Identification of Tomato Transgenic Offspring with Red Fluorescence Marker of Seeds
Zhang Li
Wei Kai
Li Shanshan
Ning Yu
Lu Feifei
Wang Xiaoxuan
Guo Yanmei
Liu Lei
Li Xin
Du Yongchen
Li Junming
Huang Zejun
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.
Keywords：TomatoSeedRed fluorescence proteinOleosin proteinTransgenic identification
Publication Date：2024-04-28
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：8( 1-8 )