Mycoviruses widely exist in filamentous fungi and sometimes cause phenotypic changes in hosts. Trichoderma harzianum hypovirus 1 (ThHV1) and its defective RNA ThHV1-S were found in T. harzianum and exhibited high transmissibility. In our previous study, ThHV1 and ThHV1-S were transferred to an excellent biological control agent T. koningiopsis T-51 to form a derivative strain 51-13. In this study, we assessed the metabolic changes in strain 51-13 and antifungal activity of its culture filtrate (CF) and volatile organic compounds (VOCs). The antifungal activity of CF and VOCs of T-51 and 51-13 was different. Compared with the CF of T-51, that of 51-13 exhibited high inhibitory activity against B. cinerea, Sclerotinia sclerotiorum, and Stagonosporopsis cucurbitacearum but low inhibitory activity against Leptosphaeria biglobosa and Villosiclava virens. The VOCs of 51-13 exhibited high inhibitory activity against F. oxysporum but low inhibitory activity against B. cinerea. The transcriptomes of T-51 and 51-13 were compared; 5531 differentially expressed genes (DEGs) were identified in 51-13 with 2904 up- and 2627 downregulated genes. In KEGG enrichment analysis, 1127 DEGs related to metabolic pathways (57.53%) and 396 DEGs related to biosynthesis of secondary metabolites (20.21%) were clearly enriched. From the CF of T-51 and 51-13, 134 differential secondary metabolites (DSMs) were detected between T-51 and 51-13 with 39 up- and 95 downregulated metabolites. From these, 13 upregulated metabolites were selected to test their antifungal activity against B. cinerea. Among them, indole-3-lactic acid and p-coumaric acid methyl ester (MeCA) exhibited strong antifungal activity. The IC₅₀ of MeCA was 657.35 μM and four genes possibly related to the synthesis of MeCA exhibited higher expression in 51-13 than in T-51. This study revealed the mechanism underlying the increase in antifungal activity of T-51 because of the mycovirus and provided novel insights in fungal engineering to obtain bioactive metabolites via mycoviruses.

Drought stress reduces water availability in plant cells and influences rapeseed yield. Currently, key genetic regulators that contribute to rapeseed response to drought remain largely unexplored, which limits breeding of drought-resistant rapeseed. In this study, we found that Brassica napus JASMONATE ZIM-DOMAIN 5 (BnA.JAZ5), one of the transcriptional repressors functioning in the jasmonate (JA) signaling pathway, was triggered by drought treatment in rapeseed, and drought-susceptibility increased in BnA.JAZ5-overexpressing rapeseed plants as compared to wild-type plants, resulting in a lower survival rate after recovery from dehydration. After recovery for 3 days, 22–40% of p35S::BnA.JAZ5 transgenic plants survived, while approximately 61% of wild-type plants survived. Additionally, seed germination of BnA.JAZ5-overexpressing rapeseed was hyposensitive to abscisic acid (ABA). The germination rate of five transgenic lines was 32~42% under 9 µM ABA treatment, while the germination rate of wild-type plants was 14%. We also found that the average stomatal density of five overexpressing lines was 371~446/mm², which is higher than that of wild-type (232/mm²) plants under normal conditions. These results indicate that BnA.JAZ5 regulated drought response in an ABA-dependent manner, possibly by affecting stomatal density. Interestingly, methyl jasmonate (MeJA) treatment rescued the ABA-hyposensitive seed germination, revealing crosstalk between JAZ5-meidated JA and the ABA signaling pathway. Taken together, our results suggest that BnA.JAZ5 attenuated drought resistance through the ABA-dependent pathway, which could represent important genetic loci for drought-resistant rapeseed breeding.

Previous research has shown that miR398 contributed to plant thermotolerance by silencing its target gene COPPER/ZINC SUPEROXIDE DISMUTASE1 (CSD1) in Arabidopsis thaliana. However, the phylogenesis of miR398 and CSD1 in Brassica crop and their role in regulating leaf cell death under heat stress remains unexplored. Here, we characterized the homologous genes of miR398a and CSD1 in Brassica rapa ssp. pekinensis (Chinese cabbage) and found miR398a abundance was accumulated under heat stress (38 °C and 46 °C for 1 h) in Chinese cabbage, while the expression level of its targets BraCSD1-1 and BraCSD2-1 were downregulated. To further explore their role in heat response, we constructed the transgenic plants overexpressing artificial miR398a (aBra-miR398a), Bra-miR398a target mimic (Bra-MIM398a), and BraCSD1-1 in Chinese cabbage for genetic study. Under high temperatures, p35S::aBra-miR398a lines reduced the areas of leaf cell death and delayed the leaf cell death. By contrast, p35S::Bra-MIM398a and p35S::BraCSD1-1 plants enlarged the areas of leaf cell death and displayed the earliness of leaf cell death. Finally, we found that the expression level of stress-responsive genes BraLEA76, BraCaM1, BraPLC, BraDREB2A, and BraP5CS increased in transgenic plants overexpressing aBra-miR398a, which may contribute to their resistance to heat-induced leaf cell death. Taken together, these results revealed the function of Bra-miR398a in attenuating leaf cell death to ensure plant thermotolerance, indicating that the miR398-CSD1 module could be potential candidates for heat-resistant crop breeding.

