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主要功能
測量光合作用、蒸騰作用、呼吸作用、葉綠素熒光(可選)
氣體交換和熒光參數的光響應曲線(xiàn)和 CO2 響應曲線(xiàn)
同步測量 CO2 氣體交換與其它光合指標
測量參數
凈光合速率,呼吸速率,蒸騰速率,氣孔導度,胞間二氧化碳濃度,葉綠素熒光參數(可選)等
應用領(lǐng)域
植物生理、植物生態(tài)、農學(xué)、林學(xué)、園藝學(xué)等。
全方位的PAR測量設計
GFS-3000 系統具備全方位的 PAR 測量設計:三個(gè) PAR 傳感器(下圖紅圈部分),分別測量環(huán)境 PAR,葉室內部葉片正面 PAR,以及葉室內部葉片背面 PAR。本設計的優(yōu)點(diǎn)在于不會(huì )忽略葉片背面的光合作用測量。
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全方位的溫度測量設計
獨特的四溫度測量設計:
Tleaf:熱電偶,測量葉片溫度
Tcuv:Pt-100 熱敏電阻,測量葉室溫度
Tamb:Pt-100 熱敏電阻,測量環(huán)境溫度
Ttop:Pt-100 熱敏電阻,測量葉室上部溫度
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多種葉室可供選擇
GFS-3000 標準葉室的設計允許快速、簡(jiǎn)便的更換各種滿(mǎn)足特殊需要的葉室。紅藍 LED 光源 3040-L 可與所有葉室連接使用。
標準葉室
適合多數葉片,標準測量面積 8 cm2,可更換配件滿(mǎn)足 1~12.5 cm2 的測量面積。
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柱狀葉室
適合地衣、苔蘚、土壤樣品和小動(dòng)物等。
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針葉葉室
適合各種針葉植物葉片或小枝條。
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擬南芥植株葉室
大容積,可放進(jìn)(盆)直徑 55 cm~70 mm 的盆栽擬南芥或其它小植株。
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同步測量植物 CO2 氣體交換與其它光合指標的解決方案
光合作用是地球上重要的化學(xué)反應之一,是整個(gè)生物圈物質(zhì)循環(huán)與能量流動(dòng)的基礎。測量生物的光合作用一直是科研界的熱點(diǎn)。
傳統的光合作用測量主要包括調制葉綠素熒光(PAM 技術(shù))、CO2 氣體交換和光合放氧三大技術(shù),幾十年來(lái)在國際科研界均得到了廣泛應用。由于調制葉綠素熒光和 CO2 氣體交換都可以做到無(wú)損、原位、活體測量,對同一個(gè)樣品可以進(jìn)行長(cháng)期的脅迫處理研究(光合放氧需要破碎葉片),因此應用更廣泛一些。
此外,還有一種差式吸收技術(shù),可以通過(guò)測量光合組分在氧化還原(或加亞基、去亞基)過(guò)程中的差式吸收來(lái)反映他們的活性。如通過(guò)測量光系統 I 反應中心葉綠素 P700 的差式吸收來(lái)測量光系統I的活性(DUAL-PAM-100),通過(guò)測量 P515/535 的差式吸收來(lái)測量跨膜質(zhì)子梯度 ΔpH 和玉米黃素(Zea)的變化(DUAL-PAM-100 的 P515/535 模塊),通過(guò)測量 500-570 nm 的差式吸收來(lái)測量 C550、Cyt b559、Cyt b563、Cyt c556、Cyt c6、Cyt f 等的活性變化(KLAS-100)。這種技術(shù)信號弱、難度高,但也具有無(wú)損、原位、活體測量的特點(diǎn)。隨著(zhù)雙通道 PAM-100 測量系統 DUAL-PAM-100 的大規模商業(yè)化生產(chǎn),差式吸收技術(shù)已在國際光合作用學(xué)界得到廣泛應用。
更加可喜的是,可以同步測量 C550、Cyt b559、Cyt b563、Cyt c556、Cyt c6、Cyt f、P515、Scatt(散射信號)、Zea(玉米黃素)等活性的動(dòng)態(tài) LED 陣列差示吸收光譜儀 KLAS-100 也已研發(fā)成功,大大拓展了差示吸收技術(shù)在光合作用研究領(lǐng)域的應用。
