The changes of DHN1 expression and subcellular distribution in A. delicisoa cells under osmotic stress were studied by using GFP as a reporter molecule. Through creating the Xba I and BamH I restriction sites at the e...The changes of DHN1 expression and subcellular distribution in A. delicisoa cells under osmotic stress were studied by using GFP as a reporter molecule. Through creating the Xba I and BamH I restriction sites at the ends of dhn1 by PCR, the expression vector for the fusion protein DHN1-mGFP4 was constructed by cloning dhn1 into plasmid pBIN-35SmGFP4. Then the DHN1-mGFP4 expression vector was transformed into A. delicisoa suspension cells by microprojectile bombardment method. Bright green fluorescence of GFP which shows the high-level expression of DHN1-mGFP4 was visualized after culture for 10 h. However, the green fluorescence was only located within the nucleus. By increasing the culture medium osmotic potential, the green fluorescence was visualized in the cytoplasm (mainly around the plasma membranes). The generation of GFP fluorescence in the cytoplasm was also promoted by increasing the medium osmotic potential. Moreover, GFP green fluorescence was abolished by protein synthesis inhibitor dicyclohexylcarbodiimid, indicating that the cytoplasmic DHN1 was newly synthesized under osmotic stress. Furthermore, ABA promoted the presence of green fluorescence in the cytoplasm, and the GFP fluorescence was visualized within a shorter time under a higher osmotic potential.展开更多
DHN(1,8-dihydroxynaphthalene)melanin is a significant secondary metabolite found in phytopathogenic fungi,predominantly localized in mature aerial hyphae,conidia,and appressoria.It performs various biological function...DHN(1,8-dihydroxynaphthalene)melanin is a significant secondary metabolite found in phytopathogenic fungi,predominantly localized in mature aerial hyphae,conidia,and appressoria.It performs various biological functions,including responding to extreme environments and regulating growth,development,and pathogenicity.Key biosynthetic enzymes and transcription factors directly involved in the DHN melanin synthesis pathway include melanin synthesis transcription factors,polyketide synthase,1,3,6,8-tetrahydroxynaphthalene reductase,scytalone dehydratase,1,3,8-trihydroxynaphthalene reductase,and a-hydrolase.The regulatory network controlling the DHN melanin biosynthesis is complex and multifaceted,involving various transcription factors,signaling pathways,and metabolic routes.This review provides a comprehensive summary of current research on the abundant biological functions of DHN melanin.Understanding the molecular mechanisms of DHN melanin synthesis in phytopathogenic fungi could facilitate the development of effective disease control strategies and novel therapeutic targets.展开更多
基金This work was supported by the Major State Basic Research Development Program of the People's Republic of China, the National Natural Science Foundation of China (Grant No. 39770077) the Science and Technology Program of the Ministry of Education of
文摘The changes of DHN1 expression and subcellular distribution in A. delicisoa cells under osmotic stress were studied by using GFP as a reporter molecule. Through creating the Xba I and BamH I restriction sites at the ends of dhn1 by PCR, the expression vector for the fusion protein DHN1-mGFP4 was constructed by cloning dhn1 into plasmid pBIN-35SmGFP4. Then the DHN1-mGFP4 expression vector was transformed into A. delicisoa suspension cells by microprojectile bombardment method. Bright green fluorescence of GFP which shows the high-level expression of DHN1-mGFP4 was visualized after culture for 10 h. However, the green fluorescence was only located within the nucleus. By increasing the culture medium osmotic potential, the green fluorescence was visualized in the cytoplasm (mainly around the plasma membranes). The generation of GFP fluorescence in the cytoplasm was also promoted by increasing the medium osmotic potential. Moreover, GFP green fluorescence was abolished by protein synthesis inhibitor dicyclohexylcarbodiimid, indicating that the cytoplasmic DHN1 was newly synthesized under osmotic stress. Furthermore, ABA promoted the presence of green fluorescence in the cytoplasm, and the GFP fluorescence was visualized within a shorter time under a higher osmotic potential.
基金“Innovation Star”Program of Excellent Postgraduates in Gansu Province(2025CXZX-789)National Natural Science Foundation of China(32372411)National Key R&D Program of China(2021YFD2100502-3).
文摘DHN(1,8-dihydroxynaphthalene)melanin is a significant secondary metabolite found in phytopathogenic fungi,predominantly localized in mature aerial hyphae,conidia,and appressoria.It performs various biological functions,including responding to extreme environments and regulating growth,development,and pathogenicity.Key biosynthetic enzymes and transcription factors directly involved in the DHN melanin synthesis pathway include melanin synthesis transcription factors,polyketide synthase,1,3,6,8-tetrahydroxynaphthalene reductase,scytalone dehydratase,1,3,8-trihydroxynaphthalene reductase,and a-hydrolase.The regulatory network controlling the DHN melanin biosynthesis is complex and multifaceted,involving various transcription factors,signaling pathways,and metabolic routes.This review provides a comprehensive summary of current research on the abundant biological functions of DHN melanin.Understanding the molecular mechanisms of DHN melanin synthesis in phytopathogenic fungi could facilitate the development of effective disease control strategies and novel therapeutic targets.
