An Efficient System for Ds Transposon Tagging in Brachypodium distachyon

• Published in: Plant physiology (Bethesda) 2019-05, Vol.180 (1), p.56-65
• Main Authors: Wu, Hongyu, Xue, Xiaodong, Qin, Caihua, Xu, Yi, Guo, Yuyu, Li, Xiang, Lv, Wei, Li, Qinxia, Mao, Chuangxue, Li, Luzhao, Zhao, Suzhen, Qi, Xiaoquan, An, Hailong
• Format: Article
• Language: English
• Subjects: Brachypodium - genetics; DNA Transposable Elements; Mutagenesis, Insertional - methods; Plants, Genetically Modified; Zea mays - genetics
• ISSN: 0032-0889; 1532-2548
• DOI: 10.1104/pp.18.00875

Transposon tagging is a powerful tool that has been widely applied in several species for insertional mutagenesis in plants. Several efforts have aimed to create transfer-DNA (T-DNA) insertional mutant populations in , a monocot plant used as a model system to study temperate cereals, but there has been a lack of research aimed at using transposon strategies. Here, we describe the application of a maize ( ) ( ) transposon tagging system in The :: cassette and element were constructed within the same T-DNA and transformed into plants. The element was readily transposed to other chromosomes or to the same chromosome under the function of ( ) transposase. Through homologous chromosome synapsis, recombination, and segregation, the element separated from the element. We selected stable -only plants using G418 and GFP assays and analyzed 241 T lines, some of which were highly efficient at producing -only progeny. Through thermal asymmetric interlaced PCR, we isolated 710 independent flanking sequences from -only plants. Furthermore, we identified a large collection of mutants with visible developmental phenotypes via this transposon tagging system. The system is relatively simple and rapid in comparison to traditional T-DNA insertion strategies, because once efficiency lines are obtained they can be reused to generate more lines from nontransposed plants without the use of time-consuming tissue culture steps.