The mechanism leading to color differences between purple-red and green partridge tea leaves.

BACKGROUND: Partridge tea (Mallotus oblongifolius) is used as an important beverage and medical plant in Hainan province of China. Although some information about the morphology, cytology, and genetics of partridge tea has been reported in the literature, knowledge about this plant is still very limited. The leaves are the most important part for every tea plant, with a major role in nutrition and other functions. The leaves of different cultivars of partridge tea are different in colors and functions. The molecular mechanism of color formation of partridge tea leaf is still unclear. We reveal the molecular mechanism of the color difference between purple-red and green partridge tea leaves through metabolome and transcriptome analysis. RESULTS: We identified 665 compounds in the two partridge tea cultivars through metabolome analysis. Among these compounds, the content of 324 differed between the two cultivars. We also annotated 50 042 unigenes in the two cultivars by transcriptome analysis; 9665 unigenes were expressed differently between the two cultivars. Using an integrated analysis of the metabolome and transcriptome data, we found that the compounds and genes involved in anthocyanin biosynthesis were up-regulated in the purple-red leaves, compared with the green leaves. CONCLUSION: Our results showed that the anthocyanin biosynthesis pathway genes were up-regulated, which resulted in the up-regulation of the anthocyanin, making the leaf color purple-red. Our study reveals the molecular mechanism of the color difference between purple-red and green partridge tea, and lays a foundation for the genetic breeding of partridge tea genetic and the utilization of its volatile components. © 2022 Society of Chemical Industry.

The RavA/VemR two-component system plays vital regulatory roles in the motility and virulence of Xanthomonas campestris.

Xanthomonas campestris pv. campestris (Xcc) can cause black rot in cruciferous plants worldwide. Two-component systems (TCSs) are key for bacterial adaptation to various environments, including hosts. VemR is a TCS response regulator and crucial for Xcc motility and virulence. Here, we report that RavA is the cognate histidine kinase (HK) of VemR and elucidate the signalling pathway by which VemR regulates Xcc motility and virulence. Genetic analysis showed that VemR is epistatic to RavA. Using bacterial two-hybrid experiments and pull-down and phosphorylation assays, we found that RavA can interact with and phosphorylate VemR, suggesting that RavA is the cognate HK of VemR. In addition, we found that RpoN2 and FleQ are epistatic to VemR in regulating bacterial motility and virulence. In vivo and in vitro experiments demonstrated that VemR interacts with FleQ but not with RpoN2. RavA/VemR regulates the expression of the flagellin-encoding gene fliC by activating the transcription of the rpoN2-vemR-fleQ and flhF-fleN-fliA operons. In summary, our data show that the RavA/VemR TCS regulates FleQ activity and thus influences the expression of motility-related genes, thereby affecting Xcc motility and virulence. The identification of this novel signalling pathway will deepen our understanding of Xcc-plant interactions.

Transcriptome Analysis of Ceriops tagal in Saline Environments Using RNA-Sequencing.

Identification of genes involved in mangrove species' adaptation to salt stress can provide valuable information for developing salt-tolerant crops and understanding the molecular evolution of salt tolerance in halophiles. Ceriops tagal is a salt-tolerant mangrove tree growing in mudflats and marshes in tropical and subtropical areas, without any prior genome information. In this study, we assessed the biochemical and transcriptional responses of C. tagal to high salt treatment (500 mmol/L NaCl) by hydroponic experiments and RNA-seq. In C. tagal root tissues under salt stress, proline accumulated strongly from 3 to 12 h of treatment; meanwhile, malondialdehyde content progressively increased from 0 to 9 h, then dropped to lower than control levels by 24 h. These implied that C. tagal plants could survive salt stress through biochemical modification. Using the Illumina sequencing platform, approximately 27.39 million RNA-seq reads were obtained from three salt-treated and control (untreated) root samples. These reads were assembled into 47,111 transcripts with an average length of 514 bp and an N50 of 632 bp. Approximately 78% of the transcripts were annotated, and a total of 437 genes were putative transcription factors. Digital gene expression analysis was conducted by comparing transcripts from the untreated control to the three salt treated samples, and 7,330 differentially expressed transcripts were identified. Using k-means clustering, these transcripts were divided into six clusters that differed in their expression patterns across four treatment time points. The genes identified as being up- or downregulated are involved in salt stress responses, signal transduction, and DNA repair. Our study shows the main adaptive pathway of C. tagal in saline environments, under short-term and long-term treatments of salt stress. This provides vital clues as to which genes may be candidates for breeding salt-tolerant crops and clarifying molecular mechanisms of salt tolerance in C. tagal. The expression levels of 20 candidate genes measured by RNA-Seq were validated via qRT-PCR. Eighteen genes showed consistent expression patterns in RNA-Seq and qRT-PCR results, suggesting that the RNA-seq dataset was dependable for gene expression pattern analysis.

