Elicitor-induced transcription factors for metabolic reprogramming of secondary metabolism in Medicago truncatula

<p>Abstract</p> <p>Background</p> <p>Exposure of <it>Medicago truncatula </it>cell suspension cultures to pathogen or wound signals leads to accumulation of various classes of flavonoid and/or triterpene defense molecules, orchestrated via a complex signalling network in which transcription factors (TFs) are essential components.</p> <p>Results</p> <p>In this study, we analyzed TFs responding to yeast elicitor (YE) or methyl jasmonate (MJ). From 502 differentially expressed TFs, <it>WRKY </it>and <it>AP2/EREBP </it>gene families were over-represented among YE-induced genes whereas <it>Basic Helix-Loop-Helix </it>(<it>bHLH</it>) family members were more over-represented among the MJ-induced genes. Jasmonate ZIM-domain <it>(JAZ</it>) transcriptional regulators were highly induced by MJ treatment. To investigate potential involvement of <it>WRKY </it>TFs in signalling, we expressed four <it>Medicago WRKY </it>genes in tobacco. Levels of soluble and wall bound phenolic compounds and lignin were increased in all cases. <it>WRKY W109669 </it>also induced tobacco <it>endo-1,3-β-glucanase </it>(<it>NtPR2</it>) and enhanced the systemic defense response to tobacco mosaic virus in transgenic tobacco plants.</p> <p>Conclusion</p> <p>These results confirm that <it>Medicago WRKY </it>TFs have broad roles in orchestrating metabolic responses to biotic stress, and that they also represent potentially valuable reagents for engineering metabolic changes that impact pathogen resistance.</p

34. 上肢のentrapment neuropathy手術例について(第520回千葉医学会例会 整形外科例会)

<p>No.8 nozzle assembly; flow rate 15 L/s; ambient pressure 30 MPa; ambient temperature = jet temperature = 373 K. In cross sections, dimensionless radius is the distance from the inner wall to the measure point divided by the total length between inner and outer wall. The tangential velocity has large gradients near both sides of inner and outer walls.</p

Association between chronic disease multimorbidity and leisure-time physical activity: Evidence from the China Multiethnic Cohort study

ObjectiveTo reveal the associations between multimorbidity and leisure-time physical activity (LTPA) by ethnicities in China.Materials and methodsSelf-reported information on a range of occupational, household, transport, and LTPA was collected by interviewer-administered questionnaire. A total of 17 chronic diseases were assessed based on self-reported lifetime diagnoses or medical examinations. Multivariable logistic regression models were used to assess the associations between multimorbidity and the risks of low LTPA.ResultsThe mean age of all participants was 51.2 years old. Of all, 61.4% were women and 57.9% were from the Han population. A significantly negative association (OR = 0.92, 95% CI = 0.89–0.95) was found between multimorbidity and low LTPA, with a stronger association among minority populations (OR = 0.86, 95% CI = 0.82–0.91) than among the Han population (OR = 0.96, 95% CI = 0.92–1.01). For both the minority population and the Han population, digestive system multimorbidity and digestive-metabolic system multimorbidity had a significantly negative association with low LTPA. For the Han population, the association of intersystem multimorbidity for the circulatory-respiratory system (OR = 1.17, 95% CI = 1.04–1.31) with low LTPA was stronger than that of intrasystem multimorbidity for the circulatory (OR = 1.12, 95% CI = 1.01–1.25) and respiratory systems (OR = 1.14, 95% CI = 1.04–1.25).ConclusionThere are significant associations between multimorbidity and low LTPA based on this large multiethnic population. Our findings suggest that LTPA-tailored interventions should be designed for specific ethnic groups according to different types of multimorbidity

Photochromic Fluorescent Probe Strategy for the Super-resolution Imaging of Biologically Important Biomarkers

Here, we report a β-galactosidase (β-Gal)-responsive photochromic fluorescent probe, NpG, that was designed to prebind to human serum albumin (HSA) to form the probe/protein hybrid, NpG@HSA. The formation of NpG@HSA led to an increase in fluorescence emission (520 nm) corresponding to the binding of the fluorescent naphthalimide unit with HSA. In addition, this enabled visualization of the spiropyran fluorescence emission in aqueous media. Our probe/protein hybrid approach afforded a unique imaging platform with enhanced cell permeability and solubility that was capable of visualizing the cellular uptake of NpG@HSA before its activation by β-Gal. The β-Gal-mediated cleavage of the galactose unit within the NpG@HSA hybrid resulted in the formation of NpM@HSA and an increase in red fluorescence emission (620 nm). The resultant merocyanine unit was then able to undergo photoisomerization (merocyanine ↔ spiropyran) to facilitate STORM (i.e., stochastic optical reconstruction microscopy) imaging with minimal phototoxicity and excellent photostability/reversibility. Using STORM, NpG@HSA was able to determine the subcellular distribution of β-Gal activity between cell lines with nanoscale precision. We believe that this system represents a versatile imaging platform for the design of photochromic fluorescent probes suitable for illuminating the precise location of disease-specific biomarkers in various cellular processes.</p

