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使用荧光染料对叶绿体外被膜进行活细胞成像。

Live-cell imaging of the chloroplast outer envelope membrane using fluorescent dyes.

作者信息

Ichikawa Shintaro, Ishikawa Kazuya, Miyakawa Hitoshi, Kodama Yutaka

机构信息

Center for Bioscience Research and Education Utsunomiya University Tochigi Japan.

Graduate School of Regional Development and Creativity Utsunomiya University Tochigi Japan.

出版信息

Plant Direct. 2022 Nov 15;6(11):e462. doi: 10.1002/pld3.462. eCollection 2022 Nov.

DOI:10.1002/pld3.462
PMID:36398034
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9666008/
Abstract

UNLABELLED

Chloroplasts are organelles composed of sub-organellar compartments-stroma, thylakoids, and starch granules-and are surrounded by outer and inner envelope membranes (OEM and IEM, respectively). The chloroplast OEM and IEM play key roles not only as a barrier separating the chloroplast components from the cytosol but also in the interchange of numerous metabolites and proteins between the chloroplast interior and the cytosol. Fluorescent protein markers for the chloroplast OEM have been widely used to visualize the outermost border of chloroplasts. However, the use of marker proteins requires an established cellular genetic transformation method, which limits the plant species in which marker proteins can be used. Moreover, the high accumulation of OEM marker proteins often elicits abnormal morphological phenotypes of the OEM. Because the OEM can currently only be visualized using exogenous marker proteins, the behaviors of the chloroplast and/or its OEM remain unknown in wild-type cells of various plant species. Here, we visualized the OEM using live-cell staining with the fluorescent dyes rhodamine B and Nile red in several plant species, including crops. We propose rhodamine B and Nile red as new tools for visualizing the chloroplast OEM in living plant cells that do not require genetic transformation.

SIGNIFICANCE STATEMENT

We established a live-cell imaging method to visualize the chloroplast outer envelope membrane by staining living cells with fluorescent dyes. This method does not require genetic transformation and allows the observation of the chloroplast outer envelope membrane in various plant species.

摘要

未标记

叶绿体是由亚细胞器区室——基质、类囊体和淀粉粒——组成的细胞器，被外膜和内膜（分别为OEM和IEM）包围。叶绿体的OEM和IEM不仅作为将叶绿体成分与细胞质分隔开的屏障发挥关键作用，而且在叶绿体内部与细胞质之间众多代谢物和蛋白质的交换中也发挥关键作用。用于叶绿体OEM的荧光蛋白标记已被广泛用于可视化叶绿体的最外层边界。然而，标记蛋白的使用需要成熟的细胞遗传转化方法，这限制了可使用标记蛋白的植物物种。此外，OEM标记蛋白的高积累常常引发OEM的异常形态表型。由于目前只能使用外源标记蛋白来可视化OEM，因此在各种植物物种的野生型细胞中，叶绿体和/或其OEM的行为仍然未知。在这里，我们使用罗丹明B和尼罗红荧光染料对几种植物物种（包括作物）进行活细胞染色，从而可视化OEM。我们提出将罗丹明B和尼罗红作为可视化活植物细胞中叶绿体OEM的新工具，这些方法不需要遗传转化。

意义声明

我们建立了一种活细胞成像方法，通过用荧光染料对活细胞进行染色来可视化叶绿体的外膜。该方法不需要遗传转化，并且可以观察各种植物物种中的叶绿体外膜。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/9a5856985cd8/PLD3-6-e462-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/ef1d613d2de0/PLD3-6-e462-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/a01c09f8332d/PLD3-6-e462-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/eb1b82ff9a2e/PLD3-6-e462-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/9a5856985cd8/PLD3-6-e462-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/ef1d613d2de0/PLD3-6-e462-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/a01c09f8332d/PLD3-6-e462-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/eb1b82ff9a2e/PLD3-6-e462-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffbd/9666008/9a5856985cd8/PLD3-6-e462-g001.jpg

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3
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Annu Rev Plant Biol. 2020 Apr 29;71:217-245. doi: 10.1146/annurev-arplant-050718-100241. Epub 2020 Feb 19.
4
Selective Gene Delivery for Integrating Exogenous DNA into Plastid and Mitochondrial Genomes Using Peptide-DNA Complexes.利用肽-DNA 复合物将外源 DNA 选择性递送至质体和线粒体基因组中。
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