Pouvreau Benjamin, Blundell Cheryl, Vohra Harpreet, Zwart Alexander B, Arndell Taj, Singh Surinder, Vanhercke Thomas
Agriculture and Food, CSIRO, Canberra, ACT, Australia.
Synthetic Biology Future Science Platform, CSIRO, Canberra, ACT, Australia.
Front Plant Sci. 2020 Mar 17;11:288. doi: 10.3389/fpls.2020.00288. eCollection 2020.
Traditional functional genetic studies in crops are time consuming, complicated and cannot be readily scaled up. The reason is that mutant or transformed crops need to be generated to study the effect of gene modifications on specific traits of interest. However, many crop species have a complex genome and a long generation time. As a result, it usually takes several months to over a year to obtain desired mutants or transgenic plants, which represents a significant bottleneck in the development of new crop varieties. To overcome this major issue, we are currently establishing a versatile plant genetic screening platform, amenable to high throughput screening in almost any crop species, with a unique workflow. This platform combines protoplast transformation and fluorescence activated cell sorting. Here we show that tobacco protoplasts can accumulate high levels of lipid if transiently transformed with genes involved in lipid biosynthesis and can be sorted based on lipid content. Hence, protoplasts can be used as a predictive tool for plant lipid engineering. Using this newly established strategy, we demonstrate the major role of in plant lipid accumulation. We anticipate that this workflow can be applied to numerous highly valuable metabolic traits other than storage lipid accumulation. This new strategy represents a significant step toward screening complex genetic libraries, in a single experiment and in a matter of days, as opposed to years by conventional means.
传统的作物功能基因研究耗时、复杂且难以轻易扩大规模。原因在于，需要培育突变体或转基因作物来研究基因修饰对特定目标性状的影响。然而，许多作物物种基因组复杂且世代周期长。因此，通常需要数月至一年多的时间才能获得所需的突变体或转基因植物，这成为新作物品种开发的一个重大瓶颈。为克服这一主要问题，我们目前正在建立一个通用的植物遗传筛选平台，该平台适用于几乎任何作物物种的高通量筛选，且具有独特的工作流程。这个平台结合了原生质体转化和荧光激活细胞分选技术。在此我们表明，烟草原生质体如果用参与脂质生物合成的基因进行瞬时转化，就能积累高水平的脂质，并且可以根据脂质含量进行分选。因此，原生质体可作为植物脂质工程的一种预测工具。利用这一新建立的策略，我们证明了[具体基因或物质]在植物脂质积累中的主要作用。我们预计，这一工作流程可应用于除储存脂质积累之外的众多高价值代谢性状。这一新策略朝着在单个实验中、在短短几天内筛选复杂遗传文库迈出了重要一步，而传统方法则需要数年时间。
