopic and biochemical analyses revealed that PNP-02 adversely affects the cell wall and cell membrane of MRSA. Mechanistic investigations, including proteomic sequencing analyses, Western blotting, and RT-qPCR assays, indicated that PNP-02 compromises cell membrane integrity by inhibiting arginine biosynthesis and pyrimidine metabolism pathways, thereby increasing membrane permeability and inducing bacterial death. In an in vivo mouse model of skin wound healing, PNP-02 exhibited antibacterial efficacy similar to vancomycin. The compound demonstrated low toxicity to cultured human cells and in hemolysis assays and remained stable during serum incubation. These findings suggest that PNP-02 possesses promising bioactivity against MRSA and represents a potential novel antibacterial agent.

Phosphate (Pi) is an essential nutrient that frequently limits plant growth because of its low availability in soils, especially during early seedling development in Arabidopsis. Under Pi-deficient conditions, etiolated seedlings exhibit elongated hypocotyls, shortened roots, and small, pale cotyledons-a morphological adaptation that enhances light-foraging capacity. However, how Pi is strategically distributed among these organs to optimize seedling establishment remains unclear. We here identify a PHR1-EIN3/EIL1 regulatory module that directs minimal Pi resources to suppress root elongation and cotyledon expansion or to accelerate greening, promoting Pi conservation. This mechanism prioritizes hypocotyl elongation to improve light acquisition. In contrast, under sufficient Pi supply, hypocotyl and root growth is promoted along with cotyledon enlargement, while greening is delayed. Thus, the PHR1-EIN3/EIL1 module enables efficient allocation of limited Pi to maximize hypocotyl elongation for light foraging during early seedling establishment while strategically restricting Pi usage in other organs to enhance overall survival.

Colorectal cancer (CRC) is one of the deadliest cancers globally, ranking as the third most prevalent and second most lethal malignancy worldwide. The standard treatment for CRC typically involves a combination of surgery, radiotherapy, and chemotherapy. Despite advancements in CRC treatment, the prognosis remains unsatisfactory, primarily due to unclear mechanisms underlying tumorigenesis and the aggression of CRC. The aberrant activation of the PI3K/AKT pathway is frequently implicated in the initiation, progression, and metastasis of CRC. Studies have demonstrated that shikonin (SK) exerts anti-cancer effects. In this study, we evaluated the anti-tumor activities of a series of semi-synthesized SK derivatives against CRC cells. Our findings revealed that the SK derivative (M12) significantly inhibited the proliferation and colony formation of CRC cells, reduced cell migration, and induced apoptosis. Mechanistically, M12 enhanced the production of reactive oxygen species and downregulated the mitochondrial membrane potential, ultimately leading to mitochondrial apoptosis. Furthermore, M12 exhibited anti-CRC effects by modulating the PI3K/AKT signaling pathway and significantly suppressed tumorigenicity without causing notable adverse effects in mice. Therefore, targeting the PI3K/AKT pathway could be a promising treatment for CRC. M12 appears to be a promising candidate for the effective and safe treatment of CRC.

Acidic soils, covering 40 to 50% of arable land, hinder productivity due to phosphorus limitation and aluminum toxicity. Using a synthetic community (SynCom) approach, we tested a native Rhizobium (Rh) and a non-native Ensifer fredii (Ef) strain, both combined with the mycorrhizal fungus Rhizophagus intraradices (Ri), on soybean growth in acidic soil. The native RhRi SynCom outperformed EfRi, significantly improving soybean growth, yield, and soil health. Metagenomics and metabolomics revealed that RhRi significantly enhanced beneficial microorganisms (AD3, Gemmataceae) and metabolites (putrescine, stearic acid), hence improving nutrient cycling, stress tolerance, and membrane integrity. RhRi also enhanced soil enzyme activity (urease, phosphatases), which resulted in an increase in nitrogen and phosphorus availability and a decrease in rhizosphere toxicity. These alterations enhanced plant resilience, soil structure, and microbial diversity. RhRi activated metabolic pathways (amino acids, lipids, ABC transporters, and secondary metabolites) that are involved in nutrient acquisition and stress response. Conversely, EfRi provided minimal advantages, emphasizing the significance of native microbial compatibility. Our findings show that native SynComs boost crop resilience and production in acidic soils through synergistic microbial interactions and metabolic reprogramming, thereby offering a sustainable agricultural strategy in harsh environments.

