By sequencing and analyzing the transcriptome of Picea wilsonii, the potential drought-related gene PwRhomboid was screened out, and its functional characteristics of drought resistance in Picea wilsonii were analyzed. Through bioinformatics analysis, the amino acid sequence and homologous gene of PwRhomboid were compared. The responses of PwRhomboid to different abiotic stresses and hormones were analyzed by RT-qPCR. Instantaneous transformation of tobacco leaves was conducted to detect the subcellular localization of PwRhomboid protein. PwRhomboid transgenic strains of Arabidopsis and potato were obtained and their drought phenotypes were verified. The results showed that the amino acid sequence of PwRhomboid has significant difference with those of its homologous species at N-terminus, but shows high similarity and good conservation at C-terminus. PwRhomboid protein is mainly localized on cell membrane. The expression level of PwRhomboid was significantly induced by drought, low temperature and abscisic acid, with the highest expression level observed in mature leaves.PwRhomboid can improve the drought resistance in transgenic Arabidopsis and potato. After drought treatment, compared with the control group Arabidopsis WT and pCM1205 strains, PwRhomboid overexpression strains PwRhomboid-L1 and PwRhomboid-L2 had higher survival rates, chlorophyll fluorescence Fv/Fm and ΦPSⅡ. After polyethyleneglycol simulated drought treatment, the overexpression strains PwRhomboid-L3 and PwRhomboid-L4 resulted in higher potato seedling heights compared to the wild-type. The results showed that the expression level of PwRhomboid is affected by drought and hormones, and overexpression of PwRhomboid could improve the drought resistance of Arabidopsis and potato.
Diabetic nephropathy (DN) is a major complication of diabetes mellitus and a major causative agent of chronic kidney disease.UBE2V1, as the only E2 enzyme involved in K63 ubiquitination, is involved in the development of diabetic nephropathy, while UHRF2 acts as E3 ubiquitin ligase in the K63 ubiquitination pathway. Real-time fluorescence quantification, gene silencing and overexpression, scratching test, western blot analysis, K63 ubiquitination protein isolation and immunoprecipitation were used to explore the mechanism of UHRF2 and UBE2V1 in the process of epithelial-mesenchymal transition of renal tubular epithelial cells induced by high glucose. It was found that high glucose could increase the expression of UHRF2 and UBE2V1 in renal tubular epithelial cells, leading to the development of epithelial-mesenchymal transition. The expression of epithelial-mesenchymal transition marker proteins α-SMA and ZEB1 was significantly up-regulated in renal tubular epithelial cells cultured with high glucose, while E-cadherin expression was significantly down-regulated. Knocking down of UHRF2 or UBE2V1 inhibited the up-regulation of α-SMA and ZEB1, and weakened the down-regulation of E-cadherin.UBE2V1 may mediate the up-regulation of UHRF2 transcription in renal tubular epithelial cells cultured with high glucose and may increase the ubiquitination of K63 in UHRF2.The results reflect that both UHRF2 and UBE2V1 are involved in the epithelial-mesenchymal transition of renal tubular epithelial cells under the condition of high glucose, which may synergistic promote the fibrosis of diabetic nephropathy. These findings provide a new idea for the treatment of diabetic nephropathy.
Aegilops tauschii, the D genome donor of wheat (Triticum aestivum), has a variety of genes resistant to biotic and abiotic stresses, and is the main gene pool for wheat genetic improvement. Heavy metal-associated isoprene plant protein (HIPP) plays an important role in regulating metal ion balance and detoxification in plants. In this study, the transgenic Arabidopsis thaliana overexpressing AetHIPP28 was obtained by Agrobacterium-mediated method, and the response of transgenic Arabidopsis thaliana to cadmium (Cd) stress was studied by a series of physiological and biochemical analyses.The results showed that AetHIPP28 was localized in the nucleus and plasma membrane. Overexpression of AetHIPP28 significantly enhanced Arabidopsis thaliana tolerance to Cd, and promoted the root growth of Arabidopsis thaliana under Cd stress. The ROS accumulation of transgenic Arabidopsis thaliana under Cd stress was significantly lower than that of the wild type, while the activity of antioxidant enzymes was significantly higher than that of the wild type. Meanwhile, overexpressing AetHIPP28 inhibited Cd2+influx and increased Cd accumulation in roots. These results suggest that overexpression of AetHIPP28 improves Cd tolerance in Arabidopsis thaliana by regulating ROS accumulation, antioxidant enzyme activity, Cd2+ influx and distribution in roots, which provide a reference for investigating the response mechanism of AetHIPP28 to Cd stress.
