藻胆体的光谱特性光能传递

1．对嗜热蓝藻层理鞭枝藻(Mastigocladus laminosus)藻胆体的光谱特性和光能传递进行了研究。其完整藻胆体的吸收峰位于622 nm，室温荧光发射峰位于673 nm。在77K荧光发射光谱中，完整藻胆体的荧光峰只有一个，位于685 nm，是末端发射体1的荧光。在低浓度磷酸缓冲液中发生严重解离的藻胆体，其77K荧光发射光谱中有二个发射峰和一个发射肩。两个荧光发射峰分别位于644 nm和683 nm。前者为主峰，属于C-藻蓝蛋白的荧光，后者是次峰，属于末端发射体2的荧光。荧光发射肩位于660 nm附近，属于别藻蓝蛋白的荧光。据此，提出层理鞭枝藻藻胆体光能传递途径如下： 藻红蓝蛋白→c—藻蓝蛋白→别藻蓝蛋白→端发射体l、末端发射体2： 2．对嗜热蓝藻层理鞭枝藻藻胆体—类囊体膜的光谱特性和光能传递进行了研究。在吸收光谱中，其藻胆体——类囊体膜在可见光区域有5个峰，它们分别位于420 nm、438 nm、490 nm、624 nm和678 nm。420 nm、438 nm和678 nm为叶绿素a的吸收峰位置。490 nm是类胡罗卜素的吸收峰，624 nm是藻胆体的吸收峰。对藻胆体——类囊体膜用580 nm波长的光激发藻胆蛋白时，在室温荧光发射光谱中有一个发射峰和一个发射肩，分别位于657 nm和690 nm，前者属于藻胆蛋白的荧光，后者属于叶绿素a的荧光。这说明藻胆蛋白能将捕获的光能传递给类囊体膜上的叶绿素a。在77K荧光发射光谱中有4个峰，它们分别位于649 nm、660 nm、688 nm和730 nm。前二者属于藻胆蛋白的荧光，后二者属于叶绿素a的荧光。这同样说明藻胆蛋白能将捕获的光能传递给类囊体膜上的叶绿素a。当用436 nm波长光激发叶绿素a时，藻胆体——类囊体膜的室温荧光发射光谱中有两个荧光峰出现，位于685 nm的峰来源于光系统Ⅱ，位于713 nm的峰来源于系统I。这说明叶绿素a捕获的光能不能逆传递给藻胆体中的藻胆蛋白。在77K荧光发射光谱中也只有叶绿素a的荧光峰，位于695 nm的峰来源于光系统Ⅱ，位于730 nm的峰来源于光系统l。此结果同样说明叶绿素a捕获的光能不能逆传递给藻胆蛋白． 3．我们以多变鱼腥藻(Anabaena variabilis)为材料，对其藻胆体核心和藻蓝蛋白进行了重组实验，得到了具有光能传递效率的藻胆体核心——藻蓝蛋白复合物。在吸收光谱中，藻胆体核心有一吸收峰和一个吸收肩，分别位于654 nm和600 nm。藻蓝蛋白的吸收光谱中只有一个峰，位于620 nm．重组样品的吸收光谱有一吸收峰和一吸收肩，分别位于654 nm和620 nm．由于620 nm与654 nm的吸收比远大于核心的600 nm与654 nm的吸收比，因此，可以认为部分藻蓝蛋白已与核心重组。在室温荧光发射光谱中，藻胆体核心只有一个峰，位于676 nm。藻蓝蛋白只有一个峰，位于653 nm。重组样品有一荧光发射峰和一荧光发射肩，分别在669 nm和650 nm附近。669 nm荧光来源于核心，650 nm荧光来源于藻蓝蛋白。重组后的核心的650 nm荧光显著大于未重组的核心，这也说明部分藻蓝蛋白与核心已重组．在77K荧光发射光谱中，藻蓝蛋白只有一个峰，位于655 nm。藻胆体核心有二个峰，分别位于666 nm和686 nm。重组样品有两个荧光发射峰和一荧光发射肩，分别位于666 nm、683 nm和648 nm附近．重组的核心的别藻蓝蛋白的荧光(F666)和藻蓝蛋白的荧光(F648)都强于未重组的核心。这一结果同样说明有藻胆体——藻蓝蛋白复合物生成。 除以上研究工作之外，我们还对多变鱼腥藻藻胆体在解离过程中的光谱特性及光能传递、藻胆体——类囊体膜的光谱特性及光能传递、藻胆体解离重组、藻胆体核心在低浓度磷酸缓冲液中的光谱特性、以及温度对藻胆体核心的影响等进行了研究。研究结果有待整理。 本文编写：PBS：藻胆体;PEB：藻红胆素;PE：藻红蛋白;PUB:藻尿胆素;PEC：藻红蓝蛋白;PCB:藻蓝胆素;PC：藻蓝蛋白;PSⅡ：光系统Ⅱ;APC:别藻蓝蛋白;PS I：光系统I;TE:末端发射体

