本文選題：甘藍型油菜 + BnaSOT基因　； 參考：《湖南農業(yè)大學》2016年碩士論文

【摘要】：磺酸基轉移酶(SOT, EC 2.8.2.-)是在動植物都廣泛存在的一類重要的蛋白家族,能夠催化磺酸基團從供體到底物的轉移反應,使底物發(fā)生磺酸化。研究表明,擬南芥中有22個編碼磺酸基轉移酶(SOT)的基因(AtSOT),其中,AtSOT12基因的表達能夠被鹽、滲透壓力和激素誘導而大大加強。并且AtSOT12基因的T-DNA插入突變體(sot12)在發(fā)芽率上表現(xiàn)出對NaCl和脫落酸(ABA)的高度敏感性。本研究對SOT蛋白家族在白菜、甘藍和甘藍型油菜中的分布和分類情況進行了系統(tǒng)的生物信息學分析。在此基礎上結合蛋白和基因結構的分析,選取了3個甘藍型油菜BnaSOT12基因作為試驗研究對象。構建了BnaSOT12的過表達載體,并對擬南芥和甘藍型油菜進行了遺傳轉化實驗。獲得了如下試驗結果：1.白菜基因組的分析表明：白菜中存在56個編碼BraSOT蛋白的基因,其中53個基因能夠分別被定位在白菜的染色體上�；蛟谌旧w上的分布并不均勻：A07號和A09號染色體含BraSOT基因最多,A05號染色體不包含BraSOT基因。56個基因能夠較好的與擬南芥一致被分類為9個亞家族。多數BraSOT基因表現(xiàn)出沒有內含子的特征。2.甘藍基因組的分析表明：甘藍基因組存在33個編碼BolSOT蛋白的基因,分別定位到甘藍的9條染色體上,并且在C03號和C06號染色體上分別最為密集。33個BolSOT蛋白的分類很不均勻,第7類亞家族成員最多,沒有與擬南芥第4和第9類亞家族蛋白對應的同源蛋白。多數BolSOT基因不含有內含子,且各基因長度相差較大。3.甘藍型油菜基因組的分析表明：甘藍型油菜基因組存在有115個編碼BnaSOT蛋白的基因。這些基因在A07、A09、C06和C09號染色體上的分布最多。其中AtSOT12基因的同源基因有13個,表明該基因在甘藍型油菜中進行了復制和倍增,暗示其具有重要的生物學功能。進而通過基因結構和蛋白結構的對比,確定了在結構上與AtSOT12相似度最高的三個基因：BnaC02g35150D、BnaA09g53490D、BnaA09g19440D,分別命名為BnaSOT12a、BnaSOT12b、BnaSOT12c,作為試驗研究對象。4.分別構建了BnaSOT12a、BnaSOT12b、BnaSOT12c基因的過表達載體,獲得了BnaSOT12a的過表達植株。對突變體的抗鹽性實驗表明,甘藍型油菜BnaSOT12a基因在擬南芥中的過表達,能提高轉基因植株對NaCl脅迫的耐受能力。5.利用pFGC5941：:35S:BnaSOT12a載體對甘藍型油菜進行了遺傳轉化,分別應用了子葉柄農桿菌浸染法和下胚軸農桿菌浸染法兩種方法。子葉柄農桿菌浸染法轉化湘油-15甘藍型油菜獲得9株抗性植株,轉化率為0.77%；下胚軸農桿菌浸染法中使用了湘油-15甘藍型油菜和中雙-6甘藍型油菜兩種材料,總共獲得92株分化的抗性苗,在產生愈傷組織和分化出不定芽的過程中,中雙-6相對于湘油-15甘藍型油菜能表現(xiàn)出更強的分化能力。
[Abstract]:Sulfonic acid transferase (EC 2.8.2.-) is an important protein family which exists widely in animals and plants. It can catalyze the transfer reaction of sulfonic groups from the donor to the end and make the substrates sulfonate. In Arabidopsis thaliana, there are 22 genes encoding SOT, in which the expression of AtSOT12 gene can be greatly enhanced by salt, osmotic pressure and hormone induction. Moreover, the T-DNA insertion mutant of AtSOT12 gene was highly sensitive to NaCl and abscisic acid (Aba) in germination rate. The distribution and classification of SOT protein family in Chinese cabbage, cabbage and Brassica napus were systematically analyzed by bioinformatics. Based on the analysis of protein and gene structure, three BnaSOT12 genes of Brassica napus were selected. The overexpression vector of BnaSOT12 was constructed and genetic transformation of Arabidopsis thaliana and Brassica napus was carried out. The following results are obtained: 1: 1. The genome analysis of Chinese cabbage showed that 56 genes encoding BraSOT protein existed in Chinese cabbage, 53 of which could be located on the chromosomes of Chinese cabbage. The distribution of the genes on chromosomes was not uniform. Chromosome A05 contained the most BraSOT genes, and chromosome A05 did not contain BraSOT genes. 56 genes could be classified into 9 subfamilies in good agreement with Arabidopsis thaliana (Arabidopsis thaliana). Most BraSOT genes showed no intron characteristics. 2. The analysis of cabbage genome showed that there were 33 genes encoding BolSOT protein in cabbage genome, which were located on 9 chromosomes of cabbage, and were the most dense on chromosome C03 and C06, respectively. The classification of 33 BolSOT proteins was very uneven. The 7 th subfamily had the most members and had no homologous proteins corresponding to the 4 and 9 subfamily proteins of Arabidopsis thaliana. Most BolSOT genes do not contain introns, and the length of each gene varies by 3. 3. The genome analysis of Brassica napus showed that there were 115 genes encoding BnaSOT protein in Brassica napus genome. These genes were most distributed on chromosomes A 07, A 09, C 06 and C 09. There were 13 homologous genes of AtSOT12 gene, which indicated that the gene was duplicated and multiplied in Brassica napus, suggesting that it had important biological function. Through the comparison of gene structure and protein structure, three genes, BnaC02g35150D, BnaA09g53490Dand BnaA09g19440D, were identified as BnaSOT12bnBnaSOT12BnaSOT12c. The overexpression vectors of BnaSOT12b BnaSOT12b BnaSOT12c gene were constructed, and the overexpression plants of BnaSOT12a were obtained. The salt-tolerance experiment of mutant showed that the overexpression of BnaSOT12a gene in Arabidopsis thaliana could improve the tolerance of transgenic plants to NaCl stress. The genetic transformation of Brassica napus was carried out by pFGC5941::35S:BnaSOT12a vector. Two methods, Agrobacterium tumefaciens soaking in cotyledon petiole and Agrobacterium tumefaciens in hypocotyls, were used. Nine resistant plants were obtained by Agrobacterium tumefaciens soaking method, and the transformation rate was 0.77. Two kinds of materials, Xiangyou -15 Brassica napus and medium double -6 Brassica napus were used in Hypocotyl Agrobacterium tumefaciens soaking method. A total of 92 differentiated resistant seedlings were obtained. In the process of callus formation and adventitious bud differentiation, medium and double -6 showed stronger differentiation ability than Xiangyou-15 Brassica napus.
【學位授予單位】：湖南農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S565.4

