四通道動態LED陣(zhen)列(lie)近(jin)紅外光譜儀
DUAL-KLAS-NIR
同步(bu)測量PSII活性（葉綠素熒光）和PSI活性（P700）
PC（質(zhi)體藍(lan)素(su)）Fd（鐵氧(yang)還蛋白）的氧(yang)化(hua)還(huan)原(yuan)變化(hua)
2016年2月Photosynthesis Research雜志發表了Schreiber博士團隊的研究文章Deconvolution of ferredoxin, plastocyanin, and P700 transmittance changes in intact leaves with a new type of kinetic LED array spectrophotometer，隆重介紹了DUAL-KLAS-NIR四通道動態LED陣列近紅外光譜儀。之后2016年4月，2017年3月Schreiber博士團隊再次發表文章，進一步闡述DUAL-KLAS-NIR的實際應用。
作為PSI的(de)(de)(de)(de)電(dian)子供體和電(dian)子受體，PC（質體藍素）和Fd（鐵氧(yang)還(huan)蛋白）對PSI的(de)(de)(de)(de)氧(yang)化還(huan)原起著至關重要(yao)的(de)(de)(de)(de)調(diao)控作用。但一(yi)直缺乏科學(xue)便(bian)捷的(de)(de)(de)(de)手段對其運轉狀(zhuang)(zhuang)態(tai)進(jin)行(xing)檢測(ce)。集成以DUALl-PAM-100為標志(zhi)的(de)(de)(de)(de)第二代(dai)PAM的(de)(de)(de)(de)基本功能，采用先進(jin)的(de)(de)(de)(de)去卷積技(ji)(ji)術(shu)(shu)（一(yi)種根據(ju)來(lai)源不同(tong)對信號進(jin)行(xing)分離的(de)(de)(de)(de)技(ji)(ji)術(shu)(shu)），WALZ公司推出了可以測(ce)量PC和Fd氧(yang)化還(huan)原狀(zhuang)(zhuang)態(tai)的(de)(de)(de)(de)新一(yi)代(dai)PAM熒光儀—DUAL-KLAS-NIR四通道(dao)動(dong)態(tai)LED陣(zhen)列近紅外(wai)光譜儀。
DUAL-KLAS-NIR不但集成了Dual-PAM-100的基本功能，可以同時測量PSP和PSI，而且能夠測量4組不同波段（780-820nm，820-870nm，840-965nm，870-965nm）的信號，實現對P700（PSI反應中心）、PC和Fd的氧化還原狀態分別測量。另外，它還可以測量由540nm和460nm光化光激發的葉綠素熒光。利用DUAL-KLAS-NIR四通道動態LED陣列近紅外光譜儀，可以準同步地測量各種不同的信號，不(bu)僅在(zai)馳豫動力下，還可持續地(di)在(zai)自然穩態下同(tong)時獲取(qu)各組分的信息(xi)。
突出特點
? 可測量活體葉片(pian)或懸(xuan)浮液，對P700、PC和Fd分別進行(xing)連續的實(shi)時(shi)的去卷積分析。
? 同時測量(liang)分別由540nm（整個(ge)葉片）和(he)460nm（表層細(xi)胞層）波(bo)段激(ji)發的兩種(zhong)葉綠(lv)素熒光。
? 通過集(ji)成發光(guang)(guang)(guang)二極管技術，獨創高度緊湊的固態照(zhao)明系(xi)統，提供635nm，460nm的光(guang)(guang)(guang)化光(guang)(guang)(guang)和(he)740nm波段(duan)遠紅光(guang)(guang)(guang)，以(yi)及635nm單周轉(zhuan)和(he)多周轉(zhuan)飽和(he)閃光(guang)(guang)(guang)。
? 擁有和DUAL-PAM-100相似的光學部件幾何結構，可與3010-DUAL兼容，結合GFS-3000光合儀，在可控條件（光照，溫度，濕度，CO2濃度）下，同步測量氣體交換和電子傳遞相關的氧化還原。
