阿拉伯芥乙烯反應子#019 對防禦反應的功能性 分析

Functional Analysis of Apetalla/Ethylene Response Factor #019 in Arabidopsis Defense Response

曾敏原(Min-Yuan Tseng)

指導教授 ： 金洛仁

國立臺灣大學/生命科學院/植物科學研究所/碩士(2014年)

https://doi.org/10.6342/NTU.2014.10177

摘要

自從ERF1 (ERF#092) 在２ ０ ０ ２ 年被發現後， 許多
Apetalla2/Ethylene response factors (AP2/ERFs)的轉錄因子已經被證
實參與阿拉伯芥(Arabidopsis thaliana)對病原菌的抵抗機制。但是，
有其他的ERF 轉錄因子還沒被證實參與抵抗機制。從大量篩選帶有
HA 標記的ERF 大量表現植株中，我們發現四個ERF (ERF#019,
ERF#023, ERF#096, and ERF#112)可能會影響抵抗機制。我們決定對
ERF#019 做進一步的研究。與野生型阿拉伯芥相比，其他獨立的
ERF#019 的大量表現植株皆顯現出對細菌性斑點病病原菌
(Pseudomonas syringae pv tomato DC3000)，細菌性軟腐病病原菌
(Pectobacterium carotovorum subsp carotovorum SCC1)以及灰黴病病
原菌(Botrytis cinerea)較易感病。根據定量RT-PCR 的結果指出在細菌
性軟腐病病原菌以及灰黴病病原菌感染後ERF#019 對兩種防禦基因
PDF1.2 與PR3 基因有負向的關係。原生質體的實驗結果顯示
ERF#019 是一種在細胞核中可以負向調節GCC box 控制的基因表現
的轉錄因子。根據Bifluorescence complementation
與
Coimmunoprecipitation 的實驗結果指出ERF#019 與HDA5 聯結。此
外，HDA5 與ERF#019 相似，可負向調控阿拉伯芥的防禦機制。在
我的論文中，我推論ERF#019 與HDA5 共同負向調控帶有GCC box
的PDF1.2 的基因表現。

關鍵字

阿拉伯芥 ； 灰黴病病原菌防禦機制 ； 細菌性軟腐病病原菌 ； 細菌性斑點病病原菌 ； 轉錄因子

並列摘要

Many transcription factors (TF) of Apetalla2/Ethylene response factors
(AP2/ERFs) have been shown involved in defense mechanisms against
pathogens in Arabidopsis thaliana (Arabidopsis) since ERF1 (ERF#092)
was found in year 2002. However, there are other ERF members have not
been proved to play roles in the defense mechanisms. From the
large-scale screening of ERF over-expressing lines HA-tagged, we found
four ERF (ERF#019, ERF#023, ERF#096, and ERF#112) could affect
the defense mechanisms. We decided to do further research on ERF#019.
Independent over-expression lines of ERF#019 showed serious
susceptible to Botrytis cinerea (B. cinerea), Pseudomonas syringae pv.
tomato DC3000 (Pst DC3000), and Pectobacterium carotovorum subsp
carotovorum SCC1 (Pcc SCC1) compared to Arabidopsis ecotype
Columbia (Col-0) wild-type control. Quantitative RT-PCR results
revealed that two defense related genes, PDF1.2 and PR3, are in a
negative relationship with ERF#019 after B. cinerea and Pcc SCC1
inoculation. Localization and transient expression in protoplast revealed
that ERF#019 is a transcription factor negatively regulating the GCC box
mediated gene expression in the nucleus. Bifluorescence
complementation (BiFC) and coimmunoprecipitation (CoIP) revealed
that ERF#019 is associated with HDA5. Surprisingly, HDA5 is similar to
ERF#019 which negatively modulates Arabidopsis plant defenses. In my
thesis, I suggest that ERF#019 negatively regulate the GCC box
containing gene, PDF1.2, by co-working with HDA5.

並列關鍵字

Arabidopsis ； Botrytis cinerea ； Defense mechanisms ； ERFs ； HDAs ； Pectobacterium carotovorum ； Pseudomonas syringae ； Transcription factors

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