Development And Application Of Transposon Display Based On NGS Technology

With the deepening of genomics research,whole genome molecular marker detection technology has become a commonly used method in Crop Science and Breeding Science.PCR-based molecular marker technologys and Genotyping By Sequencing(GBS)provide mature tools for target gene detection and genetic background detection.However,breeders still need a tool that can simultaneously detect both genetic background and presence of target genes in breeding materials.To solve this problem,we developed a Transposon display by sequencing(TD-seq)technology,it can be used to detect foreground and background simultaneously in different crops.TD-seq makes use of the character of the transposon distributing in genome to realize the detection of whole genome.TD-seq makes use of the character of high copy number of Transposon in genome,all the PCR products obtained by Transposon display were sequenced by Highthroughput sequencing(Next-generation sequencing,NGS)technology.NGS has the advantages of high sensitivity,it can detect the low content PCR products caused by the low amplification efficiency.Using the advantage of NGS,traditional gel electrophoresis detect the presence or absence of PCR products was converted to detect SNP / indel in PCR products by NGS.TD-seq uses TD-primer located on transposon to realize the detection of background markers(background detection),and also use specific primers of target gene to achieve the detection of the specific gene(foreground detection).Only one step of Tn5 interruption reaction and two PCR amplification are needed to achieve the detection of foreground and background simultaneously.This paper has summarized the detection method and development process of TD-seq technology.TD-seq was used to detect the target gene and genetic background of backcross progeny,the important application value of TD-seq in backcross breeding and near isogenic line construction was revealed.