Himuro Yasuyo, Ishiyama Kanako, Mori Fumie, Gondo Takahiro, Takahashi Fuminori, Shinozaki Kazuo, Kobayashi Masatomo, Akashi Ryo
Biomass Research Platform Team, Biomass Engineering Program Division, RIKEN Center for Sustainable Resource Science, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.
RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan.
J Plant Physiol. 2014 Aug 15;171(13):1127-31. doi: 10.1016/j.jplph.2014.04.007. Epub 2014 Apr 26.
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.
二穗短柄草(紫羊茅)是一种草本植物,属于禾本科早熟禾亚科,该亚科还包括小麦、大麦、燕麦和黑麦等主要作物。该物种已被确立为用于理解和改良谷类作物及温带禾本科植物的实验模式生物。二穗短柄草的群体标准品系Bd21的全基因组已被测序,农杆菌介导转化的方案也已发表。仍需要对实验平台进行进一步改进,包括建立更好的转基因植物评估系统。在此,我们描述了能产生高度敏感的未成熟胚(可生成用于转化的胚性愈伤组织)的Bd21植株的生长条件。延长至20小时的光周期可产生具有优良未成熟胚的种子。此外,对胚性愈伤组织进行渗透处理可提高粒子轰击转染的效率。在这些实验中,我们培育出了表达拟南芥半乳糖醇合酶2(AtGolS2)的转基因植物。表达AtGolS2的转基因植物表现出显著提高的耐旱性,且随着AtGolS2表达量的增加而增强。这些结果表明AtGolS2可赋予单子叶植物耐旱性,并证实二穗短柄草是进一步探索理解和改良主要单子叶作物物种方法的有用模式植物。