本文選題：番茄 + WRKY　； 參考：《東北農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：番茄具有豐富的營(yíng)養(yǎng)價(jià)值、較強(qiáng)的適應(yīng)能力、栽培很容易以及很高的產(chǎn)量等特點(diǎn),因此,番茄是一種重要的世界性蔬菜,在蔬菜生產(chǎn)中占有主要地位。但是番茄在生長(zhǎng)的過(guò)程中,通常會(huì)受惡劣自然環(huán)境的影響,導(dǎo)致產(chǎn)量的下降,因此研究逆境脅迫對(duì)番茄生長(zhǎng)的影響,有著重要的意義。本文主要對(duì)番茄與抗逆相關(guān)的轉(zhuǎn)錄因子WRKY家族進(jìn)行了全基因組挖掘及生物信息學(xué)分析,并且通過(guò)對(duì)部分基因在逆境脅迫下的表達(dá)模式進(jìn)行研究,獲得了該家族中與番茄抗逆應(yīng)答相關(guān)的基因。1通過(guò)生物信息學(xué)分析,鑒定出了番茄WRKY家族包含81個(gè)成員,按其WRKY結(jié)構(gòu)域及鋅指結(jié)構(gòu)分為3族,第二族按其氨基酸順序又可以分為5個(gè)亞族。除番茄的11號(hào)染色體沒(méi)有WRKY轉(zhuǎn)錄因子,其他染色體上均有分布。僅Sl WRKY19,Sl WRKY20倆個(gè)家族成員不含內(nèi)含子,其他成員都含有2~5個(gè)內(nèi)含子。番茄WRKY結(jié)構(gòu)域是高度保守的,僅有10個(gè)WRKYGQK發(fā)生了變異,占WRKY家族的10%。2對(duì)6個(gè)典型WRKY轉(zhuǎn)錄因子進(jìn)行組織特異性表達(dá)模式分析,發(fā)現(xiàn)這些基因在根、莖、葉、花、果中均有表達(dá),但不同基因在同一組織中的表達(dá)量存在差異,同一基因在不同組織中的表達(dá)量也存在差異,Sl WRKY13和Sl WRKY24在根和花中的表達(dá)量較高,在莖中幾乎不表達(dá);Sl WRKY31在根、莖、葉中的表達(dá)量大于花和果;Sl WRKY50在根、葉、花中的表達(dá)量較高,在莖中幾乎不表達(dá);Sl WRKY62在根、葉、果中的表達(dá)量較高,在莖中幾乎不表達(dá);Sl WRKY63在根中的表達(dá)量最高,在莖和果中幾乎不表達(dá)。3在鹽脅迫處理中,SlWRKY13、Sl WRKY31、Sl WRKY50、Sl WRKY62和Sl WRKY63基因的表達(dá)水平呈現(xiàn)先上升,然后逐漸下降的趨勢(shì),而Sl WRKY24基因的表達(dá)則顯著下調(diào);在干旱脅迫處理中,Sl WRKY13、Sl WRKY31、Sl WRKY62和Sl WRKY63基因表達(dá)上調(diào),Sl WRKY24和Sl WRKY50基因表達(dá)下調(diào);在低溫脅迫處理中,Sl WRKY13、Sl WRKY31、Sl WRKY50、Sl WRKY62和Sl WRKY63基因表達(dá)上調(diào),Sl WRKY24基因表達(dá)下調(diào)。4對(duì)逆境脅迫應(yīng)答分析中表達(dá)量變化較顯著的基因Sl WRKY50進(jìn)行了基因沉默分析,發(fā)現(xiàn)該基因沉默后,植株在低溫脅迫逆境條件下的MDA指標(biāo)與相同條件下的對(duì)照植株相比顯著上升,而Pro和SOD指標(biāo)顯著下降,說(shuō)明該基因的下調(diào)表達(dá)可以降低植株的抗寒性。
[Abstract]:Tomato has rich nutritional value, strong adaptability, easy cultivation and high yield. Therefore, tomato is an important vegetable in the world and plays a major role in vegetable production. But in the process of tomato growth, it is usually affected by the adverse natural environment, which leads to the decline of the yield. Therefore, it is of great significance to study the effects of stress on the growth of tomato. In this paper, the whole genome mining and bioinformatics analysis of WRKY family of tomato transcription factors related to stress resistance were carried out, and the expression patterns of some genes under stress were studied. By bioinformatics analysis, 81 members of tomato WRKY family were identified and divided into 3 families according to their WRKY domain and zinc finger structure. The second group can be divided into 5 subfamilies according to its amino acid sequence. Except for tomato chromosome 11, there were no WRKY transcription factors, but all other chromosomes were distributed. Only Sl WRKY19Sl WRKY20 family members do not contain introns, while the other members contain 2 ~ 5 introns. The WRKY domain of tomato was highly conserved, and only 10 WRKYGQK were mutated. The tissue specific expression patterns of 6 typical WRKY transcription factors were analyzed in 10.2% of the WRKY family. The results showed that these genes were expressed in root, stem, leaf, flower and fruit. However, there were differences in the expression levels of different genes in the same tissue, and there were also differences in the expression levels of SL WRKY13 and SL WRKY24 in different tissues. The expression of SL WRKY13 and SL WRKY24 in roots and flowers was higher than that in stems, and there was almost no expression of SL WRKY31 in roots and stems. The expression of Sl WRKY50 in leaves was higher than that in flowers, leaves and flowers, and the expression of Sl WRKY62 in roots, leaves and fruits was higher than that in stems, and the highest expression of Sl WRKY63 in roots was found in stems. In stem and fruit, the expression level of Sl WRKY62 and SL WRKY63 gene increased at first, then decreased gradually, while the expression of SL WRKY24 gene decreased significantly in the treatment of salt stress, while the expression level of Sl WRKY13 Sl WRKY31nSl WRKY50 Sl WRKY63 gene increased at first and then decreased gradually in both stem and fruit. The expression of Sl WRKY13 Sl WRKY31 WRKY62 and SL WRKY63 genes in drought stress was up-regulated, and the expression of Sl WRKY24 and SL WRKY50 genes were down-regulated. Under low temperature stress, Sl WRKY13 + Sl WRKY31Sl WRKY50Sl WRKY62 and SL WRKY63 down-regulated the down-regulation of Sl WRKY24 gene expression. The gene silencing of the gene SL WRKY50, which changed significantly in stress response analysis, was found after the gene silencing. The MDA index of the plant under low temperature stress was significantly higher than that of the control plant under the same condition, while the Pro and SOD indexes were significantly decreased, which indicated that the down-regulation of the gene expression could reduce the cold resistance of the plant.
【學(xué)位授予單位】：東北農(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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