Identification of Tomato Transgenic Offspring with Red Fluorescence Marker of Seeds
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.
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