Among various abiotic stresses, secondary soil salinization poses a significant threat to plant productivity and survival. Cultivated chrysanthemums (Chrysanthemum morifolium), widely grown as ornamental crops, are highly susceptible to salt stress, and their complex polyploid genome complicates the identification of stress resistance genes. In contrast, C. indicum, a native diploid species with robust stress tolerance, serves as a valuable genetic resource for uncovering stress-responsive genes and improving the resilience of ornamental chrysanthemum cultivars. In this study, we cloned, overexpressed (OE-CiHY5), and silenced (RNAi-CiHY5) the CiHY5 gene in C. indicum. OE-CiHY5 plants exhibited larger leaves, sturdier stalks, and higher chlorophyll content compared to wild-type plants, while RNAi-CiHY5 plants displayed weaker growth. Under salt stress, OE-CiHY5 plants demonstrated significantly improved growth, enhanced osmotic adjustment, and effective ROS scavenging. In contrast, RNAi-CiHY5 plants were more sensitive to salinity, showing higher electrolyte leakage and impaired osmotic regulation. Transcriptomic analyses revealed that CiHY5 regulates key hormonal pathways such as zeatin (one of cytokinins), abscisic acid and jasmonic acid, as well as metabolic pathways, including photosynthesis, carbohydrate metabolism, which collectively contribute to the enhanced stress resilience of OE-CiHY5 plants. Promoter-binding assays further confirmed that CiHY5 directly interacts with the CiABF3 promoter, highlighting its critical role in ABA signaling. Evolutionary analyses showed that HY5 is conserved across plant lineages, from early algae to advanced angiosperms, with consistent responsiveness to salt and other abiotic stresses in multiple Chrysanthemum species. These findings establish CiHY5 as a key regulator of salt tolerance in C. indicum, orchestrating a complex network of hormonal and metabolic pathways to mitigate salinity-induced damage. Given the conserved nature of HY5 and its responsiveness to various stresses, HY5 gene provides valuable insights into the molecular mechanisms underlying stress adaptation and represents a promising genetic target for enhancing salt stress resilience in chrysanthemums.

To address the issue of poor phytoremediation in Cd-contaminated saline soil caused by the biotoxicity of Cdsalinity, we constructed a symbiotic system of arbuscular mycorrhizal fungi (AMF) and the hyperaccumulator Solanum nigrum, and systematically elucidated the response strategies of Solanum nigrum and the enhancement mechanism of AMF for plant tolerance through cytological, physiological, and transcriptomic methods. The findings showed that Cd-salinity stress had synergistic aggravated Cd/Na enrichment, ultrastructural damage, photosynthetic inhibition, water loss, and reactive oxygen species (ROS) accumulation in plants. In response to the heterogeneity of Cd/salinity stress, AMF smartly regulated the Cd/salinity tolerance of host plants: AMF decreased intercellular CO2 concentration (Ci) under Cd stress to alleviate non-stomatal limitation induced by Cd, but increased Ci under salinity stress to alleviate the stomatal limitation induced by salinity; the role of AMF in strengthening the osmoregulation system was more prominent under salinity stress, thereby alleviated the more severe osmotic imbalance induced by salinity. AMF also enhanced signal transduction to consolidate resistance defense, upregulated antioxidant genes to activate antioxidant enzymes, and strengthened the AsAGSH cycle to mitigate oxidative damage. The enhancement of tolerance improved plant growth and Cd enrichment. Under high Cd-high salinity combined stress, Cd concentrations in shoots and roots increased by 14.28 % and 38.85 %, respectively, and the biomass also increased by over 30.00 % after AMF inoculation. In summary, inoculation with AMF serves as an effective and sustainable phytoremediation enhancement strategy that improves the host plants' stress resistance through multiple pathways, thereby increasing the phytoremediation potential.

Hexafluoropropylene oxide dimer acid (HFPO-DA), an emerging perfluoroalkyl substance (PFAS) that is replacing traditional PFASs, has a wide range of industrial applications and has been detected globally in the environment. However, it remains unclear whether HFPO-DA, is genuinely less toxic than perfluorooctanoic acid (PFOA) in terms of soil environmental hazards. Therefore, this study aimed to compare differences in toxicity between PFOA and its substitute, HFPO-DA, in a common species of earthworm, Eisenia fetida. Our findings revealed that both HFPO-DA and PFOA caused oxidative damage, apoptosis, reproductive disorders, and neurotoxicity in E. fetida at a concentration of 0.2 mg/kg following exposure for 28 d. Specifically, at the molecular level, PFOA resulted in a significant decline in total antioxidant capacity and lipid peroxidation, whereas HFPO-DA did not have the same effect. The Integrated Biomarker Response (IBR) index, based on the indicators studied, showed that HFPO-DA exhibited lower toxicity than PFOA. The transcriptomic results suggest that HFPO-DA can induce neurotoxicity, similar to PFOA; however, the specific mechanisms differ. Although HFPODA appears to be less toxic than PFOA to E. fetida, its potential hazards at the transcriptional level, affecting different pathways, require further investigation. This study provided new insights into the safety of HFPO-DA as a novel substitute for PFOA.

Key messageSaline-alkali stress induces oxidative damage and photosynthesis inhibition in H. citrina, with a significant downregulation of the expression of photosynthesis- and antioxidant-related genes at high concentration. Soil salinization is a severe abiotic stress that impacts the growth and development of plants. In this study, Hemerocallis citrina Baroni was used to investigate its responsive mechanism to complex saline-alkali stress (NaCl:Na2SO4:NaHCO3:Na2CO3 = 1:9:9:1) for the first time. The growth phenotype, photoprotective mechanism, and antioxidant system of H. citrina were studied combining physiological and transcriptomic techniques. KEGG enrichment and GO analyses revealed significant enrichments of genes related to photosynthesis, chlorophyll degradation and antioxidant enzyme activities, respectively. Moreover, weighted gene co-expression network analysis (WGCNA) found that saline-alkali stress remarkably affected the photosynthetic characteristics and antioxidant system. A total of 29 key genes related to photosynthesis and 29 key genes related to antioxidant enzymes were discovered. High-concentration (250 mmol L-1) stress notably inhibited the expression levels of genes related to light-harvesting complex proteins, photosystem reaction center activity, electron transfer, chlorophyll synthesis, and Calvin cycle in H. citrina leaves. However, most of them were insignificantly changed under low-concentration (100 mmol L-1) stress. In addition, H. citrina leaves under saline-alkali stress exhibited yellow-brown necrotic spots, increased cell membrane permeability and accumulation of reactive oxygen species (ROS) as well as osmolytes. Under 100 mmol L-1 stress, ROS was eliminate by enhancing the activities of antioxidant enzymes. Nevertheless, 250 mmol L-1 stress down-regulated the expression levels of genes encoding antioxidant enzymes, and key enzymes in ascorbate-glutathione (AsA-GSH) cycle as well as thioredoxin-peroxiredoxin (Trx-Prx) pathway, thus inhibiting the activities of these enzymes. In conclusion, 250 mmol L-1 saline-alkali stress caused severe damage to H. citrina mainly by inhibiting photosynthesis and ROS scavenging capacity.