除了利用上述幾種技術(shù)進(jìn)行單獨測量外,從上世紀 80 年代后期,逐漸開(kāi)始了兩種技術(shù)的同步測量,如同步測量調制葉綠素熒光與 CO2 氣體交換、同步測量調制葉綠素熒光與光合放氧等。
隨著(zhù)技術(shù)的進(jìn)步,有越來(lái)越多的指標可以同步測量,而且即使是兩種指標的同步測量(如調制葉綠素熒光與 CO2 氣體交換),也可以有多種測量模式可供選擇。
德國 WALZ 公司擁有 CO2 氣體交換、調制葉綠素熒光和差式吸收三種核心技術(shù)。為了方便廣大科研工作者更深入的了解各種光合作用的同步測量技術(shù),澤泉生態(tài)開(kāi)放實(shí)驗室(Zealquest Laboratory for Ecological Research)總結出了一套 CO2 氣體交換與其它光合指標的同步測量解決方案,希望能為相關(guān)單位提供參考。
方案功能與設備
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CO2 氣體交換與調制葉綠素熒光、差式吸收等技術(shù)的同步測量,有很多模式可供選擇。下面將根據技術(shù)難度從低到高的順序,分 5 方面進(jìn)行介紹。
同步測量一:同步測量 CO2 氣體交換與葉綠素熒光
CO2氣體交換與葉綠素熒光的同步測量,主要有以下幾種模式:
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模式一(GFS-3000/FL):便攜式光合儀 GFS-3000 連接熒光附件 3057-FL,在儀器提供的人工光下同步測量氣體交換與葉綠素熒光。3057-FL 不能單獨使用。 | 模式二(GFS-3000/F):便攜式光合儀 GFS-3000 連接熒光附件 3050-F,在自然光下或人工光下同步測量氣體交換與葉綠素熒光。3050-F 不能單獨使用。 | |
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模式三(GFS-3000/M):便攜式光合儀 GFS-3000 連接超便攜式調制熒光儀 MINI-PAM,在自然光下或人工光下同步測量氣體交換與葉綠素熒光。MINI-PAM 可單獨使用。 | 模式四(GFS-3000/J):便攜式光合儀 GFS-3000 連接基礎型調制熒光儀 JUNIOR-PAM,在自然光下或人工光下同步測量氣體交換與葉綠素熒光。JUNIOR-PAM 可單獨使用。 |
上述 4 種模式可以根據需要靈活選擇,特別是模式三,既可同步測量,也可分開(kāi)測量。由于光合儀比較沉重,在許多條件苛刻的場(chǎng)合就可以攜帶極便攜的 MINI-PAM 進(jìn)行測量。
同步測量二:同步測量 CO2 氣體交換與葉綠素熒光成像
CO2 氣體交換與葉綠素熒光成像的同步測量,主要有一下幾種模式【技術(shù)文獻見(jiàn)“代表文獻”部分】。
模式一:與 MINI-IMAGING-PAM 聯(lián)用 測量面積 2 x 3.2 cm | 模式二:與 MAXI-IMAGING-PAM 聯(lián)用 測量面積 10 x 13 cm | |
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GFS-3000/IM-MINI 連接標準測量葉室,適合大多數樣品測量。 | ||
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GFS-3000/IM-MINI-Arabidopsis 連接擬南芥整株葉室,適合小植株的整株測量。 | GFS-3000/IM-MAXI 連接特制大葉室,適合大型葉片的測量。 |
由于 M 系列 IMAGING-PAM 可以共用一個(gè)主機分別連接多個(gè)測量面積不同的探頭(MAXI-,MINI-,MICROSCOPY-),而 GFS-3000 又有多種葉室可供選擇,這就極大豐富了同步測量 CO2 氣體交換與葉綠素熒光成像的模式。如分別與 IMAGING-PAM 的 MAXI- 和 MINI- 探頭連接,就可在不同的測量面積上同步測量。另外,與擬南芥整株葉室結合,就可測量小植株的整株氣體交換和熒光成像。
