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Arabidopsis galactinol synthase AtGolS2 improves drought tolerance in the monocot model Brachypodium distachyon
Authors:Yasuyo Himuro  Kanako Ishiyama  Fumie Mori  Takahiro Gondo  Fuminori Takahashi  Kazuo Shinozaki  Masatomo Kobayashi  Ryo Akashi
Affiliation:1. Biomass Research Platform Team, Biomass Engineering Program Division, RIKEN Center for Sustainable Resource Science, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan;2. RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan;3. Frontier Science Research Center, University of Miyazaki, Miyazaki 889-2192, Japan;4. Faculty of Agriculture, University of Miyazaki, Miyazaki 889-2192, Japan
Abstract:Brachypodium distachyon (purple false brome) is a herbaceous species belonging to the grass subfamily Pooideae, which also includes major crops like wheat, barley, oat and rye. The species has been established as experimental model organism for understanding and improving cereal crops and temperate grasses. The complete genome of Bd21, the community standard line of B. distachyon, has been sequenced and protocols for Agrobacterium-mediated transformation have been published. Further improvements to the experimental platform including better evaluation systems for transgenic plants are still needed. Here we describe the growth conditions for Bd21 plants yielding highly responsive immature embryos that can generate embryogenic calli for transformation. A prolonged 20-h photoperiod produced seeds with superior immature embryos. In addition, osmotic treatment of embryogenic calli enhanced the efficiency of transfection by particle bombardment. We generated transgenic plants expressing Arabidopsis thaliana galactinol synthase 2 (AtGolS2) in these experiments. AtGolS2-expressing transgenics displayed significantly improved drought tolerance, increasing with increased expression of AtGolS2. These results demonstrate that AtGolS2 can confer drought tolerance to monocots and confirm that Brachypodium is a useful model to further explore ways to understand and improve major monocot crop species.
Keywords:2,4-D, 2,4-dichlorophenoxyacetic acid   GUS, β-glucuronidase   hpt, hygromycin phosphotransferase   MS, Murashige and Skoog   PCR, polymerase chain reaction
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