The PI3K-Akt axis is abnormally activated in KRAS-mutated colorectal cancer and is considered to be a potential therapeutic target. A novel series of phenoxyacetic acid (4-aminophenoacetic acid) shikonin esters was designed by computer-aided drug design (CADD) and synthesized as Akt allosteric inhibitors. Most compounds exhibited greater anti-proliferative activity compared to the positive control MK2206, while also demonstrating lower cytotoxicity against normal cells than shikonin. One of the promising candidates, L8, was selected for further biological evaluation. Docking studies indicated that L8 effectively bound to the allosteric site of Akt through hydrophobic and hydrogen interactions. Enzyme activity and kinetics assessments revealed that L8 bound to Akt with a Kd of 2.07 × 10-6 M and inhibited its activity. Further intracellular assays, including western blotting, enzyme activity assay, flow cytometry, etc., verified that L8 mediated the death of two KRAS-mutant colon cancer cell lines HCT116 (KRASG13D) and HCT-8 (KRASG12A) cells by inactivating Akt, causing tumor cell apoptosis, cell cycle arrest, and interfering with tumor cell invasion and metabolism. A 3D-QSAR model was constructed to understand the relationship between the structure of the shikonin derivatives and their anti-proliferative activity. The in silico ADMET and toxicity prediction studies revealed a few undesired pharmacokinetic attributes of our compounds.

Cutaneous malignant melanoma (CMM) ranks among the deadliest forms of cancer. Abnormalities in lipid metabolism may have a connection with the risk of CMM progression. Lipid metabolism-related genes were selected in MSigDB. A lipid metabolism-related predictive model was constructed in The Cancer Genome Atlas-skin cutaneous melanoma (TCGA-SKCM) using univariate Cox regression analysis, non-negative matrix factorization (NMF) clustering analysis, Weighted correlation network analysis (WGCNA), least absolute shrinkage and selection operator regression (LASSO) analysis, and outcomes-related genes were identified. The roles of candidate genes in CMM were determined using in vivo and in vitro experiments, and the pathway mechanism of the candidate gene was studied using transcriptomics, proteomics, lipid metabolomics, and other molecular biological methods. A predictive model was established, Risk Score = -0.009 * UBE2L6 + 0.033 * PLEKHA5 + 0.024 * LHB + 0.036 * CARM1 + 0.016 * PRXL2B + 0.131 * PLA2G4D. The pleckstrin homology domain-containing A5 (PLEKHA5) was identified as an essential gene and positively correlated with poor outcomes in CMM. Fc receptor-like A (FCRLA) is the downstream gene of PLEKHA5, upregulated in CMM, and tumor necrosis factor alpha (TNFα) is also an essential cytokine that promotes CMM proliferation and metastasis. Lipid metabolomics showed that PLEKHA5 knockdown increased ceramide and sphingosine levels while decreasing cholesterol ester and triglyceride levels in CMM cells, possibly related to disease progression. The predictive model of CMM related to lipid metabolism was constructed. TNFα activates the PLEKHA5-FCRLA axis to enhance neutral lipid storage and energy metabolism in CMM cells, promoting malignant behavior.

