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Efficient identification of genomic insertions and flanking regions through whole-genome sequencing in three transgenic soybean events

文献类型： 外文期刊

作者： Niu, Lu 1 ; He, Hongli 2 ; Zhang, Yuanyu 1 ; Yang, Jing 1 ; Zhao, Qianqian 1 ; Xing, Guojie 1 ; Zhong, Xiaofang 1 ; Yang, 1 ;

作者机构： 1.Jilin Acad Agr Sci, Jilin Prov Key Lab Agr Biotechnol, Changchun 130033, Peoples R China

2.Jilin Normal Univ, Sch Life Sci, Siping 136000, Peoples R China

关键词： Insertion site; Flanking sequence; Whole-genome sequencing; Virus-resistant transgenic soybean; Event-specific detection

期刊名称：TRANSGENIC RESEARCH （ 影响因子：2.788； 五年影响因子：2.377 ）

ISSN： 0962-8819

年卷期： 2021 年 30 卷 1 期

页码：

收录情况： SCI

摘要： Genomic insertions and flanking regions of transgenes in host genomes constitute a critical component of precise molecular characterization and event-specific detection, which are required in the development and assessment for regulatory approval of genetically modified (GM) crops. Previously, we reported three transgenic soybean events harboring the inverted repeats of the soybean mosaic virus NIb (nuclear inclusion b) gene, exhibiting significantly enhanced resistance to multiple Potyvirus strains. To facilitate safety assessment and event-specific detection, we identified the transgene insertion sites and flanking sequences of the events L120, L122, and L123 using whole-genome sequencing. More than 14.48 Gb sequence data (13 x coverage) were generated using the Illumina HiSeq Xten platform for each event. The sequence reads corresponding to boundaries of inserted T-DNA, and associated native flanking sequences were identified by bioinformatic comparison with the soybean reference genome (Wm82.a2.v1) and the transformation vector sequence. The results indicated that two T-DNA insertions occurred in L120, on Chr07 and Chr13, while L122 and L123 showed single insertions, on Chr02 and Chr06, respectively. Based on the flanking sequences of the inserted T-DNA, the event-specific detection for each event was established using specific PCR primers, and PCR amplification followed by sequencing of PCR products further confirmed the putative insertion loci and flanking regions in the transgenic lines. Our results demonstrate the efficacy and robustness of whole-genome sequencing in identifying the genomic insertions and flanking regions in GM crops. Moreover, the characterization of insertion loci and the establishment of event-specific detection will facilitate the application and development of broad-spectrum virus-resistant transgenic soybean cultivars.

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