开花植物在孢子体发生时期形成小孢子母细胞并分化为多细胞的雄性配子；绒毡层属于体细胞组织，与小孢子母细胞、小孢子相邻，并为配子体发育提供营养和信号分子。进年来人们揭示了LRR类-受体蛋白激酶在控制绒毡层细胞特征方面具有重要作用，但其信号途径不清楚。本项目在前期工作基础上，以两个花药绒毡层发育异常的LRR受体蛋白激酶突变体msp1-3和atd1开展研究。msp1-3花药没有绒毡层、小孢子母细胞异常增多；atd1绒毡层细胞降解异常。我们拟开展MSP1以及ATD1的受体蛋白激酶活性等生化分析；并利用酵母双杂交、磷酸化蛋白质组等对MSP1以及ATD1的细胞内结构域互作蛋白、磷酸化底物进行分析，确定其下游作用蛋白。并利用CRISPR等技术研究这些下游蛋白的生物功能；从而提出这两个LRR类-受体蛋白激酶在控制绒毡层细胞发育中的信号传导途径和作用机制。

Flowering plants form male reproductive cells (microsporocytes) during sporophytic generation, which subsequently differentiate into multicellular male gametes in the gametophytic generation. The tapetum is a somatic helper tissue neighboring microsporocytes and supporting gametogenesis. The mechanism controlling the specification of the tapetal cell fate and development within the anther has long been a mystery in biology. Recent investigations revealed molecular switches and signaling pathways including Leucine-Rich Repeat Receptor-Like Kinases (LRR-RLKs) underlying the establishment of tapetal cell fate and differentiation in plants. However, the underlying mechnism of LRR-RLKs controlling tapetal cell development remains unclear. In this proposal we plan to employ two mutants of LRR-RLKs, mutiple sporocyte 1-3 (msp1-3) and abnormal tapetal degeneration1 (atd1) to investigate the downstream signalling pathways of MSP1 and ATD1 using bioehcmical and genetic approaches. The protein kinase activitiies and their crutial residues of MSP1 and ATD1 will be analyzed. Their downsteam interacting proteins will be revealed by yeast two hybrid method using the kinase domains of MSP1 and ATD1 as the bait, respectively. In addition, the msp1-3 and atd1 anthers are to be used for identifying downstream components using phosphoproteomics. The function of the confirmed downstream proteins will be analyzed by genetic approaches such as CRISPR and expression analysis. Thus this work may fill the gap between LRR-RLK sginalling and the specification of tapetal development in rice.