Development of the VIGS System in the Dioecious Plant Silene latifolia
Development of the VIGS System in the Dioecious Plant Silene latifolia
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DOI：
10.3390/ijms20051031
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发表时间：
2019-03-01
影响因子：
5.6
通讯作者：
Komatsu, Ken
中科院分区：
文献类型：
--
作者：
Fujita, Naoko;Kazama, Yusuke;Komatsu, Ken
(1)背景：宽叶蝇子草是雌雄异株植物，其性别由XY型性染色体决定。Lychnidis-dioicae微孢霉是一种侵染番茄黑穗病菌的黑穗病真菌。这种植物能在雌花中引起雄性化，就好像微孢霉是一种决定性别的基因。最近的大规模测序工作有望提供参与植物性别决定机制的候选基因。将对这些候选基因进行功能表征分析。方法：在S.结果：通过对苹果潜伏球形病毒（ALSV）接种方法的优化，获得了苹果潜伏球形病毒（ALSV）的最佳接种条件。阔叶植物在上部叶中以高比率感染ALSV。原位杂交分析表明，ALSV可以迁移到S.阔叶植物在S.中成功实现VIGS（病毒诱导的基因沉默）。宽叶植物被证明具有八氢番茄红素去饱和酶基因的敲低。最后，将该方法应用于花器官基因的分析，以评价其在花中的应用价值。阔叶植物，可以揭示基因功能和网络的S。阔叶植物，如性别决定和真菌诱导的雄性化的机制。
(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.