光系统II反应中心吸能和转能的分子机理研究

全文分两部分，(1)．PsⅡ反应中心色素分子光破坏的分子机理研究;(2)．PSⅡ反应中心原初反应的动力学机理研究。 在第一部分中，在分离纯化的光系统Ⅱ反应中心Dl/D2/Cyt b559复合物中，采用高效液相色谱技术，首次发现PSⅡ反应中心去镁叶绿素分子的光照破坏，研究了去镁叶绿素的光破坏机理，观察到PsⅡ反应中心内部存在一个与光化学活性无关的去镁叶绿素分子，从而提供了PSⅡ反应中心存在两条电子传递链的第一个实验证据，提出了去镁叶绿素对PsⅡ反应中心的光保护假说和光合作用反应中心第二条电子传递支路的光保护假说。用高效液相色谱技术还观察到PSⅡ反应中心的6个叶绿素a分子，有三种不同的存在状态，认为PSl反应中心的最小色素组成为每个反应中心含有4个叶绿素a和2个去镁叶绿素。用光破坏的方法证明PsⅡ原初电子供体P680是由两个叶绿素n分子组成，认为P680是以一个二聚体形式存在，首次发现P680的光破坏过程包含失去中心镁原子的反应。 在第二部分中，用皮秒和飞秒时间分辨光谱技术，在PsⅡ颗粒、PsⅡ核心复合物和PSⅡ反应中心三个层次上，研究了PsⅡ原初反应的动力学性质，着重研究电荷分离和PsⅡ反应中心内部的能量传递过程。结果表明，B-胡萝卜素和P680之间的能量传递时间常数为350p8左右，去镁叶绿素a与P680之间的能量传递时间为lOOp8左右，提出了可能的动力学模型。 在目前分歧最大的原初电荷分离时间常数测定这一焦点问题上，得到的初步结果表明PsⅡ反应中心电荷分离时间为3-3.5pa左右，这一结论与文献上报道的21pa不同，丽倾向于支持国际上3p8的观点。

