Interaction Transcriptome Analysis Identifies Magnaporthe oryzae BAS1-4 as Biotrophy-Associated Secreted Proteins in Rice Blast Disease
Interaction Transcriptome Analysis Identifies Magnaporthe oryzae BAS1-4 as Biotrophy-Associated Secreted Proteins in Rice Blast Disease
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DOI：
10.1105/tpc.107.055228
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发表时间：
2009-04-01
期刊：
影响因子：
11.6
通讯作者：
Valent, Barbara
中科院分区：
文献类型：
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作者：
Mosquera, Gloria;Giraldo, Martha C.;Valent, Barbara
当稻瘟病菌 Magnaporthe oryzae 的生物营养型侵入菌丝 (IH) 连续侵入活的水稻 (Oryza sativa) 细胞时，它们会分泌效应子来改变宿主的防御和细胞过程。然而，目前已发现的爆炸效应器却很少。事实上，了解导致生物营养入侵的真菌和水稻基因一直很困难，因为在最早的感染阶段遇到 IH 的植物细胞很少。我们开发了一种用于分离受感染水稻鞘 RNA 的强大程序，其中： 20% 的 RNA 源自首次入侵细胞中的 IH。我们使用米霉寡核苷酸分析了这些 IH RNA 与对照菌丝体 RNA 的关系。通过 10 倍差异表达阈值，我们鉴定了已知的效应子 PWL2 和 58 个候选效应子。其中四个候选蛋白被证实是真菌生物营养相关分泌（BAS）蛋白。荧光标记的 BAS 蛋白以相容但不不相容的相互作用以不同的模式分泌到水稻细胞中。 BAS1 和 BAS2 蛋白与已知的无毒效应子一起优先积聚在生物营养界面复合物中，BAS3 在细胞壁交叉点附近显示出额外的定位，而 BAS4 则均匀地勾画出生长的 IH。用水稻寡阵列分析相同的感染组织 RNA，鉴定出假定的效应子诱导的水稻易感基因，这些基因高度富集传感器转导成分，而不是通常鉴定的防御反应基因。
Biotrophic invasive hyphae (IH) of the blast fungus Magnaporthe oryzae secrete effectors to alter host defenses and cellular processes as they successively invade living rice (Oryza sativa) cells. However, few blast effectors have been identified. Indeed, understanding fungal and rice genes contributing to biotrophic invasion has been difficult because so few plant cells have encountered IH at the earliest infection stages. We developed a robust procedure for isolating infected-rice sheath RNAs in which; 20% of the RNA originated from IH in first-invaded cells. We analyzed these IH RNAs relative to control mycelial RNAs using M. oryzae oligoarrays. With a 10-fold differential expression threshold, we identified known effector PWL2 and 58 candidate effectors. Four of these candidates were confirmed to be fungal biotrophy-associated secreted (BAS) proteins. Fluorescently labeled BAS proteins were secreted into rice cells in distinct patterns in compatible, but not in incompatible, interactions. BAS1 and BAS2 proteins preferentially accumulated in biotrophic interfacial complexes along with known avirulence effectors, BAS3 showed additional localization near cell wall crossing points, and BAS4 uniformly outlined growing IH. Analysis of the same infected-tissue RNAs with rice oligoarrays identified putative effector-induced rice susceptibility genes, which are highly enriched for sensor-transduction components rather than typically identified defense response genes.