Soil salinisation has become one of the major abiotic stresses limiting crop growth in the world. To enhance soybean productivity on saline lands, understanding its salt-stress response and underlying mechanisms is necessary. In this study, the salt-tolerant soybean Kefeng 1 and the salt-sensitive soybean Qihuang 1 were used to elucidate the synergistic regulatory networks underlying soybean salt tolerance. After 12 days of 150 mM NaCl treatment, both varieties were subjected to phenotypic evaluation, physiological measurements, and integrated transcriptomic and metabolomic analysis. The results showed that the salt tolerance in Kefeng 1 primarily originated from its root. Under salt stress, Kefeng 1 maintained Na⁺/K⁺ ion homeostasis by up-regulating Cation/H⁺ Exchanger 15 (CHX15) and Cation Exchanger 3 (CAX3), and down-regulating Cyclic Nucleotide-Gated Channel 13 (CNGC13). Furthermore, Kefeng 1 stabilised auxin (IAA) homeostasis by inhibiting IAA biosynthesis and regulating concentrations through PIN-FORMED 3 (PIN3)-mediated efflux. It also scavenged reactive oxygen species (ROS) by employing enhanced enzymatic antioxidant systems, specifically aldo-keto reductase 1 (AKR1), glutathione S-transferase (GST), and catalase (CAT), alongside non-enzymatic antioxidants like the isoflavone genistein. Gene–metabolite correlation network analysis identified Glyma.09G117900 (PIN3) and Glyma.19G244200 (AKR1) as two hub genes. These two genes were specifically up-regulated in Kefeng 1 root under NaCl stress, and the proteins they encoded played important roles in salt tolerance in Kefeng 1 root as described above. Accordingly, these two genes were identified as candidate genes for salt tolerance in Kefeng 1. This study offered a theoretical framework and genetic resources for developing salt-tolerant soybean cultivars.

Watermelon is one of the most important edible plants worldwide. Owing to its special cultivation conditions, watermelon is exposed to many biological and abiotic stresses during its development. Lectin receptor-like kinases (LecRLKs) are plant-specific membrane proteins that play important roles in sensing and responding to environmental stimuli. Although the LecRLK gene family has been identified in a variety of plants, a comprehensive analysis has not yet been undertaken in watermelon. In this study, 61 putative LecRLK genes were identified in watermelon, consisting of 36 G-type, 24 L-type, and 1 C-type LecRLK genes. They were distributed in clusters on chromosomes, and members from the same subfamily were mostly clustered together. The analysis of the phylogenetic tree and conserved motif indicated that there were obvious differences among three ClaLecRLK subfamilies, and there was also rich diversity in the C-terminal within subfamilies. A collinear analysis revealed that the evolution of the ClaLecRLK gene family in different Cucurbitaceae crops was asynchronous. Furthermore, the analysis of the ClaLecRLK protein structure showed that not all proteins contained signal peptides and a single transmembrane domain. A subcellular localization assay confirmed that the number and position of transmembrane domains did not affect ClaLecRLK protein localization in cells. Transcriptome data revealed distinct expression patterns of LecRLK genes of watermelon in various tissues, and their responses to different fungi infection were also significantly different. Finally, the potential binding sites of the ClaLecRLK genes targeted by miRNA were predicted. This study enhances the understanding of the characteristics and functions of the LecRLK gene family in watermelon and opens up the possibility of exploring the roles that LecRLK genes may play in the life cycle of Cucurbitaceae plants.

Shanghai-native vegetable soybean (Glycine max [L.] Merril) landraces are valuable germplasms, but their systematic evaluation for agronomic and nutritional traits remains insufficient. This study aimed to assess their phenotypic and nutritional diversity to explore their potential for breeding and meeting dietary needs. Twenty-nine local landraces and one control cultivar (‘Qingsu 7’) were evaluated for key agronomic traits, yield components, nutritional traits, and isoflavone profiles, using hierarchical clustering, principal component analysis (PCA), and correlation analysis. Substantial phenotypic diversity was found, with the germplasm classified into four groups. First pod height and effective pods per plant were highly variable. Nutritional traits showed low variability for crude protein but high diversity for crude fat, soluble sugars (dominated by sucrose), vitamin C, and free amino acids. Total isoflavone content in dry seeds varied widely, with genistin, daidzin, and daidzein as the main forms. ‘Xiangshui Maodou’ had high free amino acids and vitamin C, ‘Heiyan Susudou’ showed superior soluble sugar content, and two landraces exceeded 1500 μg/g DW total isoflavones. The landraces possess rich phenotypic diversity and nutritional diversity. This germplasm represents a valuable resource for breeding programs to enhance crop quality and address global nutritional demands.