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Weeping forsythia is an important ornamental, ecological, and medicinal plant. Brown leaf spots reduce the large-scale production of weeping forsythia as a medicinal crop. Alternaria alternata is a pathogen causing brown leaf spots in weeping forsythia; however, its pathogenesis and the immune response mechanisms of weeping forsythia remain unclear. In this study, we identified the two mechanisms based on morphological anatomy, physiological indexes, and gene expression analyses. Our results showed that A. alternata induced leaf stomata to open, invaded the mesophyll, dissolved the cell wall, destroyed the cell membrane, and decreased the number of chloroplasts by up-regulating the expression of auxin-activated signaling pathway genes. A. alternata also down-regulated iron ion homeostasis and binding-related genes, which caused an increase in the levels of iron ions and reactive oxygen species in leaves. These processes eventually led to programmed cell death, destroying palisade and spongy tissues and causing the formation of iron rust spots. A. alternata also caused defense and hypersensitive responses in weeping forsythia through signaling pathways mediated by flg22-like and elf18-like polypeptides, ethylene, H2O2, and bacterial secretion systems. Our study provides a theoretical basis for the control of brown leaf spots in weeping forsythia.
Weeping forsythia is an important ornamental, ecological, and medicinal plant. Brown leaf spots reduce the large-scale production of weeping forsythia as a medicinal crop. Alternaria alternata is a pathogen causing brown leaf spots in weeping forsythia; however, its pathogenesis and the immune response mechanisms of weeping forsythia remain unclear. In this study, we identified the two mechanisms based on morphological anatomy, physiological indexes, and gene expression analyses. Our results showed that A. alternata induced leaf stomata to open, invaded the mesophyll, dissolved the cell wall, destroyed the cell membrane, and decreased the number of chloroplasts by up-regulating the expression of auxin-activated signaling pathway genes. A. alternata also down-regulated iron ion homeostasis and binding-related genes, which caused an increase in the levels of iron ions and reactive oxygen species in leaves. These processes eventually led to programmed cell death, destroying palisade and spongy tissues and causing the formation of iron rust spots. A. alternata also caused defense and hypersensitive responses in weeping forsythia through signaling pathways mediated by flg22-like and elf18-like polypeptides, ethylene, H2O2, and bacterial secretion systems. Our study provides a theoretical basis for the control of brown leaf spots in weeping forsythia.
Aralia elata (Miq.) Seem., is grown for its medicinal and nutritional properties in northeastern China. The tender shoots are used as wild vegetables. The plant saponin components have antioxidant and neuroprotective activities, and are used for the treatment of chronic disease (Xia et al. 2021). In July 2021, root rot disease was observed in five-year-old A. elata plants in Qingyuan County (41°91′ N, 124°59′ E), Liaoning Province, China. The incidence of roots rot was approximately 50% in old fields, with the leaves of the infected plants appearing chlorotic and wilting. The lesions on the taproots were dark brown and soft, with degraded internal organization. Leading edge of necrotic tissue from symptomatic roots was cut 5×5×3 mm, placed in 75% ethanol for 30 s, and then in 3% sodium hypochlorite for 2 min. After three rinses in sterile distilled water, the samples were dried on sterile filter paper before plating on potato dextrose agar (PDA) and incubation at 25℃. Monosporic cultures were obtained by the collection of single spores from individual isolates. After 7 days on PDA, mycelia in the colonies appeared cottony and pink, white, or purple in color, while their undersides were pink and white. Spore characteristics were evaluated after transfer to carnation leaf agar (CLA) and incubation for 20 days (Zhang et al. 2021). The macroconidia were falciform, slightly curved or straight, two to five septate, and 20.57 to 33.75 × 3.62 to 6.11 μm (n=40). The microconidia were ovoid or oval, zero to one septate, and 5.12 to 13.53 × 3.04 to 4.79 μm (n=40). Chlamydospores were globose to subglobose, intercalary or terminal, with an average diameter of 13.76 μm (n=40).To identify the pathogen, the internal transcribed spacer (ITS) region, large subunit (LSU) ribosomal DNA, and translation elongation factor 1 alpha (TEF-1α) gene were amplified using the respective primer pairs ITS1/ITS4, LR0R/LR7, and EF1-728F/EF1-986R (Cheng et al. 2020; Fu et al. 2019). Comparisons with GenBank, the sequences of ITS, LSU, and TEF-1 had 99 to 100% homology with Fusarium oxysporum (accessions numbers- MH707084, OQ380519, and GU250609, respectively). The sequences were deposited in GenBank: OP482273 (ITS), OP491955 (LSU), and OP503498 (TEF-1α). Maximum likelihood phylogeny of the identified sequences using MEGA-X software indicated that the isolate represented F. oxysporum. The taproots of 30 one-year-old A. elata were washed and inoculated with 1×106/ml of the conidial suspension for two hours, and another 30 used as controls with sterile water. After planting in sterilized forest soil in flowerpots (36×30 cm), the plants were grown in a greenhouse for two weeks at 25℃ with 14 h of light. It was found that 50% of the roots showed typical root rot symptoms, while the controls were asymptomatic. The pathogenicity test was repeated three times, and reisolation of F. oxysporum from the roots fulfilled Koch’s postulates. This is the first report of root rot in A. elata caused by F. oxysporum in China and indicates the necessity for suitable management strategies to protect A. elata production. References: Cheng, Y., et al. 2020. Plant Dis. 104:3072. Fu, R., et al. 2019. Plant Dis. 103:1426. Xia, W., et al. 2021. Mini-Rev Med Chem. 21:2567. Zhang, X. M., et al. 2021. Plant Dis. 105:1223.
Velvet proteins, as well as the epigenetic regulator LaeA, are conserved in numerous fungal species, where, in response to environmental cues, they control several crucial cellular processes, including sexual and asexual morphogenesis, secondary metabolism, response to oxidative stress, and virulence. During the last two decades, knowledge of their mechanism of action as well as understanding their functional roles, has greatly increased, particularly in Aspergillus species. Research efforts from multiple groups followed, leading to the characterization of other Velvet and LaeA homologs in species of other fungal genera, including important opportunistic plant and animal pathogens. This review focuses mainly on the current knowledge of the role of Velvet and LaeA function in fungal pathogenesis. Velvet proteins and LaeA are unique to fungi, and for this reason, additional knowledge of these critical regulatory proteins will be important in the development of targeted control strategies to decrease the detrimental impact of fungal pathogens capable of causing disease in plants and animals.
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