חיפוש מתקדם
Biocontrol Science and Technology
Ezra, D., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Skovorodnikova, J., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Kroitor-Keren, T., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel, Department of Plant Pathology and Microbiology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel
Denisov, Y., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel, Department of Plant Pathology and Microbiology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel
Liarzi, O., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Symbiotic endophytes, unlike plant pathogens, do not usually induce visible host response. This may constraint the researcher's decision whether a plant has been successfully infected by the endophyte. In order to properly study the establishment, development and progress of an endophyte in the host plant and hostendophyte interactions, methods for the identification and localization of endophytic microorganisms are needed. Towards this aim, we focused at two levels: (A) We constructed M. albus-specific primers for polymerase chain reaction (PCR). In vitro, these primers specifically detected only M. albus strains and not isolates of related fungi (such as Daldinia sp. and a Xylariaceae sp.). (B) For direct visualization of the fungi, we inserted a reporter gene (gfp) into M. albus hyphae using Agrobacterium-mediated transformation. Since M. albus is a sterile fungus (i.e., without spores or fungal fruiting bodies), we used chopped fungal mycelium for the transformation procedure. We transformed three different isolates of M. albus using Agrobacterium-mediated transformation. Fifty-nine different transformants were collected with a transformation efficacy of 0.0004-0.0026%. Although PCR-based detection and direct visualization of the transformants in planta were unsuccessful, all tested transformants (with one exception) exhibited similar biological activity to their cognate wild type. This work provides a significant step forward in molecular research of the relationships between this endophytic genus and their hosts. © 2010 Taylor & Francis.
פותח על ידי קלירמאש פתרונות בע"מ -
הספר "אוצר וולקני"
אודות
תנאי שימוש
Development of methods for detection and Agrobacterium-mediated transformation of the sterile, endophytic fungus Muscodor albus
20
Ezra, D., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Skovorodnikova, J., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Kroitor-Keren, T., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel, Department of Plant Pathology and Microbiology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel
Denisov, Y., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel, Department of Plant Pathology and Microbiology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel
Liarzi, O., Department of Plant Pathology and Weed Research, ARO, The Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Development of methods for detection and Agrobacterium-mediated transformation of the sterile, endophytic fungus Muscodor albus
Symbiotic endophytes, unlike plant pathogens, do not usually induce visible host response. This may constraint the researcher's decision whether a plant has been successfully infected by the endophyte. In order to properly study the establishment, development and progress of an endophyte in the host plant and hostendophyte interactions, methods for the identification and localization of endophytic microorganisms are needed. Towards this aim, we focused at two levels: (A) We constructed M. albus-specific primers for polymerase chain reaction (PCR). In vitro, these primers specifically detected only M. albus strains and not isolates of related fungi (such as Daldinia sp. and a Xylariaceae sp.). (B) For direct visualization of the fungi, we inserted a reporter gene (gfp) into M. albus hyphae using Agrobacterium-mediated transformation. Since M. albus is a sterile fungus (i.e., without spores or fungal fruiting bodies), we used chopped fungal mycelium for the transformation procedure. We transformed three different isolates of M. albus using Agrobacterium-mediated transformation. Fifty-nine different transformants were collected with a transformation efficacy of 0.0004-0.0026%. Although PCR-based detection and direct visualization of the transformants in planta were unsuccessful, all tested transformants (with one exception) exhibited similar biological activity to their cognate wild type. This work provides a significant step forward in molecular research of the relationships between this endophytic genus and their hosts. © 2010 Taylor & Francis.
Scientific Publication
You may also be interested in