基于FRET原理的生物传感器: 小分子荧光探针在植物中的研究进展

doi:10.11983/CBB24056

摘要/Abstract

摘要： 生物小分子是指生物体内分子量较小的单体物质, 植物小分子种类繁多, 包括离子、植物激素和代谢物等。了解植物体内这些小分子的动态变化, 有助于解析相关的生理功能和调控网络, 并为植物细胞学的精确观察创造新的机遇。基于Förster共振能量转移(Förster resonance energy transfer, FRET)原理设计的基因编码荧光生物传感器/探针, 为活体内观察这些小分子的动态变化提供了强有力的工具。通过FRET传感器/探针, 能够可视化细胞内特定小分子化合物的浓度, 并实时获取高分辨率图像。这一技术因其独特的优势而被广泛应用于植物生理学、发育生物学和环境科学等研究领域。该文总结了近年来植物学研究中使用的FRET传感器/探针, 概述了它们的主要设计思路, 并阐述了其在检测离子、植物激素及代谢物方面的应用与研究进展, 旨在为植物中生物小分子的功能研究提供实用的技术手段和可能的研究方向。

关键词: 生物传感器, FRET, 生物小分子, 植物细胞

Abstract: Biological small molecules, also known as monomeric compounds with relatively low molecular weight found in organisms, encompass a wide array of substances in plants, such as ions, plant hormones and metabolites. Studying the dynamic fluctuations of these small molecules in plants is crucial for analyzing their corresponding physiological functions, regulatory networks, and enhancing the precision of botanical research. Genetically encoded fluorescent biosensors/probes utilizing Förster resonance energy transfer (FRET) technology serves as a valuable tool for real-time monitoring of these small molecules within living organisms. These FRET biosensors/probes allow for the non-invasive visualization of specific small molecule concentrations, providing detailed information at a high resolution. Because of these unique advantages, this technique has been extensively applied in various research fields, including plant physiology, developmental biology, and environmental science. This review provides a comprehensive overview of FRET sensors/probes utilized in plant research in recent years, outlines the key design concepts, and highlights their applications and advances in detecting ions, plant hormones, and metabolites. Furthermore, this review demonstrates practical technological tools and potential research directions for elucidating the functions of small biomolecules in plants.

Key words: biosensors, FRET, biological small molecules, plant cell

吕加一, 李乐攻, 侯聪聪. 基于FRET原理的生物传感器: 小分子荧光探针在植物中的研究进展. 植物学报, 2025, 60(2): 283-293.

Jiayi Lü, Legong Li, Congcong Hou. FRET-based Biosensors: Application of Small Molecule Fluorescence Probes in Plants. Chinese Bulletin of Botany, 2025, 60(2): 283-293.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB24056

https://www.chinbullbotany.com/CN/Y2025/V60/I2/283

图/表 2

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生物传感器/探针	目标小分子	感知元件	FRET供体	FRET受体	参考文献
YC	Ca²⁺	CaM (calmodulin)-M13	eCFP (enhanced cyan fluorescent protein)	eYFP (enhanced yellow fluorescent protein)	Miyawaki et al., 1997
CALWY	Zn²⁺	ATOX1和WD4	eCFP	eYFP	van Dongen et al., 2007
eCALWY	Zn²⁺	ATOX1和WD4	Cerulean	Citrine	Vinkenborg et al., 2009
NiTrac1	NO₃^-	NRT1.1 (nitrate transporter 1.1)	mCerulean	Aphrodite	Ho and Frommer, 2014
NitraMeter3.0	NO₃^-	NasR蛋白中的硝酸盐和亚硝酸盐结合结构域	edeCFP	edAFP	Chen et al., 2022
FLIPPi	Pi	PiBP (phosphate-binding protein)	eCFP	eYFP/cpVenus	Gu et al., 2006; Mukherjee et al., 2015
ABAleon	脱落酸	全长PYR1 (pyrabactin resistance 1)和截短的ABI1 (ABA insensitive 1)	mTurquoise	cpVenus73	Waadt et al., 2014
ABACUS	脱落酸	全长PYL1 (PYR-like protein 1)和ABI1催化结构域	edCerulean	edCitrine	Jones et al., 2014
GPS1	赤霉素	截短的GAI (gibberellic acid insensitive)和全长GID1C (GA insensitive dwarf 1c)	edCerulean	edAFP	Rizza et al., 2017
AuxSen	生长素	TrpR (tryptophan repressor)	Aquamarine	mNeonGreen	Herud-Sikimić et al., 2021
FLIPglus	葡萄糖	GGBP (glucose/galactose-binding protein)	eCFP	eYFP	Fehr et al., 2003
Ateam	ATP	F_oF₁-ATP合酶的ε亚基	mseCFP	cp173-mVenus	Imamura et al., 2009
-	谷氨酰胺	QBP (glutamine binding protein)	CFP	YFP	Yang et al., 2010

生物传感器/探针	目标小分子	感知元件	FRET供体	FRET受体	参考文献
YC	Ca²⁺	CaM (calmodulin)-M13	eCFP (enhanced cyan fluorescent protein)	eYFP (enhanced yellow fluorescent protein)	Miyawaki et al., 1997
CALWY	Zn²⁺	ATOX1和WD4	eCFP	eYFP	van Dongen et al., 2007
eCALWY	Zn²⁺	ATOX1和WD4	Cerulean	Citrine	Vinkenborg et al., 2009
NiTrac1	NO₃^-	NRT1.1 (nitrate transporter 1.1)	mCerulean	Aphrodite	Ho and Frommer, 2014
NitraMeter3.0	NO₃^-	NasR蛋白中的硝酸盐和亚硝酸盐结合结构域	edeCFP	edAFP	Chen et al., 2022
FLIPPi	Pi	PiBP (phosphate-binding protein)	eCFP	eYFP/cpVenus	Gu et al., 2006; Mukherjee et al., 2015
ABAleon	脱落酸	全长PYR1 (pyrabactin resistance 1)和截短的ABI1 (ABA insensitive 1)	mTurquoise	cpVenus73	Waadt et al., 2014
ABACUS	脱落酸	全长PYL1 (PYR-like protein 1)和ABI1催化结构域	edCerulean	edCitrine	Jones et al., 2014
GPS1	赤霉素	截短的GAI (gibberellic acid insensitive)和全长GID1C (GA insensitive dwarf 1c)	edCerulean	edAFP	Rizza et al., 2017
AuxSen	生长素	TrpR (tryptophan repressor)	Aquamarine	mNeonGreen	Herud-Sikimić et al., 2021
FLIPglus	葡萄糖	GGBP (glucose/galactose-binding protein)	eCFP	eYFP	Fehr et al., 2003
Ateam	ATP	F_oF₁-ATP合酶的ε亚基	mseCFP	cp173-mVenus	Imamura et al., 2009
-	谷氨酰胺	QBP (glutamine binding protein)	CFP	YFP	Yang et al., 2010