Fructose-1,6-bisphosphate aldolase (FBA) is a versatile metabolic enzyme involved in multiple important processes of glycolysis, gluconeogenesis, and Calvin cycle. Despite its significance in plant biology, the identity of this gene family in oil crops is lacking. Here, we performed genome-wide identification and characterization of FBAs in an allotetraploid species, oilseed rape Brassica napus. Twenty-two BnaFBA genes were identified and divided into two groups based on integrative analyses of functional domains, phylogenetic relationships, and gene structures. Twelve and ten B. napus FBAs (BnaFBAs) were predicted to be localized in the chloroplast and cytoplasm, respectively. Notably, synteny analysis revealed that Brassica-specific triplication contributed to the expansion of the BnaFBA gene family during the evolution of B. napus. Various cis-acting regulatory elements pertinent to abiotic and biotic stresses, as well as phytohormone responses, were detected. Intriguingly, each of the BnaFBA genes exhibited distinct sequence polymorphisms. Among them, six contained signatures of selection, likely having experienced breeding selection during adaptation and domestication. Importantly, BnaFBAs showed diverse expression patterns at different developmental stages and were preferentially highly expressed in photosynthetic tissues. Our data thus provided the foundation for further elucidating the functional roles of individual BnaFBA and also potential targets for engineering to improve photosynthetic productivity in B. napus.

DNA methylation is a process through which methyl groups are added to the DNA molecule, thereby modifying the activity of a DNA segment without changing the sequence. Increasing evidence has shown that DNA methylation is involved in various aspects of plant growth and development via a number of key processes including genomic imprinting and repression of transposable elements. DNA methylase and demethylase are two crucial enzymes that play significant roles in dynamically maintaining genome DNA methylation status in plants. In this work, 22 DNA methylase genes and six DNA demethylase genes were identified in rapeseed (Brassica napus L.) genome. These DNA methylase and DNA demethylase genes can be classified into four (BnaCMTs, BnaMET1s, BnaDRMs and BnaDNMT2s) and three (BnaDMEs, BnaDML3s and BnaROS1s) subfamilies, respectively. Further analysis of gene structure and conserved domains showed that each sub-class is highly conserved between rapeseed and Arabidopsis. Expression analysis conducted by RNA-seq as well as qRT-PCR suggested that these DNA methylation/demethylation-related genes may be involved in the heat/salt stress responses in rapeseed. Taken together, our findings may provide valuable information for future functional characterization of these two types of epigenetic regulatory enzymes in polyploid species such as rapeseed, as well as for analyzing their evolutionary relationships within the plant kingdom.

Alfalfa (Medicago sativa L.), a vital perennial legume forage, has been widely cultivated owing to a variety of favorable characteristics, including comprehensive ecological resilience, superior nutritive value, digestibility, and nitrogen fixation capacity. The productivity traits of alfalfa, particularly its biomass yield and forage quality, are profoundly influenced by a range of abiotic stress conditions. As a common abiotic stress, drought adversely impacts growth and photosynthetic efficiency, accompanied by increased oxidative damage and stomatal closure as a mechanism to minimize water loss; meanwhile, transgenic approaches have been employed to enhance drought resilience by improving antioxidant activity and water-use efficiency. Salinity stress disturbs ionic balance, resulting in sodium (Na⁺) toxicity and the generation of oxidative damage; however, alfalfa cultivars exhibit salinity tolerance through mechanisms such as Na⁺ exclusion, K⁺ retention, activation of antioxidant defenses, hormonal regulation, and the upregulation of stress-responsive genes. In addition, heavy metals pose a significant challenge to alfalfa production, as they impair plant development and disrupt symbiotic nitrogen fixation, but recent studies have highlighted the potential of microbial-assisted phytoremediation in mitigating these detrimental effects. By integrating recent findings, this review highlights the intricate physiological, biochemical, and molecular mechanisms involved in alfalfa’s responses to key abiotic stressors specifically drought, salinity, and heavy metal toxicity. Breakthroughs in genetic modification, notably the development of transgenic lines exhibiting altered expression of stress-responsive genes, offer valuable potential for improving stress resilience. Future research should employ omics approaches, advanced gene-editing and de novo gene synthesis to target key regulatory elements responsible for stress adaptation.

