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標題: | 大岩桐乙烯反應因子 SsERF-1 對花瓣生長的調控 The Role of Sinningia speciosa Ethylene Response Factor-1 (SsERF-1) in Petal Growth |
作者: | 朱苡晴 Yii-Chyng Chu |
指導教授: | 王俊能 Chun-Neng Wang |
關鍵字: | 大岩桐,兩側對稱性,CYCLOIDEA,乙烯,乙烯反應因子,花瓣大小,細胞延長, Sinningia speciosa,Floral zygomorphy,CYCLOIDEA,Ethylene,Ethylene response factor,Petal size,Cell expansion, |
出版年 : | 2023 |
學位: | 碩士 |
摘要: | 為了適應環境並有效的吸引傳粉者以達成繁殖之目的,花朵豐富的多樣性便是植物演化的重要一環,其中,兩側對稱花被認為是目前被子植物主要的演化趨勢,其不同花瓣會沿單一對稱軸向兩側呈鏡面對稱分佈,使傳粉者得以自特定的角度進入花朵中,促使更為精確的花粉傳播和柱頭接收,進而提升傳粉效率與繁殖成功率。而花朵的對稱性的建立已在金魚草 (Antirrhinum majus) 中被廣泛研究,TCP 轉錄因子中的 CYCLOIDEA (CYC) 基因會局限在背側花瓣高度表現,透過調節花瓣細胞增殖及細胞延長,使背側花瓣發育出不同於腹側及兩側花瓣的獨特性狀,然而,是否有未知的 CYC 潛在下游基因或其他調控路徑參與其中,以及它們是如何共同調節不同物種花朵中這些龐大的形態差異,都仍有許多尚未明瞭之處。為了找尋 CYC 的潛在下游,我們選用著名的園藝觀賞植株大岩桐 (Sinningia speciosa) 作為研究材料,並透過背腹側花瓣的轉錄體分析及雙螢光素酶報告基因檢測實驗等過往研究,篩選出目標基因 SsERF-1很可能為SsCYC之潛在下游,參與大岩桐背腹側花瓣的差異性生長。
為了探討 SsERF-1 與乙烯對大岩桐花朵生長發育的關聯性,我們首先建立了 SsERF-1 與模式物種阿拉伯芥 ERFs 的親緣關係樹,確立其屬於 Group IXa 子群中,具有保守的 AP2/ERF domain 與 CMIX-3 motif;接著透過 qRT-PCR 檢驗 SsERF-1 在大岩桐第 3、5、8 期花苞背腹側花瓣中的表現量,確實與 SsCYC 的表現模式一致,皆在背側花瓣中有較高的表現;而將大岩桐施加乙烯前驅物 ACC 處理後會在短時間內輕微促進 SsERF-1 的表現,且會透過抑制細胞延長使花苞生長較小;此外,更透過建立 SsERF-1 的阿拉伯芥異位表達轉殖株進行功能性分析,發現 SsERF-1 會延遲植株自營養期過渡至繁殖期的時間,且有抑制株高與節間的趨勢,而在花朵方面,SsERF-1 則會透過抑制細胞延長使轉殖株的花瓣較小。以上結果顯示乙烯前驅物 ACC 確實會抑制大岩桐花苞的大小,而 SsERF-1 亦會受到 ACC 促進,且會透過調控細胞延長抑制轉殖株花瓣大小的生長。 In order to adapt to and attract pollinators for reproductive success, flower diversity plays an important role in plant evolution. Floral zygomorphy (bilateral symmetry), in which different petals are divided into two equal halves along a single axis of symmetry, has been considered the major trend in angiosperm evolution. Zygomorphic flowers allow pollinators to enter the flower at a specific angle to improve the accuracy of pollen deposition onto stigma, increasing pollination efficiency and reproductive success. The establishment of floral symmetry has been widely studied in Antirrhinum majus. CYCLOIDEA (CYC), which belongs to TCP tracncription factor family, is highly expressed in dorsal petal and create dorsal identity by regulating cell proliferation and expansion. However, which downstream gene or regulatory network are associated with CYC and how they operate to establish the morphological differences are still unknown. To search for downstream of CYC, we use the famous horticultural plant Sinninigia speciosa as experimental material and select SsERF-1 as target gene via RNA-seq data analysis and dual-luciferase assay from previos work. These indicate that SsERF-1 might be the potential downstream of SsCYC and participate in floral development and dorsiventral differential gowth in S. speciosa. To uncover the phylogenetic relationship of SsERF-1, the phylogenetic tree reconstructed from Arabidopsis ERFs showed that SsERF-1 belongs to Group IXa, containing conserved AP2/ERF domain and CMIX-3 motif. Next, qRT-PCR analysis revealed that the expression pattern of SsERF-1 was consistent with SsCYC in which it expressed higher in dorsal petals than that in ventral petal. Moreover, the expression of SsERF-1 could be briefly induced by ethylene precursor ACC within a short time. Also, ACC treatment in S. speciosa resulted in smaller flower buds as a result of reduced petal cell expansion. On the other hand, ectopic expression of SsERF-1 in transgenetic Arabidopsis plants showed that SsERF-1 would delay the transition from vegetative to reproductive phase, and slightly inhibit plant height and internode length. Furthermore, ectopic expression of SsERF-1 caused smaller petals, regulated by inhibition of cell expansion. These results demonstrated that ethylene precursor ACC would inhibit flower bud size in S. speciosa, and induce the expression of SsERF-1. Also, SsERF-1 would inhibit petal size in transgenetic plants by regulating cell expansion. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/88605 |
DOI: | 10.6342/NTU202302061 |
全文授權: | 同意授權(全球公開) |
顯示於系所單位: | 生命科學系 |
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