植物(wu)光(guang)合熱釋(shi)光(guang)測(ce)量(liang)系統(tong)

植(zhi)物(wu)光(guang)合熱釋(shi)光(guang)測(ce)量(liang)系統(tong)是研(yan)究光(guang)合作用(yong)的(de)強有(you)力的(de)工具，是研(yan)究PSII電(dian)子(zi)傳輸的(de)有(you)效(xiao)探(tan)針，廣泛應(ying)用(yong)於除草(cao)劑(ji)對(dui)PSII的(de)受體效(xiao)應(ying)、Q_A與Q_B穩定(ding)性、光(guang)合放氧復(fu)合物（OEC）穩定(ding)性及PSII總體完整(zheng)性等，還用(yong)於光(guang)合突變個體的(de)篩(shai)選(xuan)等。

是針(zhen)對(dui)研(yan)究PSII能(neng)量(liang)水(shui)平(ping)結構(gou)而設計的(de)。PSII反應(ying)中心(xin)光(guang)誘(you)導(dao)電(dian)荷(he)分離(li)導致儲存(cun)了吸收光(guang)能(neng)的(de)激(ji)發(fa)電(dian)子(zi)對(dui)的(de)累(lei)積。加熱誘(you)導這(zhe)些(xie)激發(fa)電(dian)子(zi)對(dui)的(de)重組，從而引發(fa)光(guang)釋(shi)放(fang)，並(bing)在壹(yi)定溫(wen)度(du)範圍(wei)內形(xing)成(cheng)特異性熱釋(shi)光(guang)曲(qu)線。根據不(bu)同釋(shi)光(guang)曲(qu)線的(de)形(xing)狀、峰位(wei)和(he)峰值(zhi)，可以(yi)研(yan)究分析(xi)關(guan)於特定激(ji)發(fa)電(dian)子(zi)對(dui)的(de)能(neng)量(liang)穩定(ding)性及PSII反應(ying)中心(xin)功能(neng)等(deng)。TL200系列熱釋(shi)光(guang)測(ce)量(liang)儀的(de)測(ce)量範圍為(wei)-90°C到+190°C，使用(yong)範圍(wei)更(geng)寬，可以(yi)對(dui)低溫(wen)、高(gao)溫(wen)段熱釋(shi)光(guang)進行研(yan)究。

應(ying)用(yong)領域(yu)

1. 光(guang)合機(ji)理研(yan)究——捕光(guang)色(se)素復合體，PSII反應(ying)中心(xin)，放氧(yang)復合物研(yan)究；PSII能(neng)級(ji)分析(xi)；原初(chu)反應(ying)階(jie)段(duan)的(de)內在過程探(tan)測
2. 植物(wu)脅迫(po)生理的(de)早期(qi)檢測(ce)與(yu)診(zhen)斷
3. 植物(wu)病蟲(chong)害(hai)相(xiang)關(guan)研(yan)究
4. 除草(cao)劑(ji)影響(xiang)
5. 對(dui)植物(wu)光(guang)合研(yan)究的(de)完善補(bu)充

典(dian)型樣品(pin)   

植物(wu)碎片(pian)

各種(zhong)微(wei)藻

葉(ye)綠體懸浮(fu)液(ye)

類囊(nang)體懸浮(fu)液(ye)

工作原理

熱釋(shi)光(guang)(Thermoluminescence，縮(suo)寫TL)是晶(jing)體受到輻(fu)射照射後(hou)，會(hui)產生自由(you)電(dian)子(zi)，這(zhe)些(xie)電(dian)子(zi)被(bei)晶(jing)格缺陷(xian)俘(fu)獲而積攢(zan)起(qi)來，在加熱過程中以(yi)光(guang)形(xing)式釋放出(chu)來。其(qi)基(ji)本(ben)的(de)實(shi)驗過程是(shi)將葉(ye)片(pian)快(kuai)速冷凍(dong)到某(mou)壹(yi)溫(wen)度(du)，之後(hou)給葉(ye)片(pian)壹(yi)個足夠(gou)強，但時(shi)間盡量短（壹(yi)般<5µs）的(de)單(dan)翻(fan)轉(zhuan)光(guang)（single turn-over ?ash），用(yong)於誘導(dao)每個(ge)PSII反映中心(xin)發(fa)生僅(jin)壹(yi)次的(de)電(dian)荷(he)分離(li)；然後(hou)逐(zhu)漸(jian)升溫(wen)，同時(shi)測(ce)量葉(ye)片(pian)放出(chu)的(de)熱釋(shi)光(guang)，繪(hui)制(zhi)TL譜(pu)帶。

