Integrated mass spectrometry imaging and single-cell transcriptome atlas strategies provide novel insights into taxoid biosynthesis and transport in Taxus mairei stems.

Taxol, which is a widely used important chemotherapeutic agent, was originally isolated from Taxus stem barks. However, little is known about the precise distribution of taxoids and the transcriptional regulation of taxoid biosynthesis across Taxus stems. Here, we used MALDI-IMS analysis to visualize the taxoid distribution across Taxus mairei stems and single-cell RNA sequencing to generate expression profiles. A single-cell T. mairei stem atlas was created, providing a spatial distribution pattern of Taxus stem cells. Cells were reordered using a main developmental pseudotime trajectory which provided temporal distribution patterns in Taxus stem cells. Most known taxol biosynthesis-related genes were primarily expressed in epidermal, endodermal, and xylem parenchyma cells, which caused an uneven taxoid distribution across T. mairei stems. We developed a single-cell strategy to screen novel transcription factors (TFs) involved in taxol biosynthesis regulation. Several TF genes, such as endodermal cell-specific MYB47 and xylem parenchyma cell-specific NAC2 and bHLH68, were implicated as potential regulators of taxol biosynthesis. Furthermore, an ATP-binding cassette family transporter gene, ABCG2, was proposed as a potential taxoid transporter candidate. In summary, we generated a single-cell Taxus stem metabolic atlas and identified molecular mechanisms underpinning the cell-specific transcriptional regulation of the taxol biosynthesis pathway.

Microbial fertilizers improve soil quality and crop yield in coastal saline soils by regulating soil bacterial and fungal community structure.

Salinization is a global problem affecting agricultural productivity and sustainability. The application of exogenous microbial fertilizer harbors great potential for improving saline-alkali soil conditions and increasing land productivity. Yet the responses to microbial fertilizer application rate in terms of rhizosphere soil biochemical characteristics, soil microbial community, and crop yield and their interrelationships and underlying mechanisms are still unclear. Here, we studied changes to rhizosphere soil-related variables, soil enzyme activity (catalase, sucrase, urease), microbial community diversity, and sweet sorghum (Sorghum bicolor (L.) Moench) yield under four fertilization concentration levels (0, 0.12, 0.24, and 0.36 kg m-2) in a saline-alkali ecosystem (Shandong, China). Our results showed that the best improvement effect on soil when the microbial fertilizer was applied at a rate of 0.24 kg m-2. Compared with the control (sweet sorghum + no fertilizer), it significantly increased soil organic carbon (21.50 %), available phosphorus (26.14 %), available potassium (36.30 %), and soil urease (38.46 %), while significantly reducing soil pH (2.21 %) and EC (12.04 %). Meanwhile, the yield of sweet sorghum was increased by 24.19 %. This is mainly because microbial fertilizers enhanced the diversity and the network complexity of bacterial and fungal communities, and influenced catalase (CAT), urease (UE), and sucrase (SC), thereby facilitating nutrient release in the soil, enhancing soil fertility, and indirectly influencing sweet sorghum productivity. Among them, Gemmatimonadota and Verrucomicrobiota may be the key microbial factors affecting sweet sorghum yield, while available potassium, soil urease and available phosphorus are the main soil factors. These findings provide valuable theoretical insights for preserving the health of coastal saline-alkali soils and meeting the agricultural demand for increased yield per unit of land area.

Co-application of biochars and Piriformospora indica improved the quality of coastal saline soil and promoted the growth of forage.

Soil quality is defined as the ability of soil to maintain the soil environment and the biosphere. Due to the limitation of salt and alkali stress, soil quality can be reduced, which in turn affects agricultural production. Biochar is widely used in saline-alkali land improvement because of its special pore structure and strong ion exchange ability, while Piriformospora indica is widely used in saline-alkali land improvement because it can symbiose with plants and improve plant stress resistance. However, the synergistic effect of combined biochar application and inoculation of P. indica on the quality of saline-alkali soil and plant development is uncertain. Hence, we investigated the combined influences of biochar and P. indica on the soil physicochemical characteristics, as well as the growth and chlorophyll florescence of sorghum-sudangrass hybrids (Sorghum bicolor × Sorghum sudane) in our study. The results indicated that after applying biochar and P. indica together, there was a considerable drop in soil pH, conductivity, Na+, and Cl- concentrations. Meanwhile, the soil organic matter (SOM), available phosphorus (AP), and alkaline hydrolyzable nitrogen (AN) increased by 151.81%, 50.84%, and 103.50%, respectively, when the Bamboo biochar was combined with 120 ml/pot of P. indica. Eventually, sorghum-sudangrass hybrid biomass, transpiration rate, and chlorophyll content increased by 111.69%, 204.98%, and 118.54%, respectively. According to our findings, using P. indica and biochar together can enhance soil quality and plant growth. The results also provide insights to enhance the quality of saline-alkali soils and the role of microorganisms in nutrient cycling.

A biodegradable multifunctional pectin-montmorillonite fertilizer coating: Controlled-release, water-retention and soil-cementation.

