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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫 | |
| dc.contributor.author | CHUAN-REN LIN | en |
| dc.contributor.author | 林川仁 | zh_TW |
| dc.date.accessioned | 2021-06-17T07:38:16Z | - |
| dc.date.available | 2024-03-27 | |
| dc.date.copyright | 2019-03-27 | |
| dc.date.issued | 2019 | |
| dc.date.submitted | 2019-03-19 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/73496 | - |
| dc.description.abstract | 印度梨型孢真菌，為一內生性共生真菌且能與許多不同種類之植物建立共生，在實驗室過去的研究中發現，當小白菜與Piriformospora. indica建立共生後，會促進植物的生長及提高其對於生物與非生物性逆境之抵抗力。雖然已有許多報導指出P. indica與植物共生後對於植物具有多種效益，但對於其中的分子機制尚不瞭解，有鑑於此，本研究利用過去實驗室所建立之效應蛋白生物資訊篩選方法，從中鎖定了4個候選基因PIIN_04020, PIIN_04363, PIIN_07152及PIIN_08445，並利用小白菜毛狀根及過表現轉殖阿拉伯芥的方式，試圖找出效應蛋白可能之功能，另外亦透過酵母菌雙雜交試驗找到PIIN_04363及PIIN_07152的交互作用蛋白，分別為銅伴護蛋白(BrCCH)及穀胱苷肽S-轉移酶(BrGSTU24)。本研究發現，在P.indica與小白菜共生階段的早期便會大幅提高效應蛋白基因PIIN_04363之表現，並且將PIIN_04363過表現於毛狀根及阿拉伯芥中，可使水楊酸SA路徑的指標基因PR-1下降，且因而提高了P. indica與植物的共生效率。而在PIIN_07152方面，則在共生階段的晚期才提高表現，並且PIIN_07152於毛狀根及阿拉伯芥中過表現時， JA路徑指標基因LOX2及VSP2的表現量被提高，而氧化還原相關基因DHAR及MDAR之表現量亦有顯著的提升。最後透過觀察PIIN_04363、PIIN_07152及PIIN_08445的過表現轉殖阿拉伯芥7天大幼苗發現，PIIN_07152 OE的生長勢有較為遲緩的趨勢，推論可能是因為植株中JA路徑較高的表現抑制了植株的生長，另外在PIIN_08445 OE方面，發現其轉殖株幼苗時期的生長勢相較於EV control似乎有些許提升，推論該現象或許是因為生長素相關基因的提升導致。 | zh_TW |
| dc.description.abstract | Piriformospora indica, an endophytic root-colonizing fungus, interacts with various plants species. From our previous researches, Chinese cabbage root colonization by P. indica results in an increase in plant growth, root propagation and adaptation to abiotic stress. Although P. indica has been reported to colonize and cause various beneficial function on plants, little is known about its early colonizeal mechanism. Previously, we screened several putative effector genes of P. indica by the bioinformatics screening system established from previous researchers. Further to characterize the function of P. indica effector proteins, four candidate genes PIIN_04020, PIIN_04363, PIIN_07152 and PIIN_08445 were chosen to establish Chinese cabbage transgenic hairy-root system、transgenic Arabidopsis thaliana for further investigation. Yeast two hybrid screening were used to screen the target interacting proteins against the effector proteins. B. rapa’s copper chaperone (CCH) against PIIN_04363 and glutathione S-transferase U24 (GSTU24) against PIIN_07152 were found. In this study, we found that PIIN_04363 gene expression was upregulated in the early colonized stages. Transgenic hairy-root system and transgenic Arabidopsis thaliana showed that PIIN_04363 suppress the expression level of SA pathway marker gene, PR-1 and cause a promotion of colonization efficiency. PIIN_07152 enhances the expression level of JA pathway marker genes, LOX2 and VSP2. Redox status relative genes DHAR and MDAR are also upregulated. From the observation of 7days-old transgenic Arabidopsis thaliana, the growth rate of PIIN_07152 OE was retard. It suggests that it is caused by the up-regulation of JA pathway. On the other hand, 7days-old transgenic Arabidopsis thaliana of PIIN_08445 OE show the higher growth rate in seedling stage, suggesting the effects of the up-regulation of auxin-related genes. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T07:38:16Z (GMT). No. of bitstreams: 1 ntu-108-R05b42025-1.pdf: 5450423 bytes, checksum: b180104724ca8314fdfed37708dcfd7a (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 口試委員會審定書 i
致謝 ii 摘要 iii Abstract iv 目錄 v 圖表目錄 vii 附錄圖表目錄 ix 第一章 前言 1 第一節 印度梨型孢真菌簡介 1 第二節 植物的生長與對環境逆境之調控及反應 4 第三節 效應蛋白(effector proteins) 7 第四節 銅伴護蛋白(copper chaperone, CCH) 10 第五節 穀胱苷肽S-轉移酶(glutathione S-transferase, GST) 13 第六節 研究目的與方向 14 第二章 材料與方法 15 第一節 實驗材料 15 第二節 P. indica與小白菜之共生 15 第三節 基因表現量測定 18 第四節 全長基因序列釣取 20 第五節 酵母菌雙雜合及次細胞定位之載體構築 22 第六節 阿拉伯芥基因轉殖與農桿菌注射 24 第七節 小白菜轉殖毛狀根之建立 27 第八節 酵母菌雙雜合篩選實驗 29 第九節 原生質體次細胞定位分析(subcellular localizatioon) 34 第三章 結果 37 第一節 P. indica效應蛋白候選基因之篩選 37 第二節 P. indica效應蛋白之功能探討 39 第三節 以酵母菌雙雜合實驗篩選效應蛋白之交互作用蛋白 45 第四章 討論 49 第一節 效應蛋白的次細胞定位及其基因之核苷酸序列特性 49 第二節 PIIN_04363與PIIN_07152對植物氧化還原態相關基因、防禦反應相關基因與逆境抵抗之影響 49 第三節 PIIN_04363對植物防禦反應相關基因與共生效率之影響 51 第四節 PIIN_08445對植物生長素反應相關基因之影響 52 第五節 PIIN_04363、PIIN_07152與BrCCH、BrGSTU24間之交互作用 53 第六節 未來展望 53 參考文獻 55 圖表 66 附錄圖表 93 | |
| dc.language.iso | zh-TW | |
| dc.title | 印度梨型孢真菌的效應蛋白對其宿主小白菜之功能性探討 | zh_TW |
| dc.title | Functional characterization of putative effector proteins of Piriformospora indica during colonization with Brassica rapa cv. Chinensis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 107-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王啟正,楊藹華,王淑珍,靳宗洛 | |
| dc.subject.keyword | 印度梨型孢真菌,效應蛋白,銅伴護蛋白,穀胱??S-轉移?,共生, | zh_TW |
| dc.subject.keyword | Piriformospora. indica,effector proteins,copper chaperone,glutathione S-transferase,symbiotic, | en |
| dc.relation.page | 109 | |
| dc.identifier.doi | 10.6342/NTU201900644 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2019-03-20 | |
| 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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