The low phosphorus (P) availability of acidic soils severely limits leguminous plant growth and productivity. Improving the soil P nutritional status can be achieved by increasing the P-content through P-fertilization or stimulating the mineralization of organic P via arbuscular mycorrhizal fungi (AMF) application; however, their corresponding impacts on plant and soil microbiome still remain to be explored. Here, we examined the effects of AMF-inoculation and P-fertilization on the growth of soybean with different P-efficiencies, as well as the composition of rhizo-microbiome in an acidic soil. The growth of recipient soybean NY-1001, which has a lower P-efficiency, was not significantly enhanced by AMF-inoculation or P-fertilization. However, the plant biomass of higher P-efficiency transgenic soybean PT6 was significantly increased by 46.74%-65.22% through AMF-inoculation. Although there was no discernible difference in plant biomass between PT6 and NY-1001 in the absence of AMF-inoculation and P-fertilization, PT6 had approximately 1.9-2.5 times the plant biomass of NY-1001 after AMF-inoculation. Therefore, the growth advantage of higher P-efficiency soybean was achieved through the assistance of AMF rather than P-fertilization in available P-deficient acidic soil. Most nitrogen (N)-fixing bacteria and some functional genes related to N-fixation were abundant in endospheric layer, as were the P-solubilizing Pseudomonas plecoglossicida, and annotated P-metabolism genes. These N-fixing and P-solubilizing bacteria were positive correlated with each other. Lastly, the two most abundant phytopathogenic fungi species accumulated in endospheric layer, they exhibited positive correlations with N-fixing bacteria, but displayed negative interactions with the majority of the other dominant non-pathogenic genera with potential antagonistic activity.

LeBAHD56 is preferentially expressed in tissues where shikonin and its derivatives are biosynthesized, and it confers shikonin acylation in vivo. Two WRKY transcriptional factors might regulate LeBAHD56’s expression. Shikonin and its derivatives, found in the roots of Lithospermum erythrorhizon, have extensive application in the field of medicine, cosmetics, and other industries. Prior research has demonstrated that LeBAHD1(LeSAT1) is responsible for the biochemical process of shikonin acylation both in vitro and in vivo. However, with the exception of its documented in vitro biochemical function, there is no in vivo genetic evidence supporting the acylation function of the highly homologous gene of LeSAT1, LeBAHD56(LeSAT2), apart from its reported role. Here, we validated the critical acylation function of LeBAHD56 for shikonin using overexpression (OE) and CRISPR/Cas9-based knockout (KO) strategies. The results showed that the OE lines had a significantly higher ratio of acetylshikonin, isobutyrylshikonin or isovalerylshikonin to shikonin than the control. In contrast, the KO lines had a significantly lower ratio of acetylshikonin, isobutyrylshikonin or isovalerylshikonin to shikonin than controls. As for its detailed expression patterns, we found that LeBAHD56 is preferentially expressed in roots and callus cells, which are the biosynthesis sites for shikonin and its derivatives. In addition, we anticipated that a wide range of putative transcription factors might control its transcription and verified the direct binding of two crucial WRKY members to the LeBAHD56 promoter’s W-box. Our results not only confirmed the in vivo function of LeBAHD56 in shikonin acylation, but also shed light on its transcriptional regulation.

Triple negative breast cancer (TNBC) is associated with a poor prognosis and limited response to traditional chemotherapy, necessitating the exploration of novel treatment approaches. Recent researches have highlighted the interconnected roles of the PI3K/AKT pathway and MAPK pathway in TNBC cells, contributing to the efficacy of treatments. Therefore, the concurrent inhibition of both pathways presents a potential new therapeutic strategy for TNBC patients. This study aimed to evaluate the antitumor efficacy of M17, an AKT allosteric inhibitor and a new synthesized shikonin derivative, both alone and in combination with the MEK inhibitor trametinib. We applied various cellular assays and a subcutaneous 4T1 tumor bearing BALB/c mice model were utilized to assess the in vitro and in vivo antitumor effects. Computational docking and Bio-Layer Interferometry (BLI) were employed to investigate the binding of M17 with AKT. Additionally, flow cytometry, transwell assays, western blotting, and tumor xenograft assays were conducted to explore the potential synergistic mechanisms of the combined therapy. The results demonstrated that M17 exhibited moderate antitumor activity against TNBC cells, but significantly enhanced the apoptotic effects and inhibited proliferation and migration when combined with trametinib. Furthermore, the combination of M17 and trametinib showed even more pronounced antitumor activity in vivo. Mechanistically, the dual therapy synergistically suppressed TNBC by targeting the AKT/mTOR and MEK/ERK signaling pathways and inhibiting epithelial-mesenchymal transition. In conclusion, the findings suggested that the combination of M17 and trametinib holds promise as a synergistic treatment option for TNBC patients.