? 測(ce)量光頻率范圍廣（1 - 400 kHz），允許連續評估Fo，可以在(zai)高時間分辨率下記錄(lu)快速動(dong)態瞬變（如多相熒光上升動(dong)力學或脈沖弛豫動(dong)力學）。
主(zhu)要功能
? 測定(ding)質體藍素(su)（PC），PS I反(fan)應中心（P700）和鐵(tie)氧(yang)還蛋白（Fd）的氧(yang)化還原變化。
? 通過應(ying)用創新的分析方法獲得PC，P700和Fd光譜特征(zheng)。在線監測P700，PC和Fd的氧化還(huan)原(yuan)變化，并確(que)定PC / P700和Fd / P700的比值。
? 可以通(tong)過綠(lv)(lv)色或藍(lan)色PAM測量(liang)光(guang)(guang)來激發(fa)熒(ying)光(guang)(guang)。綠(lv)(lv)光(guang)(guang)比藍(lan)光(guang)(guang)更深入到(dao)葉子中。因此(ci)，綠(lv)(lv)色激發(fa)的熒(ying)光(guang)(guang)包括(kuo)來自(zi)更深葉層的信息，因此(ci)非常適合與整個(ge)葉子的NIR吸收測量(liang)進行對比分析。
? 專業數(shu)據(ju)記錄軟(ruan)件，入門特別簡單。可(ke)使用DUAL-KLAS-NIR軟(ruan)件的自動測(ce)量(liang)程序實驗，也可(ke)以編輯腳本（Script）或者保存手動測(ce)量(liang)程序（Trigger），輕松(song)執行復雜的測(ce)量(liang)協議。可(ke)自定義測(ce)量(liang)動作用于特殊誘導過(guo)程動力學(xue)曲線(xian)數(shu)據(ju)獲(huo)取(qu)和分析。
? 兼具慢速動力學曲(qu)線（飽和(he)脈沖(chong)(chong)分析、誘導曲(qu)線和(he)光響應(ying)曲(qu)線）和(he)快速動力學曲(qu)線（飽和(he)脈沖(chong)(chong)動力學曲(qu)線、高達30μs分辨率的馳豫動力學曲(qu)線）。
DUAL-KLAS-NIR軟(ruan)件近紅外測量光設(she)置 |
同步測量Fluo, P700, PC, Fd慢速誘(you)導動力(li)學曲線 |
應(ying)用(yong)領域
光合(he)(he)作(zuo)用電子傳遞過(guo)程各復(fu)合(he)(he)體(ti)(ti)的(de)氧化還原狀態深入剖析，類囊體(ti)(ti)膜(mo)蛋白組分功能研究。
可(ke)廣泛應用于光合合成(cheng)生物(wu)學研究相關的植物(wu)學，植物(wu)生理學，分子生物(wu)學，農學，林學的領域。
應用案例
DUAL-KLAS-NIR為光合(he)作(zuo)用開辟了一個全新的研究領域，實時顯示P700，PC和Fd在活體(ti)(ti)材料中(zhong)的氧化還原狀態，在線解(jie)卷積氧化還原信(xin)號。完美(mei)實現PS I及其供體(ti)(ti)側和受體(ti)(ti)側氧化還原動力(li)學(xue)的同(tong)步測量(liang)，從而(er)了解(jie)它們圍(wei)繞(rao)光系統(tong)I的復(fu)雜相互作(zuo)用，另外還可以(yi)探(tan)究PS I周圍(wei)的循(xun)環電子(zi)傳遞(di)的信(xin)息。
在DUAL-KLAS-NIR出現之前，測量光系統I的有效量子產量，P700信號總是會摻雜Fd的貢獻和PC的變量。上圖中圖C顯示了不同光強梯度下甘藍型油菜葉片PSI的有效PSI量子產量Y(I)，PSII的有效量子產量Y(II)和經PSI熒光修正后的PSII的有效量子產率Y(II)corr。經過修正后，Y(II)corr和Y(I)在低光強下相似（小于500μmol m-2 s-1）。然而，當光強大于500μmol m-2 s-1時，Y(I)明顯高于Y(II)，Y(I)/Y(II)最高可達1.45.
