High-efficiency generation of fertile transplastomic Arabidopsis plants

First author: Stephanie Ruf; Affiliations: Max-Planck-Institut für Molekulare Pflanzenphysiologie (马克斯-普朗克分子植物生理研究所): Potsdam, Germany

Corresponding author: Ralph Bock

The development of technologies for the stable genetic transformation of plastid (chloroplast) genomes has been a boon to both basic and applied research. However, extension of the transplastomic technology to major crops and model plants has proven extremely challenging, and the species range of plastid transformation is still very much limited in that most species currently remain recalcitrant to plastid genome engineering. Here, we report an efficient plastid transformation technology for the model plant Arabidopsis thaliana that relies on root-derived microcalli as a source tissue for biolistic transformation. The method produces fertile transplastomic plants at high frequency when combined with a clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated protein 9 (Cas9)-generated knockout allele of a nuclear locus that enhances sensitivity to the selection agent used for isolation of transplastomic events. Our work makes the model organism of plant biology amenable to routine engineering of the plastid genome, facilitates the combination of plastid engineering with the power of Arabidopsis nuclear genetics, and informs the future development of plastid transformation protocols for other recalcitrant species.

基于植物叶绿体进行稳定的遗传转化对于基础研究和应用研究都有着很大的便利。然而，在主要作物和模式植物中叶绿体转基因技术还很困难，目前大多数的植物物种都不能进行叶绿体转基因，因此该技术的应用大大受到了局限。本文，作者开发了一个高效的拟南芥叶绿体转基因技术，该技术通过基因枪转化方法对来自于根的微愈伤组织进行遗传转化。结合CRISPR-Cas9技术敲除一个核等位基因，增强植株对于筛选药剂敏感性以用于分离转基因植株，该方法能够高频生产可育的叶绿体遗传转化植株。本文的工作标志着适用于质体基因组常规工程的植物生物学模式生物的产生，有利于促进拟南芥质体工程与核遗传学研究的结合，并且有助于未来其它物种质体遗传转化技术流程的开发。

通讯：Ralph Bock (http://as.nyu.edu/content/nyu-as/as/faculty/michael-purugganan.html)

个人简介：1993年，哈雷大学，学士；1996年，弗莱堡大学，博士。

doi: https://doi.org/10.1038/s41477-019-0359-2

Journal: Nature Plants

Published date: February 18, 2019