光系统Ⅱ核心天线复合物CP43和CP47的分离纯化及其光谱学特性的研究

应用改进DEAE-Toyopearl 650S阴离子交换柱层析从高等植物菠菜(Spinacia oleracea)中分离纯化了核心天线复合物CP43和CP47。并对它们的纯度和完整性色素种类和含量，以及色素分子的结合状态进行了研究并对色素分子间的能量传递机制进行了讨论。结果如下： 1、HPLC检测结果表明：纯化的CP43和CP47均只含Chla和β-Car两种色素分子，并且，平均每分子CP43多肽含19-20分子Chla和4-5分子β-Car;而平均每分CP47则含20-21分子Chla和3-4分子β-Car。 2、以436nm和480nm激发光激发样品得到的CP43和CP47的低温荧光发射光谱的最大荧光发射峰分别位于683nm和693nm。进一步发现，CP43和CP47，在相同条件下分别以436nm和480nm激发光激发样品得到的低温荧光发射光谱经归一化后几乎完全重叠，而且400-500nm波长范围内的激发光扫描得到的三维低温荧光发射光谱沿激发轴具有较好的对应关系，表明纯化的CP43和CP47都具有较高的完整性。 3、纯化的CP43和CP47的吸收光谱的红区最大吸收峰分别位于671nm和674nm。该光区的导数光谱均分辨出偏蓝区和偏红区两个子峰，CP43的这两个子峰分别位于669nm和682nm;而CP47的两个子峰则分别位于669nm和680nm。进一步用包含这两个子峰的高斯解析参数对红区最大吸收峰进行拟合，结果证明，拟合的曲线与实测曲线几乎完全吻合，这表明，CP43和CP47均至少包含两种不同状态的Chla分子。 3．1应用不同的变性温度处理CP43，发现随变性温度的不断提高，其红区最大吸收峰的峰值逐渐减小，四阶导数光谱分辨出的两个子峰同时减小，但差光谱显示：随处理温度的不断提高，这两个组分峰值的变化并不同步进行，较低温度范围内(55℃以下)682nm吸收峰下降明显，而较高温度范围内(55℃以上)，669nm吸收峰下降明显。 同时，随处理温度不断提高CP43脱辅基蛋白的结构也在不断发生变化，其变化过程明显表现出两个跃变阶段。这两个跃变阶段分别出现在40～50℃范围内和55～60℃范围内，恰与吸收光谱两个组分峰变化的转变过程相一致。这证明，CP43中分别位于669nm和682nm的不同的色谱组分即代表两种不同结合态的Chla分子，分别简称为“CP43-669”和“CP43-682”。它们在色素蛋白复合物中所处的环境不同，因而对蛋白质结构的依赖性不同，前者更高地依赖于蛋白复合物的整体构象，而后者则主要依赖于蛋白质的二级结构。 3.2 经不同的变性温度处理的CP47，其红区最大吸收峰的峰位逐渐蓝移，而吸收峰值无明显的变化，只有当处理温度提高到65℃以后，蓝移后的吸收峰值(669nm)才开始明显减小;四阶导数光谱表现为680nm吸收峰的信号逐渐下降669nm的吸收信号逐渐明显;处理减对照差光谱只观察到680nm吸收值的逐渐减少，而几乎观察不到669nm吸收值的变化。同时，随变性温度的不断提高，CP47的脱辅基蛋白的结构也发生相应的变化与CP43不同，蛋白结构变化最大的温度范围为60℃～65℃之间，但同CP47的峰位蓝移、导数光谱中680nm信号的减小，以及差光谱中680nm吸收值的减小相一致。由此认为，同CP43一样，CP47的吸收光谱中分辨出的分别位于669nm和680nm处的两个不同光谱组分亦分别代表两种不同结合状态的Chla分子，分别简称为“CP47-669”和“CP47-680”，与CP43中的相应组分对应，它们处于不同的蛋白环境中，从而对蛋白质结构变化的依赖性不同。 3.3 CP43和CP47的CD光谱表现出明显的正负双峰，表明色素分子间存在较强的激子相互作用。随变性温度的不断提高，正负CD双峰的信号逐渐减弱，变化过程与脱辅基蛋白结构的变化以及CP43-682的变化相一致，表明色素分子间的激子相互作用更高依赖于CP43-682和CP47-680。并认为CP43-682和CP47-680可能以二聚体或多聚体的形式存在，并且二聚体或多聚体的形成依赖于蛋白天然构象。而CP43-669和CP47-669则以单体的形式位于蛋白结构中相对伸展的区域。并提出：在CP43-682以CP47-680分子之间，激发能主要以激子偶合机制进行而在CP43-669，CP47-669分子间及CP43-669至CP43-682间，CP47-669至CP47-680之间激发能则主要以Foster机制进行。 4、以488nm激发光得到的CP43和CP47的共振拉曼光谱都具有全反式构型类胡萝卜素分子的四个典型特征峰由此认为CP43和CP47中的β-Car分子亦具有全反式构型;与溶于丙酮抽体物中的β-Car分子相比较，CP43和CP47中的β-Car分子的共振拉曼光谱中具有较强的960cm~(-1)的拉曼峰，表明，CP43和CP47中的β-Car分子具有扭曲的构象。 应用经归一化后的吸收光谱与荧光激发光谱相比较的办法发现CP43和CP47中存在β-Car分子和Chla分子间的能量传递其能量传递效率分别为29.8～29.9%和52.3～56.9%。这表明，在正常条件下，CP47中β-Car分子和Chla分子间的能量传递效率远大于CP43。此外，当选用蛋白结构变化最明显的热变性温度处理样品后，发现，不论CP43还是CP47中β-Car与Chla分子间的能量传递效率大大降低，表明，这两种色素分子间的能量传递严格依赖于蛋白复合物的天然构象，并认为，正常条件下，CP43和CP47内β-Car与Chla分子间的空间距离较近，可能不大于10A，CP43和CP47相比较，CP47内这两种色素分子间的距离更近。并进一步提出，在CP43和CP47中，β-Car到Chla分子间的能量传递最大可能以Dexter的电子交换机制进行。

