Open AccessVirus‐induced gene silencing as a tool for functional studies in Cleome violacea
Author(s) -
Carey Shane,
HigueraDíaz Mónica,
Mankowski Peter,
Rocca Alexandra,
Hall Jocelyn C.
Publication year - 2021
Publication title -
applications in plant sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 23
ISSN - 2168-0450
DOI - 10.1002/aps3.11435
Subject(s) - biology , phytoene desaturase , gene silencing , heterologous , gene knockdown , gene , endogeny , arabidopsis thaliana , genetics , computational biology , botany , biochemistry , mutant
Premise Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C 4 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 Cleome violacea , bolstering Cleomaceae as a model system. Methods We leveraged the sister relationship of Cleomaceae and Brassicaceae by using constructs readily available for Arabidopsis thaliana to provide initial information about the feasibility of VIGS in C . violacea . We then developed endogenous constructs to optimize VIGS efficiency and viability for fruit development. Results PHYTOENE DESATURASE was successfully downregulated in C . violacea using both heterologous and endogenous constructs. The endogenous construct had the highest degree of downregulation, with many plants displaying strong photobleaching. FRUITFULL ‐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 C . violacea enables functional gene analyses at different developmental stages. Additionally, C . violacea 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.