Abstract
Chrysanthemum is one of the most economically important flowers globally due to its high ornamental value. In recent years, a large percentage of the Chrysanthemum seticuspe genome has been determined, making this species useful as a model chrysanthemum plant. To fully utilize the genome’s information, efficient and rapid gene functional analysis methods are needed. In this study, we optimized the tomato aspermy virus (TAV) vector for virus-induced gene silencing (VIGS) in C. seticuspe. Conventional plant virus inoculation methods, such as the mechanical inoculation of viral RNA transcripts and agroinoculation into leaves, did not achieve successful TAV infections in C. seticuspe, but vacuum infiltration into sprouts was successful without symptoms. The TAV vector harboring 100 nucleotides of the phytoene desaturase (PDS) gene caused photobleaching phenotypes and a reduction in CsPDS expression in C. seticuspe. To our knowledge, this is the first report of VIGS in chrysanthemums.
Highlights
Chrysanthemum belongs to the Asteraceae family, which is the largest family of angiosperms [1]
Efficient tomato aspermy virus (TAV) virus-induced gene silencing (VIGS) in N. benthamiana was achieved by the attenuation of the viral suppressor of RNA silencing (VSR) activity of the 2b protein [13], we demonstrated that the TAV vector with the wild-type 2b protein harboring the partial C. seticuspe phytoene desaturase (CsPDS) sequence caused photobleaching phenotypes in C. seticuspe
Five- or six-leaf-stage C. seticuspe plants were used for conventional inoculation methods, such as the mechanical inoculation of TAV RNA transcripts or agroinoculation into leaves, and germinated seeds were used for sprout vacuum infiltration
Summary
Chrysanthemum belongs to the Asteraceae family, which is the largest family of angiosperms [1]. Cultivated chrysanthemum has a large genome and high polyploidy [3,4]; the genetic transformation of chrysanthemum is difficult and inefficient compared with that of other major model plants. (hereafter referred to as C. seticuspe) is a wild diploid chrysanthemum closely related to the cultivated chrysanthemum. These properties are preferable for a model plant; 89–97% of C. seticuspe genomes have been determined by next-generation sequencing [5,6]. To fully utilize this genome information, efficient gene functional analysis methods are needed instead of laboriously inefficient conventional plant transformations
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