【參考文獻】

相關期刊論文 前10條

1 蔡曉鋒;胡體旭;葉杰;張余洋;李漢霞;葉志彪;;植物鹽脅迫抗性的分子機制研究進展[J];華中農業(yè)大學學報;2015年03期

2 夏惠;林玲;高帆;呂秀蘭;;植物脫水素對多種逆境的響應[J];干旱地區(qū)農業(yè)研究;2014年04期

3 楊靚;吳祖建;;蛋白激酶與植物滲透脅迫耐受研究進展[J];武夷科學;2012年00期

4 陳全求;詹先進;藍家樣;黃云;符家平;;湖北棉花育種現(xiàn)狀與發(fā)展[J];安徽農學通報(上半月刊);2009年19期

5 李榮華;夏巖石;劉順枝;孫莉麗;郭培國;繆紳裕;陳健輝;;改進的CTAB提取植物DNA方法[J];實驗室研究與探索;2009年09期

6 吳永美;呂炯章;王書建;李潤植;;植物抗旱生理生態(tài)特性研究進展[J];雜糧作物;2008年02期

7 邴雷;趙寶存;沈銀柱;黃占景;葛榮朝;;植物耐鹽性及耐鹽相關基因的研究進展[J];河北師范大學學報(自然科學版);2008年02期

8 高銀;;植物抗逆機制與基因工程研究進展[J];內蒙古農業(yè)科技;2007年05期

9 楊貴春,高繼國,邢少辰;植物抗非生物脅迫基因工程的研究進展[J];吉林農業(yè)科學;2005年04期

10 曾華宗,羅利軍;植物抗旱、耐鹽基因概述[J];植物遺傳資源學報;2003年03期

相關碩士學位論文 前1條

1 劉兆華;擬南芥中與非生物脅迫相關基因UGT-LIKE、UF1和UF2的初步研究[D];山東農業(yè)大學;2012年

，

本文編號：1780154