The MADS-box family is one of the largest transcription factor families in plants, and plays an important role in adversity stress and flowering response. The function of AGL12-like subgroup in Vitis vinifera remains unclear. The VvAGL12 promoter (proVvAGL12) was cloned from the Pinot Noir grape genome, and the sequence elements of the promoter were analyzed. It was found that there were several cis-acting elements related to adversity stress in the promoter region. GUS expression vector driven by proVvAGL12 was constructed and transformed in Arabidopsis thaliana and tobacco. It was found that proVvAGL12 promoter fragment had activation activity in Arabidopsis thaliana, and it can drive GUS expression in leaves, stem segments, flower organs, roots and fruit pods in transgenic Arabidopsis thaliana plants.The expression activity can last the whole growth cycle. Transgenic Arabidopsis thaliana and tobacco stress treatment showed that the activity of proVvAGL12-driven GUS was regulated by gibberellin, abscisic acid, polyethylene glycol and low temperature.
The floral organs of Ranunculales show complex and diverse characteristics, which are ideal plant group to study the related problems of morphology and genetic evolution of floral organs. This paper reviewed the floral organ morphological diversity of Ranunculales. Combining the genome and transcriptome of Ranunculales as well as the flower development researches of representative species including Thalictrum thalictroides, Aquilegia coulela, Nigella damascena and Eschscholzia californica, the current research results on the function of MADS-box gene family A, B, C, D, E genes and their downstream target genes were elaborated.Besides, the breakthrough points and possible challenges of flower development research in Ranunculales in the future were forecasted. The aim is to enrich the understanding on the molecular pathways of floral organ development in basal eudicots.
PsaA and PsaB are central proteins in photosystem Ⅰ, and play important roles in plant photosynthesis and stress regulation. Tobacco (Nicotiana tabacum L.) is a model crop for plant functional genomics research, while there are few studies on the functions of PsaA and PsaB genes. In order to understand the response of tobacco PsaA/PsaB genes to stress, the PsaA/PsaB gene family in tobacco genome were identified, and their physicochemical properties, subcellular localization, phylogenetic evolution, motifs and promoter regulatory elements were analyzed. The results showed that there are 27 PsaA/PsaB genes in tobacco. Compared with Arabidopsis thaliana, the tobacco PsaA/PsaB gene family undergone significant expansion and can be classified into three subfamilies based on their phylogenetic and structural features. The promoters of tobacco PsaA/PsaB genes contain numerous cis-regulatory elements responsive to light, low temperature, drought and plant hormones. RT-qPCR analysis revealed that most tobacco PsaA/PsaB genes were up-regulated under low temperature stress (8 up-regulated genes) and down-regulated under polyethylene glycol and jasmonic acid methyl ester stresses(8 and 7 down-regulated genes respectively). After potato virus Y(PVY) and tobacco mosaic virus (TMV) infection, the expression of most tobacco PsaA/PsaB genes were down-regulated. The down-regulated genes in the lower mesophyll and vein tissues were 19 and 17 after PVY infection, and 9 genes were down-regulated after TMV infection. In conclusion, tobacco PsaA/PsaB genes are involved in the response process of tobacco to various abiotic stresses, and the study results provide a reference for further exploring the role of PsaA/PsaB genes in regulating plant response to stress.