发状念珠藻光合特征及其在复水－失水过程中能量传递变化的研究

摘要 "发状念珠藻（Nostoc flagelliforme Born. Et Flah.），俗名发菜，是生长于干旱、半干旱土壤表面的陆生蓝藻，具有极强的抗旱能力。发菜光合作用方面的研究大多处于整体细胞水平，且研究手段非常有限。本实验对发菜光合特征进行深入研究，并探讨了发菜在干湿交替过程中能量传递的变化情况。 叶绿素和藻胆素是发菜细胞中两种主要的光合色素。发菜复水后光合活性完全恢复时，在室温（20℃）或低温（77K）下，其绝大部分的荧光是由于藻胆素被激发而产生。在室温下，大部分荧光来自藻胆体;当叶绿素被激发后，产生的荧光非常微弱。在低温下，藻胆素被激发后，荧光发射光谱中可分辨出藻胆蛋白、光系统Ⅰ和光系统Ⅱ的发射峰;叶绿素被激发后，荧光发射光谱包括光系统Ⅰ和光系统Ⅱ的荧光。相比之下，室温荧光发射光谱不适于用做发菜细胞光合作用方面的研究。 我们设计了一种新方法，从野生发菜细胞中分离得到类囊体膜及细胞质膜，并对其性质进行分析。发菜细胞外复杂的胶质结构使得现有破碎其它蓝藻细胞的方法无法破碎发菜细胞。通过实验发现，联合使用细胞破碎仪和毛地黄皂甙（0.3%）可有效破碎发菜细胞;并且毛地黄皂甙在低浓度下（≦0.5%），对色素与蛋白的结合不会造成破坏作用。随后，通过蔗糖密度梯度离心可将细胞质膜与类囊体膜分离。发菜类囊体膜的光谱性质与其它蓝藻相似。细胞质膜除结合有类胡萝卜素外，还结合有少量叶绿素前体。类囊体膜和细胞质膜膜脂及脂肪酸组成相似。其中，十六碳烯酸[16：1(9)]和亚麻酸[18：3(9,12,15)]是含量最高的两种脂肪酸，分别占总脂肪酸含量的三分之一左右。高含量的多不饱和脂肪酸可能和发菜极强的抗旱能力有关。 我们首次对发菜捕光色素蛋白复合物－藻胆体的组成和结构进行分析。发菜藻胆体为“3核＋6杆”的半圆盘结构。组成藻胆体的藻胆蛋白包括藻蓝蛋白和别藻蓝蛋白。两个藻蓝蛋白六聚体通过连接肽组成藻胆体的“杆”结构。在“杆”结构中等量分布着两条连接肽（分子量分别为29kDa和34kDa）为杆连接肽和核杆连接肽。而“核”结构中核膜连接肽的分子量为103kDa。 发菜在无霜期，几乎每天经历一次复水－干燥过程：夜间的结露使发菜在黑暗中复水，而清晨太阳升起后，在光照下迅速失水进入休眠状态。我们研究了发菜在黑暗中的复水过程及在光照下失水过程中藻胆体与光系统能量传递的变化情况。发菜在黑暗中复水后，光系统Ⅱ活性无法恢复。藻胆体内及藻胆体与光系统Ⅰ的能量传递在5分钟内恢复;而藻胆体与光系统Ⅱ的能量传递只能部分恢复。我们设想，发菜在复水过程中通过双扳机－水和光－控制光合活性的恢复，以及在黑暗中部分恢复藻胆体与光系统Ⅱ的能量传递，将减少不必要的能量消耗与通过光合作用储备尽可能多的化学能－这两个生存策略有机的结合起来。发菜在光照下的失水过程中，光合活性在含水量降至90％前基本保持稳定，随后迅速下降。而在含水量达到150％后，藻胆体内的能量传递便开始受到抑制，并且随着含水量的降低，该抑制现象逐步加剧。这样，发菜在干燥过程中，通过抑制藻胆体内的能量传递，减少了传递到光系统Ⅱ反应中心的能量，从而避免了在光合活性下降过程中过剩光能对光系统Ⅱ产生的破坏作用。"

