Analyses of the functions of Magnaporthe oryzae ATR gene with an efficient vector set for AtMT transgenesis of filamentous fungus.
M. NARUKAWA-NARA (1), Y. Hirato (2), K. Baba (2), T. Yasuda (2), T. Kamakura (2) (1) Tokyo University of Science, Japan; (2) Tokyo University of Science, Japan

Rice blast fungus Magnaporthe oryzae (Pyricularia oryzae) causes one of the most devastating fungal diseases of plant. In previous study, we showed that the Nijmegen breakage syndrome protein (NBS1) of M. oryzae is involved in DNA repair and normal development in M. oryzae. To investigate another genes that are involved in DNA repair in M. oryzae, we identified the gene sequence of ataxia telangiectasia and Rad3-related protein (ATR) homolog of M. oryzae. We tried to create the MoATR null mutants, however, we could not generate the mutants by polyethylene glycol-mediated protoplast transformation. Agrobacterium tumefaciens-mediated transformation (AtMT) is an efficient method of gene knockout in filamentous fungi. Therefore, we constructed a set of shuttle vectors for AtMT derived from the pBI121 T-DNA cloning vector using four drug-resistant genes which confer resistance to hygromycin, blasticidin S, geneticin and bialaphos. We constructed 40 vectors for gene knockout, gene silencing, gene overexpression and fluorescent protein tagging. Finally, we successfully generated the MoATR null mutants by using our new vector. The MoATR null mutants exhibited high sensitivity to DNA damage-inducing agents such as methyl methanesulfonate. This result suggests that MoATR plays (an) important role(s) in DNA repair of M. oryzae. We also estimated that our vector set for AtMT is very useful for fungal molecular genetical studies.

Abstract Number: P7-196
Session Type: Poster