Functional analysis of the homoserine O-acetyltransferase gene and its identification as a selectable marker in Gibberella zeae.
We used restriction enzyme-mediated integration (REMI) to identify a methionine auxotrophic mutant of Gibberella zeae, an important cereal pathogen. In addition to its methionine requirement, the G. zeae REMI mutant designated Z43R3912 showed pleiotropic phenotypes, including reduced virulence on host plants and lack of sexual development. Outcrossing of Z43R3912 with a mat1-1 deletion strain confirmed that the mutation of Z43R3912 was tagged with the hygromycin B resistance marker. The vector insertion site in Z43R3912 was identified within the ORF designated GzmetE, encoding a putative homoserine O-acetyltrasferase (HOA). Gene disruption analyses confirmed that GzmetE was responsible for the pleiotropic phenotypes of Z43R3912. Genetic complementation of the G. zeae methionine auxotroph with an intact copy of the Aspergillus nidulans metE and GzmetE genes suggests that the HOA gene can be used as a selectable marker for transformation of G. zeae.
Han
You-Kyoung
YK
School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University, Seoul 151-742, Korea.
Lee
Theresa
T
Han
Kap-Hoon
KH
Yun
Sung-Hwan
SH
Lee
Yin-Won
YW
D000123:Acetyltransferases / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D017353:Gene Deletion; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005819:Genetic Markers; D005874:Gibberella / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D001467:Hordeum / Q000382:microbiology; D010641:Phenotype; D003313:Zea mays / Q000382:microbiology
D017931:DNA Primers; D005819:Genetic Markers; D000123:Acetyltransferases; C021369:homoserine O-acetyltransferase