分析棉花MAPKs基因家庭并研究MAPK对黄萎病抗性的调控
发布时间：2019-06-21 12:13
【摘要】：在真核生物中,MAP激酶(促分裂原活化蛋白激酶)级联途径是高度保守的重要的信号传导途径,三种逐级磷酸化的蛋白激酶MAPKKK、MAPKK和MAPK组成了MAP激酶级联信号传导系统(MAPKs)。植物的生长发育、胁迫响应以及抗病反应等过程的调控都有MAPKs信号传导途径的参与。鉴于目前尚不全面了解棉花MAPKs各家族成员的组成和特征,并且对棉花MAPKs信号传导系统在抗黄萎病中的功能和作用也缺乏细致的研究。为此,本文利用现已公布的雷蒙德氏棉和亚洲棉的基因组数据,并从NCBI数据库中收集陆地棉的MAPKs氨基酸序列,运用同源比对的方法对三种棉花(陆地棉、雷蒙德氏棉、亚洲棉)的MAPK、MAPKK和MAPKKK的氨基酸序列进行了聚类分析。结果显示:棉花中MAPK家族具有特征性结构TEY磷酸化位点或TDY磷酸化位点,依据其氨基酸序列可将其划分为A、B、C和D族4个族,其中TEY类包含A、B和C族,TDY类只包含D族;棉花MAPKK家族具有特征性保守区域S/T-X5-S/T和活性部位基序D(I/L/V)K,依据其氨基酸序列可将其划分为A、B、C和D 4个族;棉花MAPKKK家族具有MEKK的特征性保守区域G(T/S)PX(W/Y)MAPEV和Raf的特征性保守区域GTXX(W/Y)MAPE(L/V),可将其分为MEKK类和Raf类两组。依据以上聚类分析的结果,从MAPK家族的A、B、C和D各族中选择部分成员。通过采用VIGS(virus induced gene silencing)技术,即病毒诱导的基因沉默技术,来研究以上这些MAPK基因在调控棉花抗黄萎病中的功能和作用。所选编码MAPK的基因分别为: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。研究结果表明: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,棉花对黄萎病菌的抗性未受到明显的影响;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,棉花对黄萎病菌表现更为敏感,也表明了这13个MAPK基因调控的信号传导通路可能在棉花抗黄萎病中起重要调控作用。以上这些基础工作为开拓棉花抗黄萎病信号传导途径的研究提供了可参考的基础数据。
[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.
【学位授予单位】：山西师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.62
本文编号：2504065
[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.
【学位授予单位】：山西师范大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.62
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