同步測量PSII活性(葉綠素熒光)和PSI活性(P700氧化還原)

擴展P515/535模塊可測量質(zhì)子動(dòng)力勢(pmf)組分：跨膜質(zhì)子梯度DpH和跨膜電位ΔΨ

擴展NADPH/9-AA模塊可以測量NADPH熒光和9-AA熒光

      標準版的雙通道調制葉綠素熒光儀DUAL-PAM-100可同時(shí)測量光系統II(PSII)葉綠素熒光(Fluo)和光系統I(PSI)氧化還原導致的近紅外差示吸收(P700/P700+)。自2006年DUAL-PAM-100問(wèn)世以來(lái)，它就成為了PSII和PSI活性同步測量的黃金標準。

      雙通道調制葉綠素熒光儀DUAL-PAM-100可以用于植物葉片原位活體測量，只需要將葉片夾在激發(fā)和檢測單元中間即可，幾乎不需要任何前處理。此外，它也可以測量提取的完整葉綠體或類(lèi)囊體懸浮液，微藻懸浮液。只需將裝有樣品(1.4mL)的石英杯放置在特制光學(xué)單元中間的孔內，蓋上蓋子即可。如果您在測量過(guò)程中需要往懸浮液內加藥劑，可以從蓋子中間的微小開(kāi)孔插入注射器針頭，添加藥劑。對于易沉降的樣品可以使用磁力攪拌器攪拌。

      雙通道調制葉綠素熒光儀DUAL-PAM-100除了標準激發(fā)和檢測單元配置外，還有3個(gè)擴展選項：

1、擴展P515/535模塊，可測量跨類(lèi)囊體膜的質(zhì)子梯度ΔpH和跨膜電位ΔΨ，分析與電子傳遞耦合的跨類(lèi)囊體膜質(zhì)子轉移，質(zhì)子動(dòng)力勢pmf形成，質(zhì)子導度(gH+)。質(zhì)子通量(vH+)。

2、擴展NADPH/9-AA模塊，可測量NADP+的還原程度和9AA熒光。

3、擴展3010-DUAL聯(lián)用葉室，可與GFS-3000光合儀組合，在可控條件(光照，溫度，濕度，CO2濃度)下同步測量氣體交換相關(guān)的CO2同化和電子傳遞相關(guān)的氧化還原。

      雙通道調制葉綠素熒光儀DUAL-PAM-100發(fā)表文獻數量多，僅光合作用文獻數據庫收錄的就有1680多篇，近五年，每年都有近200篇文獻發(fā)表。發(fā)文質(zhì)量高，其中不乏Molecular Plant，Nature Plants，Nature Communications，The Plant Cell，PNAS，New Phytologist，Plant Physiology，The Plant Journal等植物學(xué)領(lǐng)域的專(zhuān)業(yè)高分雜志文章(詳見(jiàn)附錄)。

主要功能

• 單獨或同步測量活體植物葉片或葉綠體/類(lèi)囊體/藻類(lèi)懸浮液的葉綠素熒光和P700差示吸收

• 單獨或同步測量?jì)蓚€(gè)光系統的誘導動(dòng)力學(xué)曲線(xiàn)（包括快相和慢相）

• 單獨或同步測量?jì)蓚€(gè)光系統的快速光曲線(xiàn)和光響應曲線(xiàn)

• 淬滅分析、暗馳豫分析

• 典型的P700氧化還原曲線(xiàn)

• 通過(guò)葉綠素熒光和/或P700的同步測量可獲知兩個(gè)光系統的電子傳遞動(dòng)力學(xué)、電子載體庫(PQ)的大小、圍繞PSI的環(huán)式電子傳遞動(dòng)力學(xué)等

• 通過(guò)測量P515/535信號變化測量跨膜質(zhì)子動(dòng)力勢pmf及其組分跨膜質(zhì)子梯度DpH和跨膜電位ΔΨ

• “P515 Flux”信號能原位反映活體樣品處于穩態(tài)的偶聯(lián)電子和質(zhì)子的流動(dòng)速率

• 通過(guò)測量NADPH熒光估算NADP+的還原程度

• 通過(guò)測量9-AA熒光來(lái)估算跨膜質(zhì)子梯度DpH

• 可編編輯腳本，自定義完成復雜的測量過(guò)程。

• 所有動(dòng)力學(xué)曲線(xiàn)均可導出Excel，實(shí)現光合反應過(guò)程動(dòng)力學(xué)曲線(xiàn)再現

測量數據

• 葉綠素熒光(PSII)數據：Fo、Fm、F、Fm’、Fv/Fm、Y(II)、Fo’、F/Fm、Fm’/Fm、qP、qL、qN、NPQ、Y(NPQ)、Y(NO)、ETR(II)、F(I)、F(I/)/Fo、α、Ik、ETRmax、Post-Illumination、Qa_Decay、Poly_300ms等

