STRUCTURE FUNCTION ANALYSIS OF INFORMATION PROCESSING IN SENSORY SYSTEMS

植物细胞信号转导的分子基础仍然是 很大程度上是未知的。虽然钙离子被认为是第二种 植物细胞中的信使，胞内钙的时空变化 离子对各种环境刺激的反应一直不是很好 由于成像中的技术困难而具有特征性。我们有 开发了一种新的方法来加载钙指示剂 钙绿-1和Indo-1进入植物细胞的细胞质，包括 气孔保卫细胞。气孔控制气体交换包括 H2O损失(蒸腾)和CO2吸收。理解规则 气孔性对今后的遗传改良具有重要意义 气体交换。激素脱落酸(ABA)导致气孔孔 在脱水的条件下。尽管ABA信号中的大部分 途径尚不清楚，已有研究提出， 胞质中的钙离子是已知的最早的ABA信号事件。使用 新开发的方法将钙指示剂加载到 蒲公英保卫细胞和表皮细胞的细胞质 Commis，我们监测了钙离子的时空变化 激光荧光共聚焦检测ABA对浓度的响应 成像技术。此外，我们已经扩展了这一方法，并 正在进行分子遗传学研究，以确定ABA是如何诱导 拟南芥ABA信号突变体对钙离子浓度的影响 塔里安娜。我们还开发了比率聚焦成像技术 包括拟南芥在内的植物细胞的S技术。这些研究是 允许对克隆的遗传基因座的功能进行细胞分析 用于拟南芥的ABA信号转导。我们还计划将这些应用于 识别和响应其他植物细胞的技术 用木糖激发子阐明其分子机制 计划特异性密度反应的信号转导。我们有 一直在使用共聚焦显微镜和图像分析设备在 NCMIR，包括在NCMIR开发的计划B Track。

Molecular bases for signal transduction in plant cells are still largely unknown. Though calcium ion was proposed as a second messenger in plant cells, spatio-temporal changes in cytosolic calcium ion in response to various environmental stimuli have not been well characterized due to the technical difficulties in imaging. We have developed a novel method to load the calcium indicators such as calcium green-1 and indo-1 into the cytoplasm of plant cells including stomatal guard cells. Stomatal pores control gas exchange including H2O loss (transpiration) and CO2 uptake. Understanding the regulation of stomatal pores is important for the future genetic improvement of gas exchange. The hormone absisic acid (ABA) causes stomatal pores to close uder dehydrated conditions. Although much of the ABA signaling pathway is unknown, it has been proposed that an increase in cytoplasmic CA2+ is the earlies known ABA signaling event. Using the newly-developed method to load the calcium indicatiors into the cytoplasm of guard cells as well as epidermal cells of Commelina communis, we monitored the spatio-temporal changes in Ca2+ concentration in response to ABA by using laser fluorescence confocal imaging techniques. Furthermore, we have extended this approach and are pursuing molecular genetic studies to determine how ABA-induced CA2+ increases are affected in ABA signaling mutants of Arabidopsis thaliana. We have also developed ratiometric onfocal imaging technique s for plant cells including Arabidopsis. These studies are allowing a cellular analysis of the function of cloned genetic loci for ABA signaling in Arabidopsis. We are also planning to apply these techniques to other plant cells that recognizes and respons to ligosaccharide elicitors to elucidate the molecular mechanisms of signal transduction for plan-specific denfense responses. We have been using the confocal microscopy and image analysis facilites at the NCMIR, including the program B track developed at the NCMIR.