假根羽藻细胞色素b6f蛋白复合体中α-胡萝卜素的结构与功能

细胞色素b6f蛋白复合体（Cyt b6f）是光合链中连接两个光系统（PSII 和PSI）的中间电子载体蛋白复合物，其主要的生理功能是催化电子传递和质子跨膜转移，形成跨膜质子电化学梯度，为ATP的合成提供能量，在光合作用光能转化过程中占有很重要的地位。细菌和莱茵衣藻Cyt b6f的晶体结构已于2003年底获得了近原子水平的解析，但有关该复合物中两种色素（Chl a和β-Car）的生理功能及其机理尚无明确的解释。预计它们将成为今后几年的研究热点,因为揭示Cyt b6f蛋白复合体中Chl a和β-Car分子的生理功能对于进一步阐明光合作用高效转能及其调控的分子机理具十分重要的意义。鉴于目前尚未见到海洋绿藻Cyt b6f的报道，本文以海洋绿藻—假根羽藻（Bryopsis corticulans）类囊体膜上的Cyt b6f蛋白复合体为对象，对其中的类胡萝卜素的分子结构与生理功能进行了比较系统地研究。 首先，我们改进了原用于菠菜类囊体膜Cyt b6f的分离、纯化流程，在原流程的基础上增加了一次2 mol/L NaBr洗膜，彻底地去除了膜表面的杂蛋白;还调整了第二次硫酸铵分级沉淀时的饱和度，并将38-45%饱和度下的沉淀物确定为需要收集的Cyt b6f制剂。采用此改进的流程，我们首次从假根羽藻类囊体膜中分离纯化了高活性、高纯度的Cyt b6f制剂。SDS-PAGE分析的结果显示该制剂的4个多肽亚基 （Cytf 、Cyt b6 、Rieske[Fe-s]及亚基IV）的表观分子量分别为34.8、24.0、18.7和16.7 kD;Cyt b6 / f 比值接近2.0， 其纯度值为9.9 nmol cyt f/mg;其催化电子传递的活性 （C10-PQH2→PC）为73 e/s。HPLC 和共振拉曼光谱分析表明，假根羽藻Cyt b6f中的类胡萝卜素为α-胡萝卜素分子，它是一种在Cyt b6f中尚未报道过的类胡萝卜素。定量分析表明，每个假根羽藻Cyt b6f单体中全反式(all-trans)和9顺式（9-cis）α-胡萝卜素的含量分别为0.2和0.7个分子，另外还含有1.2分子的Chl a。CD光谱分析表明该9-cis-α-胡萝卜素处在一个不对称的蛋白环境中。TLC分析表明该制剂是一种缺脂的Cyt b6f蛋白复合体。 采用稳态荧光激发光谱，时间分辨吸收光谱及Chl a的光破坏实验对假根羽藻Cyt b6f中α-胡萝卜素的功能进行了研究。结果表明，Cyt b6f中α-胡萝卜素可以将它吸收的光能传递给Chl a，其能量传递效率为62.4%，提出α-胡萝卜素分子与Chl a分子之间的单线态能量传递是遵从Föster 机制进行的;α-胡萝卜素分子对Chl a分子有一定的光保护作用，这种保护作用是通过清除单线态氧来实现的。另外还发现Cyt b6f中的Chl a分子可能与其周围的氨基酸残基存在相互作用，认为这是其进行自我光保护的一种方式。 此外，还采用HPLC研究了光和暗交替对假根羽藻Cyt b6f中α-胡萝卜素构型的影响，并对假根羽藻Cyt b6f选择结合α-胡萝卜素的原因进行了初步的分析。

