918博天堂(中国)

四通道动态LED阵列近红外光谱仪

DUAL-KLAS-NIR

同步测量PSII活性（叶绿素荧光）和PSI活性（P700）

PC（质体蓝素）Fd（铁氧还蛋白）的氧化还原变化

相较于经典的双通道叶绿素荧光仪DUAL-PAM-100测量系统，新一代四通道动态LED阵列近红外光谱仪DUAL-KLAS-NIR，在光合作用电子传递链组分质体蓝素(PC)、光系统I反应中心(P700)及铁氧还蛋白(Fd)的氧化还原测量方面实现了重大技术突破。它创新性的采用了四波长对近红外探测技术，成功解决了围绕光系统I供体侧和受体侧电子传递精准解析的难题，为光合作用研究开辟了一条崭新的道路。

作为PSI的电子供体和电子受体，PC(质体蓝素)和Fd(铁氧还蛋白)对PSI的氧化还原起着至关重要的调控作用。但一直缺乏科学便捷的手段对其运转状态进行检测。DUAL-KLAS-NIR采用先进的去卷积技术(一种根据来源不同对信号进行分离的技术)，能够同时测量4组不同波长对(780-820nm，820-870nm，840-965nm，870-965nm)的信号，实现对P700(PSI反应中心)、PC和Fd的氧化还原状态的同步分析。另外，它还可以测量由540nm和460nm光化光激发的叶绿素荧光。还有， DUAL-KLAS-NIR四通道动态LED阵列近红外光谱仪也可以扩展P515/535模块，测量跨类囊体膜的质子梯度ΔpH和跨膜电位ΔΨ，分析与电子传递耦合的跨类囊体膜质子转移，质子动力势pmf形成。可以扩展NADPH/9-AA模块，测量NADP+的还原程度。最后，DUAL-KLAS-NIR也可以通过联用叶室3010-DUAL与GFS-3000光合仪联用，同步测量光反应电子传递和暗反应CO2同化，系统全面的研究光合作用机理。

从2016年2月Photosynthesis Research杂志发表Schreiber博士团队标题为“Deconvolution of ferredoxin, plastocyanin, and P700 transmittance changes in intact leaves with a new type of kinetic LED array spectrophotometer”的研究论文，隆重介绍了最新设计的DUAL-KLAS-NIR四通道动态LED阵列近红外光谱仪。时至今日，四通道动态LED阵列近红外光谱仪DUAL-KLAS-NIR已累计发表论文超过80篇。其中不乏Nature Plants，Nature Communications，The Plant Cell，New Phytologist，Plant Physiology，The Plant Journal等植物学领域的专业高分杂志文章(详见附录)。

突出特点：

1. 可测量活体植物叶片或叶绿体/类囊体/藻类悬浮液，对P700、PC和Fd分别进行连续实时的分析。

2. 蓝光460nm和绿光540nm双波长调制叶绿素荧光测量，可分别测量叶片表层和深层细胞的光能转换。

3. 通过板载芯片LED技术设计了高度紧凑的固态照明系统，可提供635nm，460nm的光化光和740nm的远红光，以及635nm单周转和多周转饱和闪光。

4. 光学部件的几何结构完美兼容3010-DUAL联用叶室，可与GFS-3000光合仪组合，在可控条件(光照，温度，湿度，CO2浓度)下同步测量气体交换相关的CO2同化和电子传递相关的氧化还原。

5. 测量光频率范围广(1- 400 kHz)，允许连续评估Fo，也可以在高时间分辨率下记录叶绿素荧光快速动态瞬变(如多相荧光上升动力学或脉冲弛豫动力学)。

6. 专业数据记录软件，入门特别简单。

主要功能：

1. 测量暗适应样品的PC，P700和Fd最大氧化/还原量，根据光谱特征可计算PC/P700和Fd/P700的比值，评估PSI及其与供体侧和受体侧的氧化还原平衡，用于PSI复合体组装中间体功能的研究。

2. 测量并记录光适应条件下光合电子传递过程中质体蓝素(PC)，PS I反应中心(P700)和铁氧还蛋白(Fd)的氧化还原比例，评估PSI及其与供体侧和受体侧的相互关系和协调性。

3. 可以通过蓝色和/或绿色PAM荧光测量叶片表层和深层细胞的光能转换，非常适合与整个叶子的NIR吸收测量进行对比分析。

4. 完整的慢速诱导动力学曲线和快速诱导动力学曲线测量功能，慢速诱导动力学可进行饱和脉冲分析、淬灭分析，诱导曲线，光合控制，快速光曲线和暗弛豫曲线测量；快速诱导动力学可进行Qa_Decay，Poly300ms等十几种程序测量。

5. 可使用软件的自动测量程序实验，也可以编辑脚本(Script)或者保存手动测量程序(Trigger)，轻松执行复杂的测量协议。可自定义测量动作用于特殊诱导过程动力学曲线数据获取和分析，如状态转换和波动光曲线等。

6. 扩展P515/535模块，可测量跨类囊体膜的质子梯度ΔpH和跨膜电位ΔΨ，分析与电子传递耦合的跨类囊体膜质子转移，质子动力势pmf形成。

7. 扩展NADPH/9-AA模块，可测量NADP+的还原程度。

软件界面：

扩展模块：

1、扩展P515/535模块，可测量跨类囊体膜的质子梯度ΔpH和跨膜电位ΔΨ，分析与电子传递耦合的跨类囊体膜质子转移，质子动力势pmf形成。

2、扩展NADPH/9-AA模块，可测量NADP+的还原程度。

3、扩展3010-DUAL联用叶室，可与GFS-3000光合仪组合，在可控条件(光照，温度，湿度，CO2浓度)下同步测量气体交换相关的CO2同化和电子传递相关的氧化还原。

