鉴定转基因小麦中荧光蛋白的新技术应用研究  

Observation and Identification of Transgenic Wheat Based on Fluorescent Protein Labeling

作　　者：薛晓东 李勤霞 XUE Xiaodong;LI Qinxia(Shangluo University,Shangluo,Shaanxi 726000,China;Collaborative Innovation Center for Resource and Biological Comprehensive Development in Qinba Mountain Area,Hanzhong,Shaanxi 723000,China)

机构地区：[1]商洛学院,陕西商洛726000 [2]陕南秦巴山区资源生物综合开发协同创新中心,陕西汉中723000

出　　处：《陕西农业科学》2025年第1期4-8,共5页Shaanxi Journal of Agricultural Sciences

基　　金：陕西省教育厅重点科学研究计划项目(21JY008);商洛学院科学与技术研究基金项目(20SKY010)。

摘　　要：荧光显微成像技术由于特异性好、对比度高等优点,被广泛应用于动物学、神经科学、细胞生物学、分子生物学等研究领域。荧光显微成像最常用的检测仪器是荧光显微镜和活体荧光成像仪,目前活体荧光成像仪用于植物领域的检测分析报道较少。本研究以已获得的含有红色荧光蛋白标记(DsRed)和绿色荧光蛋白标记(GFP)的小麦种子为材料,通过荧光显微镜和活体成像仪观察及与PCR的对比进行分析,结果表明:利用荧光显微镜观察到的荧光表达范围主要是根尖及幼芽着生处,可以针对单个样品精细观察,但是由于视野所限,难以大批量分析,而活体荧光成像仪可同时观察的植株数量较多,通量高,观察GFP和RFP时,都能与野生型明显的区分开,特异性较好,不会出现背景噪音,且不同于荧光显微镜的是RFP的标准值更高,也更明亮。经过PCR鉴定分析,两种设备均能正确筛选出阳性植株,且结果较为一致。本研究可以为表达荧光蛋白基因植株的鉴别和筛选提供理论依据和技术参考。Fluorescent microscopic imaging technology is widely used in zoology,neuroscience,cell biology,molecular biology,and other research fields due to its specificity and contrast.The most commonly used detection instruments for fluorescence microscopy imaging are fluorescence microscopy and in vivo imagers,although in vivo imagers have not been extensively reported for plant detection and analysis.This study analyzed the wheat seeds containing red fluorescent markers(DsRed)and green fluorescent markers(GFP)using fluorescence microscopy,in vivo imaging,and PCR comparative observation.The results showed that the fluorescence expression observed with fluorescence microscopy was mainly located at the root tip and young bud growth regions,allowing detailed observation of individual samples.However,due to the limited field of view of the microscope,conducting large-scale analysis was challenging.The in vivo fluorescence imaging instrument could simultaneously observe a large number of plants with high throughput.When observing GFP and RFP,these markers could be clearly distinguished from the wild-type exhibiting high specificity and minimal background noise.Moreover,unlike fluorescence microscopy,the fluorescence intensity of RFP was higher and brighter.After PCR identification and analysis,both devices were able to correctly screen positive plants,and the results were consistent.This study provides theoretical basis and technical support for the identification and screening of transgenic plants.

关 键 词：活体成像仪 荧光显微镜 小麦 荧光观察 

分 类 号：S512[农业科学—作物学]