title: cDNA Subtractive Cloning of Genes Expressed during Early Stage of Appressorium Formation by Magnaporthe grisea.
abstract: The conidial germ tube of the fungus Magnaporthe grisea differentiates an infection-specific structure, an appressorium, for penetration into the host plant. Formation of the appressorium is also observed on synthetic solid substrata such as polycarbonate. We found that a plant lectin, concanavalin A, specifically suppressed the appressorium formation without affecting the germling adhesion if it was applied within 2-3 hours after germination. Standing on the result, we constructed a cDNA library that represents the early stage of germ tube development and/or appressorium formation from the 2.5-hour-old germ tubes using a cDNA subtraction strategy by the combination of the biotin labeled driver method and adapter-primed PCR method. Out of 686 colonies of the library, 158 distinct clones' nucleotide sequences were partially analyzed. Some clones' expression patterns were detected by RT-PCR and from those results, our library seemed to well represent the objective developmental stage of M. grisea.
authors.affiliation: a Microbial Toxicology Laboratory, The Institute of Physical and Chemical Research (RIKEN).
authors.forename: T
authors.initials: T
authors.lastname: Kamakura
authors.affiliation: a Microbial Toxicology Laboratory, The Institute of Physical and Chemical Research (RIKEN).
authors.forename: J Z
authors.initials: JZ
authors.lastname: Xiao
authors.affiliation: a Microbial Toxicology Laboratory, The Institute of Physical and Chemical Research (RIKEN).
authors.forename: W B
authors.initials: WB
authors.lastname: Choi
authors.affiliation: b Tokyo University of Agriculture and Technology.
authors.forename: T
authors.initials: T
authors.lastname: Kochi
authors.affiliation: b Tokyo University of Agriculture and Technology.
authors.forename: S
authors.initials: S
authors.lastname: Yamaguchi
authors.affiliation: b Tokyo University of Agriculture and Technology.
authors.forename: T
authors.initials: T
authors.lastname: Teraoka
authors.affiliation: a Microbial Toxicology Laboratory, The Institute of Physical and Chemical Research (RIKEN).
authors.forename: I
authors.initials: I
authors.lastname: Yamaguchi
keywords: Magnaporthe grisea; Pyricularia oryzae; appressorium formation; cDNA subtraction; differential library