熱釋(shi)光(guang)研(yan)究中的(de)壹(yi)個主要(yao)工具是單(dan)次(ci)翻(fan)轉(zhuan)光(guang)閃(shan)，要求(qiu)足夠(gou)強（光(guang)源強度(du)高(gao)達(da) 150 000 µmol(photons).m^－2.s^－1）和(he)足夠(gou)短（典(dian)型 < 5 微(wei)秒(miao)）來誘(you)導每壹(yi)個PSII反應(ying)中心(xin)發(fa)生壹(yi)次，且僅(jin)壹(yi)次的(de)電(dian)荷(he)分離(li)。光(guang)閃(shan)的(de)飽(bao)和(he)效(xiao)果(guo)可以(yi)通(tong)過Q_B段的(de)強度(du)來檢(jian)查，它應(ying)該(gai)在*光(guang)閃(shan)後(hou)達(da)到zui大，在2次光(guang)閃(shan)後(hou)不(bu)再(zai)增加。當前(qian)許多實(shi)驗中使(shi)用(yong)的(de)氙(xian)燈光(guang)閃(shan)具有(you)明(ming)顯缺陷(xian)——壹(yi)個長(chang)的(de)持(chi)續發(fa)光(guang)，或(huo)者(zhe)“閃(shan)光(guang)拖(tuo)尾(wei)”，這(zhe)會在某(mou)些(xie)PSII反應(ying)中心(xin)中產生兩(liang)次(ci)的(de)電(dian)荷(he)分離(li)（連擊）。雖然激光(guang)閃(shan)光(guang)能(neng)夠(gou)有(you)效(xiao)降(jiang)低連擊，但(dan)不(bu)能(neng)消(xiao)除。

TL200系列熱釋(shi)光(guang)測(ce)量(liang)儀使(shi)用(yong)能(neng)量(liang)足夠(gou)強的(de)LED光(guang)源，所(suo)釋放5-10µs的(de)方(fang)波脈沖(chong)能(neng)夠(gou)飽和(he)所有(you)的(de)PSII反應(ying)中心(xin)，其(qi)溫(wen)度(du)控制(zhi)單(dan)元(yuan)可以(yi)在降(jiang)溫(wen)後(hou)，再(zai)使樣品(pin)的(de)溫(wen)度(du)以(yi)0.1℃/sec到 2℃/sec的(de)速(su)率(lv)線性增加。不(bu)同的(de)閃(shan)光(guang)序(xu)列及樣品(pin)處(chu)理能(neng)夠(gou)使樣品(pin)處(chu)於不(bu)同的(de)能(neng)量(liang)狀態，不(bu)同的(de)溫(wen)度(du)下釋(shi)放的(de)光(guang)能(neng)源自光(guang)合機(ji)構(gou)的(de)不(bu)同結構(gou)。分析(xi)釋光(guang)曲(qu)線的(de)形(xing)狀、峰位(wei)和(he)峰值(zhi)，可以(yi)研(yan)究分析(xi)關(guan)於特定激(ji)發(fa)電(dian)子(zi)對(dui)的(de)能(neng)量(liang)穩定(ding)性及PSII反應(ying)中心(xin)功能(neng)等(deng)。

熱釋(shi)光(guang)與(yu)光(guang)合機(ji)構(gou)間關(guan)系

 TL200系列熱釋(shi)光(guang)測(ce)量(liang)儀三(san)種(zhong)型號(hao)的(de)控溫(wen)方式與範圍

系統(tong)組成(cheng)

