促丝裂原活化蛋白激酶(Mitogen-activated protein kinase,MAPK)级联途径在植物的多种发育和生理过程中发挥重要作用,并应对各种生物和非生物胁迫.本研究中利用农杆菌介导的遗传转化方法获得了SlMAPKKK43过表达植株和CRISPR-Cas9介导的SlMAPKKK43敲除突变体,用灰葡萄孢接种转基因株系的离体叶片和果实,分析发现SlMAPKKK43正调控番茄对灰霉病的抗性.为进一步解析SlMAPKKK43调控番茄灰霉病抗性的分子机制,利用Pull down-MS技术筛选到已知的灰霉病调控因子S1MKK2和S1MKK4可能作为S1MAPKKK43的底物,利用体外磷酸化试验初步验证S1MAPKKK43可以磷酸化S1MKK2和S1MKK4.
SlMAPKKK43 Regulates Tomato Resistance to Gray Mold
The mitogen-activated protein kinase(MAPK)cascade plays an important role in multiple developmental and physiological processes in plants and in response to various biotic and abiotic stresses.In this study,SlMAPKKK43 overexpressed plants and CRISPR-Cas9-mediated SlMAPKKK43 knockout mutant using Agrobacterium infection genetic transformation were obtained.Through the inoculation of Botrytis cinerea with transgenic lines,we found that SlMAPKKK43 positively regulated the tolerance of tomato to Botrytis cinerea.In order to further elucidate the molecular mechanism of SlMAPKKK43 regulating the resistance of gray mold,the known gray mold regulators S1MKK2 and S1MKK4 were screened by pull down-MS technology,and in vitro phosphorylation assay was used to preliminarily verify that S1MAPKKK43 could phosphorylate S1MKK2 and SlMKK4.
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