B-D-GLUCURONIDASE GENE (GRUSA) AND GREEN FLUORESCENT PROTEINS (GFP) AS A TOOL FOR MONITORING THE SPATIAL DISTRIBUTION OF FUSARIUM OXYSPORLIM IN BANANA RHIZOSPHEREP.S. WONG AND M. SARIAH
The nonpathogenic fungus FUSL7r7.1-1171 oxysporum (Fo4) was successfully transformed with P-Dglucuronidase gene (gus4) (Gus system) from Escherichia coli and green fluorescent proteins (gfp) via (Agrobacterium tuniefactens-mediated transformation method with hygromycin B resistance gene as selectable marker. The GUS expression was detected by 5-bromo-4-chloro-3-indolyl-g-D- glucuronide (XGluc) and gfp expression detected by observation using the fluorescence microscopy. The transformants were stable in culture over a period of five conidiation cycles based on the cultural and morphological descriptions, antagonistic activity towards F. oxysporum f sp. race 4 (FocR4) and DNA polymorphism. Expression of the gusA was also detected during growth and colonization of the fungus in the soil and rhizosphere of banana, indicating that GUS and gfp fusion system can be used to develop a direct system for monitoring the spatial distribution of potential antagonist in the banana rhizosphere.
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