TL200系列熱釋(shi)光(guang)測(ce)量(liang)系統(tong)由(you)3部分組成(cheng)：多功(gong)能(neng)控制(zhi)單(dan)元(yuan)、溫(wen)度(du)調節(jie)器控制(zhi)單(dan)元(yuan)及測量(liang)室 。

多功(gong)能(neng)控制(zhi)單(dan)元(yuan)（Multipurpose Control Unit）根據用(yong)戶定(ding)義方(fang)案(an)或熱發(fa)光(guang)向(xiang)導(dao)提(ti)供(gong)的(de)實(shi)驗程序(xu)來執(zhi)行(xing)實驗(yan)過程，有(you)兩(liang)個(ge)輸入(ru)頻(pin)道，壹(yi)個用(yong)於測量(liang)熱發(fa)光(guang)信號(hao)（TL信號(hao)），另壹(yi)個用(yong)於測量(liang)溫(wen)度(du)。 測量(liang)曲線以(yi)兩種(zhong)格式顯示：時(shi)間/溫(wen)度(du)和(he)時(shi)間/TL信號(hao)，或溫(wen)度(du)/TL信號(hao)。

溫(wen)度(du)調節(jie)器控制(zhi)單(dan)元(yuan)（Thermoregulator Control Unit）可以(yi)在-90°C 到 +190°C範圍(wei)內以(yi)0.1°C的(de)精(jing)確(que)度(du)控制(zhi)樣品(pin)的(de)溫(wen)度(du)。 系統(tong)前(qian)面板(ban)可以(yi)顯示實(shi)際的(de)溫(wen)度(du)，溫(wen)度(du)調節(jie)可以(yi)通(tong)過手(shou)動或(huo)程序(xu)控制(zhi)（軟(ruan)件）來實(shi)現(xian)。有(you)兩(liang)種(zhong)工作模(mo)式：恒溫(wen)模(mo)式和(he)溫(wen)度(du)梯(ti)度(du)模(mo)式，在恒(heng)溫(wen)模(mo)式下儀器將維持(chi)樣品(pin)在恒(heng)定(ding)的(de)溫(wen)度(du)，而在溫(wen)度(du)梯(ti)度(du)模(mo)式下，可以(yi)使樣品(pin)的(de)溫(wen)度(du)以(yi)0.1°C/sec到 2°C/sec的(de)速(su)率(lv)線性變化(hua)。

AC-88水(shui)冷單(dan)元(yuan)可將系統(tong)溫(wen)度(du)降低(di)到4°C，包含(han)壹(yi)個電(dian)子(zi)控制(zhi)的(de)抽(chou)水(shui)泵和(he)內部可以(yi)儲水(shui)的(de)制(zhi)冷器，用(yong)於降低(di)測(ce)量(liang)室(shi)的(de)環(huan)境溫(wen)度(du)。

CryoFab液(ye)氮(dan)罐通(tong)過管(guan)路(lu)連(lian)接(jie)到測(ce)量(liang)室，通(tong)過電(dian)子(zi)控制(zhi)的(de)低(di)溫(wen)輸出(chu)閥可以(yi)將系統(tong)溫(wen)度(du)控制(zhi)在 -20°C 到 -90°C。

由(you)TFPE單(dan)元(yuan)控制(zhi)的(de)輔(fu)助加熱模(mo)塊可以(yi)將系統(tong)溫(wen)度(du)加熱到+190°C。

測量(liang)室（Measuring Chamber）又(you)包括(kuo)四(si)個關(guan)鍵組成(cheng)部分：光(guang)源、 光(guang)電(dian)倍(bei)增(zeng)器、A/D 轉換(huan)器、具有(you)溫(wen)度(du)控制(zhi)器的(de)樣品(pin)盤(pan)：

