Analysis of T-DNA integration events in transgenic rice

文献类型： 外文期刊

第一作者： Gong, Wankui

作者： Gong, Wankui;Zhou, Yun;Wang, Rui;Wei, Xiaoli;Zhang, Lei;Dai, Yan;Zhu, Zhen

作者机构：

关键词： Transgenic rice; T-DNA integration; Microhomology; Filler DNA; Translocation; Cut; paste mechanism

期刊名称：JOURNAL OF PLANT PHYSIOLOGY （ 影响因子：3.549； 五年影响因子：4.164 ）

ISSN： 0176-1617

年卷期： 2021 年 266 卷

页码：

收录情况： SCI

摘要： Agrobacterium-mediated plant transformation has been widely used for introducing transgene(s) into a plant genome and plant breeding. However, our understanding of T-DNA integration into rice genome remains limited relative to that in the model dicot Arabidopsis. To better elucidate the T-DNA integration into the rice genome, we investigated extensively the T-DNA ends and their flanking rice genomic sequences from two transgenic rice plants carrying Cowpea Trypsin Inhibitor (CpTI)-derived gene Signal-CpTI-KDEL (SCK) and Bacillus thuringiensis (BT) gene, respectively, by TAIL-PCR method. Analysis of the junction sequences between the T-DNA ends and rice genome DNA indicated that there were three joining patterns of microhomology, filler DNA sequences, and exact joining, and both the T-DNA ends tend to adopt identical manner to join the rice genome. After T-DNA integration, there were several variations of rice genomic sequences, including small deletions at the integration sites, superfluous DNA inserted between T-DNA and genome, and translocation of genomic DNA in the flanking regions. The translocation block could be from a noncontiguous region in the same chromosome or different chromosomes at the integration sites, and the originating position of the translocated block resulted in comparable deletion based on a cut/paste mechanism rather than a replication mechanism. Our study may lead to a better understand of T-DNA integration mechanism and facilitate functional genomic studies and further crop improvement.

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