Coated fertilizers have been widely used to improve fertility in barren land. However, improving soil structure and water-retention capacity is also essential for arid and semi-arid areas with sandy soils to promote crop growth. Most currently available coated fertilizers rarely meet these requirements, limiting their application scope. Therefore, this study "tailored" pectin-montmorillonite (PM) multifunctional coatings for arid areas, featuring intercalation reactions and nanoscale entanglement between pectin and montmorillonite via hydrogen bonding and electrostatic and van der Waals forces. Notably, PM coatings have demonstrated an effective "relay" model of action. First, the PM-50 coating could act as a "shield" to protect urea pills, increasing the mechanical strength (82.12 %). Second, this coating prolonged the release longevity of urea (<0.5 h to 15 days). Further, the remaining coating performed a water-retention function. Subsequently, the degraded coating improved the soil properties. Thus, this coating facilitated the growth of wheat seedlings in a simulated arid environment. Moreover, the cytotoxicity test, life cycle assessment, and soil biodegradation experiment showed that the PM coating exhibited minimal environmental impact. Overall, the "relay" model of PM coating overcomes the application limitations of traditional coated fertilizers and provides a sustainable strategy for developing coating materials in soil degradation areas.

A metabolic constraint in the kynurenine pathway drives mucosal inflammation in IBD.

Inflammatory bowel disease (IBD) is associated with perturbed metabolism of the essential amino acid tryptophan (Trp). Whether increased degradation of Trp directly fuels mucosal inflammation or acts as a compensatory attempt to restore cellular energy levels via de-novo nicotinamide adenine dinucleotide (NAD + ) synthesis is not understood. Employing a systems medicine approach on longitudinal IBD therapy intervention cohorts and targeted screening in preclinical IBD models, we discover that steady increases in Trp levels upon therapy success coincide with a rewiring of metabolic processes within the kynurenine pathway (KP). In detail, we identify that Trp catabolism in IBD is metabolically constrained at the level of quinolinate phosphorybosyltransferase (QPRT), leading to accumulation of quinolinic acid (Quin) and a decrease of NAD + . We further demonstrate that Trp degradation along the KP occurs locally in the inflamed intestinal mucosa and critically depends on janus kinase / signal transducers and activators of transcription (JAK/STAT) signalling. Subsequently, knockdown of QPRT in-vitro induces NAD + depletion and a pro-inflammatory state, which can largely be rescued by bypassing QPRT via other NAD + precursors. We hence propose a model of impaired de-novo NAD + synthesis from Trp in IBD. These findings point towards the replenishment of NAD + precursors as a novel therapeutic pathway in IBD.

Investigation of the role of TmMYB16/123 and their targets (TmMTP1/11) in the tolerance of Taxus media to cadmium.

The toxicity and stress caused by heavy metal contamination has become an important constraint to the growth and flourishing of trees. In particular, species belonging to the genus Taxus, which are the only natural source for the anti-tumor medicine paclitaxel, are known to be highly sensitive to environmental changes. To investigate the response of Taxus spp. to heavy metal stress, we analyzed the transcriptomic profiles of Taxus media trees exposed to cadmium (Cd2+). In total, six putative genes from the metal tolerance protein (MTP) family were identified in T. media, including two Cd2+ stress inducible TMP genes (TmMTP1, TmMTP11 and Taxus media). Secondary structure analyses predicted that TmMTP1 and TmMTP11, which are members of the Zn-CDF and Mn-CDF subfamily proteins, respectively, contained six and four classic transmembrane domains, respectively. The introduction of TmMTP1/11 into the ∆ycf1 yeast cadmium-sensitive mutant strain showed that TmMTP1/11 might regulate the accumulation of Cd2+ to yeast cells. To screen the upstream regulators, partial promoter sequences of the TmMTP1/11 genes were isolated using the chromosome walking method. Several myeloblastosis (MYB) recognition elements were identified in the promoters of these genes. Furthermore, two Cd2+-induced R2R3-MYB TFs, TmMYB16 and TmMYB123, were identified. Both in vitro and in vivo assays confirmed that TmMTB16/123 play a role in Cd2+ tolerance by activating and repressing the expression of TmMTP1/11 genes. The present study elucidated new regulatory mechanisms underlying the response to Cd stress and can contribute to the breeding of Taxus species with high environmental adaptability.

Mass spectrometry imaging and single-cell transcriptional profiling reveal the tissue-specific regulation of bioactive ingredient biosynthesis in Taxus leaves.