Functional Characterization of Proanthocyanidin Pathway Enzymes from Tea and Their Application for Metabolic Engineering

Tea (Camellia sinensis) is rich in specialized metabolites, especially polyphenolic proanthocyanidins (PAs) and their precursors. To better understand the PA pathway in tea, we generated a complementary DNA library from leaf tissue of the blister blight-resistant tea cultivar TRI2043 and functionally characterized key enzymes responsible for the biosynthesis of PA precursors. Structural genes encoding enzymes involved in the general phenylpropanoid/flavonoid pathway and the PA-specific branch pathway were well represented in the library. Recombinant tea leucoanthocyanidin reductase (CsLAR) expressed in Escherichia coli was active with leucocyanidin as substrate to produce the 2R,3S-trans-flavan-ol (+)-catechin in vitro. Two genes encoding anthocyanidin reductase, CsANR1 and CsANR2, were also expressed in E. coli, and the recombinant proteins exhibited similar kinetic properties. Both converted cyanidin to a mixture of (+)-epicatechin and (−)-catechin, although in different proportions, indicating that both enzymes possess epimerase activity. These epimers were unexpected based on the belief that tea PAs are made from (−)-epicatechin and (+)-catechin. Ectopic expression of CsANR2 or CsLAR led to the accumulation of low levels of PA precursors and their conjugates in Medicago truncatula hairy roots and anthocyanin-overproducing tobacco (Nicotiana tabacum), but levels of oligomeric PAs were very low. Surprisingly, the expression of CsLAR in tobacco overproducing anthocyanin led to the accumulation of higher levels of epicatechin and its glucoside than of catechin, again highlighting the potential importance of epimerization in flavan-3-ol biosynthesis. These data provide a resource for understanding tea PA biosynthesis and tools for the bioengineering of flavanols

miR-K12-7-5p Encoded by Kaposi's Sarcoma-Associated Herpesvirus Stabilizes the Latent State by Targeting Viral ORF50/RTA

Seventeen miRNAs encoded by Kaposi's sarcoma-associated herpesvirus (KSHV) have been identified and their functions have begun to be characterized. Among these miRNAs, we report here that miR-K12-7 directly targets the replication and transcription activator (RTA) encoded by open reading frame 50. We found that miR-K12-7 targeted the RTA 3′ untranslated region (RTA3′UTR) in a seed sequence-dependent manner. miR-K12-7-5p derived from miR-K12-7 mediates the inhibition of RTA expression, and the mutation of the seed match site totally abrogated the inhibitory effect of miR-K12-7 on RTA3′UTR. The inhibition of RTA expression by miR-K12-7 was further confirmed in the latently KSHV-infected 293/Bac36 cell line through transient transfection of miR-K12-7 expression plasmid or specific inhibitor of miR-K12-7-5p, respectively. The transient transfection of miR-K12-7 into 293/Bac36 cells reduced RTA expression and the expression of the downstream early genes regulated by RTA, and also the production of progeny virus was significantly reduced after treatment with chemical inducers. Our study revealed that another miRNA, miR-K12-7-5p, targets the viral immediate early gene RTA and that this miRNA contributes to the maintenance of viral latency

Construction of a cross-species cell landscape at single-cell level.

Individual cells are basic units of life. Despite extensive efforts to characterize the cellular heterogeneity of different organisms, cross-species comparisons of landscape dynamics have not been achieved. Here, we applied single-cell RNA sequencing (scRNA-seq) to map organism-level cell landscapes at multiple life stages for mice, zebrafish and Drosophila. By integrating the comprehensive dataset of > 2.6 million single cells, we constructed a cross-species cell landscape and identified signatures and common pathways that changed throughout the life span. We identified structural inflammation and mitochondrial dysfunction as the most common hallmarks of organism aging, and found that pharmacological activation of mitochondrial metabolism alleviated aging phenotypes in mice. The cross-species cell landscape with other published datasets were stored in an integrated online portal-Cell Landscape. Our work provides a valuable resource for studying lineage development, maturation and aging