Transient Expression Assay and Microscopic Observation in Kumquat Fruit

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-04-05 DOI:10.21769/BioProtoc.4968

Jinli Gong, Xuepeng Sun

引用次数: 0

Abstract

Citrus fruits encompass a diverse family, including oranges, mandarins, grapefruits, limes, kumquats, lemons, and others. In citrus, Agrobacterium tumefaciens–mediated genetic transformation of Hongkong kumquat (Fortunella hindsii Swingle) has been widely employed for gene function analysis. However, the perennial nature of woody plants results in the generation of transgenic fruits taking several years. Here, we show the procedures of Agrobacterium-mediated transient transformation and live-cell imaging in kumquat (F. crassifolia Swingle) fruit, using the actin filament marker GFP-Lifeact as an example. Fluorescence detection, western blot analysis, and live-cell imaging with confocal microscopy demonstrate the high transformation efficiency and an extended expression window of this system. Overall, Agrobacterium-mediated transient transformation of kumquat fruits provides a rapid and effective method for studying gene function in fruit, enabling the effective observation of diverse cellular processes in fruit biology.

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金桔果实中的瞬时表达测定和显微观察

柑橘类水果种类繁多，包括橙子、柑橘、葡萄柚、酸橙、金橘、柠檬等。在柑橘中，农杆菌介导的香港金桔（Fortunella hindsii Swingle）基因转化已被广泛用于基因功能分析。然而，由于木本植物的多年生特性，转基因果实的生成需要数年时间。在这里，我们以肌动蛋白丝标记 GFP-Lifeact 为例，展示了农杆菌介导的金橘（F. crassifolia Swingle）果实瞬时转化和活细胞成像的过程。荧光检测、Western 印迹分析和共聚焦显微镜下的活细胞成像证明了该系统的高转化效率和扩展的表达窗口。总之，农杆菌介导的金橘果实瞬时转化为研究果实中的基因功能提供了一种快速有效的方法，可有效观察果实生物学中的各种细胞过程。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.