Abstract
(1) Background: Silene latifolia is a dioecious plant, whose sex is determined by XY-type sex chromosomes. Microbotryum lychnidis-dioicae is a smut fungus that infects S. latifolia plants and causes masculinization in female flowers, as if Microbotryum were acting as a sex-determining gene. Recent large-scale sequencing efforts have promised to provide candidate genes that are involved in the sex determination machinery in plants. These candidate genes are to be analyzed for functional characterization. A virus vector can be a tool for functional gene analyses; (2) Methods: To develop a viral vector system in S. latifolia plants, we selected Apple latent spherical virus (ALSV) as an appropriate virus vector that has a wide host range; (3) Results: Following the optimization of the ALSV inoculation method, S. latifolia plants were infected with ALSV at high rates in the upper leaves. In situ hybridization analysis revealed that ALSV can migrate into the flower meristems in S. latifolia plants. Successful VIGS (virus-induced gene silencing) in S. latifolia plants was demonstrated with knockdown of the phytoene desaturase gene. Finally, the developed method was applied to floral organ genes to evaluate its usability in flowers; (4) Conclusion: The developed system enables functional gene analyses in S. latifolia plants, which can unveil gene functions and networks of S. latifolia plants, such as the mechanisms of sex determination and fungal-induced masculinization.
Highlights
Silene latifolia is a dioecious plant model system, where sex is determined by the XY sex chromosomes
Our results demonstrated that Apple latent spherical virus (ALSV) infected S. latifolia plants at a rate of up to 100%, the infected plants had no symptoms, and virus vectors were detected throughout the flower meristems
Apple latent spherical virus (ALSV) has favorable characteristics as a virus vector, since it infects a variety of plant species, including legume, cucurbits, and Rosaceae fruit trees, and that its infection is asymptomatically and systemically dispersed throughout the plant, including the floral organs [18]
Summary
Silene latifolia is a dioecious plant model system, where sex is determined by the XY sex chromosomes. This plant has been applied for various studies, including those on plant sex chromosome evolution, sex determination, dosage compensation, and DNA methylation [1,2,3,4,5,6]. Previous studies suggested that there are two sex-determining genes located on the Y chromosome, designated as stamen-promoting function (SPF) and gynoecium-suppressing function (GSF) [7]. S. latifolia female flowers develop stamens when an anther smut fungus, Microbotryum lychnidis-dioicae, infects female plants, as if Microbotryum acts as an SPF (shown in the cover figure). The detailed molecular mechanisms of this phenomenon have largely remained a mystery, the stamens produced by Microbotryum infection are true stamens that have originated from the ordinary suppressed stamen primordia in female flowers [8]
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