拟南芥Argonaute10调控不定芽再生的分子机制研究

植物不定芽再生机制研究已成热点，但有关转录后调控因子的作用仍不清楚。我们发现RNA沉默复合效应子突变体ago10（敲除系）和miR166a激活标签突变体men1的不定芽再生能力高于野生型，推测AGO10通过调控miR165/6表达影响不定芽再生。在此基础上，本研究拟利用Col-0、ago10、35S::AGO10、MIM165/6（敲除系）及men1进行以下研究：1）分析上述材料不定芽形成过程，确定AGO10和miR165/6对不定芽形成不同阶段的作用大小，并利用植物杂交互补，确定二者的遗传学关系；2）分析不定芽再生相关基因（如干细胞相关、激素途径等）在ago10和men1背景下的表达变化，确定它们的协调作用关系；3）利用miRNA芯片，筛选受AGO10调控的候选miRNA，分析其突变体不定芽再生能力，确定目标miRNA及可能参与的调控网络，最终揭示AGO10调控离体苗再生的分子机制。

The study of shoot regeneration mechanism has become a research focus these years. However, the post-transcriptional control of shoot regeneration is not clear now. In our previous studies, the knockout mutant ago10 and activation-tagged mutant men1 exhibited increased shoot regeneration capabilities. We presume that AGO10 controls shoot regeneration by regulating the expression of miR165/6. In this study, the following experiments will be carried out with Col-0, ago10, 35S::AGO10, MIM165/6 and men1: 1) determine which step was influenced by AGO10 and miR165/6 through analyzing the shoot regeneration processes and analyze the genetic relationship of AGO10 and miR165/6 by plant crossing; 2) analyze the expression of key genes of shoot regeneration in ago10 and men1 background; 3) screen the candidate genes regulated by AGO10 and finally illustrate the molecular mechanism of AGO10 in regulating shoot regeneration.