• P700(PSI)數據：P700、Pm、Pm’、P700ox、Y(I)、Y(ND)、Y(NA)、ETR(I)、P700+-Reduction、PQ Poolsize等

• P515/535數據：質(zhì)子動(dòng)力勢pmf、跨膜質(zhì)子梯度ΔpH、跨膜電位ΔΨ、質(zhì)子導度(gH+)、質(zhì)子通量(vH+)、P515_Fast、“P515 Flux”等

• NADPH/9-AA數據：NADP+的還原程度，ΔpH等

應用領(lǐng)域

• 光合作用光系統光能轉換效率，過(guò)剩激發(fā)能耗散，狀態(tài)轉換，光合控制等機制研究。

• 電子傳遞速率，包括線(xiàn)性電子傳遞速率和環(huán)式電子傳遞速率，及系統間電子載體庫研究。

• 人工光合作用和能源相關(guān)領(lǐng)域，如生物光伏研究。

• 其他與植物光合作用結構解析、復合體組裝，色素合成與降解，逆境脅迫與抗逆生理相關(guān)的植物學(xué)，植物生理學(xué)，分子生物學(xué)，合成生物學(xué)，農學(xué)，林學(xué)、園藝，水生生物學(xué)、環(huán)境科學(xué)等領(lǐng)域。

軟件界面

可選附件

1、擴展P515/535模塊，可測量跨類(lèi)囊體膜的質(zhì)子梯度ΔpH和跨膜電位ΔΨ，分析與電子傳遞耦合的跨類(lèi)囊體膜質(zhì)子轉移，質(zhì)子動(dòng)力勢pmf形成。

2、擴展NADPH/9-AA模塊，可測量NADP+的還原程度。

3、擴展3010-DUAL聯(lián)用葉室，可與GFS-3000光合儀組合，在可控條件(光照，溫度，濕度，CO2濃度)下同步測量氣體交換相關(guān)的CO2同化和電子傳遞相關(guān)的氧化還原。

4、針對控制溫度測量的特殊實(shí)驗要求，您可以為DUAL-PAM-100懸浮樣品光學(xué)單元配置控溫模塊：A、循環(huán)水浴，需要用戶(hù)自備恒溫水浴裝置，只需將其通過(guò)軟件連接到控溫模塊即可；B、Pelletier控溫模塊，兼具加熱和制冷兩種模式，可直接裝在懸浮樣品光學(xué)單元頂部，探頭插入樣品即可。

5、對于低濃度的葉綠體/類(lèi)囊體/藻類(lèi)懸浮液，熒光信號也會(huì )特別低。為了保證信號質(zhì)量需要選用靈敏度更高的檢測器來(lái)記錄信號。A，光電二極管探測器單元 DUAL-DPD的靈敏度比標準探頭提高了約10倍?？梢赃M(jìn)行樣品濃度低至5 μg Chl/L 的測量；B、光電倍增管探測器單元 DUAL-DPM的靈敏度比DUAL-DPD更高，可以非?？煽康販y量葉綠素濃度低至 0.5 μg/L 的懸浮液。

DUAL-PAM-100與GFS-3000聯(lián)用

DUAL-PAM-100	3010-DUAL	GFS-3000

3010-DUAL是專(zhuān)們?yōu)镈UAL-PAM-100與GFS-3000聯(lián)用而設計的特制氣體交換葉室。葉室由線(xiàn)性定位支架、溫度和PAR傳感器、風(fēng)扇、導光桿、電子盒構成。同步測量時(shí)，葉片夾在葉室中間，DUAL-PAM-100提供測量需要的所有廣源。氣體交換由GFS-3000的紅外氣體分析器(IRGA)檢測，P700和葉綠素熒光由DUAL-PAM-100的檢測器測量。

