Virus-induced gene silencing as a tool for functional studies in .

Premise: Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C photosynthesis and floral diversity. However, functional techniques are lacking for elucidating this diversity. Herein, we establish virus-induced gene silencing (VIGS) as a method of generating functional data for , bolstering Cleomaceae as a model system.

Methods: We leveraged the sister relationship of Cleomaceae and Brassicaceae by using constructs readily available for to provide initial information about the feasibility of VIGS in . . We then developed endogenous constructs to optimize VIGS efficiency and viability for fruit development.

Results: was successfully downregulated in . using both heterologous and endogenous constructs. The endogenous construct had the highest degree of downregulation, with many plants displaying strong photobleaching. -treated plants were also successfully downregulated, with a high rate of survival but less effective silencing; only a small percentage of survivors showed a strong phenotype.

Discussion: Our optimized VIGS protocol in . enables functional gene analyses at different developmental stages. Additionally, . is amenable to heterologous knockdown, which suggests that a first pass using non-endogenous constructs is a possible route to test additional species of Cleomaceae.