冷胁迫响应玉米miRNA的靶基因预测及RACE鉴定.docxVIP

PAGE \* MERGEFORMAT16 摘 要 玉米（Zea mays L.）为禾本科，玉蜀黍属一年生草本C4植物，起源于热带或亚热带地区。玉米用途十分广泛，是重要的饲料作物，又是食品、化工、燃料及医药等行业的重要原料，因此全球玉米种植面积居粮食作物首位，我国的玉米种植面积也于2008年超过水稻成为第一大粮食作物。玉米在我国国家粮食安全中起着极其重要的作用，生产生活中对玉米的需求不断增加，因而向高纬度、高海拔地区拓展种植区域的需求不断增加。但是起源于热带、亚热带地区的玉米，在生长周期中表现出对低温冷害较为敏感，在整个生长周期过程中极易遭受低温冷害的影响而导致减产。 研究表明，逆境胁迫不仅会诱导植物蛋白质编码基因的表达，也会诱导一些非蛋白质编码基因的表达，这类非蛋白质编码基因的表达产物在植物的生长、发育和应对逆境胁迫等过程中发挥重要的调控作用。microRNA（miRNA）是一类内源性小分子的非编码RNA，它通过对其靶基因mRNA的降解或抑制翻译等进行基因表达调控，进而参与调控植物相关生理活动。miRNA在植物体的新陈代谢、组织器官发育以及分化中起着重要的作用，同时可以响应多种胁迫，能够在植物感受逆境胁迫过程中，做出相应的适应性调整，在植物抵御逆境胁迫过程屮发挥着非常重要的作用。 本次研究主要是对冷胁迫响应玉米miRNA的靶基因预测与5’RACE鉴定。通过对玉米三叶一心期幼苗不同处理，提取玉米幼苗二、三叶总RNA，并对提取的总RNA进行反转录、巢式PCR、构建载体转化、培养大肠杆菌等，获得单菌落并测序对比，最终得到玉米中miR171n-3p的剪切产物和剪切位点。 关键词：玉米；冷胁迫；microRNA；RACE；靶基因  Abstract Maize （Zea mays L.） is a C4 plant species of gramineous and originated in tropical or subtropical regions.Growth and productivity of maize are severely constrained by chilling stress. Because maize is widely used as forage crop, and is an important raw material for chemical, food, fuel and medicine industry, the planting area of maize ranks first world wide, and it became the first major grain crops in China in 2008. Maize plays an extremely important role in the national food security in our country. The boosting requirement for maize demand of expanding the planting area to the high latitude and high altitude region is increasing. As originated in tropical and subtropical areas, maize showed sensitive to low temperature during the entire growth season. Studies have shown that stress can not only induce the expression of plant protein coding genes, but also induce the expression of some nonprotein-encoding genes.The expression products of these nonprotein-encoding genes play an important regulatory role in the process of plant growth, development and stress. MicroRNA （miRNA） is a kind of non-coding RNA of endogenous small molecule. It is involved in the regulation of plant-related physiological activities by regulating gene expression of its