GFS-3000 設計的一個(gè)重要特點(diǎn)就是,標準葉室、柱狀葉室、針葉/簇狀葉室、擬南芥整株葉室的上表面是相同的,都可以與 MINI-IMAGING-PAM 結合使用,更進(jìn)一步擴大了同步測量的應用范圍。
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同步測量三:同步測量 CO2 氣體交換、P700 與葉綠素熒光
光合儀 GFS-3000 與 DUAL-PAM-100 測量系統聯(lián)用,完美實(shí)現了氣體交換、葉綠素熒光與差式吸收同步測量。同步測量 CO2 氣體交換與 葉綠素熒光和P700 。
系統組成
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DUAL-PAM-100 | 3010-DUAL | GFS-3000 |
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GFS-DUAL |
主要功能
同步測量 P700、葉綠素熒光與氣體交換
同步測量 P700、葉綠素熒光與氣體交換的暗-光誘導曲線(xiàn)
同步測量 P700、葉綠素熒光與氣體交換的光響應曲線(xiàn)和 CO2 響應曲線(xiàn)
典型的氣體交換測量,如光合作用、蒸騰作用、呼吸作用
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、暗馳豫等
典型的 P700 曲線(xiàn)測量
葉綠素熒光與 P700 的快速誘導動(dòng)力學(xué)等
編程進(jìn)行復雜的同步或獨立測量
測量參數
PS II 參數:Fo, Fm, F, Fm’, Fv/Fm, Y(II), Fo’, qP, qL, qN, NPQ, Y(NPQ), Y(NO) 和 ETR(II) 等
PS I 參數:P700, Pm, Pm’, P700red, Y(I), Y(ND), Y(NA) 和 ETR(I) 等
氣體交換參數:參比室和樣品室的 CO2 絕對值(CO2abs,CO2sam),參比室和樣品室的 H2O 絕對值(H2Oabs,H2Osam),流速(gas flow),環(huán)境氣壓(Pamb),葉室溫度(Tcuv),葉片溫度(Tleaf),環(huán)境溫度(Tamb),環(huán)境 PAR(PARamb),葉室內葉片正面 PAR(PARtop),葉室內葉片背面 PAR(PARbot),葉室相對濕度(rH),蒸騰速率(E),水氣壓飽和虧(VPD),葉片氣孔導度(GH2O),凈光合速率(A),胞間 CO2 濃度(Ci),環(huán)境 CO2 濃度(Ca),植物水分利用效率,CO2 響應曲線(xiàn),光響應曲線(xiàn)等
DUAL-PAM 氣體交換葉室——3010-DUAL
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3010-DUAL | 3010-DUAL 與 DUAL-PAM-100 的測量頭連接 |
專(zhuān)為 DUAL-PAM-100 與 GFS-3000 的同步測量設計,由特制葉室(帶溫度和 PAR 傳感器)、風(fēng)扇、導光桿、電子盒與支架構成。同步測量時(shí),光源完全由 DUAL-PAM-100 的測量頭提供,氣體交換由 GFS-3000 的紅外分析器檢測,P700和葉綠素熒光由 DUAL-PAM-100 的檢測器測量。
需要注意的是,3010-DUAL 可以連接 DUAL-PAM-100 的 DUAL-DB 測量頭,但不能連接 DUAL-DR 測量頭。DUAL-DR 的光學(xué)單元太復雜,連接 3010-DUAL 容易損傷 DUAL-DR。
測量實(shí)例
下面的兩個(gè)圖是以洋常春藤(Hedera helix)為材料,利用本系統同步測量的 P700、葉綠素熒光和氣體交換的誘導曲線(xiàn)。
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洋常春藤(Hedera helix)的 P700(藍色)和葉綠素熒光(紅色)的誘導曲線(xiàn) |
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洋常春藤(Hedera helix)的凈光合速率(紅色)和氣孔導度(藍色)的誘導曲線(xiàn) |
