A Versatile High Throughput Screening Platform for Plant Metabolic Engineering Highlights the Major Role of ABI3 in Lipid Metabolism Regulation

Abstract

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 ABI3 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.

High oil accumulation in tuber of yellow nutsedge compared to purple nutsedge is associated with more abundant expression of genes involved in fatty acid synthesis and triacylglycerol storage

“…Collectively, these results reinforce the fact that WRI1 and ABI3, as the master regulators of oil accumulation, not only regulate gene expressions involved in fatty acid synthesis, but also control the expression of genes encoding for seed-specific oil-body storage proteins [ 53 , 63 , 72 , 73 ]. Therefore, WRI1-related regulation network of oil production in yellow nutsedge is most likely to differ either from oil seeds or oil fruits, possibly tuber-specific.…”

“…In this respective, however, it is unclear whether ABI3 is the upstream regulator of WRI1 and controls WRI1 expression in the nutsedge tubers as in plant oilseeds. Recent study indicated that ABI3 played an important role in regulating plant oil accumulation, which might be independent from WRI1 and LEC2 regulation networks occurred in oil-rich seed tissues [ 63 ].…”

High Oil Accumulation in Tuber of Yellow Nutsedge Compared to Purple Nutsedge is Associated with More Abundant Expression of Genes Involved in Fatty Acid Synthesis and Triacylglycerol Storage

“…Collectively, these results reinforce the fact that WRI1 and ABI3, as the master regulators of oil accumulation, do not only regulate gene expressions involved in fatty acid synthesis, but also control the expression of genes encoding for seed-speci c oil-body storage proteins [58,66,75,76]. Therefore, WRI1related regulation network of oil production in yellow nutsedge is most likely to differ either from oil seeds or oil fruits, possibly tuber-speci c.…”

“…In this respective, however, it is unclear whether ABI3 is the upstream regulator of WRI1 and controls WRI1 expression in the nutsedge tubers as in plant oilseeds. Recent study indicated that ABI3 played an important role in regulating plant oil accumulation, which might be independent from WRI1 and LEC2 regulation networks occurred in oil-rich seed tissues [66].…”

A synthetic switch based on orange carotenoid protein to control blue light responses in chloroplasts

“…Synthetic biology endeavours in plants are complicated by non‐negligible hurdles associated with genetic engineering, such as long timeframes for the generation of transgenic individuals, low rates of homologous recombination and, in some cases, cumbersome procedures for plant genetic transformation. However, populations of isolated cells, obtained from the enzymatic digestion of cell walls from plant tissues (typically young, poorly lignified leaves), can be adopted as an easy and convenient alternative to implement fast, combinatorial and high‐throughput tests for newly designed synthetic devices (Yoo, Cho and Sheen, 2007; Pouvreau et al , 2020). We therefore opted for this strategy to characterize the modules of split‐NlucOCP2.…”