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Plant Physiol. 2019 Mar 13. pii: pp.00875.2018. doi: 10.1104/pp.18.00875. [Epub ahead of print]

An efficient system for Ds transposon tagging in Brachypodium distachyon.

Author information

1
Shandong Agricultural University CITY: Taian STATE: Shandong China [CN].
2
Shan dong Agriculture University CITY: tai an China [CN].
3
Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences CITY: Beijing China [CN].
4
Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences . CITY: Beijing POSTAL_CODE: . China [CN].
5
Shandong Agricultural University CITY: Taian STATE: Shandong POSTAL_CODE: 271018 China [CN] hlan@sdau.edu.cn.

Abstract

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 T-DNA insertional mutant populations in Brachypodium distachyon L., 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 Ds transposon tagging system in B. distachyon. The 35S::AcTPase cassette and Ds element were constructed within the same T-DNA and transformed into B. distachyon plants. The Ds element was readily transposed to other chromosomes or to the same chromosome under the function of Ac transposase. Through homologous chromosome synapsis, recombination, and segregation, the Ds element separated from the Ac element. We selected stable Ds-only plants using G418 and GFP assays and analyzed 241 T0 lines, some of which were highly efficient at producing Ds-only progeny. Through thermal asymmetric interlaced PCR, we isolated 710 independent Ds flanking sequences from Ds-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 re-used to generate more lines from non-transposed plants without the use of time-consuming tissue culture steps.

PMID:
30867334
DOI:
10.1104/pp.18.00875

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