1. 光(guang)源由(you)8個超(chao)亮的(de)發(fa)光(guang)二(er)極管(guan)(λmax=630 nm)組成(cheng)，發(fa)射的(de)光(guang)閃(shan)強度(du)高(gao)達(da)150,000 µmol(photons). m^－2.s^－1以(yi)上，光(guang)閃(shan)持(chi)續時(shi)間zui長(chang)為(wei)150 µs（典(dian)型5-10us），光(guang)強和(he)光(guang)閃(shan)持(chi)續時(shi)間通(tong)過軟件控制(zhi)。
2. 光(guang)電(dian)倍(bei)增(zeng)器可以(yi)探(tan)測從300到900nm範圍(wei)的(de)光(guang)量(liang)子(zi)，從而測量(liang)熱發(fa)光(guang)信號(hao)和(he)緩(huan)發(fa)熒光(guang)。光(guang)電(dian)倍(bei)增(zeng)器包括(kuo)自己的(de)電(dian)源。
3. A/D轉換(huan)器用(yong)於光(guang)電(dian)倍(bei)增(zeng)器的(de)電(dian)流(liu)放(fang)大、軟件控制(zhi)增(zeng)益(yi)和(he)數字化(hua)，放(fang)大器的(de)時(shi)間反應(ying)固(gu)定在50ms，以(yi)確(que)定zui小取(qu)樣周期(qi)到100ms。

技(ji)術(shu)參數

• 溫(wen)度(du)範圍(wei)：

TL200/PMT標(biao)準版(ban)：-25 ℃ 到+70 ℃

TL 300/HT高(gao)溫(wen)版：-25 ℃ 到+190 ℃

TL 400/LT液(ye)氮(dan)版：-90 ℃ 到+70 ℃

• 控溫(wen)模(mo)式：恒溫(wen)；線性變化(hua)（0.1ºC/sec - 2ºC/sec）
• 過熱保護：提(ti)供(gong)
  環(huan)境光(guang)保護：提(ti)供(gong)
• 控制(zhi)模(mo)式：手(shou)動（恒(heng)溫(wen)）；程序(xu)設定(ding)溫(wen)度(du)曲線
• 樣品(pin)盤(pan)：直徑(jing)½英寸(cun)鍍(du)金銅盤
• 測量(liang)樣品(pin)：藻類(lei)、藍(lan)細(xi)菌、葉(ye)綠體懸浮(fu)液(ye)，葉(ye)片(pian)碎片等
• 光(guang)源：波(bo)長(chang)lmax=625nm，光(guang)源強度(du)高(gao)達(da) 150 000 µmol(photons). m^－2.s^－1以(yi)上
• 探(tan)測系統(tong)：傳感(gan)器為可以(yi)通(tong)過軟件靈(ling)敏(min)控制(zhi)的(de)光(guang)電(dian)倍(bei)增(zeng)器，光(guang)譜(pu)響(xiang)應(ying)為(wei)300nm-900nm，zui小取(qu)樣周期(qi)100ms，時(shi)間響(xiang)應(ying)50ms，接通(tong)延(yan)遲100ms
• 控制(zhi)：用(yong)戶可通(tong)過語言(yan)自定(ding)義程序(xu)控制(zhi)儀器測量(liang)過程
• 通(tong)訊(xun)：USB
• 軟件：FluorWin 3.6
• 電(dian)源：90V-240V

操(cao)作(zuo)軟(ruan)件與實(shi)驗(yan)結果

典(dian)型應(ying)用(yong)

