Identification of a novel small bioactive molecule and its effect on stomata development

商业上可获得的小生物活性分子，如抗生素和独特的杀虫剂，已被用于不同的目的，以增加全球范围内的公共卫生。此外，生物活性分子在基础研究中也变得越来越重要，因为它们通过调节而不是整体改变生物系统，为迄今为止已知的经典遗传方法提供了一种替代方法。Torii实验室与名古屋的转化生物分子研究所（ITbM）合作，对拟南芥野生型幼苗进行了基于表型的化学筛选，以鉴定影响气孔发育的新型生物活性小化合物。一种名为Kc9的小生物活性分子已被鉴定出以剂量依赖的方式增加气孔数量。基于初步表型和生化分析，目前的假设是Kc9通过直接或间接靶向MPK3/MPK6在气孔发育和植物免疫之间起着分子开关作用。因此，在1D-DART检测中，胞质丙酮酸激酶被确定为Kc9的潜在靶标。Walter Benjamin项目让我有机会更详细地研究这种新的生物活性小分子及其在气孔发育中的作用。在这项研究的范围内，我计划在体外和植物中确认并进一步表征Kc9的潜在靶点。在此基础上，进一步研究了气孔分化下游关键效应因子MPK3/MPK6的分子调控机制、分子网络及其串扰。

Commercially available, small bio active molecules such as antibiotics and distinctive pesticides have been used for different purposes to increase public health on a global scale. Besides for the common usage, bio active molecules become more and more important for basic research, since they provide an alternative to the so far known classical genetic methods, through modulation but not overall changes within a biological system. In collaboration with the Institute of Transformative Bio-Molecules (ITbM) in Nagoya, the Torii laboratory performed a phenotype-based chemical screen with Arabidopsis wildtype seedlings to identify novel bioactive small compounds, influencing stomata development. A small bioactive molecule named Kc9 has been identified to increase the number of stomata in a dose dependent manner. Based on preliminary phenotypical and biochemical analyses, the current hypothesis is that Kc9 acts as molecular switch between stomata development and plant immunity by either direct or indirect targeting of MPK3/MPK6. Thus, cytosolic pyruvate kinases have been identified to be the potential target of Kc9 in a 1D-DART assay. The Walter Benjamin Programme would give me the opportunity to investigate this new small bio active molecule and its role during stomata development in more detail. Within this scope of this fellowship I plan to confirm and further characterize the potential target of Kc9 both in vitro as well in planta. Furthermore, I try to decipher the molecular regulation behind key downstream effectors of stomatal differentiation such as MPK3/MPK6 as well the molecular network and their crosstalk more closely.