P700氧化还原动力学的测量方法及原理

doi:10.11983/CBB20064

摘要/Abstract

摘要： P700氧化还原动力学技术可快速且无损地检测植物光系统I (PSI)的活性, 是光合研究领域中广泛使用的一种技术。该文系统归纳了P700氧化还原动力学的主要测量方法, 详细阐述其原理并探讨该技术的局限性, 旨在为深入研究光合作用机理提供技术支持。

关键词: P700氧化还原动力学技术, PSI功能

Abstract: P700 redox technique is referred to examine plant photosystem I (PSI) function quickly and non-intrusively, and widely used in the field of photosynthesis research. In this paper, we summarize the main measurement methods of the P700 redox kinetics systematically, expound its principles, and discuss the limitations. The aim is to provide a technical support for in-depth study of photosynthesis mechanisms.

Key words: P700 redox kinetics technique, PSI function

张春艳. P700氧化还原动力学的测量方法及原理. 植物学报, 2020, 55(6): 740-748.

Chunyan Zhang. The Measurement Methods and Principles of P700 Redox Kinetics. Chinese Bulletin of Botany, 2020, 55(6): 740-748.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB20064

https://www.chinbullbotany.com/CN/Y2020/V55/I6/740

图/表 4

图1 活化光诱导的P700氧化还原动力学(改自Alric et al., 2010) 620 nm的红光光强为5 μmol·m-2·s-1 (黑色)、20 μmol·m-2·s-1 (红色)、50 μmol·m-2·s-1 (绿色)、200 μmol·m-2·s-1 (蓝色)、500 μmol·m-2·s-1 (紫色)。莱茵衣藻野生型加入10 μmol·L-1二氯苯基二甲脲(DCMU)和1 mmol·L-1羟胺(HA)。

Figure 1 The curves of P700 redox kinetics induced by actinic lights (modified from Alric et al., 2010) The red light intensity (620 nm) is approximately 5 μmol·m-2·s-1 (black), 20 μmol·m-2·s-1 (red), 50 μmol·m-2·s-1 (green), 200 μmol·m-2·s-1 (blue), 500 μmol·m-2·s-1 (purple). Wild type algae is incubated with 10 μmol·L-1 3-(3,4-dichloro- phenyl)-1,1-dimethylurea (DCMU) and 1 mmol·L-1 hydroxylamine (HA).

图2 饱和脉冲法计算PSI光化学量子产量Y(I) (改自Yang et al., 2017) AL: 活化光(80 μmol·m-2·s-1); FR: 远红光(720 nm, 20 μmol·m-2·s-1); SP: 饱和脉冲(3000 μmol·m-2·s-1); ΔA: 活化光诱导的部分氧化的P700; ΔAmax: 最大光氧化的P700; ox.: 氧化状态的P700; red.: 还原状态的P700

Figure 2 The saturation pulse method for determination of Y(I) (modified from Yang et al., 2017) AL: Actinic light (80 μmol·m-2·s-1); FR: Far-red light (720 nm, 20 μmol·m-2·s-1); SP: Saturation pulse (3000 μmol·m-2·s-1); ΔA: Partial photo-oxidizable P700 induced by AL; ΔAmax: Maximum photo-oxidizable P700; ox.: P700 oxidation; red.: P700 reduction

图3 利用饱和脉冲法计算PSI能量转化 (A)利用饱和脉冲法计算PSI能量转化的原理示意图(改自Klughammer and Schreiber, 2008); (B) 利用饱和脉冲法计算PSI能量转化的测量曲线。AL: 活化光; SP: 饱和脉冲(10000 μmol·m-2·s-1); FR: 远红光(720 nm); Po: 远红光和饱和脉冲关闭后, 全部PSI反应中心处于完全还原状态, 此时P700信号是0; P: 活化光照光过程中, 部分PSI反应中心被氧化的P700信号; Pm': 活化光关闭, 具有开放、有活性的PSI反应中心被饱和脉冲完全氧化的P700信号; Pm: 远红光关闭, 全部PSI反应中心被饱和脉冲完全氧化的P700信号; a P700+ A: 由于PSI电子供体侧限制导致的关闭的PSI反应中心; A: PSI的下游电子受体; b P700 A: 开放的PSI反应中心; c P700 A-: 由于PSI电子受体侧限制导致的关闭的PSI反应中心; a、b、c是PSI反应中心的3个部分, b是开放的、有活性的, 其光化学量子产量定义为1, 而a和c是关闭的、没有活性的, 其光化学量子产量定义为0。

Figure 3 The saturation pulse method for determination of efficiency of energy conversion in PSI (A) Principle of saturation pulse method for determination of efficiency of energy conversion in PSI (modified from Klughammer and Schreiber, 2008); (B) Curve of saturation pulse method for determination of efficiency of energy conversion in PSI. AL: Actinic light; SP: Saturation pulse (10000 μmol·m-2·s-1); FR: Far-red light (720 nm); Po: Complete reduction is induced after the cessation of SP and far-red light with the zero P700 signal; P: In the presence of AL, a part of the PSI centers are oxidized by the AL resulting in an intermediate P700 signal; Pm': A part of the PSI centers are oxidized completely by the SP with the maximal P700 signal after the AL is turned off; Pm: All the PSI centers are oxidized completely by the SP with the maximal P700 signal after FR pre-illumination; a P700+ A: Donor-side limited closed centers; A: The effective ensemble of PSI acceptors, the same as below; b P700 A: Open centers P700A; c P700 A-: Acceptor-side limited closed centers. a, b, c are the three parts of all PSI centers, b is open and active, and the photochemical quantum yield is unity, while a and c are closed and inactive, and the photochemical quantum yield is 0.

图4 光系统间的电子载体库 FR: 远红光(720 nm, 10 μmol·m-2·s-1); ST: 单周转饱和脉冲(200000 μmol·m-2·s-1); MT: 多周转饱和脉冲(20000 μmol·m-2·s-1)

Figure 4 The capacity of the intersystem electron carrier pool FR: Far-red light (720 nm, 10 μmol·m-2·s-1); ST: Single turnover flash (200000 μmol·m-2·s-1); MT: Multiple turnover flash (20000 μmol·m-2·s-1)

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