【摘要】：在真核生物中,MAP激酶(促分裂原活化蛋白激酶)級(jí)聯(lián)途徑是高度保守的重要的信號(hào)傳導(dǎo)途徑,三種逐級(jí)磷酸化的蛋白激酶MAPKKK、MAPKK和MAPK組成了MAP激酶級(jí)聯(lián)信號(hào)傳導(dǎo)系統(tǒng)(MAPKs)。植物的生長發(fā)育、脅迫響應(yīng)以及抗病反應(yīng)等過程的調(diào)控都有MAPKs信號(hào)傳導(dǎo)途徑的參與。鑒于目前尚不全面了解棉花MAPKs各家族成員的組成和特征,并且對棉花MAPKs信號(hào)傳導(dǎo)系統(tǒng)在抗黃萎病中的功能和作用也缺乏細(xì)致的研究。為此,本文利用現(xiàn)已公布的雷蒙德氏棉和亞洲棉的基因組數(shù)據(jù),并從NCBI數(shù)據(jù)庫中收集陸地棉的MAPKs氨基酸序列,運(yùn)用同源比對的方法對三種棉花(陸地棉、雷蒙德氏棉、亞洲棉)的MAPK、MAPKK和MAPKKK的氨基酸序列進(jìn)行了聚類分析。結(jié)果顯示:棉花中MAPK家族具有特征性結(jié)構(gòu)TEY磷酸化位點(diǎn)或TDY磷酸化位點(diǎn),依據(jù)其氨基酸序列可將其劃分為A、B、C和D族4個(gè)族,其中TEY類包含A、B和C族,TDY類只包含D族;棉花MAPKK家族具有特征性保守區(qū)域S/T-X5-S/T和活性部位基序D(I/L/V)K,依據(jù)其氨基酸序列可將其劃分為A、B、C和D 4個(gè)族;棉花MAPKKK家族具有MEKK的特征性保守區(qū)域G(T/S)PX(W/Y)MAPEV和Raf的特征性保守區(qū)域GTXX(W/Y)MAPE(L/V),可將其分為MEKK類和Raf類兩組。依據(jù)以上聚類分析的結(jié)果,從MAPK家族的A、B、C和D各族中選擇部分成員。通過采用VIGS(virus induced gene silencing)技術(shù),即病毒誘導(dǎo)的基因沉默技術(shù),來研究以上這些MAPK基因在調(diào)控棉花抗黃萎病中的功能和作用。所選編碼MAPK的基因分別為:A族的ADI52626.1G.hirsutum、Gorai.3g139900.1、ADT65136.1G.hirsutum、Gorai.5g011100.1;B族的Gorai.1g158800.5、Gorai.1g158800.1、A 32714、ADI52627.1G.hirsutum;C族的ABA00652.1G.hirsutum、Gorai.5g109500.1、Gorai.7g050000.1、ADI52622.1G.hirsutum;D族的ADI52623.1G.hirsutum、A 01430、A37089、A 25334、A 24541、A 22593、AHW85276.1G.hirsutum、AET36534.1G.hirsutum、A 22570、A 07602。研究結(jié)果表明:基因ADI52623.1G.hirsutum(D族)、A 01430(D族)、A 37089(D族)、A 25334(D族)、A 24541(D族)、AET36534.1G.hirsutum(D族)、ADI52627.1G.hirsutum(B族)、ADI52626.1G.hirsutum(A族)和Gorai.3g139900.1(A族)沉默后,棉花對黃萎病菌的抗性未受到明顯的影響;而基因AHW85276.1G.hirsutum(D族)、A 22593(D族)、A 07602(D族)、A 22570(D族)、Gorai.7g050000.1(C族)、ADI52622.1G.hirsutum(C族)、Gorai.5g109500.1(C族)、ABA00652.1G.hirsutum(C族)、Gorai.1g158800.5(B族)、Gorai.1g158800.1(B族)、A 32714(B族)、ADT65136.1G.hirsutum(A族)和Gorai.5g011100.1(A族)沉默后,棉花對黃萎病菌表現(xiàn)更為敏感,也表明了這13個(gè)MAPK基因調(diào)控的信號(hào)傳導(dǎo)通路可能在棉花抗黃萎病中起重要調(diào)控作用。以上這些基礎(chǔ)工作為開拓棉花抗黃萎病信號(hào)傳導(dǎo)途徑的研究提供了可參考的基礎(chǔ)數(shù)據(jù)。
[Abstract]:In eukaryotes, the cascade approach of MAP kinase (mitogen-activated protein kinase) is a highly conserved signaling pathway, and three levels of phosphorylated protein kinase MAPKKK, MAPKK and MAPK form a MAP kinase cascade signal transduction system (MAPKs). The development of the plant, the response of stress and the control of the disease-resistant reaction have the participation of the signal transduction pathway of MAPKs. In view of the fact that there is no comprehensive understanding of the composition and characteristics of the members of the various family members of the cotton MAPKs, the function and function of the cotton MAPKs signal transduction system in the anti-verticillium wilt are not studied in detail. To this end, this paper, using the published genome data of Raymond's cotton and Asian cotton, and collecting the MAPKs amino acid sequence of Gossypium hirsutum from the NCBI database, and using the method of homologous alignment to the MAPK of three kinds of cotton (Gossypium