本文選題：番茄　切入點(diǎn)：SlSINA1　出處：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：果實(shí)大小作為番茄重要的商品性狀,也是生物學(xué)研究和番茄育種的重要表型性狀。通過(guò)分子生物學(xué)發(fā)掘調(diào)控果實(shí)大小的基因,闡釋調(diào)控番茄果實(shí)大小的分子機(jī)理,不僅對(duì)番茄分子育種具備關(guān)鍵的指導(dǎo)性作用,也為新品種的鑒定提供了重要的科學(xué)依據(jù)。番茄中關(guān)于SINA(Seven in absentia)的研究很少,SINA家族含有高度保守的Sina和RING功能域,其RING功能域能夠行使E3泛素連接酶活性。通過(guò)功能研究發(fā)現(xiàn),在番茄中超量表達(dá)SlSINA1導(dǎo)致番茄的果實(shí)變小。主要研究結(jié)果如下:1.通過(guò)組織表達(dá)譜分析表明,SlSINA1在所有組織器官中均有表達(dá),在花中表達(dá)量最高,在花蕾中表達(dá)量次之,在根和紅熟期(RR)果實(shí)中表達(dá)量最低。隨著果實(shí)的發(fā)育,SlSINA1在17DPA(Days post-anthesis)時(shí)的果實(shí)中表達(dá)量達(dá)到峰值,然后表達(dá)量不斷降低,在紅熟期降至最低。2.與對(duì)照(AC,Ailsa Craig)相比,SlSINA1超量轉(zhuǎn)基因材料T1代和T3代果實(shí)縱徑、橫徑和果實(shí)重量顯著減小。3.細(xì)胞生物學(xué)觀察發(fā)現(xiàn),0DPA、7DPA、24DPA和35DPA的超量轉(zhuǎn)基因株系番茄果實(shí)的果皮厚度、果皮細(xì)胞層數(shù)與對(duì)照相比顯著減小,7DPA、24DPA和35DPA的果皮細(xì)胞平均面積較對(duì)照顯著的減小。隨著時(shí)間的推移,果皮的厚度、細(xì)胞層數(shù)和細(xì)胞平均面積均不斷增加,并且在7DPA至24DPA期間這三者增加了至少10-15倍,其他時(shí)間內(nèi)這三者雖然有所增加,但是變化較小。因此推測(cè)SlSINA1可能通過(guò)影響細(xì)胞的大小和數(shù)量影響果皮厚度而調(diào)控番茄果實(shí)大小的發(fā)育。4.酵母雙雜交實(shí)驗(yàn)表明,SlSINA1能夠與RAP30、SPG和FW2.2蛋白互作,SlSINA1可能與這些蛋白形成復(fù)合體發(fā)揮功能。5.在SlSINA1超量轉(zhuǎn)基因系中進(jìn)行表達(dá)量分析結(jié)果顯示,較對(duì)照而言,SlSINA1表達(dá)量顯著增加,而RAP30、SPG和FW2.2的表達(dá)量均顯著下降。6.通過(guò)Western-blot實(shí)驗(yàn),未能檢測(cè)到SPG和FW2.2蛋白的表達(dá),Co-IP驗(yàn)證未發(fā)現(xiàn)SlSINA1與RAP30在植物體內(nèi)存在互作關(guān)系。
[Abstract]:Fruit size, as an important commodity trait of tomato, is also an important phenotypic trait in biological research and tomato breeding. It not only has a key guiding role in tomato molecular breeding, but also provides an important scientific basis for the identification of new varieties. Few studies on SINA(Seven in absentia in tomato show that Sina family contains highly conserved Sina and RING functional domains. Its RING domain can exercise the activity of E3 ubiquitin ligase. The main results were as follows: 1.The results of tissue expression analysis showed that SlSINA1 was expressed in all tissues and organs, the highest in flower and the second in flower bud, the main results were as follows: 1.The main results were as follows: 1. Through tissue expression analysis, SINA1 was expressed in all tissues and organs, the highest in flower, and the second in flower bud. The expression level of SlSINA1 reached its peak value in the fruit of 17DPADDays post-anthesism, and then decreased, with the development of the fruit, the expression of SlSINA1 was the lowest in the root and red mature fruit, and the expression of SlSINA1 reached its peak value in the fruit of 17DPADDays post-anthesism. The fruit diameter, transverse diameter and fruit weight of T _ 1 and T _ 3 generations decreased significantly compared with those of the control, Ailsa Craigi. The fruit skin thickness of transgenic tomato lines were observed by cell biology observation, and the results showed that the fruit skin thickness of the transgenic lines was 7DPA-7DPA-24DPA and 35DPA, and the fruit diameter was significantly lower than that of the control (Ailsa Craigg), and the fruit diameter, transverse diameter and fruit weight of T _ 1 and T _ 3 generations were significantly decreased. The average area of pericarp cells of 7DPA-24DPA and 35DPA decreased significantly compared with the control, and the thickness of pericarp, the number of cell layers and the average area of cells increased with the passage of time. And between 7DPA and 24DPA, these three increased by at least 10-15 times, but at other times these three increased, But the change is small. Therefore, it is speculated that SlSINA1 may regulate the development of tomato fruit size by affecting the cell size and the number of the fruit peel thickness. 4. Yeast two-hybrid experiment shows that SlSINA1 can interact with RAP30SINA1 and FW2.2 protein and SlSINA1 may interact with this. The expression of these protein forming complexes in SlSINA1 superabundant transgenic lines was analyzed, and the results showed that, Compared with the control, the expression of SlSINA1 was significantly increased, while the expression of SPG and FW2.2 was decreased significantly. 6. The expression of SPG and FW2.2 protein was not detected by Western-blot test. Co-IP verification showed that there was no interaction between SlSINA1 and RAP30 in plants.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S641.2

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本文編號(hào)：1590875