PSII色素蛋白复合物的脂质体重组及反应中心的功能研究

光系统II（PSII）是存在于类囊体膜中的多亚基色素蛋白复合物，是吸收光能、催化光诱导水裂解释放氧气、质子和电子的重要机构。它在体内的基本单位是由外周天线蛋白（LHCII）与PSII核心复合物结合形成的PSII-LHCII超分子复合物，这一结构保证了LHCII吸收的能量能够快速有效的传递到PSII反应中心（RC），进行原初光化学反应。 本论文分为两部分：1、利用捕光色素蛋白复合物（LHCII）与PSII核心复合物在以DGDG、PG、SQDG三种类囊体膜脂形成的脂质体中重组的方法，研究了LHCII与PSII在脂膜上结构与功能的相互作用;2、通过研究光破坏和色素置换对PSII RC的影响，探讨了RC中不同色素的功能。主要结果如下： 1、LHCII与PSII核心复合物的蛋白脂质体研究： 将OECC（粗提核心复合物）、pdOE（纯化核心复合物）、LHCII（大量天线）制剂分别与脂质体重组并研究了其光谱性质。LHCII在与脂质体重组前表现出典型的聚集态光谱特征，重组后吸收和荧光发射峰发生明显蓝移;LHCII、OECC和pdOE三种蛋白脂质体与重组前的样品相比荧光发射强度增加;表明脂环境影响了色素蛋白复合物的聚集状态以及色素和蛋白之间的相互作用。 OECC和pdOE分别与LHCII在脂质体中重组，得到两种重组蛋白（LHCII-OECC和LHCII-pdOE）脂质体，用冰冻蚀刻电镜技术和低温荧光光谱的方法研究其结构和功能特征。LHCII和核心复合物（OECC或pdOE）结合形成PSII-LHCII重组颗粒，并在脂质体中均匀排布，阻止了LHCII晶格状结构的形成。重组蛋白脂质体的吸收光谱既有LHCII的吸收特征，又有核心复合物的特征吸收峰，但低温荧光光谱的主要发射峰是核心复合物的特征峰（684 nm-685 nm），而不是LHCII的特征峰（680 nm）;而且激发不同色素得到的荧光发射光谱基本一致，这些结果证明LHCII吸收的能量传递到了核心复合物中，在重组蛋白脂质体中不同色素蛋白复合物在结构和功能上都实现了相互偶联。 通过对OECC或pdOE与LHCII重组形成的蛋白脂质体放氧或DCPIP光还原活性的检测研究了PSII光化学活性特征。LHCII和核心复合物（OECC或pdOE）的重组蛋白脂质体与单独核心脂质体相比，在强光和弱光下光化学活性都明显提高。这从另一个角度证明了核心复合物与LHCII的功能偶联，LHCII的结合使捕光截面积增大，从而使PSII光化学活性增加。 用77K飞秒时间分辨荧光光谱分析了几种蛋白脂质体的能量传递和捕获情况。LHCII、OECC和pdOE三种蛋白脂质体的主要荧光衰减组分分别是670 ps（发射峰在680 nm）、650 ps（发射峰在690 nm）和570 ps（发射峰在685 nm）。LHCII-OECC和LHCII-pdOE脂质体的主要衰减组分分别是940 ps（发射峰在690 nm）和840 ps（发射峰在685 nm），并且出现了一个在核心复合物脂质体和LHCII脂质体中没有的40 ps组分，可以推测，这是LHCII和核心复合物之间达到平衡的时间组分，比激发态衰减的平均寿命要快得多，因此支持了PSII的trap-limited激发能衰减动力学模型。此外，可以看到天线的增大使Chl a荧光衰减的寿命延长，这一特性可能与PSII的光保护机制有关。 LHCII和OECC、LHCII和pdOE在脂质体中都成功的实现了重组，而且在结构和功能上没有明显差异;表明小天线以及23 kDa、17 kDa蛋白可能不是LHCII和核心复合物结合及能量传递所必需的。 2、受体侧光破坏和色素置换对PSII RC的影响： 在800 μmol.m-2 .s-1光照和无外加电子受体、供体的情况下，研究了PSII RC色素的受体侧光破坏情况。Chl a、Pheo和β-Car的光漂白几乎同时发生，其中在680 nm吸收的色素破坏最为显著，670 nm吸收的外周Chl比其他色素更加稳定。荧光发射强度呈先升高后降低的趋势，最大发射峰位逐渐蓝移，表明色素之间的能量传递受到破坏。用β-Car的主要吸收波长488 nm和514.5 nm激发得到两组谱带峰位和强度不同的拉曼光谱，表明在PSII RC中存在两个光谱性质不同的β-Car。光破坏过程中两组谱带的位置和带宽都没有明显变化，表明β-Car的光保护机制不涉及自身构象的变化。 将PSII RC与Cu-Chl a进行色素置换，得到了与Cu-Chl重组的RC（Cu-Chl-RC），含有0.5 Cu-Chl/2Pheo。与对照RC（按同样方式与Chl a置换的RC）和天然RC相比，Cu-Chl含量增加而Chl含量减少，660 nm的吸收增加而670 nm吸收降低，因此推测是外周Chl被替换。色素置换过程对RC的多肽组分及大部分的P680活性没有影响，CD光谱的变化也很小，表明产生CD信号的色素和蛋白环境也没有受到明显影响。但是Cu-Chl-RC的荧光发射强度明显降低，最大发射峰蓝移且峰形发生变化，Cu-Chl可能在重组RC中作为激发态的淬灭剂，阻碍了色素之间的能量传递。

Wavelength-tuneable liquid crystal lasers from the visible to the near-infrared

The study of band-edge lasing from dye-doped chiral nematic liquid crystals has thus far been largely restricted to visible wavelengths. In this paper, a wide range of commercially available laser dyes are examined for their suitability as infrared emitters within a chiral nematic host. Problems such as poor solubility and reduced quantum efficiencies are overcome, and successful band-edge lasing is demonstrated within the range of 735-850 nm, using the dyes LD800, HITC-P and DOTC-P. This paper also reports on progress towards widely tuneable liquid crystal lasers, capable of emission in the region 460- 850 nm. Key to this is the use of common pump source, capable of simultaneously exciting all of the dyes (both infrared and visible) that are present within the system. Towards this aim, we successfully demonstrate near-infrared lasing (800 nm) facilitated by Förster energy transfer between the visible dye DCM, and the infra-red dye LD800, enabling pump wavelengths anywhere between 420 and 532 nm to be used. These results demonstrate that small and low-cost tuneable visible to near-infrared laser sources are achievable, using a single common pump source. Such devices are envisaged to have wide-ranging applications including medical imaging (including optical coherence tomography), point-of-care optical medical diagnostics (such as flow cytometry), telecommunications, and optical signatures for security coatings. © 2011 Copyright Society of Photo-Optical Instrumentation Engineers (SPIE).

