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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫(Kai-Wun Yeh) | |
| dc.contributor.author | Shou-Chen Hsu | en |
| dc.contributor.author | 徐守宸 | zh_TW |
| dc.date.accessioned | 2021-06-17T04:51:33Z | - |
| dc.date.available | 2020-08-02 | |
| dc.date.copyright | 2018-08-02 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-07-30 | |
| dc.identifier.citation | Akum, F. N., Steinbrenner, J., Biedenkoph, D., Imani, J., and Kogel, K.H. (2015). The Piriformospora indica effector PIIN_08944 promotes the mutualistic Sebacinalean symbiosis. Front. Plant Sci. 6: 906.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/71072 | - |
| dc.description.abstract | 印度梨形孢真菌(Piriformospora indica)為一宿主範圍極廣且具農業應用價值的共生真菌。共生性微生物能產生效應蛋白(effector protein)進而調控植物的生理機制，促進建立與宿主之共生關係。鑑於P. indica廣大的宿主範圍，對P. indica效應蛋白的研究有助於建立內共生菌(endophyte)與植物間共生機制的模型。本研究著重於進行效應蛋白的功能鑑定，根據實驗室過去研究成果，已由生物資訊方法篩選出兩個P. indica效應蛋白基因PIIN_09643與PIIN_11103，本研究利用建立小白菜轉殖毛狀根、小白菜葉部農桿菌注射的短暫過量表現與轉殖阿拉伯芥的方式，驗證水楊酸路徑指標基因PR-1之表現顯著下降。為闡明效應蛋白候選基因在共生階段的功能，本研究藉由酵母菌雙雜合篩選實驗得到與效應蛋白PIIN_11103互相作用的小白菜阿拉伯半乳聚醣蛋白Arabinogalactan protein 4 (AGP4) 。阿拉伯半乳聚醣蛋白為高度醣基化的醣蛋白，多分佈於植物細胞膜上，參與細胞增生、程序性細胞凋亡、花粉管生長、體胚再生等各種功能。本研究發現BrAGP4位於細胞膜與葉綠體的被膜，利用雙分子螢光互補實驗也證實PIIN_11103與BrAGP4可以交互作用，並且坐落於葉綠體被膜。以處理AGPs抑制劑的方式了解AGPs能夠影響生長素相關基因之表現、抑制根毛的發育，推論AGPs對於P. indica與小白菜建立共生之過程中也扮演著重要角色。 | zh_TW |
| dc.description.abstract | Piriformospora indica is a mutualistic endophyte with a broad host range and application potential in agriculture. Mutualistic microbes are able to produce effector proteins to interfere plant physiology for faciliting symbiosis with plants. In consideration of P. indica’s broad host-range, studies of P. indica effector proteins can establish a model for endophyte symbiosis with plants. This study focuses on functional characterization of effector proteins. According to lab’s previous studies, we already screen out two P. indica’s effector genes, PIIN_09643 and PIIN_11103 by bioinformatics approaches. We established overexpression lines of Chinese cabbage hairy root system and transgenic Arabidopsis thaliana. We also do agroinfiltrantion for transient overespression in Chinese cabbage leaves to confirm suppression of salicylic acid signal transduction. To investigate effectors function, we establish cDNA library to undertake yeast two-hybrid screening experiment. We found B. rapa’s Arabinogalactan protein 4 (AGP4) would interact with PIIN_11103. Arabinogalactan proteins are highy glycosylated glycoproteins mostly localized on plasma membrane, involved in cell proliferation, programmed cell death, pollen-tube development and somatic embryogenesis. This study found that BrAGP4 localized on plasma membrane and chloroplast envelope, and confirmed interaction of BrAGP4 and PIIN_11103 by BiFC. To characterize BrAGP4, we treated Chinese cabbage with AGPs inhibitor, found the inhibitory effects on root hair development but up-regulateing of auxin-relaed gene expressions. We also found that AGPs play a critical role in symbiosis process of P. indica and Chinese cabbage. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T04:51:33Z (GMT). No. of bitstreams: 1 ntu-107-R04b42002-1.pdf: 3185720 bytes, checksum: 2c74bd56a26b5b1c45d8ccb99dd99453 (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 目錄
