山东农业科学2024，Vol.56Issue(4)：1-8,8.DOI:10.14083/j.issn.1001-4942.2024.04.001
利用种子红色荧光标记鉴定转基因番茄后代
Identification of Tomato Transgenic Offspring with Red Fluorescence Marker of Seeds
摘要
Abstract
Transgenic technology has contributed to gene functional studies and genetic improvement of tomato.The identification of transgenic offspring is a cost/labor-intensive and time-consuming process.We cloned a seed-specific and highly expressed oleosin gene SlOLE1(Solyc06g034040)according to the results of sequence analysis of tomato oleosin gene family and a search in the tomato gene expression database SGN-TEA.The coding region of the red fluorescent protein gene TagRFP was inserted into the upstream of the stop codon of SlOLE1 gene to produce a chimeric gene SlOLE1-TagRFP,which was then inserted into binary vector pBI121 to construct a plant expression vector pSlOLE1-TagRFP.The expression vector was introduced into to-mato variety Money Maker by Agrobacterium-mediated method.The seeds from self-crossed T0 generation plants displayed bright red fluorescence or not under fluorescence microscope.Further verification with PCR molecu-lar marker showed that TagRFP sequence was present in the seedlings arising from fluorescent seeds,indica-ting that the accuracy of identifying transgenic tomato progeny by red fluorescence at seed stage was 100%.Therefore,this study established a simple,fast and low-cost method for identifying transgenic tomato offspring through seed red fluorescence visualization analysis using SlOLE1-TagRFP chimeric gene.关键词
番茄/种子/红色荧光蛋白/油体蛋白/转基因鉴定Key words
Tomato/Seed/Red fluorescence protein/Oleosin protein/Transgenic identification分类
农业科技引用本文复制引用
章力,杜永臣,李君明,黄泽军,魏凯,李珊珊,宁宇,路菲菲,王孝宣,国艳梅,刘磊,李鑫..利用种子红色荧光标记鉴定转基因番茄后代[J].山东农业科学,2024,56(4):1-8,8.基金项目
国家重点研发计划项目(2022YFD1200500) （2022YFD1200500）
国家大宗蔬菜产业技术体系项目(CARS-23-A06) （CARS-23-A06）
国家自然科学基金项目(31872949) （31872949）