上圖為(wei)源自擬(ni)南(nan)芥(jie)未(wei)冷凍(dong)葉(ye)片(pian)的(de)熱釋(shi)光(guang)（M.Roman ，1998）。實心(xin)符號(hao)：對(dui)照（a）；空心(xin)符號(hao)：輕(qing)度(du)脫水(shui)（b）。單(dan)閃(shan)（細(xi)線）產生75%的(de)S₂和(he)25%的(de)S₁（只有(you)S₂和(he)S₃產生熱釋(shi)光(guang)，S₁無），雙(shuang)閃(shan)（粗線）25%的(de)S₂和(he)75%的(de)S₃，3閃(shan)（點線）25%的(de)S₃。a、對(dui)照植物(wu)。單(dan)閃(shan)後(hou)，熱釋(shi)光(guang)B段與(yu)S₂Q_B-相(xiang)*（B2，見(jian)表1）且可以(yi)被單(dan)因(yin)子(zi)擬(ni)合得很好(hao)。2次閃(shan)光(guang)後(hou)，則(ze)需(xu)要3個因(yin)子(zi)，S₂Q_B-（B1），S₃Q_B-（B2）和(he)壹(yi)個剩余(yu)因子(zi)（未(wei)顯示）。b、適度(du)脫水(shui)的(de)植(zhi)物。B段下(xia)調，S₃比(bi)S₂在更(geng)大程度(du)上表明(ming)了類(lei)囊體腔內壹(yi)個暗穩態(tai)的(de)酸(suan)性pH。45攝(she)氏(shi)度(du)段（余(yu)輝）源自S_2/3Q_B中心(xin)中熱誘(you)導的(de)從基質(zhi)還原劑(ji)向Q_B的(de)電(dian)子(zi)傳遞，使它們(men)發(fa)光(guang)：它的(de)增(zeng)加表明(ming)了壹(yi)個強的(de)同化(hua)勢能(neng)NADP+ATP（Ducruet 2003）。

產地(di)：歐(ou)洲(zhou)

