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四通道动态LED阵列近红外光谱仪 DUAL-KLAS-NIR
日期:2019-05-15 00:00:00

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)PSI活性(P700

PC(质体蓝素)Fd(铁氧还蛋白)的氧化还原变化

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2016年2月Photosynthesis Research杂志发表了Schreiber博士团队的研究文章Deconvolution of ferredoxin, plastocyanin, and P700 transmittance changes in intact leaves with a new type of kinetic LED array spectrophotometer,隆重介绍了DUAL-KLAS-NIR四通道动态LED阵列近红外光谱仪。之后2016年4月2017年3月Schreiber博士团队再次发表文章,进一步阐述DUAL-KLAS-NIR的实际应用。

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。集成以DUALl-PAM-100为标志的第二代PAM的基本功能,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),WALZ公司推出了可以测量PC和Fd氧化还原状态的新一代PAM荧光仪—DUAL-KLAS-NIR四通道动态LED阵列近红外光谱仪。

DUAL-KLAS-NIR不但集成了Dual-PAM-100的基本功能,可以同时测量PSP和PSI,而且能够测量4组不同波段(780-820nm,820-870nm,840-965nm,870-965nm)的信号,实现对P700(PSI反应中心)、PC和Fd的氧化还原状态分别测量。另外,它还可以测量由540nm和460nm光化光激发的叶绿素荧光。利用DUAL-KLAS-NIR四通道动态LED阵列近红外光谱仪,可以准同步地测量各种不同的信号,不仅在驰豫动力下,还可持续地在自然稳态下同时获取各组分的信息

 

突出特点

Ø  可测量活体叶片或悬浮液,对P700、PC和Fd分别进行连续的实时的去卷积分析。

Ø  同时测量分别由540nm(整个叶片)和460nm(表层细胞层)波段激发的两种叶绿素荧光。

Ø  通过集成发光二极管技术,独创高度紧凑的固态照明系统,提供635nm,460nm的光化光和740nm波段远红光,以及635nm单周转和多周转饱和闪光。

Ø  拥有和DUAL-PAM-100相似的光学部件几何结构,可与3010-DUAL兼容,结合GFS-3000光合仪,在可控条件(光照,温度,湿度,CO2浓度)下,同步测量气体交换和电子传递相关的氧化还原。

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


主要功能

Ø  测定质体蓝素(PC),PS I反应中心(P700)和铁氧还蛋白(Fd)的氧化还原变化。

Ø  通过应用创新的分析方法获得PC,P700和Fd光谱特征。在线监测P700,PC和Fd的氧化还原变化,并确定PC / P700和Fd / P700的比值。

Ø  可以通过绿色或蓝色PAM测量光来激发荧光。绿光比蓝光更深入到叶子中。因此,绿色激发的荧光包括来自更深叶层的信息,因此非常适合与整个叶子的NIR吸收测量进行对比分析。

Ø  专业数据记录软件,入门特别简单。可使用DUAL-KLAS-NIR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。

Ø  兼具慢速动力学曲线(饱和脉冲分析、诱导曲线和光响应曲线)和快速动力学曲线(饱和脉冲动力学曲线、高达30µs分辨率的驰豫动力学曲线)。

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DUAL-KLAS-NIR软件近红外测量光设置


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同步测量Fluo, P700, PC, Fd慢速诱导动力学曲线



        

应用领域

光合作用电子传递过程各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。

可广泛应用于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,林学的领域。


 

应用案例

DUAL-KLAS-NIR为光合作用开辟了一个全新的研究领域,实时显示P700,PC和Fd在活体材料中的氧化还原状态,在线解卷积氧化还原信号。完美实现PS I及其供体侧和受体侧氧化还原动力学的同步测量,从而了解它们围绕光系统I的复杂相互作用,另外还可以探究PS I周围的循环电子传递的信息。

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在DUAL-KLAS-NIR出现之前,测量光系统I的有效量子产量,P700信号总是会掺杂Fd的贡献和PC的变量。上图中图C显示了不同光强梯度下甘蓝型油菜叶片PSI的有效PSI量子产量Y(I),PSII的有效量子产量Y(II)和经PSI荧光修正后的PSII的有效量子产率Y(II)corr。经过修正后,Y(II)corr和Y(I)在低光强下相似(小于500μmol m-2 s-1)。然而,当光强大于500μmol m-2 s-1时,Y(I)明显高于Y(II),Y(I)/Y(II)最高可达1.45.

光系统I的有效天线尺寸测量。植物样品从在黑暗条件转移到光下时,在PSI附近,首先PC被氧化,开始积累,之后才是P700被氧化。单纯的PC信号变化的初始斜率可以用作PS I的有效天线尺寸的度量。


右图是放大后的PC(红色)和P700(蓝色)初始吸光度变化,显示了他们初始斜率的巨大差异。对于黑暗适应的叶子,转到光下的短时间内,光系统I受体侧未活化,Fd还原的初始斜率也也说明了这一点。
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DUAL-KLAS-NIR软件设有一个窗口显示P700和PC氧化还原状态的相对变化。该功能可以用来计算PC和P700之间的表观平衡常数。这对研究P700与其供体侧的相互关系是非常重要的。


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 对暗适应的叶子施加饱和脉冲,测量Fd氧化还原动力学。我们不难发现,饱和脉冲产生的电子将Fd还原,饱和脉冲之后的黑暗中,Fd被缓慢再氧化。之后,PSI的受体侧的电子流被激活,再氧化动力学变得更快。在激活PSI的受体侧之后,可以通过监测脉冲后Fd再氧化的速率来研究Fd的暗灭活。这些动力学变化可以通过指数拟合程序拟合。图A给出了Fd再氧化动力学曲线指数拟合程序拟合的实例,图B显示了常春藤叶片不同暗适应时间后的PSI受体侧的暗灭活动力学差异。


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PC,P700和Fd的最大NIR透射率变化与这些复合物的在样品中的含量成比例,并且PC,P700和Fd的消光系数的比率是恒定的。这可以用于探究不同物种或不同生长条件下(例如阳生/阴生,胁迫/非胁迫)样品的PC / P700和Fd / P700比率,以及PC和Fd库的相对大小。现已观察到高PC / P700比率与高电子传递速率(ETR)值相关。上图显示,在常春藤阳生和阴生叶片中,相对于P700,它们PC和Fd含量有着显著的不同。


主要测量参数:

Ø  叶绿素荧光测量:Fo, Fm, Fm’, F, Fo’, Fv/Fm, Y(II), qP, qL, qN, NPQ, Y(NO), Y(NPQ) , ETR(II)等参数,以及各种荧光动力学曲线。

Ø  P700测量:必须能够测量Pm, Pm’, Y(I), ETR(I), Y(ND)和Y(NA)等参数,以及各种P700动力学曲线。

Ø  PC测量:PCm, PCm’, PCox, Rel PCox

Ø  Fd测量:Fdm, Fdm’, Fdred, Rel Fdred, Fd/PC

Ø  实时显示数据采集,可以连续显示数据采集过程即完整的动力学曲线过程

Ø  软件程序:慢速动力学曲线,快速动动力学曲线,曲线拟合


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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