title: Generation of β-glucuronidase reporter-tagged strain to monitor Ustilaginoidea virens infection in rice.
abstract: 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 1×105conidia. 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.
authors: Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, China.
authors: Mebeaselassie
authors: M
authors: Andargie
authors: Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, China.
authors: Chao
authors: C
authors: Yang
authors: Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou, China. Electronic address: jxli@scbg.ac.cn.
authors: Jianxiong
authors: J
authors: Li
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D001233:Aspergillus nidulans / Q000235:genetics; D018920:Coculture Techniques; D004271:DNA, Fungal; D005786:Gene Expression Regulation; D005800:Genes, Fungal / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics*; D006921:Hygromycin B; D006999:Hypocreales / Q000235:genetics* / Q000472:pathogenicity; D012275:Oryza / Q000382:microbiology*; D010641:Phenotype; D010935:Plant Diseases / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D013172:Spores, Fungal / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D004271:DNA, Fungal; D006921:Hygromycin B; D005966:Glucuronidase
keywords: Agrobacterium tumefaciens-mediated transformation (ATMT); Hygromycin B; Ustilaginoidea virens; Vector construction; β-Glucuronidase gene (GUS)