口試委員會審定書 i 致謝 ii 中文摘要 iii Abstract iv 目錄 v 第一章 前言 1 第一節 印度梨形孢真菌 (Piriformospora indica)簡介 1 1.1.1 發現、型態與分類學 1 1.1.2 Piriformospora indica與植物之交互作用與影響 2 第二節 植物與微生物間之交互作用與防禦反應 3 第三節 效應蛋白 (effector proteins) 4 1.3.1 共生真菌的效應蛋白 4 1.3.2 Piriformospora indica的效應蛋白 5 第四節 阿拉伯半乳聚醣蛋白(Arabinogalactan protein, AGP) 6 第五節 研究目的 7 第二章 材料與方法 8 第一節 實驗材料 8 第二節 P. indica與小白菜之共生 8 2.2.1 小白菜在培養基上的無菌栽培 8 2.2.3 AGP抑制劑處理 9 2.2.4 P. indica 繼代培養 9 2.2.5 P. indica 與小白菜共培養 10 第三節 小白菜根部染色 10 第四節 基因表現量測定 11 2.4.1 總體RNA萃取 11 2.4.2 RNA 膠體電泳 11 2.4.3合成第一股cDNA 12 2.4.4 反轉錄聚合酶連鎖反應 (RT-PCR) 鑑定基因表現量 12 2.4.5 即時定量聚合酶連鎖反應 (Real-Time quantitative PCR, qPCR) 13 第五節 全長基因序列選殖 13 2.5.1 製備欲構築之DNA片段 13 2.5.2 DNA膠體純化 14 2.5.3 酵母菌雜合、次細胞定位螢光蛋白表現之載體構築 15 2.5.4大腸桿菌之勝任細胞的轉型(transformation)與DNA定序 16 2.5.5 質體抽取 17 2.5.6農桿菌轉型 17 2.5.7農根菌轉型 18 第六節 阿拉伯芥基因轉殖與分析 19 2.6.1 阿拉伯芥栽種 19 2.6.2農桿菌轉殖前處理 19 2.6.3阿拉伯芥之農桿菌轉殖 20 2.6.4阿拉伯芥之轉殖株篩選 20 第七節 阿拉伯芥原生質體細胞次定位分析 (subcellular localization) 21 2.7.1阿拉伯芥葉肉原生質體之分離與轉形 21 2.7.2 次細胞定位載體構築 22 2.7.3 阿拉伯芥原生質體轉形與影像分析 23 第八節 以農根菌A4品系產生小白菜轉殖毛狀根 23 2.8.1轉殖毛狀根系使用載體構築 23 2.8.2轉殖毛狀根系之建立與鑑定 23 第九節 酵母菌轉型 24 2.9.1酵母菌勝任細胞備製 24 2.9.2酵母菌轉型 25 第十節 真菌共生量測定 26 第十一節 農桿菌注射短暫表現實驗 27 第十二節 酵母菌雙雜合篩選實驗 27 2.12.1 建立cDNA library 27 2.12.2 酵母菌雙雜合實驗 29 第三章 結果 31 第一節 P. indica效應蛋白功能探討 31 3.1.1 建立小白菜毛狀根株系的轉殖系統 31 3.1.2利用小白菜轉殖毛狀根轉殖株，以分析效應蛋白之功能 31 3.1.3利用農桿菌注射(agroinfiltration)之短暫過量表現(transient overexpression)法以驗證效應蛋白PIIN_11103功能 32 第二節 轉殖阿拉伯芥以分析效應蛋白之功能 32 第三節 以酵母菌雙雜合篩選(yeast two-hybrid screening)實驗找尋效應蛋白在共生階段的交互作用蛋白(interacting protein) 33 第四節 小白菜之 arabinogalactan protein 4 (BrAGP4)的胺基酸序列分析以及親緣關係樹(phylogeny tree) 34 第五節 交互作用蛋白BrAGP4的基因表現樣態 35 第六節 BrAGP4於植物細胞中之次細胞定位 (subcellular localization) 35 第七節 以雙分子螢光互補實驗驗證BrAGP4與PIIN_11103交互作用 35 第八節 處理AGP生合成抑制劑3,4-dehydro-L-proline探討BrAGP4的生理功能 36 第四章 討論 38 第一節 效應蛋白PIIN_09643與PIIN_11103之功能 38 第二節 小白菜BrAGP4的功能 39 第三節 PIIN_11103與BrAGP4的交互作用 40 第四節 AGPs對於P. indica與小白菜共生效率的影響 41 第五節 BrAGP4在小白菜與P. indica共生時交互作用可能的生理意義 41 第六節 未來展望 42 參考文獻 43 | |
| dc.language.iso | zh-TW | |
| dc.title | 印度梨形孢真菌與小白菜共生時之效應蛋白功能性探討 | zh_TW |
| dc.title | Functional characterization of the putative effector proteins of Piriformospora indica in symbiotic interaction with Chinese cabbage | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄭秋萍(Chiu-Ping Cheng),王淑珍(Shu-Jen Wang),詹明才(Mimg-Tsair Chan),夏凱(Kai Xia) | |
| dc.subject.keyword | 印度梨型孢真菌,效應蛋白,阿拉伯半乳聚醣蛋白,共生, | zh_TW |
| dc.subject.keyword | Piriformospora indica,Effector protein,Arabinogalactan protein,Mycorrhizal symbiosis, | en |
| dc.relation.page | 80 | |
| dc.identifier.doi | 10.6342/NTU201802198 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-07-31 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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