cDNA Subtractive Cloning of Genes Expressed during Early Stage of Appressorium Formation by Magnaporthe grisea.
cDNA Subtractive Cloning of Genes Expressed during Early Stage of Appressorium Formation by Magnaporthe grisea.
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DOI：
10.1271/bbb.63.1407
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发表时间：
1999
期刊：
影响因子：
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通讯作者：
T. Kamakura;J. Xiao;W. Choi;T. Kochi;S. Yamaguchi;T. Teraoka;I. Yamaguchi
中科院分区：
文献类型：
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作者：
T. Kamakura;J. Xiao;W. Choi;T. Kochi;S. Yamaguchi;T. Teraoka;I. Yamaguchi
稻瘟病菌的分生孢子管分化出一种侵染特有的结构，即附着胞，以便穿透到寄主植物中。附着胞的形成也可以在聚碳酸酯等合成固体底物上观察到。我们发现，一种植物凝集素刀豆蛋白A，如果在萌发后2-3小时内使用，就能特异性地抑制附着胞的形成，而不影响种子的附着。在此基础上，我们结合生物素标记的驱动器法和接头引导型聚合酶链式反应的方法，以2.5h的胚管为材料，采用消减策略，构建了一个代表胚管发育和附着胞形成早期的cDNA文库。在文库的686个菌落中，有158个不同克隆的核苷酸序列被部分分析。RT-PCR检测到一些克隆的表达模式，从这些结果来看，我们的文库似乎很好地代表了稻瘟病菌的客观发育阶段。
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.