【作者】 王明慧； 李炳辰； 殷焕顺； 艾仕云；

【Author】 Minghui Wang;Bingchen Li;Huanshun Yin;Shiyun Ai;College of Chemistry and Material Science, Shandong Agricultural University;

【机构】 山东农业大学化学与材料科学学院；

【摘要】 MicroRNA是一类重要的非编码RNA,在植物生长调控中发挥着重要的作用。本文提出了一种基于光电材料和酶催化原位生成电子供体技术的光电化学分析方法,实现对microRNA-319a的灵敏检测。以CuO-CuWO₄作为光活性材料,利用滚环扩增反应,磷酸根与Phos-tag的特异性识别以及生物素和亲和素的特异性识别作用,构建了一种灵敏的光电化学生物传感器,并将其成功的应用于检测外源植物激素对水稻幼苗叶片中microRNA-319a表达水平的影响。该生物免疫传感器对microRNA的检测呈现出高灵敏度,检出限约为0.13 fM（S/N=3）,可以灵敏地检测出microRNA-319a在植物细胞中的含量,同时可以用于检测不同激素作用后植物中microRNA表达量的变化。本文为研究microRNA参与植物激素调控水稻幼苗叶片发育的路径提供了新的研究方法和技术,同时可以为研究植物激素信号网络的基本原理做出巨大贡献。更多还原

【Abstract】 MicroRNAs play a crucial role in regulating plant growth, such as involving in the pathways of phytohormons signaling, which function in broad range of important stages of plant development and evolution.To figure out the components part of microRNA in phytohormone signaling network, a sensitive photoelectrochemical biosensor was developed to detect miRNA-319a specifically using CuO-CuWO₄ as photoactive material and in situ generation electron donor technique.Taking the advantage of rolling circle amplification, nicking endonuclease triggered exponential amplification of target molecule, specific interaction between phosphate group and phos-tag, and between biotin and avidin, the developed strategy can greatly amplify the detection signal upon trace amount of target miRNA-319a.The wide linear range was obtained from 0.1 nM to 1 fM, and the detection limit was estimated to be 0.13 fM（S/N=3）.The developed method also showed good detection specificity.The effect of five phytohormones on the expression level of miRNA-319 a in rice leaves was also investigated.This can make huge contribution for the basal principle of investigating the signaling network of phytohormones.更多还原