Pi Mengting, Gao Qi, Kang Chunying
Key Laboratory of Horticultural Plant Biology (Ministry of Education), College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, 430070, China.
Bio Protoc. 2019 Jun 5;9(11):e3249. doi: 10.21769/BioProtoc.3249.
Strawberry, including the woodland strawberry (2) and the cultivated strawberry ( × , 8), has emerged as a model system for studying fruit development and ripening. Transient expression provides a quick assay for gene functions or gene interactions. In strawberry, virus-induced gene silencing (VIGS) and mediated transformation in fruit have been widely used as the transient expression approaches. Unlike VIGS, the latter one can be utilized not only for gene knock-down, but also for overexpression and knock-out. Here, we show the procedures of transiently expressing the 35S:: construct into fruit of the white-fruited accession Yellow Wonder. As a master regulator of anthocyanin production, overexpressing will cause fruit coloration, which was observed at one week post infiltration. We also exhibit the previous results of knocking down ), encoding an anthocyanin transporter, by RNAi in fruit of the strawberry cultivar 'Sweet Charlie'. Overall, -mediated transient transformation in strawberry fruit is a quick and versatile approach for studying gene functions in fruit ripening.
草莓，包括野生草莓（2n = 2x = 14）和栽培草莓（2n = 8x = 56），已成为研究果实发育和成熟的模式系统。瞬时表达为基因功能或基因相互作用提供了一种快速检测方法。在草莓中，病毒诱导的基因沉默（VIGS）和果实中的介导转化已被广泛用作瞬时表达方法。与VIGS不同，后者不仅可用于基因敲低，还可用于过表达和基因敲除。在这里，我们展示了将35S::构建体瞬时表达至白果品种黄奇迹果实中的过程。作为花青素产生的主要调节因子，过表达将导致果实着色，这在浸润后一周即可观察到。我们还展示了之前通过RNAi在草莓品种‘甜查理’果实中敲低编码花青素转运蛋白的结果。总体而言，草莓果实中的介导瞬时转化是研究果实成熟过程中基因功能的一种快速且通用的方法。
