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内容記述 |
Plants are exposed to a variety of environmental stresses, and starvation of inorganic phosphorus could be a major constraint for crop production. In plants, in response to phosphate deficiency in soil, miR399, one kind of miRNA, is upregulated. By detecting miR399 in a simple manner, early diagnosis of phosphorus deficiency stress in plants could be accomplished. However, since general miRNA detection methods require complicated experimental manipulations, simple and rapid miRNA detection methods are needed for plant nutritional diagnosis. For the simple detection of miR399, microfluidic technology is suitable for point of care (POC) application because of its ability to detect target molecules in small amounts, in a short time, and with simple manipulation. In this study, we developed microfluidic device to detect miRNAs from rough purified plant extract for easy diagnosis of plant nutritional conditions. In this technique, target miRNAs are detected by fluorescence of sandwich hybridization in a microfluidic channel. For plant stress diagnostics, we developed a protocol for miRNA detection by microfluidic technology by validating the sample preparation buffer, rough purification by filtering,
sample injection method, and signal amplification method. With this system, endogenous sly-miR399 in tomatoes expressed in response to phosphorus deficiency was detected before stress symptoms appeared. |
