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Efficient Gene Stacking in Rice Using the GA A NTRY System
Rice ( IF 4.8 ) Pub Date : 2021-02-06 , DOI: 10.1186/s12284-021-00460-5
Leyla T. Hathwaik , James Horstman , James G. Thomson , Roger Thilmony

Genetic engineering of rice provides a means for improving rice grain quality and yield, and the introduction and expression of multiple genes can produce new traits that would otherwise be difficult to obtain through conventional breeding. GAANTRY (Gene Assembly in Agrobacterium by Nucleic acid Transfer using Recombinase technologY) was previously shown to be a precise and robust system to stably stack ten genes (28 kilobases (kb)) within an Agrobacterium virulence plasmid Transfer-DNA (T-DNA) and obtain high-quality Arabidopsis and potato transgenic events. To determine whether the GAANTRY system can be used to engineer a monocotyledonous crop, two new T-DNA constructs, carrying five (16.9 kb) or eleven (37.4 kb) cargo sequences were assembled and transformed into rice. Characterization of 53 independent transgenic events demonstrated that more than 50% of the plants carried all of the desired cargo sequences and exhibited the introduced traits. Additionally, more than 18% of the lines were high-quality events containing a single copy of the introduced transgenes and were free of sequences from outside of the T-DNA. Therefore, GAANTRY provides a simple, precise and versatile tool for transgene stacking in rice and potentially other cereal grain crops.



中文翻译:

利用GA A NTRY系统高效进行水稻基因堆叠

水稻的基因工程为提高水稻的籽粒品质和产量提供了一种手段,而多个基因的引入和表达可以产生新的性状,而这些特征通常很难通过常规育种获得。GA A NTRY(利用重组酶技术通过核酸转移在农杆菌中进行基因组装)先前被证明是一种精确而强大的系统,可在农杆菌毒力质粒Transfer-DNA(T-DNA )中稳定地堆叠十个基因(28 KB)),并获得高质量的拟南芥和马铃薯转基因事件。判断GA A是否NTRY系统可用于设计单子叶作物,将两个带有五个(16.9 kb)或十一个(37.4 kb)货物序列的新T-DNA构建体组装起来并转化为水稻。53个独立的转基因事件的特征表明,超过50%的植物带有所有所需的货物序列并表现出引入的性状。此外,超过18%的品系是高质量事件,其中包含导入的转基因的单个拷贝,并且没有来自T-DNA外部的序列。因此,GA A NTRY提供了一种简单,精确和通用的工具,可用于水稻和其他潜在谷物作物中的转基因堆叠。

更新日期:2021-02-07
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