Background: Paclitaxel is widely used in the treatment of cancer and has a good effect in the treatment of non-small cell lung cancer. The combination of TMT proteomics and bioinformatics is used to systematically analyze the molecular mechanism of paclitaxel in the treatment of lung adenocarcinoma A549 cell, which is helpful to screen new therapeutic targets. Methods: MTT assay was used to analyze the inhibitory effect of paclitaxel on the proliferation of A549 cells. The proteins were identified by TMT quantitative proteomics and the differential expression proteins (DEPs) database was constructed. The DEPs were enriched by Gene Ontology (GO) and KEGG pathway annotation. Based on the information in the STRING database, find the interaction between DEPs, and the protein–protein interaction (PPI) networks of DEPs were constructed and analyzed by using the Cytoscape software. According to the PPI network results, select the hub proteins from DEPs for WB verification. Results: A total of 5449 proteins were identified in A549 by TMT proteomics. Compared with the control group, 281 DEPs were significantly up-regulated and 218 were significantly down-regulated after paclitaxel treatment. GO functional analysis, we found that the main functions of these DEPs are binding, catalytic activity, molecular function regulator and so on. They are mainly involved in cellular process, metabolic process, biological regulation and so on. KEGG analysis showed that the three most significant signal transduction pathways of DEPs enrichment were DNA replication, steroid biosynthesis, oxidative phosphorylation. In PPI network, there are 294 nodes among which CDK1, MCM2-5 and PCNA are located at the center of proteins interaction. WB analysis confirmed that the expression of CDK1 was significantly down-regulated, consistent with the TMT results. Conclusion: Paclitaxel significantly increased the expression of tubulin, binding tubulin to promote A549 cell death. In addition, paclitaxel significantly inhibited the expression of hub proteins, DNA replication and cell cycle pathways, thus killing lung adenocarcinoma cell A549. These findings will enhance the understanding of the mechanism of paclitaxel in the treatment of lung adenocarcinoma cell A549 and provide new valuable targets.

Salt stress is widely recognized as a major abiotic constraint that limits plant growth and productivity. Alfalfa (Medicago sativa L.), a globally important forage legume, exhibits considerable variability in salt toll,erance across its diverse cultivars. The present study aims to provide a comprehensive evaluation of eight alfalfa varieties by examining their morphological, physiological, and biochemical responses at four NaCl levels (0, 60, 120, and 180 mM NaCl). Salt tolerance is quantified using a combination of the membership function method, correlation analysis, principal component analysis, and different indices of salt tolerance. High salt stress significantly reduced plant height (PH), shoot dry weight (SDW), root dry weight (RDW), and number of leaves (NOL) of all alfalfa varieties by 47.21%, 61.36%, 64.19%, and 59.91%, respectively, compared with the control. Similarly, physiological traits including relative water content (RWC) and stomatal conductance (SC) declined by 43.93% and 60.87%, respectively, whereas electrolyte leakage (EL) and malondialdehyde (MDA) increased by 267.35% and 178.48%, respectively. Salt stress also markedly altered antioxidant responses among varieties, with superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities increasing by 111.27%, 170.87%, and 125.87%, while V5 (A'erjienong) and V2 (Weinasi) consistently showed tolerance, whereas V4 (Fudina) and V8 (Qianjing) exhibited susceptibility across varying stress levels. Principal component analysis (PCA) classified the traits into three main groups, which explained 93.15% of the total variation, and identified SOD, POD, CAT, SS, and Car as crucial factors affecting salt tolerance. Comprehensive membership function analysis indicated that variety V5 (A'erjienong) was the most tolerant, while variety V4 (Fudina) remained the most sensitive. This study highlights the importance of morpho-physiological and biochemical traits, along with stress tolerance indices, as reliable indicators for evaluating salt tolerance in alfalfa. Future integration of these traits with advanced genetic and breeding approaches may facilitate the development of high-yielding, salt-tolerant alfalfa cultivars.