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Chemical stimulation of the Avabidopsis thaliana root using multi-laminar flow on a microfluidic chip

机译:在微流控芯片上使用多层流化学刺激拟南芥根

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In this article, we developed a "plant on a chip" microfluidic platform that can control the local chemical environment around live roots of Arabidopsis thaliana with high spatial resolution using multi-laminar flow. We characterized the flow profile around the Arabidopsis root, and verified that the shear forces within the device (~10 dyne cm~(-2)) did not impede growth of the roots. Our platform was able to deliver stimuli to the root at a spatial resolution of 10-800 urn. Further, the platform was validated by exposing desired regions of the root with a synthetic auxin derivative, 2,4-dichlorophenoxyacetic acid (2,4-D), and its inhibitor N-naphthylphthalamic acid (NPA). The response to the stimuli was observed using a DR5::GFP Arabidopsis line, where GFP expression is coupled to the auxin response regulator DR5. GFP expression in the root matched the position of the flow-focused stream containing 2,4-D. When the regions around the 2,4-D stimulus were exposed to the auxin transport inhibitor NPA, the active and passive transport mechanisms of auxin could be differentiated, as NPA blocks active cell-to-cell transport of auxin. Finally, we demonstrated that local 2,4-D stimulation in a ~10 μm root segment enhanced morphological changes such as epidermal hair growth. These experiments were proof-of-concept and agreed with the results expected based on known root biology, demonstrating that this "root on a chip" platform can be used to test how root development is affected by any chemical component of interest, including nitrogen, phosphate, salts, and other plant hormones.
机译:在本文中,我们开发了一种“芯片上种植”微流体平台,该平台可以使用多层流控制高分辨率的拟南芥活根周围的局部化学环境。我们对拟南芥根周围的水流特征进行了表征,并验证了装置内的剪切力(〜10达因厘米〜(-2))没有阻碍根的生长。我们的平台能够以10-800 um的空间分辨率向根部传递刺激。此外,通过用合成生长素衍生物2,4-二氯苯氧基乙酸(2,4-D)及其抑制剂N-萘基邻苯二甲酸酯(NPA)暴露根部的所需区域来验证平台。使用DR5 :: GFP拟南芥系观察到对刺激的应答,其中GFP表达与生长素应答调节剂DR5偶联。根中的GFP表达与包含2,4-D的以流量为中心的流的位置匹配。当2,4-D刺激周围的区域暴露于生长素运输抑制剂NPA时,可以区分生长素的主动和被动运输机制,因为NPA阻止了生长素的主动细胞间运输。最后,我们证明了在〜10μm根段中的局部2,4-D刺激增强了形态变化,例如表皮毛发生长。这些实验是概念验证的研究,与基于已知根生物学的预期结果相符,表明该“芯片上的根”平台可用于测试感兴趣的任何化学成分(包括氮)如何影响根发育,磷酸盐,盐和其他植物激素。

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