State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, Harbin, 150040, China.
BMC Genomics. 2019 Apr 23;20(1):311. doi: 10.1186/s12864-019-5636-y.
Integration of T-DNA into plant genomes via Agrobacterium may interrupt gene structure and generate numerous mutants. The T-DNA caused mutants are valuable materials for understanding T-DNA integration model in plant research. T-DNA integration in plants is complex and still largely unknown. In this work, we reported that multiple T-DNA fragments caused chromosomal translocation and deletion in a birch (Betula platyphylla × B. pendula) T-DNA mutant yl.
We performed PacBio genome resequencing for yl and the result revealed that two ends of a T-DNA can be integrated into plant genome independently because the two ends can be linked to different chromosomes and cause chromosomal translocation. We also found that these T-DNA were connected into tandem fragment regardless of direction before integrating into plant genome. In addition, the integration of T-DNA in yl genome also caused several chromosomal fragments deletion. We then summarized three cases for T-DNA integration model in the yl genome. (1) A T-DNA fragment is linked to the two ends of a double-stranded break (DSB); (2) Only one end of a T-DNA fragment is linked to a DSB; (3) A T-DNA fragment is linked to the ends of different DSBs. All the observations in the yl genome supported the DSB repair model.
In this study, we showed a comprehensive genome analysis of a T-DNA mutant and provide a new insight into T-DNA integration in plants. These findings would be helpful for the analysis of T-DNA mutants with special phenotypes.
通过农杆菌将 T-DNA 整合到植物基因组中可能会打断基因结构并产生大量突变体。T-DNA 引起的突变体是研究植物中 T-DNA 整合模型的有价值的材料。T-DNA 在植物中的整合是复杂的,目前仍知之甚少。在这项工作中,我们报道了在一个桦木(Betula platyphylla×B. pendula)T-DNA 突变体 yl 中,多个 T-DNA 片段导致了染色体易位和缺失。
我们对 yl 进行了 PacBio 基因组重测序,结果表明 T-DNA 的两端可以独立地整合到植物基因组中,因为这两端可以连接到不同的染色体上并导致染色体易位。我们还发现,这些 T-DNA 在整合到植物基因组之前,无论方向如何,都被连接成串联片段。此外,T-DNA 在 yl 基因组中的整合也导致了几个染色体片段的缺失。然后,我们总结了 T-DNA 在 yl 基因组中的三种整合模型。(1)T-DNA 片段连接到双链断裂(DSB)的两个末端;(2)只有 T-DNA 片段的一个末端连接到 DSB;(3)T-DNA 片段连接到不同 DSB 的末端。yl 基因组中的所有观察结果都支持 DSB 修复模型。
本研究对 T-DNA 突变体进行了全面的基因组分析,为植物中 T-DNA 的整合提供了新的见解。这些发现将有助于分析具有特殊表型的 T-DNA 突变体。