阿拉伯芥血基質蛋白 AtMAPR3 可能藉由活性氧化物
訊息路徑參與植物防禦反應

本研究室之前的研究指出，新穎阿拉伯芥血基質蛋白 AtMAPR3 (membrane associated progesterone receptor 3 in Arabidopsis thilana) 的基因表現量，會受外加的過氧化氫及茉莉花酸 (jasmonate, JA) 誘導而上升。此外，查詢微陣列資料庫，AtMAPR3 的表現量在許多逆境下，例如以葡萄灰黴菌 (Botrytis cinerea) 感染後，亦有受誘導而上升的現象。為了進一步了解AtMAPR3 的生理功能是否和 ROS 產生之逆境有關，本研究比較野生型阿拉伯芥和 atmapr3-1 突變株在各種 ROS 生成逆境下的性狀差異，發現以 B. cinerea 感染五周大之阿拉伯芥植株，atmapr3-1 突變株葉部的感染面積 (lesion area) 約為野生型的 1.7 倍。另外，感染 B. cinerea 的兩天後，野生型阿拉伯芥中的 AtMAPR3 的表現量上升至約 11 倍。進一步利用 qPCR 研究 AtMAPR3 的存在是否會影響 ROS 反應相關基因。將阿拉伯芥野生型和 atmapr3-1 突變株以 B. cinerea 感染之後，兩種病原菌誘導基因 PR5 (pathogenesis-related gene 5) 及 PDF1.2b (plant defensin 1.2b) 在野生型阿拉伯芥中，分別在 24 小時及 36 小時後，表現量分別上升約 5 倍及 3.5 倍。而在 atmapr3-1 突變株中，受誘導的表現量均有過敏感的現象 (分別達 11至 15倍)。此外，兩種 ROS 反應基因 TAT3 (tyrosine aminotransferase 3) 和 ZAT10 (salt tolerance zinc finger 10) 在以 B. cinerea 感染之後，在阿拉伯芥野生型中有和 atmapr3-1 突變株中呈現不同的表現模式。其中 TAT3 基因在野生型阿拉伯芥中，在感染後表現量逐漸上升，至 48 小時後達到最高 (約 14倍)。然而在 atmapr3-1 突變株中，感染 24 小時後表現量即達最高 (約 9 倍)。ZAT10 亦有類似的表現模式。這些結果顯示 AtMAPR3 的存在與否的確影響了許多 ROS 反應相關基因。本研究顯示，在植物遭受病原菌侵害時，與其所引發的 ROS 訊息傳遞路徑 (ROS signaling pathway)，會促進 AtMAPR3 的表現，而 AtMAPR3 可能與該種逆境的生理反應相關。

關鍵字

血基質蛋白質；活性氧化物；葡萄灰黴菌；植物防禦系統

並列摘要

It is known that the expression of AtMAPR3 (membrane associated progesterone receptor 3 in Arabidopsis thaliana), a novel heme binding protein, is induced by exogenesis hydrogen peroxide and jasmonate. As revealed from public microarray database, the expression of AtMAPR3 is upregulated under various biotic stresses, such as infection by Botrytis cinerea and Phytophthora infestans. To further understand whether the physiological role of AtMAPR3 is related to ROS generating stresses, this study compare the phenotypic difference between wild type and a knock out mutant of AtMAPR3, atmapr3-1. It was found that the lesion area of atmapr3-1 mutant was larger than wild type by 1.7 fold when infected with B. cinerea. Also, the expression of AtMAPR3 in wild type was increased to 11 fold when infected for two days, indicate AtMAPR3 is responsive to the B. cinerea infection. To clarify if the presence of AtMAPR3 affects ROS responsive genes, the expression of these genes were examined in wild type or atmapr3-1 with B. cinerea infection. The expression of two pathogen responsive genes, PR5 (pathogenesis-related gene 5) and PDF1.2b (plant defensin 1.2b) in wild type infected with B. cinerea increased by 5 and 3.5 folds respectively after 24 hours and 36 hours. However, in atmapr3-1 mutants, the induced expression of those two genes were hypersensitive (reach to 11 and 15 folds respectively) to. Moreover, the expression of two ROS related genes TAT3 (tyrosine aminotransferase 3) and ZAT10 (salt tolerance zinc finger 10) in response to B. cinerea infection showed different expression patterns between wild type and atmapr3-1 mutant. The expression of TAT3 in wild type infected by B. cinerea increased along time and reached to its peak at 48 hours (about 14 folds). However in the atmapr3-1 mutant, the expression of those genes reached to its peak at 24 hours (about 9 folds). The ZAT gene response to B. cinerea infection showed similar expression pattern. These results implied the presence of AtMAPR3 affected the expression of some ROS responsive genes. I proposed that when plant infected by pathogens, the triggered ROS signaling pathway might induce the expression of AtMAP3, and the physiological role of AtMAPR3 might relate to this stress.

並列關鍵字

heme protein ； ROS ； Botrytis ； plant defense

參考文獻

Kim, W. C., J. H. Ko, J. Y. Kim, J. Kim, H. J. Bae and K. H. Han

Mandal, S., I. Kar, A. K. Mukherjee and P. Acharya

Riechmann, J. L., J. Heard, G. Martin, L. Reuber, C. Jiang, J. Keddie, L. Adam, O. Pineda, O. J. Ratcliffe, R. R. Samaha, R. Creelman, M. Pilgrim, P. Broun, J. Z. Zhang, D. Ghandehari, B. K. Sherman and G. Yu

Navarro, L., C. Zipfel, O. Rowland, I. Keller, S. Robatzek, T. Boller and J. D. Jones

Kamiya, T., M. Borghi, P. Wang, J. M. Danku, L. Kalmbach, P. S. Hosmani, S. Naseer, T. Fujiwara, N. Geldner and D. E. Salt

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阿拉伯芥血基質蛋白 AtMAPR3 可能藉由活性氧化物訊息路徑參與植物防禦反應

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