To investigate the effect of composite nanofiber membrane prepared from isoorientin (ISO) and zein/Arabic gum (GA) on skin injury healing in mice, this study used blank spinning membrane as control, and wound dressings were applied to mice using zein/GA loaded nanofiber membrane, ISO loaded nanofiber membrane, and ISO-zein/GA loaded composite nanofiber membrane, respectively. The results showed that the nanofiber membranes loaded with zein/GA, ISO and ISO-zein/GA had promoting effects on wound healing in mice. They effectively reduced wound oxidative damage and mRNA expression levels of pro-inflammatory factors(TNF-α、IL-1β、iNOS), and promoted wound vascular reconstruction, thereby accelerating wound healing. The research results verify the promoting effect of ISO-zein/GA composite nanofiber membrane on wound healing, expand the application range of isoorientin, and provide ideas for the research of new wound dressing.
Polygonatum kingianum Coll. et Hemsl. (PK), a well-known Food and Medicine homology in China, riches in various bioactive ingredients and has been consumed for thousands of years with a wide range of applications. PK has shown great tonic effects. Previous studies have consistently shown nutritional functions of PK including liver protection. The Nine Steaming Nine Drying is a traditional processing technique that has been considered as an effective method in enriching tonic properties, but its impacts on improving efficacy of PK on liver protection remain largely unexplored. This study herein applied integrative analysis of metabolomics and bioinformatics, as well as molecular docking, to reveal the core bioactive compounds responsible for the improved effects of Nine Steaming Nine Drying processed PK extracts (PPK) on the prevention and treatment of alcoholic liver injury and its underlying mechanisms.Through constructing an alcohol induced injury model using LO2 cells, the regulatory effects of PPK on cell activity, oxidative stress, and alcohol metabolism enzymes (ethanol dehydrogenase and acetaldehyde dehydrogenase) were investigated. A total of 204 metabolites in the PPK were compared with crude PK extracts (CPK). Alcohol induced liver injury related targets were obtained by searching for DisGeNET, TTD, GeneCard, and OMIM databases, and a compound-target network was established. Results from protein-protein interaction networks and enrichment analysis identified 23 functional metabolites (including rutin, furanone, caffeic acid, gallic acid, etc.) that may be responsible for the protective effects of PPK on alcoholic liver injury via regulating 15 action targets involved in PI3K-Akt, AMPK, NF- kB signaling pathways. High affinity between the identified key constituents of PPK and their predicted acting targets was confirmed using molecular docking. Results indicate that PPK can alleviate alcoholic liver injury by regulating oxidative stress, inflammation, and ethanol metabolisms. It provides novel insights in nutritional functions of PPK, and an effective strategy for systematically elucidating the mechanisms of functional foods.
The aim of this study was to investigate the effects of Ginkgo biloba powder on growth performance, serum indexes, rumen microbiome and rumen metabolomics of Angus cattle. A total of 25 healthy Angus cattle with an average body weight of 453.8 kg aged 12~16 months were randomly divided into 5 groups. The cattle were fed the basal diet (Con, n=5) and diets containing 3% (G3, n=5), 6% (G6, n=5), 9% (G9, n=5) and 12% (G12, n=5) Ginkgo biloba powder, respectively. The pre-test lasted for 10 d and the trial lasted for 75 d. The results showed as follows: dietary Ginkgo biloba powder increased the average daily gain of Angus cattle, and significantly increased the dry matter intake of Angus cattle. Compared with Con group, serum triglyceride content in G9 group was significantly decreased,total antioxidant capacity was significantly increased,malonaldehyde concentrations in G6 and G9 groups were significantly decreased. Compared with Con group, the proportions of propionic acid in G6 and G9 groups were significantly increased, while the proportions of acetic acid were significantly decreased. There was no significant difference in the diversity and richness of rumen microbial communities among all groups. At the phylum level, compared with Con group, the relative abundances of Firmicutes in rumen of Angus cattle in G6 and G9 groups were significantly increased, while the relative abundances of Spirochaetota were significantly decreased. At the genus level, the relative abundances of F082_unclassified and Paludibacter_unclassified in G6 group were significantly decreased compared with Con group.The relative abundance of F082_unclassified in G9 group was significantly decreased, and that of Succiniclasticum was significantly increased. The results of rumen metabolomics showed that in positive ion mode, compared with Con group, the contents of fatty acids in the rumen of G6 and G9 groups were significantly decreased, and the contents of other metabolites were significantly increased. In negative ion mode, the contents of diazine and organic oxygen compounds in rumen of G6 and G9 groups were significantly decreased compared with Con group, while the contents of other metabolites were significantly increased. In conclusion, dietary supplementation of 6%~9% Ginkgo biloba powder has positive effects on the growth performance and blood indexes of Angus cattle, improves rumen fermentation parameters, and has certain effects on the composition of rumen metabolites. It is suggested that the optimal supplemental amount of Ginkgo biloba powder should be 6%~9% of the basal diet.