Impact of plankton structure on primary productivity in two beels of West Bengal, India

Planktonic community in floodplain wetlands embodies the energy transfer through this phase and indicates trophic status of lake. Originally rich bottom coupled with a conducive physicochemical environment encourages fast colonization of the plankton population. Present investigation was carried out in two floodplain wetlands having characteristics of open (Amda beel) and closed (Suguna beel) system. The physicochemical parameters of water and soil of the investigated heels were by and large conducive for planktonic growth. The density of plankton population varied between 1,346 and 2,170 u/l in Suguna bed whereas in Amda beel it ranged from 1,030 to 1,802 u/l. Seasonal fluctuations in water column were conspicuous and mostly dependent on the replenished resources and volume, A mixed and balanced population of diversified fauna constituted the plankton population of the investigated ecosystems. Mostly the diversity was observed to be maximum during winter seasons with coincidence of favorable temperature, dissolved oxygen and other physico-chemical parameters of water besides optimum solar penetration. Richness of planktonic structure in closed system (Suguna) resulted in higher fish production (1,570,05 kg/ha/yr) than that of open system (Amda) (384.4 kg/ha/yr).

Study of density, distribution and yield on benthic fauna and brown trout (Salmo trutta fario) feeding in Laar reservoirs

Lar lake, with the international UTM specification of 39S 579680 3976567 & 39S 589930 3976184 is Situated in Lar national Park with an aerial distance of 55 Km of Tehran along Haraz road. The present research is carried out as part of a comprehensives Plan for assessment of bioresearches of Lar lake & the rivers flowing into it. This research includes examination of there benthic Samplings performed in Lar lake and each of the related rivers including Delichaee, Ab-e-sefid , Alarm & Lar (Kamardasht).Tubifex and Chironomus genus were found to have the highest frequencies of occurrence in the lake with %77.117 & %21.823 respectively followed by Chironomidae and Simulidae from the Diptera order which accounted for %72.328 and %13.812 occurrences in four rivers examined in the Study. The benthic biomass at various examined Sites and the average wet weight of the benthic biomass in station No one in the lake Was 17.397g and the figure for the examined site in Alarm was 20.242 g which were the highest level among Other examination stations the index for the abundance of species in Alarm river was greater than the rest of the examined rivers with 12.57. A sum of 354 Pieces of brown trouts was caught in the course of sampling which were closely investigated in terms of their digestive tract Content. It was identified that Daphniidae and Chironomus constituted the bulk of eaten items from the lake with %17.985 and %63.973 respectively. Meanwhile, Chironomidae and Simuladae were the most frequently eanten benthos by the fish with %81.47 and %7.93 respectively.The index for the relative length of gut was recorded at 0.49± 0.08 which is well indicative of the carnivorous diet of the fish.The index for the feeding intensity amounted to 138  83 showing that the one year old fish were of more feeding intensity.The coefficient of condition (K) was estimated at 1.02  0.142 for all the caught fish. The average wet weight of the benthos was 10.348 g per square meter which if extended to 700ha surface area of the lake, the total macrobenthic production in the lake would amount to 72730Kg of wet weight or 6510 Kg of dry weight. Since the Secondary Production of macrobenthos have always been double that of their biomass, it is reasonable to assume that the Secondary Production of macrobenthos amount to 145640 Kg by their wet weight and Since the energy transfer in the food chain of the lake from benthos to fish is 10 percent, the fish production Capacity Coming from benthic resources of the lake (Lar) would be 14.5 MT, half of which (7000-8000MT) could annually be harvested. Further more, the actual fish Production Capacity might exceed the projected level Since Daphnia, Rotifers and Ostracoda which belong to Zooplanktons, play a part in the natural diet of trout. Meanwhile, rivers Play a major role in fish nutrition and the annual fish production in Delichaee river is about 4481.8Kg while the figures for Ab-e-sefid, Alerm and Lar rivers are 2370.7 4848.7 and 2586.2 Kg respectively, that further increase fish Production in the area and every year half of these resources can be exploitable from the river & the lake.Nevertheless, due to ecological & biological importance of rivers and the probability of environmental Pollution, devastation of natural fish habitats & their nursery grounds, Sport fishing is not recommended at all.

Wavelength-tuneable liquid crystal lasers from the visible to the near-infrared

The study of band-edge lasing from dye-doped chiral nematic liquid crystals has thus far been largely restricted to visible wavelengths. In this paper, a wide range of commercially available laser dyes are examined for their suitability as infrared emitters within a chiral nematic host. Problems such as poor solubility and reduced quantum efficiencies are overcome, and successful band-edge lasing is demonstrated within the range of 735-850 nm, using the dyes LD800, HITC-P and DOTC-P. This paper also reports on progress towards widely tuneable liquid crystal lasers, capable of emission in the region 460- 850 nm. Key to this is the use of common pump source, capable of simultaneously exciting all of the dyes (both infrared and visible) that are present within the system. Towards this aim, we successfully demonstrate near-infrared lasing (800 nm) facilitated by Förster energy transfer between the visible dye DCM, and the infra-red dye LD800, enabling pump wavelengths anywhere between 420 and 532 nm to be used. These results demonstrate that small and low-cost tuneable visible to near-infrared laser sources are achievable, using a single common pump source. Such devices are envisaged to have wide-ranging applications including medical imaging (including optical coherence tomography), point-of-care optical medical diagnostics (such as flow cytometry), telecommunications, and optical signatures for security coatings. © 2011 Copyright Society of Photo-Optical Instrumentation Engineers (SPIE).