The impacts of transgenic crops on soil microbiology and fertility are critical in determining their biosafety. While transgenic crops can alter soil microbes, their effects are often context-dependent; therefore, the ecological importance of these changes remains a topic of ongoing research. Using high-throughput sequencing, we investigated the effects of Bacillus thuringiensis (Bt) maize expressing the mcry1Ab and mcry2Ab genes (2A7) on soil nutrient dynamics, as well as the diversity and function of soil microbial communities, including bacteria and fungi, within different soil compartments. Our findings revealed a plant-shaped rhizosphere (RS) microbial community as a result of the selective recruitment of microorganisms from the surrounding environment. The transgene insertion had a significant impact on the RS niche, and several species eventually became associated with Z58 and 2A7 plants. For example, Neocosmospora rubicola fungal and Pantoea dispersa bacterial microorganisms were significantly decreased in the dual Bt-transgenic 2A7 rhizosphere but enriched in the Z58 rhizospheres. The activity of soil enzymes such as urease, invertase, and alkaline phosphatase was boosted by Bt-transgenic 2A7. LefSe analysis identified significant bacterial and fungal biomarker species that were responsible for the differential effects of Bt-transgenic 2A7 and control Z58 within rhizosphere soils. Mantel analysis further demonstrated that the root exudates of 2A7 altered nutrient-acquisition enzymes by influencing biomarker taxa. PICRUSt2 functional characterization revealed a significantly higher abundance of the phosphate-starvation-inducible protein in control Z58 than in Bt-transgenic 2A7. Furthermore, taxonomy, alpha (Shannon diversity), and beta diversity analyses all revealed niche-driven microbial profile differentiation. Niche partitioning also had a significant impact on N- and P-related COGs as well. Our findings suggests that Bt-transgenic 2A7 modulates rhizosphere microbial communities by affecting biomarker taxa and soil enzyme activity. These findings will promote sustainable agriculture practices by advancing our knowledge of the ecological effects of Bt crops on soil microbial communities.

The symbiosis between arbuscular mycorrhizal fungi (AMF) and plants often stimulates plant growth, increases agricultural yield, reduces costs, thereby providing significant economic benefits. AMF can also benefit plants through affecting the rhizosphere microbial community, but the underlying mechanisms remain unclear. Using Rhizophagus intraradices as a model AMF species, we assessed how AMF influences the bacterial composition and functional diversity through 16 S rRNA gene sequencing and non-targeted metabolomics analysis in the rhizosphere of aluminum-sensitive soybean that were inoculated with pathogenic fungus Nigrospora oryzae and phosphorus-solubilizing fungus Talaromyces verruculosus in an acidic soil. The inoculation of R. intraradices, N. oryzae and T. verruculosus didn’t have a significant influence on the levels of soil C, N, and P, or various plant characteristics such as seed weight, crude fat and protein content. However, their inoculation affected the structure, function and nutrient dynamics of the resident bacterial community. The co-inoculation of T. verruculosus and R. intraradices increased the relative abundance of Pseudomonas psychrotolerans, which was capable of N-fixing and was related to cry-for-help theory (plants signal for beneficial microbes when under stress), within the rhizosphere. R. intraradices increased the expression of metabolic pathways associated with the synthesis of unsaturated fatty acids, which was known to enhance plant resistance under adverse environmental conditions. The inoculation of N. oryzae stimulated the stress response inside the soil environment by enriching the polyene macrolide antifungal antibiotic-producing bacterial genus Streptomyces in the root endosphere and upregulating two antibacterial activity metabolic pathways associated with steroid biosynthesis pathways in the rhizosphere. Although inoculation of pathogenic fungus N. oryzae enriched Bradyrhizobium and increased soil urease activity, it had no significant effects on biomass and N content of soybean. Lastly, the host niches exhibited differences in the composition of the bacterial community, with most N-fixing bacteria accumulating in the endosphere and Rhizobium vallis only detected in the endosphere. Our findings demonstrate that intricate interactions between AMF, associated core fungi, and the soybean root-associated ecological niches co-mediate the regulation of soybean growth, the dynamics of rhizosphere soil nutrients, and the composition, function, and metabolisms of the root-associated microbiome in an acidic soil. AMF R. intraradices improved soybean stress resistance by recruiting specific PGPR and upregulating plant resistance promotion metabolic pathway. The pathogenic fungus N. oryzae stimulated the stress response by enriching the antifungal antibiotic production bacteria and upregulating the antibacterial reaction-associated metabolic pathways. The utility evaluation of some microbial agents on host plants should consider their potential impact on Olsen-P content in acidic soils with limited P availability. The sampling compartments (i.e., host niches of the soybean rhizosphere) exerted greater influence on the assembly and shift of the bacterial community than the application of microbial agents.

