Real-time monitoring of stromal NADPH levels in Arabidopsis using a metagenome-derived NADPH-binding fluorescent protein

IF 6.1 2区生物学 Q1 PLANT SCIENCES

Plant Physiology and Biochemistry Pub Date : 2024-11-01 DOI:10.1016/j.plaphy.2024.109260

Jin Hoon Jang , Da Been Kim , Yeonsu Choi , Roshanzadeh Amir , Dae-Eun Cheong , Hea-Jong Chung , Sun-Hee Ahn , Geun-Joong Kim , Dong Wook Lee , Ok Ran Lee , Eung-Sam Kim

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引用次数: 0

Abstract

The light irradiation to the plant chloroplasts drives NADPH and ATP synthesis in the stroma via the electron transport chains within the thylakoid membranes. Conventional methods for assessing photosynthetic light reactions are often invasive or require specific conditions. While detection markers do not significantly affect plant growth itself, developing a method for the real-time and non-invasive detection of NADPH is a highly impactful and important research area in plant physiology and biochemistry. This study introduces a genetically encoded NADPH-binding blue fluorescent protein (mBFP) targeted to the chloroplast stroma or thylakoid membrane in Arabidopsis thaliana and Nicotiana benthamiana. Using two-photon microscopy, we monitored real-time stromal NADPH levels in transgenic leaves of Arabidopsis in response to light exposure. A mutant mBFP construct targeted to the thylakoid membrane allowed us to detect the stromal NADPH levels in real time under different light conditions. This in planta biosensor provides a non-invasive tool for studying photosynthetic responses to light more quantitatively and holds potential for optimizing light conditions in controlled-environment agriculture, such as indoor vertical farms, to improve crop productivity.

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利用源于元基因组的 NADPH 结合荧光蛋白实时监测拟南芥基质 NADPH 水平

植物叶绿体受到光照射后，基质中的 NADPH 和 ATP 会通过类囊体膜内的电子传递链合成。评估光合作用光反应的传统方法通常具有侵入性或需要特定条件。虽然检测标记不会对植物生长本身产生重大影响，但开发一种实时、非侵入式检测 NADPH 的方法是植物生理学和生物化学领域极具影响力的重要研究领域。本研究引入了一种基因编码的 NADPH 结合型蓝色荧光蛋白（mBFP），该蛋白靶向拟南芥和烟草属植物的叶绿体基质或类囊体膜。利用双光子显微镜，我们实时监测了拟南芥转基因叶片中基质 NADPH 水平对光照射的响应。我们利用针对类木质膜的突变 mBFP 构建物，在不同光照条件下实时检测基质 NADPH 水平。这种植物体内的生物传感器为更定量地研究光合作用对光的反应提供了一种非侵入式工具，并有望优化室内垂直农场等受控环境农业中的光照条件，从而提高作物产量。

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来源期刊

Plant Physiology and Biochemistry 生物-植物科学

CiteScore

11.10

自引率

3.10%

发文量

410

审稿时长

33 days

期刊介绍： Plant Physiology and Biochemistry publishes original theoretical, experimental and technical contributions in the various fields of plant physiology (biochemistry, physiology, structure, genetics, plant-microbe interactions, etc.) at diverse levels of integration (molecular, subcellular, cellular, organ, whole plant, environmental). Opinions expressed in the journal are the sole responsibility of the authors and publication does not imply the editors'' agreement. Manuscripts describing molecular-genetic and/or gene expression data that are not integrated with biochemical analysis and/or actual measurements of plant physiological processes are not suitable for PPB. Also "Omics" studies (transcriptomics, proteomics, metabolomics, etc.) reporting descriptive analysis without an element of functional validation assays, will not be considered. Similarly, applied agronomic or phytochemical studies that generate no new, fundamental insights in plant physiological and/or biochemical processes are not suitable for publication in PPB. Plant Physiology and Biochemistry publishes several types of articles: Reviews, Papers and Short Papers. Articles for Reviews are either invited by the editor or proposed by the authors for the editor''s prior agreement. Reviews should not exceed 40 typewritten pages and Short Papers no more than approximately 8 typewritten pages. The fundamental character of Plant Physiology and Biochemistry remains that of a journal for original results.