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標題: | 解析番茄核因子NF-Y在叢枝菌根菌共生過程中的角色 Deciphering the role of the NF-Y transcription factors in coordinating arbuscular mycorrhizal symbiosis in tomato |
作者: | 堯景泓 Ching-Hung Yao |
指導教授: | 楊淑怡 Shu-Yi Yang |
關鍵字: | 轉錄因子,核因子YA,核因子YB,核因子YC3,CYCLOPS,叢枝菌根菌共生,番茄, transcription factor,nuclear factor Y,NF-YC3,CYCLOPS,GCC-box,arbuscular mycorrhizal (AM) symbiosis,tomato, |
出版年 : | 2023 |
學位: | 碩士 |
摘要: | 叢枝菌根菌共生(arbuscular mycorrhizal symbiosis)是球囊菌亞門 (Glomeromycotina) 真菌和植物之間的一類互利共生機制,能夠有效幫助植物吸收營養並促進其生長。然而,參與調控叢枝菌根菌共生發展的轉錄因子研究尚未完備。本實驗室團隊先前已發現番茄的轉錄因子,核因子NF-YC3可正向調節菌根菌共生,而其啟動子中的GCC-boxes也對其被共生誘導的表現十分重要。從酵母雙雜交中,我們進一步找到了三個可與NF-YC3相互作用的NF-YB (錢衡與郭庭伃,未發表)。基於這些發現,在本研究中已NF-YC3為出發,更全面的研究NF-Y家族的轉錄因子在菌根菌共生中的作用。NF-YC3座落於圓葉菸草葉片的細胞核中,暗示其可能參與轉錄調控。在nf-yc3基因靜默 (gene silencing) 之番茄根部中,叢枝 (arbuscule) 起始標記基因SbtM1的表達顯著降低,這表明NF-YC3可能在轉錄層次上參與早期叢枝發育。NF-YC3的基因表現在CYCLOPS基因靜默和過表達的根部中分別被下調和上調,表明CYCLOPS可能是NF-YC3的上游。共同表達 CYCLOPS和具活性的CCaMK 的確可在圓葉菸草葉片中活化 NF-YC3啟動子。雙分子螢光互補 (BiFC) 也證實之前找到的三個NF-YB可與NF-YC3相互作用,且三個NF-YB表現在細胞核和細胞質。然而,此研究未能利用yeast-triple-hybrid 或 BiFC找到能與NF-YC3相互作用的任何NF-YA。最後在基因靜默NF-YB的植物中,沒有觀察到任何共生型態的改變,可能是因為這些植物中的真菌量太低。總體而言,此研究強調 NF-YC3是菌根菌共生的關鍵參與者,並對調控菌根菌共生的分子機制提供了新的見解。 Arbuscular mycorrhizal (AM) symbiosis is a mutually beneficial relationship between plants and fungi from Glomeromycotina, facilitating nutrient uptake and promoting plant growth. However, the studies about transcription factors involved in mediating the development of AM symbiosis are not complete. In previous studies, we found that a transcription factor nuclear factor Y, NF-YC3, positively regulated AM symbiosis, and its AM-responsive expression may require GCC-boxes in its promoter. Moreover, three NF-YBs interacted with NF-YC3 were identified from yeast-2-hybrid (Chien and Kuo, unpublished). Based on these findings, a more comprehensively study to reveal the role of NF-Ys in AM symbiosis was performed starting with NF-YC3. NF-YC3 localized to the nucleus in N. benthamiana leaves, implying its potential role in transcriptional regulation. The expression of one arbuscule initiation marker gene, SbtM1, significantly decreased in nf-yc3 RNAi hairy roots, suggesting that NF-YC3 may play a crucial role in early arbuscule development. NF-YC3 expression was downregulated and upregulated in CYCLOPS RNAi and overexpression plants, respectively, indicating that CYCLOPS may be upstream of NF-YC3. Co-expression of CYCLOPS and autoactive CCaMK indeed transactivated NF-YC3 promoter in N. benthamiana leaves. Three NF-YBs identified from previous yeast-2-hybrid could interact with NF-YC3 in planta as shown in BiFC, and three NF-YBs were nucleocytoplasmic. However, no any NF-YA subunit that interacted with NF-YC3 could be identified by using the yeast triple hybrid assay and BiFC. Additionally, no phenotype of AM symbiosis was observed in any of the nf-yb single, double and triple RNAi plants, which may be due to the low colonization level in these composite plants. Overall, this study highlights NF-YC3 as a key player in AM symbiosis and provides new insights into the molecular mechanisms regulating this process. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/90686 |
DOI: | 10.6342/NTU202301481 |
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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