Dual-PAM-100: 一套空前强大的测量系统Schreiber教授因发明PAM系列调制叶绿素荧光仪而获得首届国际光合作用协会(ISPR)创新奖1985年开始商品化的全世界第一台调制荧光仪PAM-100被几代科学家所广泛采用。Dual-PAM-100相当于两台PAM-100的功能。一方面,它继承了PAM-100的所有优点,可以进行复杂的叶绿素荧光分析(PS II活性);另一方面,它还可以通过测量P700的吸收变化来检测PS I的活性。特别需要强调的是,Dual-PAM-100可以在完全同步的情况下测量叶绿素荧光和P700吸收变化。此外,通过特殊的激发-检测单元还可以测量叶绿体或微藻的许多重要光合参数,如跨膜质子梯度delta pH(通过9-AA荧光或吖啶黄荧光)、类囊体膜的电势(通过类胡萝卜素的差示吸收,&ldquo P515&rdquo )和NADP的氧还状态(通过NADPH荧光)等。如果需要极高的灵敏度可以通过连接光电倍增管检测器实现。主要功能 * 单独或同步测量微藻、大藻、水生植物等的叶绿素荧光(光系统II活性)和P700(光系统I活性) * 两个光系统的诱导动力学曲线(包括快相和慢相) * 两个光系统的快速光曲线和光响应曲线 * 淬灭分析、暗驰豫分析 * 典型的P700曲线测量 * 通过叶绿素荧光和P700的同步测量获知两个光系统的电子传递动力学、电子载体库的大小、围绕PSI 的环式电子传递动力学等应用领域全球最先进的测量光合作用的技术,已被成功应用于高等植物和蓝藻的P700和叶绿素荧光测量中,用于光合作用机理、胁迫生理学、生理生态学等领域。在其它微藻和大型海藻中的应用还处于起步阶段,恰恰也是比较容易出成果的领域。测量参数PS II参数: Fo, Fm, F, Fm&rsquo , Fv/Fm, Y(II)=△F/Fm&rsquo , Fo&rsquo , qP, qL, qN, NPQ, Y(NPQ), Y(NO)和ETR(II)等PS I参数: P700, Pm, Pm&rsquo , P700red, Y(I), Y(ND), Y(NA)和ETR(I)等与PAM-100相比,Dual-PAM-100的主要特点:1)Dual-PAM-100完全由电脑控制,通过专业的Windows操作软件DualPAM进行。2)软件DualPAM除了基本的系统操作外,还提供许多特定的测量程序。3)所有必需的光源(激发叶绿素荧光的红光和蓝光、测量P700的近红外光、红色和蓝色的光化光、单脉冲与多脉冲饱和闪光、远红光)均整合在基础系统中,不再需要复杂的电缆连接。4)采用了专为Dual-PAM-100设计的许多新光-电配件,使得激发-检测单元和整合式光源非常便携、非常便于安装和拆卸。5)所有的光源都可通过软件在2.5 &mu s的时间分辨率下控制。6)测量光的频率范围非常大(1 Hz~400 KHz),因此同一个测量光源既可以用于测量Fo,也可以用于诱发快速动力学(如荧光快速上升相或闪光弛豫动力学)。7)用户可将针对特殊实验/样品的仪器设置存储起来,此后可在完全相同的设置下重复实验。8)叶绿素荧光和P700的信号变化完全同步,并且是用同一个检测器检测,且不会互相干扰。9)测量蓝藻时注意:用红光激发PS II的荧光,用蓝光或远红光激发PS I。 Dual-PAM-100的这些特点开启了基础光合作用研究和应用光合作用研究的新途径。过去,同步测量PS I和PS II的量子产量需要很强的专业背景和熟练的操作技巧,只有光合作用领域的少数专家会这项技术。现在,即使是初学者,也可迅速掌握同步测量PS I和PS II活性的技术,不再需要复杂的操作技巧。主要技术参数P700双波长测量光:LED,830 nm和870 nmPSII荧光测量光:LED,460 nm(DUAL-DB)或620 nm(DUAL-DR)红色光化光:LED阵列,635 nm;最大连续光强2000 &mu mol m-2 s-1蓝色光化光:LED,460 nm;最大连续光强700 &mu mol m-2 s-1单周转饱和闪光(ST):200000 &mu mol m-2 s-1,5~50 &mu s可调多周转饱和闪光(MT):20000 &mu mol m-2 s-1,1~1000 ms可调Kramer的新荧光参数资料,这是一篇2004年发表在Photosynthesis Research 上的文章,见如下描述:Kramer提出的新参数:qL,用于代替qPY(II)=Y=(Fm&rsquo -F)/Fm&rsquo Y(NPQ) 表征PS II处过量光能耗散为热,与光保护有关Y(NO) 表征PS II处过量光能引起的光损伤Y(II)+Y(NPQ)+Y(NO)=1Schreiber教授和其学生Klughammer博士新设计的Dual-PAM-100中,借鉴测量叶绿素荧光的方法,新增了几个参数:Y(I) PS I的量子产量或PS I的光合效率Y(ND) 表征PS I处过量光能耗散为热,与光保护有关Y(NA) 表征PS I处过量光能引起的光损伤Y(I)+Y(ND)+Y(NA)=1色素分子处于氧化态和还原态时,或增加/减少亚基后,其吸收峰会有变化。基于此原理,P700吸收变化、P515吸收变化(类胡萝卜素能态)、P505吸收变化(叶黄素循环)等均可通过Dual-PAM-100测量。主机共用,只是更换激发-检测单元即可。此外,Dual-PAM-100还可用于测量NADPH荧光、9AA荧光(跨膜质子梯度)等。部分利用DUAL-PAM-100研究藻类的P700文献1.Ma W, Mi H, Shen Y: Influence of the redox state of QA on phycobilisome mobility in the cyanobacterium Synechocystis sp. strain PCC 6803 Journal of Luminescence2010:in press.[DUAL-PAM-100]2.Berná t G, Waschewski N, Rö gner M: Towards efficient hydrogen production: the impact of antenna size and external factors on electron transport dynamics in Synechocystis PCC 6803 Photosynthesis Research2009, 99(3):205-216.[DUAL-PAM-100]3.Chiu Y-F, Lin W-C, Wu C-M, Chen Y-H, Hung C-H, Ke S-C, Chu H-A: Identification and characterization of a cytochrome b559 Synechocystis 6803 mutant spontaneously generated from DCMU-inhibited photoheterotrophical growth conditions Biochimica et Biophysica Acta2009, 1787(10):1179-1188.