hirsutum, Raymond's cotton and Asian cotton), The amino acid sequences of MAPKK and MAPKKK were analyzed by cluster analysis. The results showed that the MAPK family of cotton had a characteristic structure of TEY phosphorylation site or TDY phosphorylation site, which can be divided into 4 families of group A, B, C and D according to its amino acid sequence, where the TEY class comprises group A, group B and group C, and the TDY class only contains group D; The cotton MAPKK family has a characteristic conserved region S/ T-X5-S/ T and an active site motif D (I/ L/ V) K, which can be divided into a, B, C, and D 4 families according to their amino acid sequence; the cotton MAPKKK family has a characteristic conserved region G (T/ S) PX (W/ Y) MAPEV of MEKK and a characteristic conservative region GTXX (W/ Y) MAPE (L/ V) of Raf, It can be divided into two groups of MEKK and Raf. Based on the results of the above cluster analysis, some members were selected from the group A, B, C and D of the MAPK family. The function and function of these MAPK genes in the control of cotton against verticillium wilt are studied by using the technology of VIGS (virus-induced gene silencing), that is, the gene silencing technique induced by the virus. The genes selected for coding MAPK are: ADI52626.1 G. Hirsuum, Goran.3 g139900.1, ADT65136.1 G. Hirsuum, Goran.5 g011100.1; Group B's Goran.1 g158800.5, Goran.1 g158800.1, A 32714, ADI52627.1 G. Hirsuum; Group C ABA00652.1 G. Hirsuum, Goran.5 g109500.1, Goran.7 g050000.1, ADI52622.1 G. Hirsuum; Group D's ADI52623.1 G. Hirsuum, A 01430, A37089, A 25334, A 24541, A 22593. The results of the study indicated that the resistance of the cotton to Verticillium dahliae was not significantly affected after the silencing of the genes ADI52623.1 G. Hirsuum (group D), A 01430 (D family), A 37089 (D group), A 25334 (group D), A 24541 (D family), AET36534.1 G. Hirsuum (Group D), ADI52627.1 G. Hirsuum (Group B), ADI52626.1 G. Hirsuum (Group A) and Goran.3 g 139900.1 (A), and the gene AHW8527.1 G. Hirsuum (D family), A 22593 (D family), A 07602 (D family), The cotton is more sensitive to Verticillium dahliae, after the silences of the A 22570 (Group D), Goran.7 g050000.1 (Group C), the ADI52622.1 G. Hirsuum (Group C), Goran.5 g109500.1 (Group C), ABA00652.1 G. Hirsuum (Group C), Goran.1 g158800.5 (Group B), Goran.1 g158800.1 (Group B), A 32714 (Group B), ADT65136.1 G. Hirsuum (Group A) and Goran.5 g011100.1 (Group A), It is also shown that the signal transduction pathway regulated by the 13 MAPK genes may play an important role in the resistance to verticillium wilt of cotton. The above basic work provides the reference basic data for the research of the signal transduction pathway of the cotton against the Verticillium wilt.
【學(xué)位授予單位】：山西師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S435.62

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