參考文(wen)獻：（僅(jin)2011年發(fa)表(biao)的(de)部分文(wen)獻，77篇(pian)中的(de)14篇(pian)）

• Thermoluminescence. PV Sane, et al, 2012. Photosynthesis
• Analysis of S2QA-charge recombination with the Arrhenius, Eyring and Marcus theories. S Rantamäki, et al, 2011. Journal of Photochemistry and Photobiology B: Biology
• Manganese limitation induces changes in the activity and in the organization of photosynthetic complexes in the cyanobacterium Synechocystis sp. strain PCC 6803. E Salomon, et al, 2011. Plant physiology
• Inhibition of photosynthetic oxygen evolution and electron transfer from the quinone acceptor QA− to QB by iron deficiency. N Msilini, et al, 2011. Photosynthesis research
• Chlorophyll fluorescence emission as a reporter on cold tolerance in Arabidopsis thaliana accessions. A Mishra, et al. Plant Signaling & Behavior, 2011
• Characterization of photosystem II in transgenic tobacco plants with decreased iron superoxide dismutase. Y Zhang, et al, 2011. Biochimica et Biophysica Acta
• Two functional sites of phosphatidylglycerol for regulation of reaction of plastoquinone Q< sub> B</sub> in photosystem II. S Itoh, et al, 2011. Biochimica et Biophysica Acta
• Binding Stoichiometry and Affinity of the Manganese-Stabilizing Protein Affects Redox Reactions on the Oxidizing Side of Photosystem II. JL Roose, et al, 2011. Biochemistry
• Species-dependence of the redox potential of the primary quinone electron acceptor QA in photosystem II verified by spectroelectrochemistry. T Shibamoto, et al, 2010. FEBS letters
• Flavodiiron proteins in oxygenic photosynthetic organisms: photoprotection of photosystem II by Flv2 and Flv4 in Synechocystis sp. PCC 6803. P Zhang, et al, 2009. PloS one
• Comparative analysis of leaf‐type ferredoxin‐NADP+ oxidoreductase isoforms in Arabidopsis thaliana. M Lintala, et al, 2009. The Plant Journal
• Psb28 protein is involved in the biogenesis of the photosystem II inner antenna CP47 (PsbB) in the cyanobacterium Synechocystis sp. PCC 6803. M Dobáková, et al, 2009. Plant physiology
• Functional complementation of the Arabidopsis thaliana psbo1 mutant phenotype with an N-terminally His6-tagged PsbO-1 protein in photosystem II. H Liu, et al, 2009. Biochimica et Biophysica Acta
• Interaction of methylamine with extrinsic and intrinsic subunits of photosystem II. S Hamdani, et al, 2009. Biochimica et Biophysica Acta
• Investigation of the low-affinity oxidation site for exogenous electron donors in the Mn-depleted photosystem II complexes. VN Kurashov, et al, 2009. Biochimica et Biophysica Acta
• Changes in the photosynthetic reaction centre II in the diatom Phaeodactylum tricornutum result in non‐photochemical fluorescence quenching. D Eisenstadt, et al, 2008. Environmental Microbiology
• Characterization of photosystem II in salt-stressed cyanobacterial Spirulina platensis cells. H Gong, et al, 2008. Biochimica et Biophysica Acta
• The cyanobacterial homologue of HCF136/YCF48 is a component of an early photosystem II assembly complex and is important for both the efficient assembly and Repair of Photosystem II in Synechocystis sp. PCC 6803*. J Komenda, et al, 2008. Journal of Biological chemistry.
• Early detection of bean infection by Pseudomonas syringae in asymptomatic leaf areas using chlorophyll fluorescence imaging. L Rodríguez-Moreno, et al, 2008. Photosynthesis research
• Effects of heat stress on PSII photochemistry in a cyanobacterium Spirulina platensis. B Zhao, et al, 2008. Plant Science
• The effects of simultaneous RNAi suppression of PsbO and PsbP protein expression in photosystem II of Arabidopsis. X Yi, et al, 2008. Photosynthesis research
• Functional analysis of photosystem II in a PsbO-1-deficient mutant in Arabidopsis thaliana. H Liu, et al, 2007. Biochemistry
• Efficiency and role of loss processes in light‐driven water oxidation by PSII. M Grabolle, et al, 2007. Physiologia plantarum
• Changes of Photosystem II Electron Transport in the Chlorophyll‐deficient Oilseed Rape Mutant Studied by Chlorophyll Fluorescence and Thermoluminescence. JW Guo, et al, 2007. Journal of Integrative Plant Biology
• The exposed N-terminal tail of the D1 subunit is required for rapid D1 degradation during photosystem II repair in Synechocystis sp PCC 6803. J Komenda, et al, 2007. The Plant Cell
• The chlorophyll a fluorescence induction pattern in chloroplasts upon repetitive single turnover excitations: accumulation and function of QB-nonreducing centers. W Vredenberg, et al, 2006. Biochimica et Biophysica Acta
• The polyphasic chlorophyll a fluorescence rise measured under high intensity of exciting light. D Lazár , 2006. Functional Plant Biology
• The synechocystis sp PCC 6803 oxa1 homolog is essential for membrane integration of reaction center precursor protein pD1. F Ossenbühl, et al, 2006. The Plant Cell
• Sucrose and glycerol effects on photosystem II. KM Halverson, et al, 2003. Biophysical journal
• Copper-induced inhibition of photosynthesis: limiting steps of in vivo copper chlorophyll formation in Scenedesmus quadricauda. H Küpper, et al, 2003. Functional Plant Biology
• Heavy metal‐induced inhibition of photosynthesis: targets of in vivo heavy metal chlorophyll formation1. h küpper, et al, 2002. journal of Phycology
• Experimental and theoretical studies on the excess capacity of Photosystem II. R Kaňa, et al, 2002. Photosynthesis research
• Nitrogen deprivation strongly affects Photosystem II but not phycoerythrin level in the divinyl-chlorophyll b-containing cyanobacterium Prochlorococcus marinus. C Steglich, et al, 2001. Biochimica et Biophysica Acta
• Integrity and activity of photosystem 2 complexes isolated from the thermophilic cyanobacterium Synechococcus elongatus using various detergents. E Šetlíková, et al, 1999. Photosynthetica
• The PsbY protein is not essential for oxygenic photosynthesis in the cyanobacterium Synechocystis sp. PCC 6803. M Meetam, et al, 1999. Plant physiology
• Nonphotochemical reduction of the plastoquinone pool in sunflower leaves originates from chlororespiration. TS Feild, et al, 1998. Plant physiology