Open AccessCloning, sequencing and functional analysis of Magnaporthe grisea MVP1 gene, a hex‐1 homolog encoding a putative ‘woronin body’ protein
Author(s) -
Asiegbu Frederick O.,
Choi Woobong,
Jeong Jun Seop,
Dean Ralph A.
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00858-9
Subject(s) - biology , magnaporthe grisea , neurospora crassa , southern blot , magnaporthe , gene , genetics , microbiology and biotechnology , complementary dna , untranslated region , mutant , rna , oryza sativa
A hex‐1 homolog named MVP1 was isolated from an appressoria cDNA library of the rice blast fungus Magnaporthe grisea . The transcript of ∼1.6 kb contains 546 bp of coding sequence with a 3′ untranslated region about 168 bp long and a 5′ untranslated region about 870 bp long. Southern gel blot analysis of genomic DNA following digestion with three restriction enzymes ( Bam HI, Eco RI, Hin dIII) indicated that the gene exists as a single copy in M. grisea genome. RNA gel blot analyses showed that MVP1 was highly expressed in germinating conidia and the mycelial stage compared to appressoria or non‐germinated conidia. MVP1 showed a high degree of homology to the hex‐1 gene recently described to encode a major protein in the woronin bodies of Neurospora crassa . Double homologous recombination was used to replace MVP1 with the hyg R gene. MVP1 knockout mutants showed apical swellings when grown on agar plates containing 2% sorbose but they were not impaired in any other vegetative or pathogenic properties evaluated. The pathological and other phenotypic consequences of gene disruption are discussed.