Influences of mesoporous magnesium silicate on the hydrophilicity, degradability, mineralization and primary cell response to a wheat protein based biocomposite.

A novel bioactive composite based on wheat protein (WP) and mesoporous magnesium silicate (m-MS) with a high specific surface area is presented in this study for potential bone tissue regeneration. Wheat protein (WP) is a type of a biodegradable natural polymer material. The m-MS was prepared by the sol-gel technique, which was incorporated into WP to fabricate m-MS/WP composites. The increasing amount of m-MS improved the surface hydrophilicity of m-MS/WP composites. The results showed that the degradation ratio of the m-MS/WP composites increased with an increase in the m-MS content after it was soaked in a Tris-HCl solution for 12 weeks. Moreover, the m-MS/WP composites with 40 wt% m-MS content (WP40) were able to maintain a suitable pH value over a prolonged soaking time, which might be dependent on the content of the m-MS. The WP40 showed a good apatite formation ability after it was soaked in simulated body fluid (SBF) for 7 days, indicating good bioactivity. Moreover, the WP40 with cytocompatibility stimulated the attachment, proliferation and differentiation of MC3T3-E1 osteoblast cells. Briefly, the results indicated that WP40 had good bioactivity, degradability, cytocompatibility and osteogenesis and might be a new biomaterial for bone regeneration.

MicroRNA-545 suppresses cell proliferation by targeting cyclin D1 and CDK4 in lung cancer cells.

An increasing number of reports have shown that diverse microRNAs are involved in tumorigenesis and tumor progression. We performed this study to identify novel miRNAs that may be involved in lung cancer and study on their functions. We tested the expression of 450 miRNAs in lung tumor tissues and adjacent non-cancerous tissues. We found that miRNA-545 was less abundant in cancerous lung tissues than in adjacent non-cancerous tissues. Our further studies showed that miR-545 suppressed cell proliferation in vitro and in vivo. We also found that miR-545 caused cell cycle arrest at the G0/G1 phase and induced cell apoptosis in lung cancer cells by targeting cyclin D1 and CDK4 genes. The effects of cyclin D1 and CDK4 down-regulated by miR-545 were similar to those caused by siRNAs of cyclin D1 and CDK4, and overexpression of cyclin D1 and CDK4 could abolish the miR-545-induced inhibition of cell proliferation. In conclusion, miR-545 suppressed cell proliferation by inhibiting the expression of cyclin D1 and CDK4. Our findings provide new knowledge regarding the role of miR-545 in the development of lung cancer and indicate the potential application of miR-545 in cancer therapy.

Tumor-suppressive microRNA-449a induces growth arrest and senescence by targeting E2F3 in human lung cancer cells.

MicroRNA-449a (miR-449a) was significantly downregulated in 156 lung cancer tissues (p<0.001). We found that the low expression of miR-449a was highly correlated with cancer recurrence and survival of lung cancer patients. The transient introduction of miR-449a caused cell cycle arrest and cell senescence in A549 and 95D cells. Further studies revealed that E2F3 was a direct target of miR-449a in lung cancer cells. miR-449a also suppressed tumor formation in vivo in nude mice. These results suggest that miR-449a plays an important role in lung cancer tumorigenesis and that miR-449a might predict cancer recurrence and survival of lung cancer patients.