应用领域：

1. 光合电子传递链复合体的氧化还原状态深入剖析，类囊体膜蛋白组分功能研究，如光系统I的装。

2. 光合合成生物学研究相关的植物学，植物生理学，分子生物学，农学，林学、园艺的领域。

3. 人工光合作用和能源相关领域，如生物光伏等

应用案例：

案例1. 德国Christian-Albrechts大学的科学家Jens Appel使用四通道动态LED阵列近红外光谱仪Dual-KLAS-NIR，测量蓝藻Synechocystis sp.PCC 6803围绕PSI的质体蓝素、P700和FeS簇（包括铁氧还蛋白）的氧化还原状态，首次以绝对值量化了光合生物中通过光系统I的电子流。该研究确定了线性和环式电子传递的比例：环式电子传递占通过PSI的电子流的35%。

Marius L. , et al.2020, BBA – Bioenergetics

http://doi.org/10.1016/j.bbabio.2020.148353

案例2. 德国WALZ的应用科学家Gert Schansker博士使用四通道动态LED阵列近红外光谱仪DUAL-KLAS-NIR测量33种植物的光曲线，探测和表征光合控制的光强度依赖性。研究发现， PC在光强≤400 µmolm-2s-1时完全氧化(阴生植物的叶片的光照强度低于阳生植物叶片)。qP和还原态P700之间的关系可以用于衡量光合控制的程度。除了测量光曲线，也可以使用单个中等光强度来表征叶片之间的相对状态。进一步的发现，在一些适应阴生环境的叶片中，Fd在高光强度下变得更加氧化表明从PQ库到P700的电子传递无法跟上PS I受体侧的电子流出。与光合控制的诱导相比，NPQ诱导需要较低的光强度(类囊体腔酸化程度低)。测量结果还可以用于比较qP和qL，比较的结果是qP是与光合控制更相关的叶绿素荧光参数。

Gert Schansker, 2022, Photosynthesis Research

http://doi.org/10.1007/s11120-022-00934-7

案例3. 芬兰图尔库大学的Tapio Lempiäinen等人通过对正常温度下培养的拟南芥设置5种不同的光处理：(1).不处理，(2).60% PSI光抑制，(3).85% PSI光抑制, (4) .60% PSI光抑制后在生长条件下“恢复”24小时，(5).85% PSI 光抑制后在生长条件下“恢复”24 小时。然后对不同处理样品的主要光合复合物、光合光反应的功能和调节、ATP 合酶和碳同化进行分析。研究功能性PSII和PSI之间的不平衡是否会诱导光合作用适应PSI受限的条件。探索植物短期和长期的驯化机制。研究发现，抑制后直接测量可探测短期适应机制，包括将激发能量重定向到PSI 的类囊体蛋白磷酸化，光合作用反馈调节的变化，比如放松光合作用控制(Photosynthetic Control)和激发能淬灭。处理后恢复24小时测量可以有效探测光合机构的长期适应机制，比如基质氧化还原系统的重建以及ATP合酶和细胞色素b6f丰度的增加。植物在适应了PSI限制条件后无需进行大量PSI修复即可恢复CO2同化能力。对 PSI 抑制的反应表明植物有效地适应了光合机构中发生的变化，这可能是植物适应不利环境条件的关键组成部分。

Lempiäinen, T., et al. 2022.

http://doi.org/10.1111/pce.14400

案例4. 德国亥姆霍兹环境研究中心Lai Bin团队通过四通道动态LED阵列近红外光谱仪Dual-KLAS-NIR系统地研究了在BPV系统中培养的蓝藻集胞藻的光合电子流，揭示了电子传递链中各组分的氧化还原状态，并描绘了相应的电子流向各种汇。该研究表明，EET与PSI下游的类Mehler反应竞争电子。在高浓度下，亚铁氰化物对电子传递链的影响与微量氰化物相似，突出了精心设计BPV实验的必要性。此外，该团队另外一项研究还通过Dual-KLAS-NIR测量PSI、质体蓝素和铁氧还蛋白的氧化还原变化。阐明了生物光伏蓝藻集胞藻动态切换电子源，并根据生理和环境条件，利用不同的细胞外转移途径将电流输出到外部电子汇的机理。

Jianqi Yuan., et al. 2024, http://doi.org/10.1016/j.ese.2024.100519.

Schneider, H., et al. 2025, http://doi.org/10.1111/tpj.17225

案例5. 英国谢菲尔德大学Matthew P Johnson课题组利用基于CRISPR/Cas9的基因编辑技术，在莱茵衣藻中构建了通过PSAF将FNR锚定于PSI的嵌合型突变体。使用DUAL-PAM-100双通道叶绿素荧光仪P515/535模块检测电致变色位移(ECS)，使用 DUAL-KLAS-NIR四通道动态LED阵列近红外光谱仪以类似方法测量P700氧化。研究发现，相较于野生型，嵌合突变体因NADPH还原速率降低导致光合生长受限、线性电子传递受阻，且PSI受体侧限制增强。但该突变体同时表现出增强的跨膜质子梯度(ΔpH)和非光化学淬灭(NPQ)，表明CET活性显著提升。因此，将FNR锚定于PSI并未促进光合线性电子传递，反而通过牺牲线性电子传递与CO₂固定效率优先支持环式电子传递。这一发现揭示了FNR定位对光合电子流向分配的关键调控作用。

Emrich-Mills, T. Z., et al. 2025.

http://doi.org/10.1093/plcell/koaf042

产地：德国WALZ

代表文献：

数据来源：光合作用文献Endnote数据库

原始数据来源：Google Scholar

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