2014-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/714268摘要：植物防禦基因LsGRP1 為水楊酸誘導百合抗灰黴病菌Botrytis elliptica 過程中表現量增加的葉部基因，解碼產物為植物第二型富含甘胺酸蛋白(plant class II glycine-rich protein)。LsGRP1 之泛素化(ubiquitination)及其蛋白質交互作用與百合誘導抗病性具有高度相關性。在圓葉菸草(Nicotiana benthamiana)與阿拉伯芥 (Arabidopsis thaliana)表現LsGRP1 可顯著減少灰黴病菌Botrytis cinerea 感染之病徵，而LsGRP1 C 端區段合成胜&#32957;具有生體外抑菌活性與誘發灰黴病菌類程序性細胞死亡的能力。這些結果顯示LsGRP1 可能透過促進多重作用以抵抗灰黴病菌的侵染。本研究擬藉由過度表現與基因靜默方式，在單子葉植物百合與雙子葉植物阿拉伯芥上分析LsGRP1 對病徵、病原菌量、過敏性反應、細胞死亡、活性氧分子、癒傷葡聚醣累積、防禦相關基因表現，以及寄主誘導病原菌程序性細胞死亡之影響，並以抑制劑處理方式驗證LsGRP1 開啟之防禦反應。此外，將透過比 較表現全長與區段缺失LsGRP1 對植物抗灰黴病之影響，瞭解LsGRP1 促進防禦反應的功能分區。另一方面，將鑑定LsGRP1 泛素化位點，並分析LsGRP1 泛素化對蛋白質交互作用及植物抗灰黴病之影響，探索LsGRP1 促進植物抗病性的調控機轉，以累積LsGRP1 及其衍生產物之背景資訊，利於灰黴病管理策略之整合運用。<br> Abstract: LsGRP1 is a plant defense gene increasingly expressed in lily leaves exhibiting salicylic acid-induced resistance against Botrytis elliptica and encodes a plant class II glycine-rich protein (GRP). The ubiquitination of LsGRP1 is highly correlated to protein-protein interaction of LsGRP1 and the induced disease resistance of lily. As expressed in Nicotiana benthamiana and Arabidopsis thaliana, LsGRP1 significantly reduced symptoms caused by Botrytis cinerea. Besides, a synthetic peptide corresponding to the C-terminal region of LsGRP1 showed in vitro antimicrobial activity and capable induced programmed cell death-like phenomena in Botrytis spp. These findings indicated that LsGRP1 probably increases plant disease resistance against gray mold by multiple functions. In this project, plant defenses in response to Botrytis infection as affected by LsGRP1 will be examined by overexpression and interference of LsGRP1 expression in monocot lily and dicot Arabidopsis, including symptom development, pathogen population, hypersensitive response, cell necrosis, reactive oxygen specie accumulation, callose deposition, defense-related gene expression and plant-induced fungal programmed cell death. Meanwhile, inhibitor treatments will be applied to verify the defense responses activated by LsGRP1. Additionally, the playing roles of different regions of LsGRP1 in plant defense will be investigated. On the other hand, the ubiquitination site in LsGRP1 and the inferences of LsGRP1 ubiquitination on protein-protein interaction and disease resistance against gray mold will be assessed to increase the background knowledge for the application of LsGRP1 and its derived products and facilitate their integrations in the gray mold disease management.植物第二型富含甘胺酸蛋白植物防禦植物誘導真菌程序性細胞死亡泛素化抗病性灰黴病plant class II glycine-rich proteinplant defenseplant-induced fungal programmed cell deathubiquitinationdisease resistancegray mold