Transcriptome and cell wall degrading enzyme-related gene analysis of Pestalotiopsis neglecta in response to sodium pheophorbide a.

Sodium pheophorbide a (SPA) is a new alternative fungicide with low toxicity and high efficiency, which has high fungicidal activity against Pestalotiopsis neglecta, a pathogen that causes black spot needle blight of Pinus sylvestris var. mongolica. To utilize SPA for plant disease control, understanding its antifungal mechanism is essential. Six cDNA libraries were constructed from 3 d-old P. neglecta mycelia (three SPA-infected and three untreated groups) and 29,850 expressed genes were obtained by Illumina HiSeq4000 sequencing. Compared with controls, 3268 differentially expressed genes (DEGs) were identified in SPA-treated groups, including 1879 upregulated and 1389 downregulated genes. Most DEGs were involved in the metabolism of amino acids, carbohydrates, and lipids, as well as cell structure and genetic information processing. These findings were further confirmed by decreased conductivity, RNA and protein content, and activities of nicotinamide adenine dinucleotide-dependent malate dehydrogenase, citrate synthase, isocitrate dehydrogenase, and succinate dehydrogenase. Moreover, qRT-PCR verified the reliability of the transcriptome results. After treatment with SPA at different concentrations for 60 min, the expressions of three cell wall degrading enzyme-related genes (PnEG, PnBG, and PnPG) were all suppressed. Overall, this study provided insights into the molecular mechanisms through which SPA inhibits P. neglecta, increasing the possibility of developing SPA into an effective fungicide in the future.

Key factors affecting photoactivated fungicidal activity of sodium pheophorbide a against Pestalotiopsis neglecta.

Recently, photodynamic therapy (PDT) and photoactivated pesticides have attracted considerable research attention. In the present study, we aimed to investigate the photodynamic activity of a chlorophyllous derivative, sodium pheophorbide a (SPA), and to evaluate its potential as a photoactivated fungicide. The singlet oxygen quantum yield, the photoreaction process, the anti-photobleaching ability in sterile water (H2O), the effect of light conditions on its antifungal activity, and its stability were all investigated. SPA showed significant fungicidal activity and photostability, during which Type I and Type II photodynamic reactions occurred simultaneously on Pestalotiopsis neglecta, and the influence of Type I was slightly larger than that of Type II. In addition, light promoted the antifungal activity of SPA. In particular, the antifungal activity was enhanced with increasing light intensity, and was strongest under 8000 lx conditions. Under monochromatic light sources, antifungal activity was strongest under green light s; however, the effect of monochromatic light was not as good as that of white light. From 0 to 24 h, the antifungal effect of the SPA solution was enhanced; however, the activity of the solution began to weaken after 24 h. Furthermore, our study confirmed that the antifungal activity of SPA was stable under different temperatures, pH values, and UV irradiation durations.

Sodium pheophorbide a controls cherry tomato gray mold (Botrytis cinerea) by destroying fungal cell structure and enhancing disease resistance-related enzyme activities in fruit.

Sodium pheophorbide a (SPA) is a natural photosensitizer. The present study investigated the antifungal activity and mechanism of SPA against Botrytis cinerea in vitro and in vivo. Its inhibitory effect was studied on the spore germination and mycelial growth of B. cinerea. The effects of SPA on cell wall integrity, cell membrane permeability, and mycelial morphology of B. cinerea were also determined. Additionally, how SPA effected B. cinerea in vivo was evaluated using cherry tomato fruit. The results showed that SPA effectively inhibited the spore germination and mycelial growth of B. cinerea under light conditions (4000 lx). SPA significantly affected both cell wall integrity and cell membrane permeability (P < .05). In addition, SEM analysis suggested that B. cinerea treated with SPA (12.134 mg/mL) showed abnormal mycelial morphology, including atrophy, collapse, flattening, and mycelial wall dissolution. In vivo tests showed that SPA could increase the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) significantly (P < .05); however, SPA had no significant effect on phenylalanine ammonia lyase (PAL) activity. In short, SPA could destroy the fungal cell structure and enhance disease resistance-related enzyme activity in cherry tomatoes, thereby controlling cherry tomato gray mold.

Effect of Emodin on Coxsackievirus B3m-Mediated Encephalitis in Hand, Foot, and Mouth Disease by Inhibiting Toll-Like Receptor 3 Pathway In Vitro and In Vivo.

BACKGROUND: Encephalitis in hand, foot, and mouth disease (HFMD) is a serious threat to children's health and life. Toll-like receptor 3 (TLR3) is an innate immune-recognition receptor that can recognize virus and initiate innate immune responses. Emodin has the effects of anti-inflammatory and regulating immune function, but the mechanism is not very clear. METHODS: Cells and mice were pretreated with coxsackievirus B3m (CVB3) and treated with emodin. The messenger ribonucleic acid (mRNA) and protein levels of TLR3 and downstream molecules were detected by quantitative real-time polymearse chain reaction and western blotting analysis, respectively. TLR3 expression was also downregulated by anti-TLR3 antibody (TLR3Ab) or small interfering RNA (siRNA). Pathological changes were assessed with hematoxylin and eosin staining. Immunohistochemistry was used to examine the expression of TLR3 in brain tissues. The expression of interleukin (IL)-6, nuclear factor (NF)-κB, and interferon (IFN)-ß in serum were tested with enzyme-linked immunosorbent assay. RESULTS: Emodin decreased the mRNA and protein levels of TLR3 and downstream molecules in vitro and in vivo. After downregulating TLR3 using anti-TLR3Ab or siRNA, emodin could still decrease the mRNA and protein levels of TLR3 and downstream molecules. Emodin also displayed notable effects on pathology, TLR3 protein in brain tissues, and expression of IL-6, NF-κB, IFN-ß, in serum. CONCLUSIONS: Emodin exerts a protective effect in CVB3-mediated encephalitis in HFMD by inhibiting the TLR3 pathway.