An Agrobacterium-mediated genetic transformation system for the rice false smut fungus Ustilaginoidea virens was developed using conidia as recipients. A binary vector, pCAMBIA1301-PgpdA-GUS-TtrpC, was constructed. The gpdA promoter (PgpdA) from Aspergillus nidulans was used to drive the expression of the β-glucuronidase (GUS) gene which enabled GUS activity visualization. The conidia transformation efficiency reached approximately 110 to 250 transformants per 1×105 conidia. Based on the analysis made on five successive generations of subcultures and PCR, the pCAMBIA1301-GUS cassette had integrated into the genomes of all transformants and clearly showed mitotic stability. The novel reporter vector constructed will promote the functional characterization of genes and the construction of genetically engineered strains of this important fungus.
Keywords: Agrobacterium tumefaciens-mediated transformation (ATMT); Hygromycin B; Ustilaginoidea virens; Vector construction; β-Glucuronidase gene (GUS).
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