• 同步測量P700、葉綠素熒光與氣體交換，誘導曲線(xiàn)。

• 典型的氣體交換測量，如光合速率、蒸騰速率、VPD、氣孔導度、胞間CO2濃度、光響應曲線(xiàn)和CO2響應曲線(xiàn)。

• 典型的葉綠素熒光測量，如誘導曲線(xiàn)、快速光曲線(xiàn)、淬滅分析、暗馳豫，快速誘導動(dòng)力學(xué)等

• 典型的P700氧化還原測量，

• 可擴展P515/535同步測量功能

• 可編程進(jìn)行復雜的同步或獨立測量

應用案例

       案例1：中國農業(yè)科學(xué)院作物科學(xué)研究所周文彬研究員課題組使用CRISPR-Cas9技術(shù)隨機編輯水稻中Rubisco的五個(gè)rbcS基因(OsrbcS1–5)，產(chǎn)生一系列敲除突變體。以此來(lái)研究編碼RuBisCO小亞基(RbcS)的基因突變后對水稻光合作用的影響。研究發(fā)現，水稻光合組織中最主要的 rbcS 基因 OsrbcS2-5 突變后，水稻在田間條件下的生長(cháng)受抑制、抽穗延遲和產(chǎn)量降低，同時(shí)RuBisCO含量和活性降低，光合效率顯著(zhù)降低。多個(gè)突變導致的遲緩表型更嚴重。本研究中，野生型和rbcs突變體水稻光系統I和光系統II葉綠素熒光光響應曲線(xiàn)通過(guò)DUAL-PAM-100雙通道葉綠素熒光儀完成，測量前暗適應30分鐘，設置一系列光梯度，每個(gè)光梯度照光30s，然后施加一個(gè)200ms的光強為20000μmolm-2s-1的飽和脈沖。測量Y(I)、Y(II)、ETRI、ETRII、Y(ND)、Y(NA)等參數。

Zhou, Y., et al. 2024. https://doi.org/10.1111/pbi.14535

      案例2. 英國謝菲爾德大學(xué)Matthew P Johnson課題組利用基于CRISPR/Cas9的基因編輯技術(shù)，在萊茵衣藻中構建了通過(guò)PSAF將FNR錨定于PSI的嵌合型突變體。以此來(lái)研究FNR定位對光合作用的影響。研究發(fā)現，相較于野生型，嵌合突變體因NADPH還原速率降低導致光合生長(cháng)受限、線(xiàn)性電子傳遞受阻，且PSI受體側限制增強。但該突變體同時(shí)表現出增強的跨膜質(zhì)子梯度(ΔpH)和非光化學(xué)淬滅(NPQ)，表明CET活性顯著(zhù)提升。因此，將FNR錨定于PSI并未促進(jìn)光合線(xiàn)性電子傳遞，反而通過(guò)犧牲線(xiàn)性電子傳遞與CO?固定效率優(yōu)先支持環(huán)式電子傳遞。這一發(fā)現揭示了FNR定位對光合電子流向分配的關(guān)鍵調控作用。

      本研究中，葉綠素熒光與電子傳遞，使用IMAGING-PAM葉綠素熒光成像系統測定Fm、光響應曲線(xiàn)及誘導曲線(xiàn)，400μL樣品，100 μg/mL葉綠素濃度，添加15% Ficoll(聚蔗糖)；使用DUAL-PAM-100雙通道葉綠素熒光儀P515/535模塊檢測電致變色位移(ECS)。質(zhì)子動(dòng)力勢根據紅色光化光關(guān)閉后P515信號的衰減來(lái)計算，方法是對黑暗中的前300ms進(jìn)行單指數衰減，以確定信號衰減的跨度(ECSt)；質(zhì)子導度(gH+)是根據該衰減速率常數的倒數計算。質(zhì)子通量(vH+)按ECSt×gH+計算；使用 DUAL-KLAS-NIR四通道動(dòng)態(tài)LED陣列近紅外光譜儀以類(lèi)似方法測量P700氧化。樣品葉綠素濃度100 μg/mL，添加10 μM DCMU和1 mM HA后進(jìn)行測量前的暗適應。在502 μmol·m-2·s-1的紅色光化光照射10秒后，測量P700氧化的衰減(波長(cháng)對840-965nm)。在光照結束時(shí)，使用200ms的多周轉飽和閃光(MT)來(lái)測量P700氧化的最大值(Pm)，并使用脈沖后最初的1.5秒曲線(xiàn)擬合非線(xiàn)性衰減函數來(lái)計算P700還原一階速率常數K；3、NADPH熒光，激發(fā)波長(cháng)340 nm，檢測波長(cháng)460 nm，光暗交替下測定動(dòng)態(tài)變化。

Emrich-Mills, T. Z., et al. 2025.                                                                             https://doi.org/10.1093/plcell/koaf042

產(chǎn)地：德國WALZ

代表文獻

數據來(lái)源：光合作用文獻Endnote數據庫

原始數據來(lái)源：Google Scholar

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