A Cyan Fluorescent Reporter Expressed from the Chloroplast Genome of Marchantia polymorpha.
A Cyan Fluorescent Reporter Expressed from the Chloroplast Genome of Marchantia polymorpha.
复制标题
DOI：
10.1093/pcp/pcv160
复制
发表时间：
2016-02
影响因子：
4.9
通讯作者：
Haseloff J
中科院分区：
文献类型：
--
作者：
Boehm CR;Ueda M;Nishimura Y;Shikanai T;Haseloff J
地钱作为多细胞研究的基础植物模型，近年来受到越来越多的关注。其易于处理，良好表征的质体和成熟的生物射弹质体转化方案，使M。polymorpha作为一个有吸引力的平台，研究从水到陆地的过渡过程中叶绿体的进化。此外，M.由于缺乏RNA编辑的机制，多形体基因表达的遗传元件的表征提供了一个方便的测试平台。报道基因在叶绿体基因表达的定性和定量研究中具有重要的应用价值，而绿色荧光蛋白（GFP）在M. polymorpha已经证明是有问题的。我们报道了一个密码子优化的绿色荧光蛋白变体mturq 2cp的设计，它允许在烟草psbA启动子的控制下成功地表达来自M.多形。我们证明了mturq 2cp在以下方面的效用：（i）在M.多形孢;（ii）地钱属中作为质体结构元素的小座的可视化;和（iii）用于分析启动子活性的定量显微镜。
Recently, the liverwort Marchantia polymorpha has received increasing attention as a basal plant model for multicellular studies. Its ease of handling, well-characterized plastome and proven protocols for biolistic plastid transformation qualify M. polymorpha as an attractive platform to study the evolution of chloroplasts during the transition from water to land. In addition, chloroplasts of M. polymorpha provide a convenient test-bed for the characterization of genetic elements involved in plastid gene expression due to the absence of mechanisms for RNA editing. While reporter genes have proven valuable to the qualitative and quantitative study of gene expression in chloroplasts, expression of green fluorescent protein (GFP) in chloroplasts of M. polymorpha has proven problematic. We report the design of a codon-optimized gfp varian, mturq2cp, which allowed successful expression of a cyan fluorescent protein under control of the tobacco psbA promoter from the chloroplast genome of M. polymorpha. We demonstrate the utility of mturq2cp in (i) early screening for transplastomic events following biolistic transformation of M. polymorpha spores; (ii) visualization of stromules as elements of plastid structure in Marchantia; and (iii) quantitative microscopy for the analysis of promoter activity.