同步測量四:同步測量 CO2 氣體交換與跨膜質(zhì)子動(dòng)力勢(pmf)、跨膜質(zhì)子梯度(ΔpH) 、跨膜電位(ΔΨ)和玉米黃素(Zea)
P515/535 模塊是 WALZ 公司為 DUAL-PAM-100 設計的測量模塊,可以直接連接 DUAL-PAM-100 的主機,測量 550-510 nm 的差式吸收以及 535 nm 波長(cháng)的信號變化。P515/535 模塊可以測量光合器官的跨膜質(zhì)子動(dòng)力勢(pmf)、跨膜電位(Δψ)、跨膜質(zhì)子梯度(ΔpH)和玉米黃素(Zea)變化等內容。此外,該模塊還提供一種特殊的 “P515 Flux” 操作模式,可讓光化光以光-暗脈沖形式打開(kāi)-關(guān)閉(1/1調制光/暗),原位測量活體樣品處于穩態(tài)的偶聯(lián)電子和質(zhì)子的流動(dòng)速率。
GFS-3000 通過(guò)專(zhuān)用氣體交換葉室 3010-DUAL 與 DUAL-PAM-100 以及 P515/535 模塊聯(lián)用后,做到了同步測量 CO2 氣體交換與跨膜質(zhì)子動(dòng)力勢(pmf)、跨膜質(zhì)子梯度(ΔpH) 、跨膜電位(ΔΨ)和玉米黃素(Zea)。由于 ΔpH 和 Zea 都與葉黃素循環(huán)密切相關(guān),而葉黃素循環(huán)是植物光保護的重要調節機制,再結合氣體交換指標反映的 Calvin 循環(huán)狀況,就可以非常深入的對植物光保護調節機制進(jìn)行研究。
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DUAL-PAM-100 P515/535 與 GFS-3000 聯(lián)用 |
同步測量五:同步測量 CO2 氣體交換與 C550、Cyt b559、Cyt b563、Cyt c556、Cyt c6、Cyt f、P515、Scatt(散射信號)、Zea(玉米黃素)
傳統的差式吸收技術(shù)每次只能測量一個(gè)組分的差式吸收變化,而光合機構特別復雜,很多組分是偶聯(lián)在一起發(fā)揮作用的,幾種組分分開(kāi)測量的結果難以完美表征它們在偶聯(lián)的功能體下的實(shí)際作用機制。
Schreiber 教授花費 20 年時(shí)間研制的動(dòng)態(tài) LED 陣列差式吸收光譜儀 KLAS-100 完美的解決了上述缺陷。它采用 500-570 nm的差示吸收光譜技術(shù)來(lái)同步測量光合膜上 C550、Cyt b559、Cyt b563、Cyt c556、Cyt c6、Cyt f、P515、Scatt(散射信號)、Zea(玉米黃素)等8種組分的氧化還原變化。
著(zhù)名的 DUAL-PAM-100 測量系統只有兩個(gè)測量通道,而 KLAS-100 有 10 個(gè)測量通道。因此,1 臺 KLAS-100 相當于 5 臺 DUAL-PAM-100 的功能。
GFS-3000 通過(guò) 3010-DUAL 專(zhuān)用氣體交換葉室與 KLAS-100 聯(lián)用后,做到了同步測量 CO2 氣體交換與 C550、Cyt b559、Cyt b563、Cyt c556、Cyt c6、Cyt f、P515、Scatt(散射信號)、Zea(玉米黃素)。
這些測量指標結合在一起,可以進(jìn)行非常復雜的、前人未做過(guò)的深入研究。KLAS-100 是剛剛發(fā)展成熟的一種技術(shù),在著(zhù)名的《Plant Physiology》上發(fā)表了兩篇文獻。
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KLAS-100 |
同步測量技術(shù)選購指南
利用 GFS-3000 與其它技術(shù)(調制葉綠素熒光、差式吸收)的聯(lián)用,可以實(shí)現多種光合指標的同步測量,利用無(wú)損、原位、活體的測量進(jìn)行非常復雜、深入的機理性研究。下表是實(shí)現這些測量功能的選購指南:
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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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