In order to achieve accurate and rapid detection for Staphylococcus aureus in drugs, through adding internal amplification control (IAC) and optimizing PCR reaction system, an internal reference based fluorescence quantitative PCR rapid detection method of Staphylococcus aureus that can monitor the process in real time was established, and its specificity, detection limit, reproducibility and feasibility were verified. The verification results showed that the specificity of the established method was good with only Staphylococcus aureus showed typical amplification curve. The detection limit of Staphylococcus aureus genomic DNA was 0.23 pg/μL. There was a good linear relationship between Ct values and the copy numbers of template (R2=0.99). The relative standard deviations of reproducibility experiments were all less than 3.0%.Staphylococcus aureus can be detected 10 h after the bacteria increase in artificially contaminated drugs. The method can not only shorten the detection time of Staphylococcus aureus in drugs, but can also monitor the PCR reaction process, effectively prevent the occurrence of “false negative” result, thus can be used as a supplementary method for staphylococcus aureus detection.
This study focuses on the real-time dynamic monitoring of meteorological factors and sap flow rates of Haloxylon ammodendron subjected to different pruning treatments in the northeast part of the Ulan Buh Desert. The sap flow characteristics of Haloxylon ammodendron at different time scales during the vigorous growth season (from June to August) under different pruning treatments were investigated and their responses to meteorological factors were examined. Water consumption model in relation to meteorological factors was established to reveal the patterns of water transportation in Haloxylon ammodendron after pruning. Thermal diffusion sap flow meter and automatic weather station were used to monitor the sap flow in Haloxylon ammodendron subjected to different pruning treatments and solar radiation, air temperature, relative humidity and vapor pressure deficit.Sap flow rates in Haloxylon ammodendron under different pruning treatments (120 cm pruning, 90 cm pruning, 60 cm pruning) exhibited a “broad peak” pattern on sunny days, while unpruned Haloxylon ammodendron displayed a “single-peak” pattern. On rainy days, sap flow rates fluctuated dramatically, presenting an “irregular multi-peak” curve, which are lower compared to those of sunny days. The daily average sap flow rates in unpruned, 120 cm pruned, 90 cm pruned and 60 cm pruned Haloxylon ammodendron were 2.314 6, 3.333 4, 1.333 3 and 0.767 2 cm/h, respectively. The daily water consumptions were 8.62, 13.43, 2.40 and 2.89 kg/d, respectively. On the hour scale, solar radiation was the primary factor influencing sap flow rates in both pruned and unpruned Haloxylon ammodendron.On the daily scale, solar radiation was the main factor affecting sap flow rates in 90 cm pruned and unpruned Haloxylon ammodendron, while air temperature influenced sap flow rates in 120 cm pruned Haloxylon ammodendron and wind speed affected sap flow rates in 60 cm pruned Haloxylon ammodendron.On the month scale, water consumption in 120 cm pruned Haloxylon ammodendron was mainly influenced by solar radiation and air temperature, water consumption in 90 cm pruned Haloxylon ammodendron was primarily influenced by relative humidity and solar radiation, and water consumption in 60 cm pruned Haloxylon ammodendron was mainly influenced by wind speed. With the increase in time scale, the number of meteorological factors included in the model decreased, and the explanatory power for sap flow decreased. The fitted equations in this study reached significant levels and can predict the transpiration characteristics of Haloxylon ammodendron at different time scales. The results of this study are of great significance for understanding the patterns of water transportation in Haloxylon ammodendron after pruning.