摘要
丝裂原活化蛋白激酶(mitogen-activated protein kinase, MAPK)级联途径,由三种级次磷酸化的MAPKKK (MAPK kinase kinase)、MAPKK (MAPK kinase)和MAPK蛋白激酶组成,广泛参与植物防御反应的调控。本文利用已发布的陆地棉Gossypium hirsutum L.全基因组数据,对其中的MAPK家族基因编码序列进行多序列比对和聚类分析。结果显示,53个GhMAPK大都含有植物MAPK保守的TXY基序,并且能够聚集到AtMAPK的A、B、C、D四个族中。进而利用病毒诱导的基因沉默(virus-induced gene silencing, VIGS)技术,研究GhMAPK是否参与调控棉花对链格孢Alternaria alternata抗性。链格孢是引起棉花轮纹斑病并导致棉花早衰的主要病原真菌。结果发现,沉默GhMAPK3、GhMAPK7、GhMAPK9及GhMAPK19的棉花植株,抗病性显著下降,表明这4种GhMAPK正调控棉花对链格孢的抗性。进一步利用靶向性沉默策略,验证这4种GhMAPK中各同源基因对棉花链格孢抗性的调控作用。结果显示,仅同源基因GhMAPK3b、GhMAPK7b、GhMAPK7c、GhMAPK9a、GhMAPK9b、GhMAPK19b和GhMAPK19c参与正调控棉花对链格孢的抗性,而同源基因GhMAPK3a、GhMAPK7a、GhMAPK7d和GhMAPK19a并不调控棉花对链格孢的抗性。
Mitogen-activated protein kinase(MAPK) cascades, which are comprised of the hierarchically phosphorylated MAPKKK(mitogen-activated protein kinase kinase kinase), MAPKK(mitogen-activated protein kinase kinase) and MAPK, normally involved in plant defense signal transduction and regulation processes. In this study, all MAPK family genes in upland cotton(Gossypium hirsutum L.) genome were collected, and then multi-sequence alignment and cluster analysis were performed. The results showed that most of the 53 GhMAPKs contained the conserved TXY motifs of plant MAPK. GhMAPKs could be classified into A, B, C and D groups corresponding to the classification of AtMAPKs. Virus induced gene silencing(VIGS) technology was further used to test whether the GhMAPKs were involved in regulating cotton resistance to Alternaria alternata, the major fungal pathogen causing cotton Alternaria leaf spot disease and leading cotton premature senescence. The results showed that the GhMAPK3, GhMAPK7, GhMAPK9 and GhMAPK19 silenced cotton plants were significantly decreased in resistance to A.alternata. The results indicated that these four types of GhMAPKs positively regulated cotton resistance to A.alternata. Then the cotton resistance regulation by each homologous gene of the four types of GhMAPKs was further tested by target silencing strategy. The results showed that only the homologous genes GhMAPK3b, GhMAPK7b, GhMAPK7c, GhMAPK9a, GhMAPK9b, GhMAPK19b and GhMAPK19c were involved in positive regulation of the cotton resistance to A.alternata. The homologous genes GhMAPK3a, GhMAPK7a, GhMAPK7d and GhMAPK19a were not involved in regulation of the cotton resistance to A.alternata.
引文
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