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標題: | 解析番茄核因子SlNF-YBs在叢枝菌根菌共生中扮演之角色 Deciphering the roles of tomato SlNF-YBs involved in arbuscular mycorrhizal symbiosis |
作者: | Ting-Yu Kuo 郭庭伃 |
指導教授: | 楊淑怡(Shu-Yi Yang) |
關鍵字: | 番茄,叢枝菌根菌共生,轉錄因子,核因子Y,核因子YC3,核因子YB, tomato,arbuscular mycorrhiza (AM) symbiosis,transcription factor,Nuclear Factor Y (NF-Y),NF-YC3,NF-YB, |
出版年 : | 2021 |
學位: | 碩士 |
摘要: | 叢枝菌根菌共生(arbuscular mycorrhiza symbiosis)是指大多數陸生植物與球囊菌亞門 (Glomeromycotina) 真菌建立的一種互利共生關係。為了形成以利養分交換的叢枝狀結構(arbuscule),植物勢必大幅度地改變轉錄體以重塑植物細胞內部。然而在共生進程中,植物透過什麼樣的轉錄因子聯絡網來調控轉錄體,仍不甚清楚。本研究團隊先前在番茄中發現一個核因子Y (Nuclear Factor Y) 能正向調控叢枝菌根菌共生,並將之命名為NF-YC3 (錢衡,未發表)。屬於轉錄因子的核因子Y一般由核因子YA、YB及YC三種次單元組成,以異三聚體的形式調控下游基因。但至今為止尚未有研究報導完整的異三具體核因子Y如何調控叢枝菌根菌共生,因此本研究致力於尋找能與NF-YC3結合、共同參與調控共生的核因子Y。本篇研究以酵母雙雜合蛋白系統加上雙分子螢光互補方法,發現三個會被共生調控表現量的核因子YB: NF-YB3a、NF-YB5c以及NF-YB3b能與NF-YC3結合。單獨過量表現核因子Y於菸草葉片中,觀察到三個NF-YB分佈於細胞質及核,而NF YC3則主要座落於細胞核中,而雙分子螢光互補的結果則顯現NF-YB與NF-YC3結合之後會位於細胞核中。啟動子─GUS報導基因系統揭示NF-YB3a、NF-YB5c和NF-YB3b的啟動子無論在未接種真菌或是接種真菌的情況下,皆能在根部的任何細胞組織開啟活性。然而,在透過RNA干擾方法降低目標核因子YB表現量的植物中,觀察到叢枝菌根菌的表現型態只有非常微弱的改變。再加上許多其他核因子YB在RNA干擾的目標NF-YB植物中被上調,意味著番茄中的核因子YB在調控叢枝菌根菌共生時具有功能重複性。 Most land plants engage in a mutually beneficial interaction with Glomeromycotina fungi called arbuscular mycorrhiza (AM) symbiosis. To form the nutrient exchange fungal structure, arbuscule, host plants undergo a massive transcription reprogramming to remodel the root cells intracellularly. However, how this transcriptomic alteration is governed by a network of transcription factors still remains unclear. Previously, our team found a tomato Nuclear Factor Y transcription factor, NF-YC3, can positively regulate AM symbiosis (Chien, unpublished). Nuclear Factor Y (NF-Y) families of transcription factors are often composed by NF-YA, NF-YB and NF-YC subunits, functioning as heterotrimers. However, no complete NF-Y heterotrimers have been reported to be involved in AM symbiosis until now. Therefore, this study focuses on identifying interaction and co regulating partners of NF YC3 during AM symbiosis. Here, three candidate NF-YBs, NF YB3a, NF-YB5c, and NF-YB3b, which were regulated by AM symbiosis, could interact with NF-YC3 in yeast-two-hybrid and BiFC assays. Overexpression of NF Y in tobacco leaves showed that NF-YBs were localized to the cytoplasm and nucleus, and NF-YC3 were mainly localized to the nucleus. However, the BiFC assay showed that they only interacted with each other in the nucleus. GUS assays revealed that the promoters of NF YB3a, NF-YB5c, and NF-YB3b can be activated in all cell layers of roots under both mock and mycorrhizal conditions. However, knockdown of the candidate NF YBs by RNAi only showed a minor phenotype of AM symbiosis. Together with upregulating of other NF-YBs in the RNAi lines, these results suggested that there might be functional redundancy of NF-YBs to regulate AM symbiosis in tomato. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/21188 |
DOI: | 10.6342/NTU202100274 |
全文授權: | 未授權 |
顯示於系所單位: | 植物科學研究所 |
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