Comparative transcriptome analysis reveals gene network regulation of TGase-induced thermotolerance in tomato

Transglutaminase (TGase), the ubiquitous protein in plants, catalyzes the post-translational transformation of proteins and plays a vital role in photosynthesis. However, its role and mechanism in tomato subjected to heat stress still remain unknown. Here, we carried out a transcriptomic assay to compare the differentially expressed genes (DEGs) between wild type (WT) and TGase overexpression (TGaseOE) plants employed to high-temperature at 42 °C and samples were collected after 0, 6, and 12 h, respectively. A total of 11,516 DEGs were identified from heat-stressed seedlings, while 1,148 and 1,353 DEGs were up-and down-regulated, respectively. The DEGs upon high-temperature stress were closely associated with the pathways encompassing protein processing in the endoplasmic reticulum, carbon fixation, and photosynthetic metabolism. In addition, 425 putative transcription factors (TFs) were identified, and the majority of them associated with the bHLH, HSF, AP2/ERF, MYB, and WRKY families. RNA-seq data validation further confirmed that 8 genes were linked to protein processing and photosynthesis, and the mRNA level of these genes in TGaseOE was higher than that in WT plants, which is consistent in transcriptome results. In conclusion, these results reveal the transcriptional regulation between WT and TGaseOE in tomato under heat stress and shed light on a new dimension of knowledge of TGase-mediated thermotolerance mechanism at the molecular level

Protective effect of five-flavor sophora flavescens enteric-coated capsules on inflammatory bowel disease and its molecular mechanism

Abstract This study aims to investigate the effect of five-flavor sophora flavescens enteric-coated capsules (FSEC) on TNF-α-induced inflammatory bowel disease and its molecular mechanism. Wistar Rats were divided divided into 6 groups: Normal control group (group A): normal diet, drinking water; Model group (group B): 100 μg/L TNF-α; FSEC high-dose group (group C): 100μg/L TNF-α + FSEC (432 mg/kg); FSEC medium-dose group (group D): 100 μg/L TNF-α + FSEC (216 mg/kg); FSEC low-dose group (group E): 100 μg/L TNF-α + FSEC (108 mg/kg); Positive control group (group F): 100 μg/L TNF-α + 500 mg/kg sulfasalazine (SAZ). Animals in each group were intragastrically administered twice daily for 7 days. Animals were sacrificed 24 hours after the last treatment and colon tissues were collected for subsequent experiments. The results of HE staining showed that the colonic tissue of TNF-α-fed animals appeared damage, while the colonic tissue of animals treated with FSEC was improved to various degrees, and the histological characteristics of colon were basically recovered in the high-dose group, suggesting that FSEC could be used to treat TNF-α-induced colonic tissue damage. According to the results of ELISA and immunohistochemistry, the recovery of colonic tissue structure in rats treated with different doses of FSEC might be related to the decrease of TNF-α, IL-6, IL-17, TLR-4 and NF-κB proteins expression. According to the results of Western blotting, TNF-α-pretreated IEC-6 cells cultured with medicated serum decreased the expression of TRIF and IFN-γ proteins. These results suggest that FSEC has a protective effect on ulcerative colitis (UC), and the mechanism may be through inhibiting the activation of TLR-4/NF-κB signaling pathway and preventing the release of related inflammatory factors

Proteomic and Physiological Analyses Reveal Putrescine Responses in Roots of Cucumber Stressed by NaCl

Soil salinity is a major environmental constraint that threatens agricultural productivity. Different strategies have been developed to improve crop salt tolerance, among which the effects of polyamines have been well reported. To gain a better understanding of the cucumber (Cucumis sativus L.) responses to NaCl and unravel the underlying mechanism of exogenous putrescine (Put) alleviating salt-induced damage, comparative proteomic analysis was conducted on cucumber roots treated with NaCl and/or Put for 7 days. The results showed that exogenous Put restored the root growth inhibited by NaCl. 62 differentially expressed proteins implicated in various biological processes were successfully identified by MALDI-TOF/TOF MS. The four largest categories included proteins involved in defense response (24.2%), protein metabolism (24.2%), carbohydrate metabolism (19.4%) and amino acid metabolism (14.5%). Exogenous Put up-regulated most identified proteins involved in carbohydrate metabolism, implying an enhancement in energy generation. Proteins involved in defense response and protein metabolism were differently regulated by Put, which indicated the roles of Put in stress resistance and proteome rearrangement. Put also increased the abundance of proteins involved in amino acid metabolism. Meanwhile, physiological analysis showed that Put could further up-regulated the levels of free amino acids in salt stressed-roots. In addition, Put also improved endogenous polyamines contents by regulating the transcription levels of key enzymes in polyamine metabolism. Taken together, these results suggest that Put may alleviate NaCl-induced growth inhibition through degradation of misfolded/damaged proteins, activation of stress defense, and the promotion of carbohydrate metabolism to generate more energy

Can heavy metal pollution induce bacterial resistance to heavy metals and antibiotics in soils from an ancient land-mine?

