Dynamins are large superfamily GTPase proteins that are involved in various cellular processes including budding of transport vesicles, division of organelles, cytokinesis, and pathogen resistance. Here, we characterized several dynamin-related proteins from the rice blast fungus Magnaporthe oryzae and found that MoDnm1 is required for normal functions, including vegetative growth, conidiogenesis, and full pathogenicity. In addition, we found that MoDnm1 co-localizes with peroxisomes and mitochondria, which is consistent with the conserved role of dynamin proteins. Importantly, MoDnm1-dependent peroxisomal and mitochondrial fission involves functions of mitochondrial fission protein MoFis1 and WD-40 repeat protein MoMdv1. These two proteins display similar cellular functions and subcellular localizations as MoDnm1, and are also required for full pathogenicity. Further studies showed that MoDnm1, MoFis1 and MoMdv1 are in complex to regulate not only peroxisomal and mitochondrial fission, pexophagy and mitophagy progression, but also appressorium function and host penetration. In summary, our studies provide new insights into how MoDnm1 interacts with its partner proteins to mediate peroxisomal and mitochondrial functions and how such regulatory events may link to differentiation and pathogenicity in the rice blast fungus.
This study was conducted to determine the efficacy of 4 Chinese herbal ingredients (CHI) as immune stimulators for an active vaccine in chickens using both in vitro and in vivo assays. The CHI used were Astragalus polysaccharide (APS), Isatis root polysaccharide (IRPS), Propolis polysaccharide, and Epimedium flavone at various concentrations. Two hundred 14-d-old male White Roman chickens were randomly divided into 10 groups. Chickens in groups 1 to 9 were inoculated with the New-castle disease virus (NDV) strain IV vaccine by intranasal and intraocular administration. Chickens in groups 1 to 8 were also administered subcutaneously on the dorsal region of the neck with 0.5 mL of the corresponding CHI at 2 doses: 29 and 58 mg/kg of BW for APS and IRPS and 7.25 and 14.5 mg/kg of BW for the others, once daily for 3 successive days. In group 9 (CHI-free control) and group 10 (both vaccine- and CHI-free control), chickens were injected with 0.5 mL of physiological saline. New-castle disease virus-specific serum hemagglutination inhibition antibody (Ab) production in immunized chickens was quantified using established methods. The results indicate that a majority of the CHI used at appropriate concentrations were effective in enhancing in vitro proliferation of chick embryo fibroblasts in response to the NDV infection. In vivo administration of CHI to vaccinated chickens (7.25 to 58 mg/kg of BW, depending on type) increased serum anti-NDV hemagglutination inhibition Ab titer concentrations, compared with the administration the NDV alone. For all CHI, a beneficial effect on the Ab production was observed on d 21 after the initiation of the vaccination. On the basis of the in vivo doses used, Propolis polysaccharide and Epimedium flavone were more potent than APS and IRPS in promoting the humoral immune response in the young birds (P < 0.05). Collectively, these findings suggest that appropriate doses of CHI can be used as novel, effective immune stimulators for chickens.
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