Background:Skin cutaneous melanoma (SKCM) is the deadliest dermatology tumor. Ongoing researches have confirmed that the NOD-like receptors (NLRs) family are crucial in driving carcinogenesis. However, the function of NLRs signaling pathway-related genes in SKCM remains unclear.Objective:To establish and identify an NLRs-related prognostic signature and to explore its predictive power for heterogeneous immune response in SKCM patients.Methods:Establishment of the predictive signature using the NLRs-related genes by least absolute shrinkage and selection operator-Cox regression analysis (LASSO-COX algorithm). Through univariate and multivariate COX analyses, NLRs signature's independent predictive effectiveness was proven. CIBERSORT examined the comparative infiltration ratios of 22 distinct types of immune cells. RT-qPCR and immunohistochemistry implemented expression validation for critical NLRs-related prognostic genes in clinical samples.Results:The prognostic signature, including 7 genes, was obtained by the LASSO-Cox algorithm. In TCGA and validation cohorts, SKCM patients with higher risk scores had remarkably poorer overall survival. The independent predictive role of this signature was confirmed by multivariate Cox analysis. Additionally, a graphic nomogram demonstrated that the risk score of the NLRs signature has high predictive accuracy. SKCM patients in the low-risk group revealed a distinct immune microenvironment characterized by the significantly activated inflammatory response, interferon-α/γ response, and complement pathways. Indeed, several anti-tumor immune cell types were significantly accumulated in the low-risk group, including M1 macrophage, CD8 T cell, and activated NK cell. It is worth noting that our NLRs prognostic signature could serve as one of the promising biomarkers for predicting response rates to immune checkpoint blockade (ICB) therapy. Furthermore, the results of expression validation (RT-qPCR and IHC) were consistent with the previous analysis.Conclusion:A promising NLRs signature with excellent predictive efficacy for SKCM was developed.

Colorectal cancer is a common malignancy and the second leading cause of cancer-related death. Emerging evidence suggests that the Wnt/β-catenin pathway has a significant role in cancer development. Shikonin is one of the main bioactive components of the plant, Lithospermum erythrorhizon, which possesses various biological properties. In this study, we identified a shikonin derivative, E5, as a novel suppressor of colorectal carcinoma that acted by inhibiting cell proliferation and migration, and promoting apoptosis. Mechanistically, the drug activated caspase-dependent pathways, negatively regulating the Wnt/β-catenin pathway, and inhibited epithelial–mesenchymal transition. The findings suggest the potential of β-catenin as a molecular target in colon carcinoma and propose E5 as a promising therapeutic strategy for targeting Wnt/β-catenin