Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.
Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.
复制标题
DOI：
10.1111/j.1469-8137.2012.04350.x
复制
发表时间：
2012-12
期刊：
影响因子：
--
通讯作者：
Toki S
中科院分区：
文献类型：
--
作者：
Nishizawa-Yokoi A;Nonaka S;Saika H;Kwon YI;Osakabe K;Toki S
非同源末端连接 (NHEJ) 途径参与农杆菌介导的转移 DNA (T-DNA) 整合到模式植物拟南芥基因组中的证据仍无定论。使用盾片来源的愈伤组织在水稻（Oryza sativa）中建立了快速高效的农杆菌介导的转化系统，我们在这里检查了NHEJ途径在农杆菌介导的水稻稳定转化中的参与。来自 OsKu70、OsKu80 和 OsLig4 敲低 (KD) 植物的水稻愈伤组织用农杆菌感染，该农杆菌具有敏感的翠绿荧光素酶 (LUC) 报告构建体以评估稳定表达，并感染绿色荧光蛋白 (GFP) 构建体以监测 T-DNA 的瞬时表达。在 KD 植物中，瞬时表达没有受到抑制，但稳定表达显着降低。此外，与对照愈伤组织相比，KD-Ku70 和 KD-Lig4 愈伤组织表现出同源重组 (HR) 频率的增加。此外，在用 DNA 损伤剂治疗时，OsKu70、OsKu80 和 OsLig4 的抑制诱导 HR 相关基因的表达。我们的研究结果强烈表明，NHEJ 参与了水稻中农杆菌介导的稳定转化，并且 NHEJ 和 HR 途径在水稻 DNA 双链断裂修复中存在竞争和互补关系。
Evidence for the involvement of the nonhomologous end joining (NHEJ) pathway in Agrobacterium-mediated transferred DNA (T-DNA) integration into the genome of the model plant Arabidopsis remains inconclusive. Having established a rapid and highly efficient Agrobacterium-mediated transformation system in rice (Oryza sativa) using scutellum-derived calli, we examined here the involvement of the NHEJ pathway in Agrobacterium-mediated stable transformation in rice. Rice calli from OsKu70,OsKu80 and OsLig4 knockdown (KD) plants were infected with Agrobacterium harboring a sensitive emerald luciferase (LUC) reporter construct to evaluate stable expression and a green fluorescent protein (GFP) construct to monitor transient expression of T-DNA. Transient expression was not suppressed, but stable expression was reduced significantly, in KD plants. Furthermore, KD-Ku70 and KD-Lig4 calli exhibited an increase in the frequency of homologous recombination (HR) compared with control calli. In addition, suppression of OsKu70,OsKu80 and OsLig4 induced the expression of HR-related genes on treatment with DNA-damaging agents. Our findings suggest strongly that NHEJ is involved in Agrobacterium-mediated stable transformation in rice, and that there is a competitive and complementary relationship between the NHEJ and HR pathways for DNA double-strand break repair in rice.
登录
查看更多内容
影响因子：
14.9
作者：
Bundock, P;van Attikum, H;Hooykaas, P
通讯作者：
Hooykaas, P
影响因子：
4.8
作者：
Brenkman, Arjan B.;van den Broek, Niels J. F.;Burgering, Boudewijn M. T.
通讯作者：
Burgering, Boudewijn M. T.
影响因子：
3.8
作者：
Charbonnel, Cyril;Allain, Elisabeth;White, Charles I.
通讯作者：
White, Charles I.
影响因子：
2.6
作者：
Bertolini, Luciana R.;Bertolini, Marcelo;Murray, James D.
通讯作者：
Murray, James D.
影响因子：
7.2
作者：
Friesner, J;Britt, AB
通讯作者：
Britt, AB