Microbial resistance to antibiotics is a growing challenge to human health. Recent evidence has indicated that antibiotic resistance can be co-selected for by exposure to heavy metals in agricultural soils. It remains unknown if this is a concern in other environments contaminated by metals. We here investigated soil microbial activities, composition and tolerance to heavy metals and antibiotics in a mining soil survey. We found that microbial respiration, growth, and biomass were affected by available metal concentrations. Most of the variation in microbial PLFA composition was explained by differences in heavy metal and pH. Additionally, pollution-induced bacterial community tolerance to toxicants including Cu, Pb, Zn, tetracycline and vancomycin was determined. Although only bacterial tolerance to Pb increased with higher levels of metals, the links between bacterial metal tolerance and soil metal concentrations were clear when considered together with previously published reports, suggesting that bacterial metal tolerance were universally elevated in the surveyed soils. The induced levels of heavy metal tolerance coincided with elevated levels of tolerance to vancomycin, but not to tetracycline. Our study showed that heavy metals can co-select for resistance to clinically important antibiotics also in ecosystems without manure input or antibiotic pollution

Neural-Network-Based Approach for Extracting Eigenvectors and Eigenvalues of Real Normal Matrices and Some Extension to Real Matrices

This paper introduces a novel neural-network-based approach for extracting some eigenpairs of real normal matrices of order n. Based on the proposed algorithm, the eigenvalues that have the largest and smallest modulus, real parts, or absolute values of imaginary parts can be extracted, respectively, as well as the corresponding eigenvectors. Although the ordinary differential equation on which our proposed algorithm is built is only n-dimensional, it can succeed to extract n-dimensional complex eigenvectors that are indeed 2n-dimensional real vectors. Moreover, we show that extracting eigen-pairs of general real matrices can be reduced to those of real normal matrices by employing the norm-reducing skill. Numerical experiments verified the computational capability of the proposed algorithm

Overexpression of miR-27b-3p Targeting Wnt3a Regulates the Signaling Pathway of Wnt/β-Catenin and Attenuates Atrial Fibrosis in Rats with Atrial Fibrillation

MicroRNAs (miRNAs) are regarded as a potential method for the treatment of atrial fibrillation (AF) although its molecular mechanism remains unknown. We found in our previous study that the level of peripheral blood miR-27b-3p and the expression of atrial tissue CX43 were both significantly downregulated in AF patients. In the present study, we propose and test this hypothesis that overexpression of miR-27b-3p attenuates atrial fibrosis, increases CX43 expression, and regulates the signaling pathway of Wnt/β-Catenin by targeting Wnt3a. miR-27b-3p overexpression was induced by rat tail vein injection of adeno-associated virus. Two weeks after transfection of adeno-associated virus, the rat AF model was established by tail vein injection of acetylcholine- (ACh-) CaCl2 for 7 days, and 1 ml/kg was injected daily. The incidence and duration of AF were recorded with an electrocardiogram. Cardiac function was monitored by cardiac ultrasound. Serum cardiac enzyme was detected by ELISA. The expression of atrial miR-27b-3 and Wnt3a was assayed by quantitative RT-PCR. Atrial fibrosis was determined by Masson’s trichrome staining. Expression of atrial Collagen-I and Collagen-III was tested by the immunohistochemical method. Expression of CX43 was measured by immunofluorescence. The expression of Collagen-I, a-SMA, Collagen-III, TGF-β1, CX43, Wnt3a, β-Catenin, and p-β-Catenin was assayed by western blot. Our results showed that miR-27b-3p overexpression could reduce the incidence and duration of AF, alleviate atrial fibrosis, increase atrial CX43 expression, and decrease the expression of Collagen-I, a-SMA, Collagen-III, TGF-β1, Wnt3a, and p-β-Catenin. In addition, the results of luciferase activity assay showed that Wnt3a is a validated miR-27b-3p target in HEK 293T cells. Our results provide a new evidence that miR-27b-3p regulates the signaling pathway of Wnt/β-Catenin by targeting Wnt3a, which may play an important role in the development of atrial fibrosis and AF