MicroRNA cluster 302-367 enhances somatic cell reprogramming by accelerating a mesenchymal-to-epithelial transition.

MicroRNAs (miRNAs) are emerging critical regulators of cell function that frequently reside in clusters throughout the genome. They influence a myriad of cell functions, including the generation of induced pluripotent stem cells, also termed reprogramming. Here, we have successfully delivered entire miRNA clusters into reprogramming fibroblasts using retroviral vectors. This strategy avoids caveats associated with transient transfection of chemically synthesized miRNA mimics. Overexpression of 2 miRNA clusters, 106a-363 and in particular 302-367, allowed potent increases in induced pluripotent stem cell generation efficiency in mouse fibroblasts using 3 exogenous factors (Sox2, Klf4, and Oct4). Pathway analysis highlighted potential relevant effectors, including mesenchymal-to-epithelial transition, cell cycle, and epigenetic regulators. Further study showed that miRNA cluster 302-367 targeted TGFß receptor 2, promoted increased E-cadherin expression, and accelerated mesenchymal-to-epithelial changes necessary for colony formation. Our work thus provides an interesting alternative for improving reprogramming using miRNAs and adds new evidence for the emerging relationship between pluripotency and the epithelial phenotype.

MicroRNA-192 targeting retinoblastoma 1 inhibits cell proliferation and induces cell apoptosis in lung cancer cells.

microRNAs play an important roles in cell growth, differentiation, proliferation and apoptosis. They can function either as tumor suppressors or oncogenes. We found that the overexpression of miR-192 inhibited cell proliferation in A549, H460 and 95D cells, and inhibited tumorigenesis in a nude mouse model. Both caspase-7 and the PARP protein were activated by the overexpression of miR-192, thus suggesting that miR-192 induces cell apoptosis through the caspase pathway. Further studies showed that retinoblastoma 1 (RB1) is a direct target of miR-192. Over-expression of miR-192 decreased RB1 mRNA and protein levels and repressed RB1-3'-UTR reporter activity. Knockdown of RB1 using siRNA resulted in a similar cell morphology as that observed for overexpression of miR-192. Additionally, RB1-siRNA treatment inhibited cell proliferation and induced cell apoptosis in lung cancer cells. Analysis of miRNA expression in clinical samples showed that miR-192 is significantly downregulated in lung cancer tissues compared to adjacent non-cancerous lung tissues. In conclusion, our results demonstrate that miR-192 is a tumor suppressor that can target the RB1 gene to inhibit cell proliferation and induce cell apoptosis in lung cancer cells. Furthermore, miR-192 was expressed at low levels in lung cancer samples, indicating that it might be a promising therapeutic target for lung cancer treatment.

A mesenchymal-to-epithelial transition initiates and is required for the nuclear reprogramming of mouse fibroblasts.

Epithelial-to-mesenchymal transition (EMT) is a developmental process important for cell fate determination. Fibroblasts, a product of EMT, can be reset into induced pluripotent stem cells (iPSCs) via exogenous transcription factors but the underlying mechanism is unclear. Here we show that the generation of iPSCs from mouse fibroblasts requires a mesenchymal-to-epithelial transition (MET) orchestrated by suppressing pro-EMT signals from the culture medium and activating an epithelial program inside the cells. At the transcriptional level, Sox2/Oct4 suppress the EMT mediator Snail, c-Myc downregulates TGF-beta1 and TGF-beta receptor 2, and Klf4 induces epithelial genes including E-cadherin. Blocking MET impairs the reprogramming of fibroblasts whereas preventing EMT in epithelial cells cultured with serum can produce iPSCs without Klf4 and c-Myc. Our work not only establishes MET as a key cellular mechanism toward induced pluripotency, but also demonstrates iPSC generation as a cooperative process between the defined factors and the extracellular milieu. PAPERCLIP: