本文選題：番茄　切入點：SlSINA1　出處：《華中農業(yè)大學》2017年碩士論文　論文類型：學位論文

【摘要】：果實大小作為番茄重要的商品性狀,也是生物學研究和番茄育種的重要表型性狀。通過分子生物學發(fā)掘調控果實大小的基因,闡釋調控番茄果實大小的分子機理,不僅對番茄分子育種具備關鍵的指導性作用,也為新品種的鑒定提供了重要的科學依據。番茄中關于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。通過功能研究發(fā)現,在番茄中超量表達SlSINA1導致番茄的果實變小。主要研究結果如下:1.通過組織表達譜分析表明,SlSINA1在所有組織器官中均有表達,在花中表達量最高,在花蕾中表達量次之,在根和紅熟期(RR)果實中表達量最低。隨著果實的發(fā)育,SlSINA1在17DPA(Days post-anthesis)時的果實中表達量達到峰值,然后表達量不斷降低,在紅熟期降至最低。2.與對照(AC,Ailsa Craig)相比,SlSINA1超量轉基因材料T1代和T3代果實縱徑、橫徑和果實重量顯著減小。3.細胞生物學觀察發(fā)現,0DPA、7DPA、24DPA和35DPA的超量轉基因株系番茄果實的果皮厚度、果皮細胞層數與對照相比顯著減小,7DPA、24DPA和35DPA的果皮細胞平均面積較對照顯著的減小。隨著時間的推移,果皮的厚度、細胞層數和細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,其他時間內這三者雖然有所增加,但是變化較小。因此推測SlSINA1可能通過影響細胞的大小和數量影響果皮厚度而調控番茄果實大小的發(fā)育。4.酵母雙雜交實驗表明,SlSINA1能夠與RAP30、SPG和FW2.2蛋白互作,SlSINA1可能與這些蛋白形成復合體發(fā)揮功能。5.在SlSINA1超量轉基因系中進行表達量分析結果顯示,較對照而言,SlSINA1表達量顯著增加,而RAP30、SPG和FW2.2的表達量均顯著下降。6.通過Western-blot實驗,未能檢測到SPG和FW2.2蛋白的表達,Co-IP驗證未發(fā)現SlSINA1與RAP30在植物體內存在互作關系。
[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.
【學位授予單位】：華中農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S641.2

【參考文獻】

相關期刊論文 前2條

1 胡婷麗;李魏;劉雄倫;戴良英;;泛素化在植物抗病中的作用[J];微生物學通報;2014年06期

2 宋素勝;謝道昕;;泛素蛋白酶體途徑及其對植物生長發(fā)育的調控[J];植物學通報;2006年05期

相關博士學位論文 前1條

1 歐陽波;幾種病程相關蛋白基因轉化番茄的研究[D];華中農業(yè)大學;2003年

相關碩士學位論文 前1條

1 侯政;利用酵母雙雜交系統(tǒng)篩選與番茄RIN互作蛋白的初步研究[D];華中農業(yè)大學;2009年

，

本文編號：1590875