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首页> 外文会议>Conference on multiphoton microscopy in the biomedical sciences >Quantitative 3-Dimensional Imaging of Auxin and Cytokinin Levels in Transgenic Soybean and Medicago Truncatula Roots via Two-Photon Induced Fluorescence Imaging
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Quantitative 3-Dimensional Imaging of Auxin and Cytokinin Levels in Transgenic Soybean and Medicago Truncatula Roots via Two-Photon Induced Fluorescence Imaging

机译：通过双光子诱导荧光成像对转基因大豆和and藜根中生长素和细胞分裂素水平的三维定量成像

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Industrial nitrogen fertilizers account for nearly 50% of the fossil fuel costs in modern agriculture and contribute to soil and water pollution. Therefore, significant interest exists in understanding and characterizing the efficiency of nitrogen fixation, and the biochemical signaling pathways which orchestrate the plant-microbial symbiosis through which plants fix nitrogen. Legume plant species exhibit a particularly efficient nitrogen uptake mechanism, using root nodules which house nitrogen-fixing rhizobial bacteria. While nodule development has been widely studied, there remain significant gaps in understanding the regulatory hormones' role in plant development. In this work, we produce 3-dimensional maps of auxin (AX) and cytokinin (CK) hormone concentrations within model plant root tips and nodules with respect to root architecture and cell type. Soybean and Medicago plants were transfected with a two-color fluorescent vector with AX-sensitive green fluorescent protein (GFP) and CK-sensitive TdTomato (TdT). 3D images of soybean root nodules were captured using two-photon induced fluorescence microscopy. The resulting images were computationally analyzed using the localization code first developed by Weeks and later adapted by Kilfoil, and analyzed in the context of the root architecture. Statistical analysis of the resulting 3D hormone level maps reproduce-well the known roles of AX and CK in developing plant roots, and are the first quantitative description of these regulatory hormones tied to specific plant architecture. The analytical methods used, and the spatial distribution of these key regulatory hormones in plant roots, nodule primordia and root nodules, and their statistical interpretation are presented.
机译：工业氮肥占现代农业中化石燃料成本的近50％，并造成土壤和水污染。因此，人们对理解和表征固氮效率以及协调植物与微生物共生的植物-微生物共生的生化信号传导途径有着极大的兴趣。豆科植物物种利用容纳根固定氮的根瘤菌的根瘤来表现出特别有效的氮吸收机制。虽然根瘤发育已得到广泛研究，但在了解调节激素在植物发育中的作用方面仍存在巨大差距。在这项工作中，我们在模型植物的根尖和结节内生成了关于根系结构和细胞类型的生长素（AX）和细胞分裂素（CK）激素浓度的三维图。用带有AX敏感的绿色荧光蛋白（GFP）和CK敏感的TdTomato（TdT）的双色荧光载体转染大豆和苜蓿植物。使用双光子诱导荧光显微镜捕获大豆根瘤的3D图像。使用首先由Weeks开发并随后由Kilfoil修改的本地化代码对所得图像进行计算分析，并在根体系结构的上下文中进行分析。对所得3D激素水平图的统计分析很好地再现了AX和CK在植物根系发育中的已知作用，并且是这些与特定植物结构相关的调节激素的首次定量描述。介绍了所用的分析方法，以及这些关键调节激素在植物根，根瘤原基和根瘤中的空间分布，以及它们的统计解释。

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