The WRKY transcription factor family plays a vital role in plant development and environmental response. However, the information of WRKY genes at the genome-wide level is rarely reported in Caragana korshinskii. In this study, we identified and renamed 86 CkWRKY genes, which were further classified into three groups through phylogenetic analysis. Most of these WRKY genes were clustered and distributed on eight chromosomes. Multiple sequence alignment revealed that the conserved domain (WRKYGQK) of the CkWRKYs was basically consistent, but there were also six variation types (WRKYGKK, GRKYGQK, WRMYGQK, WRKYGHK, WKKYEEK and RRKYGQK) that appeared. The motif composition of the CkWRKYs was quite conservative in each group. In general, the number of WRKY genes gradually increased from lower to higher plant species in the evolutionary analysis of 28 species, with some exceptions. Transcriptomics data and RT-qPCR analysis showed that the CkWRKYs in different groups were involved in abiotic stresses and ABA response. Our results provided a basis for the functional characterization of the CkWRKYs involved in stress resistance in C. korshinskii.

Reverse transcription quantitative PCR (RT-qPCR) is a technique widely used to investigate the expression of genes. An appropriate reference gene (RG) is essential for RT-qPCR analysis to obtain accurate and reliable results. Caragana intermedia plays an important role in afforestation as a bush. However, due to the lack of appropriate RGs, the research on development-related genes is limited. In this study, the selection for suitable RGs of different organs at various development stages to normalize the results of RT-qPCR about development-related genes was performed. To test the expression stability across all samples, we used the software algorithms such as geNorm, NormFinder, BestKeeper, and RefFinder to evaluate all the candidate RGs. Our results showed that CiEF1α was the most stable RG with little fluctuation among all samples. In addition, CiGAPDH in roots, CiSKIP1 in stems and leaves, and CiEF1α in different organs were selected as the most stable RGs. To confirm the applicability of the most stable RGs, the relative expression of CiWRKY17 was normalized using different candidate RGs. Taken together, our research laid a foundation for the study of development-related genes in C. intermedia.

Caragana intermedia (C. intermedia) is a kind of drought-tolerant leguminous shrub. WRKY transcription factors are one of the largest family of transcription factors in plants and play critical regulatory roles in stress tolerance and the development of plants. In our study, CiWRKY48 was cloned from C. intermedia, analyzed using bioinformatics software, and expressed with a prokaryotic expression system. The results showed that the open reading frame (ORF) of CiWRKY48 was 1158bp, the molecular weight (MW) was 42 kDa, and its subcellular localization was in the nucleus. Additionally, fusion protein was obtained, and confirmed by western blotting. The stress resistance of the pET30a-His-MBP-CiWRKY48 transformed Escherichia coli expression strain was reduced under mannitol and salt treatment, compared with the control. Overall, our findings provided a foundation for uncovering the function of CiWRKY48.

Background: Platycodon grandiflorus (PG) is a traditional Chinese medicinal herb widely cultivated in China. Although studies have demonstrated its effects in combating aging, the specific mechanisms underlying these effects have rarely been reported. This study utilized network pharmacology and molecular docking techniques to explore the active components of PG and its potential antiaging mechanisms. Methods: Active components of PG sourced from the traditional Chinese medicine systems pharmacology database, as well as the targets of active components and aging culled from databases like GeneCards, and their common targets were presented using a Venn diagram. Common targets were imported into the STRING database for protein-protein interaction analysis, and core targets were defined, which were enriched for Kyoto Encyclopedia of Genes and Genomes pathways. Molecular docking between active components and targets was performed using AutoDockTools software. Results: Our findings unveiled a total of 264 targets correspond to 7 active ingredients of PG, as well as 2906 potential targets related to aging, of which 152 were common targets. Among these common targets, 10 core targets have been identified as potentially playing a role in the antiaging effects of PG. Kyoto Encyclopedia of Genes and Genomes enrichment analysis of these targets revealed that PG regulates the aging process through various pathways, such as estrogen signaling pathway, lipid and atherosclerosis pathway, and apoptosis pathway. Molecular docking analysis further demonstrated a robust binding affinity between the active component acacetin and the antiaging targets CASP3 and SRC, as well as between spinasterol and the target ESR1. Conclusion: In summary, through this research, we screened information pertaining to PG’s active components and initially predicted potential targets and pathways for its antiaging effects, which required further experimental verification, however, our study provided a solid theoretical foundation for subsequent research on PG’s antiaging mechanisms and the development of related products.

Background: Codonopsis pilosula (CP), a traditional Chinese medicine, holds considerable potential in antioxidant and antiaging properties, however, the antiaging mechanism of its active components have been scarcely documented. Methods: Network pharmacology was employed to predict the active components of CP and elucidate its underlying antiaging mechanisms. Active components of CP were retrieved from traditional Chinese medicine systems pharmacology, followed by screening targets from GeneCards. Protein–protein interaction networks were constructed using String and analyzed with Cytoscape. Pathway enrichment was done using gene ontology and Kyoto encyclopedia of genes and genomes. Finally, molecular docking of active compounds with core targets was performed using Autodock and PyMOL. Results: Our study identified 21 active components of CP targeting 433 targets, and 2906 aging-related targets. By overlapping, 232 key targets emerged, including 14 core antiaging targets. CP regulated aging through targets such as AKT1, TNF, and SRC, as well as pathways including lipid metabolism, cancer progression, and estrogen signaling, and so on. Molecular docking demonstrated that both luteolin and 7-(β-xylosyl) cephalomannine_qt exhibited strong binding affinity to these key targets. Conclusion: In short, network pharmacology analyses screened active components of CP, predicted antiaging targets and pathways, thereby establishing a basis for subsequent product development and mechanistic research.