AUTHOR=Collier Ray , Bragg Jennifer , Hernandez Bryan T. , Vogel John P. , Thilmony Roger 

TITLE=Use of Agrobacterium rhizogenes Strain 18r12v and Paromomycin Selection for Transformation of Brachypodium distachyon and Brachypodium sylvaticum

JOURNAL=Frontiers in Plant Science

VOLUME=7

YEAR=2016

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2016.00716

DOI=10.3389/fpls.2016.00716

ISSN=1664-462X

ABSTRACT=<p>The genetic transformation of monocot grasses is a resource intensive process, the quality and efficiency of which is dependent in part upon the method of DNA introduction, as well as the ability to effectively separate transformed from wildtype tissue. <italic>Agrobacterium</italic>-mediated transformation of <italic>Brachypodium</italic> has relied mainly on <italic>Agrobacterium tumefaciens</italic> strain AGL1. Currently the antibiotic hygromycin B has been the selective agent of choice for robust identification of transgenic calli in <italic>Brachypodium distachyon</italic> and <italic>Brachypodium sylvaticum</italic> but few other chemicals have been shown to work as well for selection of transgenic <italic>Brachypodium</italic> cells in tissue culture. This study demonstrates that <italic>Agrobacterium rhizogenes</italic> strain 18r12v and paromomycin selection can be successfully used for the efficient generation of transgenic <italic>B. distachyon</italic> and <italic>B. sylvaticum</italic>. Additionally we observed that the transformation rates were similar to or higher than those obtained with <italic>A</italic>. <italic>tumefaciens</italic> strain AGL1 and hygromycin selection. The <italic>A</italic>. <italic>rhizogenes</italic> strain 18r12v harboring the pARS1 binary vector and paromomycin selection is an effective means of generating transgenic <italic>Brachypodium</italic> plants. This novel approach will facilitate the transgenic complementation of T-DNA knockout mutants of <italic>B. distachyon</italic> which were created using hygromycin selection, as well as aid the implementation of more complex genome manipulation strategies which require multiple rounds of transformation.</p>