[DUAL-PAM-100]4.Grouneva I, Jakob T, Wilhelm C, Goss R: The regulation of xanthophyll cycle activity and of non-photochemical fluorescence quenching by two alternative electron flows in the diatoms Phaeodactylum tricornutum and Cyclotella meneghiniana Biochimica et Biophysica Acta 2009, 1787(7):929-938.[DUAL-PAM-100]5.Kromkamp JC, Beardall J, Sukenik A, Kopeck J, Masojidek J, van Bergeijk S, Gabai S, Shaham E, Yamshon A: Short-term variations in photosynthetic parameters of Nannochloropsis cultures grown in two types of outdoor mass cultivation systems. Aquatic Microbial Ecology2009, 56:309-322.[DUAL-PAM, Flow-Through WATER-PAM]6.Lin A-P, Wang G-C, Yang F, Pan G-H: Photosynthetic parameters of sexually different parts of Porphyra katadai var. hemiphylla (Bangiales, Rhodophyta) during dehydration and re-hydration Planta2009, 229(4):803-810.[DUAL-PAM-100]7.Perreault F, Ali NA, Saison C, Popovic R, Juneau P: Dichromate effect on energy dissipation of photosystem II and photosystem I in Chlamydomonas reinhardtii. Journal of Photochemistry and Photobiology B: Biology2009, 96(1):24-29.[DUAL-PAM-100]8.Schultze M, Forberich B, Rexroth S, Dyczmons NG, Roegner M, Appel J: Localization of cytochrome b6f complexes implies an incomplete respiratory chain in cytoplasmic membranes of the cyanobacterium Synechocystis sp. PCC 6803 Biochimica et Biophysica Acta2009, 1787(12):1479-1485.[DUAL-PAM-100]9.Sukenik A, Beardall J, Kromkamp JC, Kopeck J, Masojí dek J, van Bergeijk S, Gabai S, Shaham E, Yamshon A: Photosynthetic performance of outdoor Nannochloropsis mass cultures under a widerange of environmental conditions. Aquatic Microbial Ecology2009, 56(2-3):297-308.[DUAL-PAM-100, FLOW THROUGH WATER-PAM]10.Tsunoyama Y, Berná t G, Dyczmons NG, Schneider D, Rö gner M: Multiple rieske proteins enable short- and long-term light adaptation of Synechocystis sp. PCC 6803. Journal of Biological Chemistry2009, 284:27875-27883.[DUAL-PAM-100]11.Bailey S, Melis A, Mackey KRM, Cardol P, Finazzi G, Dijken Gv, Berge GM, Arrigo K, Shrager J, Grossman A: Alternative photosynthetic electron flow to oxygen in marine Synechococcus Biochimica et Biophysica Acta 2008, 1777(3):269-276.[DUAL-PAM-100, WATER-PAM]12.Franç ois P, Nadia AA, Cyril S, Philippe J, Radovan P: Alteration of Energy Dissipation by Dichromate in Xanthophyll Deficient Mutants of Chlamydomonas reinhardtii In: Photosynthesis Energy from the Sun: 14th International Congress on Photosynthesis. Edited by Allen JF, Gantt E, Golbeck JH, Osmond B: Springer 2008: 1535-1538.13.Ma W, Chen L, Wei L, Wang Q: Excitation energy transfer between photosystems in the cyanobacterium Synechocystis 6803 Journal of Luminescence2008, 128(3):546-548.[DUAL-PAM-100]14.Ma W, Wei L, Wang Q: The response of electron transport mediated by active NADPH dehydrogenase complexes to heat stress in the cyanobacterium Synechocystis 6803 Science in China2008, 51(12):1082-1087.[dual-pam-100]15.Xu M, Berná t G, Singh A, Mi H, Rö gner M, Pakrasi HB, Ogawa T: Properties of mutants of Synechocystis sp. strain PCC 6803 lacking inorganic carbon sequestration systems. Plant Cell and Physiology2008, 49(11):1672-1677.[DUAL-PAM-100]
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