Taxus leaves provide the raw industrial materials for taxol, a natural antineoplastic drug widely used in the treatment of various cancers. However, the precise distribution, biosynthesis, and transcriptional regulation of taxoids and other active components in Taxus leaves remain unknown. Matrix-assisted laser desorption/ionization-mass spectrometry imaging analysis was used to visualize various secondary metabolites in leaf sections of Taxus mairei, confirming the tissue-specific accumulation of different active metabolites. Single-cell sequencing was used to produce expression profiles of 8846 cells, with a median of 2352 genes per cell. Based on a series of cluster-specific markers, cells were grouped into 15 clusters, suggesting a high degree of cell heterogeneity in T. mairei leaves. Our data were used to create the first Taxus leaf metabolic single-cell atlas and to reveal spatial and temporal expression patterns of several secondary metabolic pathways. According to the cell-type annotation, most taxol biosynthesis genes are expressed mainly in leaf mesophyll cells; phenolic acid and flavonoid biosynthesis genes are highly expressed in leaf epidermal cells (including the stomatal complex and guard cells); and terpenoid and steroid biosynthesis genes are expressed specifically in leaf mesophyll cells. A number of novel and cell-specific transcription factors involved in secondary metabolite biosynthesis were identified, including MYB17, WRKY12, WRKY31, ERF13, GT_2, and bHLH46. Our research establishes the transcriptional landscape of major cell types in T. mairei leaves at a single-cell resolution and provides valuable resources for studying the basic principles of cell-type-specific regulation of secondary metabolism.

Role of female-predominant MYB39-bHLH13 complex in sexually dimorphic accumulation of taxol in Taxus media.

Taxus trees are major natural sources for the extraction of taxol, an anti-cancer agent used worldwide. Taxus media is a dioecious woody tree with high taxol yield. However, the sexually dimorphic accumulation of taxoids in T. media is largely unknown. Our study revealed high accumulation of taxoids in female T. media trees using a UPLC-MS/MS method. Thereafter, many differential metabolites and genes between female and male T. media trees were identified using metabolomic and transcriptomic analyses, respectively. Most of the taxol-related genes were predominantly expressed in female trees. A female-specific R2R3-MYB transcription factor gene, TmMYB39, was identified. Furthermore, bimolecular fluorescence complementation and yeast two-hybrid assays suggested the potential interaction between TmMYB39 and TmbHLH13. Several taxol biosynthesis-related promoter sequences were isolated and used for the screening of MYB recognition elements. The electrophoretic mobility shift assay indicated that TmMYB39 could bind to the promoters of the GGPPS, T10OH, T13OH, and TBT genes. Interaction between TmMYB39 and TmbHLH13 transactivated the expression of the GGPPS and T10OH genes. TmMYB39 might function in the transcriptional regulation of taxol biosynthesis through an MYB-bHLH module. Our results give a potential explanation for the sexually dimorphic biosynthesis of taxol in T. media.

Ultra-sensitive detection of hydrogen peroxide and levofloxacin using a dual-functional fluorescent probe.

Herein, a flower-shaped fluorescent probe was proposed for hydrogen peroxide (H2O2) and levofloxacin (LVF) sensing based on MoOx QDs@Co/Zn-MOFs with porous structure. Both MoOx QDs and Co/Zn-MOFs exhibited peroxidase-like properties, and the combination of them greatly aroused the synergistic catalytic capabilities between them. In o-Phenylenediamine (OPD)-H2O2 system, MoOx QDs@Co/Zn-MOFs efficiently catalyzed H2O2 to produce •OH and then oxidized OPD to its oxidation product (OxOPD). The OxOPD could not only emit blue fluorescence, but also inhibit the fluorescent intensity of MoOx QDs through fluorescence resonance energy transfer (FRET). Moreover, when introducing LVF into the system, the fluorescent intensities of MoOx QDs increased along with the aggregation of themselves while that of OxOPD remained unchanged, which was explained by the joint behavior of FRET and photo-induced electron transfer (PET) instead of the conventional aggregation-induced emission enhancement (AIEE). With these observation, the proposed probe was employed for H2O2 and LVF determination in biological samples with the limit of detection (LOD) of 32.60 pmol/L and 0.85 µmol/L, respectively, suggesting the method holds great promises for trace H2O2 and LVF monitoring in eco-environment.

Elevated CO2 improved soil nitrogen mineralization capacity of rice paddy.

Elevated CO2 would increase rice yields and may lead to nitrogen limitation and potentially influence the sustainability of agricultural production. Blindly increasing the amount of chemical fertilizer will damage the environment and is very unwise. Therefore, clarifying the response of soil nitrogen mineralization capacity to elevated CO2 is critical for both sustainable agriculture production and environmental protection. Here, we relied on Free-Air CO2 Enrichment (FACE) platform and used a waterlogged incubation method to investigate the effects of elevated CO2 on soil nitrogen mineralization capacity under different fertilization levels when planted different rice cultivars (strong and weak-CO2 response rice). According to the first-order kinetic equation fitting, compared with Ambient, elevated CO2 increased soil potential mineralized nitrogen (Np) by 16.18%. Path analysis indicated that fertilization status, rice cultivar, soil organic carbon and soil C: N ratio might affect Np. There was a significant positive correlation between soil nitrogen mineralization rate and Np. Under different fertilization conditions and rice cultivars, the improvement degree of soil nitrogen mineralization capacity (Np and soil nitrogen mineralization rate) by elevated CO2 was different. These findings suggest that more parameters and influencing factors should be taken into account when studying soil nitrogen cycle models under the condition of global change.