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病毒诱导的虾脊兰属(苦苣苔科)基因沉默。

Virus-induced Gene Silencing in Streptocarpus rexii (Gesneriaceae).

机构信息

Royal Botanic Garden Edinburgh, 20A Inverleith Row, Edinburgh, EH3 5LR, Scotland, UK.

Kanagawa University, 2946, Tsuchiya, Hiratsuka-shi, Kanagawa, 259-1293, Japan.

出版信息

Mol Biotechnol. 2020 Jul;62(6-7):317-325. doi: 10.1007/s12033-020-00248-w.

Abstract

Many members of the family Gesneriaceae are cultivated as ornamental plants, including Cape primrose (Streptocarpus) species. The range of plant architecture found in this genus has also made it a model to study leaf and meristem development and their evolution. However, the lack of tools to study gene functions through reverse genetics in Streptocarpus has limited the exploitation of its genetic potential. To aid functional genomic studies in Streptocarpus rexii, we sought to investigate virus-induced gene silencing (VIGS). Using the broad host range Tobacco Rattle Virus (TRV) to target the PHYTOENE DESATURASE (PDS) gene of S. rexii, we show that infection with sap from Nicotiana benthamiana triggered VIGS efficiently. VIGS was most effective in the seedling leaves 8 weeks after sowing, but was limited in duration and systemic spread. This study reports the first successful use of VIGS in Streptocarpus and in the family Gesneriaceae. The inoculation of viral sap derived from N. benthamiana was able to overcome the difficulties of standard Agrobacterium-mediated transformation in this genus. Irrespective of its transient effect, this VIGS system will be useful to assess gene function at the cellular level and represent an important tool for further understanding molecular mechanisms in Streptocarpus.

摘要

许多 Gesneriaceae 科的成员被培育为观赏植物,包括海角樱草(Streptocarpus)物种。该属植物的结构范围也使其成为研究叶片和分生组织发育及其进化的模式。然而,由于缺乏通过反向遗传学研究 Streptocarpus 基因功能的工具,限制了其遗传潜力的开发。为了辅助 Streptocarpus rexii 的功能基因组研究,我们试图研究病毒诱导的基因沉默(VIGS)。我们利用广谱宿主范围的烟草脆裂病毒(TRV)来靶向 S. rexii 的 PHYTOENE DESATURASE(PDS)基因,证明了从 Nicotiana benthamiana 中提取的汁液感染可以有效地触发 VIGS。VIGS 在播种后 8 周的幼苗叶片中效果最佳,但持续时间和系统传播有限。本研究报告了 VIGS 在 Streptocarpus 和 Gesneriaceae 家族中的首次成功应用。从 N. benthamiana 中提取的病毒汁液接种能够克服该属中标准农杆菌介导转化的困难。无论其瞬时效应如何,该 VIGS 系统将有助于在细胞水平上评估基因功能,并代表进一步理解 Streptocarpus 中分子机制的重要工具。

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