光(guang)系(xi)統I的(de)有效天(tian)線尺寸測(ce)量。植物樣(yang)品從在黑暗條件轉移(yi)到光(guang)下時，在PSI附近，首先PC被(bei)(bei)氧(yang)(yang)化(hua)，開始積累，之(zhi)后才是P700被(bei)(bei)氧(yang)(yang)化(hua)。單純的(de)PC信號變化(hua)的(de)初始斜率可以用作PS I的(de)有效天(tian)線尺寸的(de)度(du)量。
| 右圖是放大后的PC（紅色(se)）和P700（藍色(se)）初(chu)始(shi)吸光度變化，顯示了他們初(chu)始(shi)斜(xie)(xie)率的巨大差異。對于(yu)黑暗適應(ying)的葉子，轉到(dao)光下(xia)的短時間內，光系統I受體(ti)側未活化，Fd還原(yuan)的初(chu)始(shi)斜(xie)(xie)率也也說明了這一(yi)點。 |  |
DUAL-KLAS-NIR軟件(jian)設有一個窗(chuang)口顯示P700和PC氧(yang)化還原(yuan)狀(zhuang)態的相(xiang)對變化。該功能(neng)可以(yi)用來計算PC和P700之間的表觀平衡常數。這對研究P700與其(qi)供(gong)體側的相(xiang)互關系是(shi)非常重(zhong)要的。 |  |
對暗(an)(an)適應的(de)(de)葉(xie)子(zi)(zi)施加飽(bao)和脈沖(chong)，測量Fd氧(yang)化(hua)還(huan)(huan)原動力(li)學。我們不(bu)(bu)難發現，飽(bao)和脈沖(chong)產生(sheng)的(de)(de)電子(zi)(zi)將Fd還(huan)(huan)原，飽(bao)和脈沖(chong)之后(hou)的(de)(de)黑(hei)暗(an)(an)中，Fd被(bei)(bei)緩慢(man)再氧(yang)化(hua)。之后(hou)，PSI的(de)(de)受體側(ce)的(de)(de)電子(zi)(zi)流被(bei)(bei)激活(huo)(huo)，再氧(yang)化(hua)動力(li)學變得更快。在激活(huo)(huo)PSI的(de)(de)受體側(ce)之后(hou)，可以通過監測脈沖(chong)后(hou)Fd再氧(yang)化(hua)的(de)(de)速率來研(yan)究Fd的(de)(de)暗(an)(an)滅(mie)活(huo)(huo)。這(zhe)些動力(li)學變化(hua)可以通過指數擬(ni)合(he)程(cheng)序擬(ni)合(he)。圖A給出了Fd再氧(yang)化(hua)動力(li)學曲線指數擬(ni)合(he)程(cheng)序擬(ni)合(he)的(de)(de)實例，圖B顯示了常(chang)春藤葉(xie)片(pian)不(bu)(bu)同暗(an)(an)適應時間后(hou)的(de)(de)PSI受體側(ce)的(de)(de)暗(an)(an)滅(mie)活(huo)(huo)動力(li)學差(cha)異。
PC，P700和(he)(he)(he)Fd的(de)(de)(de)最大NIR透射(she)率(lv)(lv)變化與(yu)這(zhe)些復合(he)物(wu)的(de)(de)(de)在(zai)樣品中的(de)(de)(de)含(han)量(liang)成比(bi)例，并且PC，P700和(he)(he)(he)Fd的(de)(de)(de)消光系數的(de)(de)(de)比(bi)率(lv)(lv)是恒(heng)定的(de)(de)(de)。這(zhe)可以用于探(tan)究不(bu)同物(wu)種或不(bu)同生(sheng)長條件下（例如陽生(sheng)/陰(yin)生(sheng)，脅迫/非脅迫）樣品的(de)(de)(de)PC / P700和(he)(he)(he)Fd / P700比(bi)率(lv)(lv)，以及PC和(he)(he)(he)Fd庫的(de)(de)(de)相(xiang)(xiang)對(dui)大小。現(xian)已(yi)觀察(cha)到高PC / P700比(bi)率(lv)(lv)與(yu)高電子傳遞速率(lv)(lv)（ETR）值相(xiang)(xiang)關。上(shang)圖(tu)顯(xian)示，在(zai)常春藤陽生(sheng)和(he)(he)(he)陰(yin)生(sheng)葉片中，相(xiang)(xiang)對(dui)于P700，它們(men)PC和(he)(he)(he)Fd含(han)量(liang)有(you)著顯(xian)著的(de)(de)(de)不(bu)同。
主要測量(liang)參數：
? 葉綠素熒光測量：Fo, Fm, Fm’, F, Fo’, Fv/Fm, Y(II), qP, qL, qN, NPQ, Y(NO), Y(NPQ) , ETR(II)等參數，以及各種熒光動力學(xue)曲線(xian)。
? P700測量：必須能夠(gou)測量Pm, Pm’, Y(I), ETR(I)， Y(ND)和Y(NA)等參數，以及各種(zhong)P700動力學(xue)曲線。
? PC測量：PCm, PCm’, PCox, Rel PCox
? Fd測(ce)量：Fdm, Fdm’, Fdred, Rel Fdred, Fd/PC
? 實時顯示數據采(cai)集，可以(yi)連續顯示數據采(cai)集過(guo)程即完整的動力學曲線過(guo)程
? 軟件(jian)程(cheng)序(xu)：慢速動(dong)力學曲(qu)(qu)線(xian)，快速動(dong)動(dong)力學曲(qu)(qu)線(xian)，曲(qu)(qu)線(xian)擬(ni)合
產(chan)地：德國WALZ
代表文獻
數據來源：光合作用(yong)文獻Endnote數據庫
原始數據來(lai)源：Google Scholar
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