Hot electron field emission via individually transistor-ballasted carbon nanotube arrays.

We present electronically controlled field emission characteristics of arrays of individually ballasted carbon nanotubes synthesized by plasma-enhanced chemical vapor deposition on silicon-on-insulator substrates. By adjusting the source-drain potential we have demonstrated the ability to controllable limit the emission current density by more than 1 order of magnitude. Dynamic control over both the turn-on electric field and field enhancement factor have been noted. A hot electron model is presented. The ballasted nanotubes are populated with hot electrons due to the highly crystalline Si channel and the high local electric field at the nanotube base. This positively shifts the Fermi level and results in a broad energy distribution about this mean, compared to the narrow spread, lower energy thermalized electron population in standard metallic emitters. The proposed vertically aligned carbon nanotube field-emitting electron source offers a viable platform for X-ray emitters and displays applications that require accurate and highly stable control over the emission characteristics.

Enhanced down conversion of photons emitted by photoexcited Er xY2-xSi2O7 films grown on silicon

Photon cutting with efficiencies up to 400% is demonstrated in Erx Y2-x Si2 O7 films grown on Si and its concentration dependence is analyzed. The cutting is the result of cross-energy-transfer processes occurring within a single rare earth (Er3+) acting as both sensitizer and activator. Similarities with upconversion are revealed and possible applications in solar cells are discussed. © 2010 The American Physical Society.

A body force-based method for prediction of multiple-pure-tone noise: Validation

Emissions, fuel burn, and noise are the main drivers for innovative aircraft design. Embedded propulsion systems, such as for example used in hybrid-wing body aircraft, can offer fuel burn and noise reduction benefits but the impact of inlet flow distortion on the generation and propagation of turbomachinery noise has yet to be assessed. A novel approach is used to quantify the effects of non-uniform flow on the creation and propagation of multiple pure tone (MPT) noise. The ultimate goal is to conduct a parametric study of S-duct inlets to quantify the effects of inlet design parameters on the acoustic signature. The key challenge is that the effects of distortion transfer, noise source generation and propagation through the non-uniform flow field are inherently coupled such that a simultaneous computation of the aerodynamics and acoustics is required to capture the mechanisms at play. The technical approach is based on a body force description of the fan blade row that is able to capture the distortion transfer and the blade-to-blade flow variations that cause the MPT noise while reducing computational cost. A single, 3-D full-wheel CFD simulation, in which the Euler equations are solved to second-order spatial and temporal accuracy, simultaneously computes the MPT noise generation and its propagation in distorted inlet flow. A new method of producing the blade-to-blade variations in the body force field for MPT noise generation has been developed and validated. The numerical dissipation inherent to the solver is quantified and used to correct for non-physical attenuation in the far-field noise spectra. Source generation, acoustic propagation and acoustic energy transfer between modes is examined in detail. The new method is validated on NASA's Source Diagnostic Test fan and inlet, showing good agreement with experimental data for aerodynamic performance, acoustic source generation, and far-field noise spectra. The next steps involve the assessment of MPT noise in serpentine inlet ducts and the development of a reduced order formulation suitable for incorporation into NASA's ANOPP framework. © 2010 by Jeff Defoe, Alex Narkaj & Zoltan Spakovszky.

Mechanics of capillary forming of aligned carbon nanotube assemblies.

Elastocapillary self-assembly is emerging as a versatile technique to manufacture three-dimensional (3D) microstructures and complex surface textures from arrangements of micro- and nanoscale filaments. Understanding the mechanics of capillary self-assembly is essential to engineering of properties such as shape-directed actuation, anisotropic wetting and adhesion, and mechanical energy transfer and dissipation. We study elastocapillary self-assembly (herein called "capillary forming") of carbon nanotube (CNT) microstructures, combining in situ optical imaging, micromechanical testing, and finite element modeling. By imaging, we identify sequential stages of liquid infiltration, evaporation, and solid shrinkage, whose kinetics relate to the size and shape of the CNT microstructure. We couple these observations with measurements of the orthotropic elastic moduli of CNT forests to understand how the dynamic of shrinkage of the vapor-liquid interface is coupled to the compression of the forest. We compare the kinetics of shrinkage to the rate of evporation from liquid droplets having the same size and geometry. Moreover, we show that the amount of shrinkage during evaporation is governed by the ability of the CNTs to slip against one another, which can be manipulated by the deposition of thin conformal coatings on the CNTs by atomic layer deposition (ALD). This insight is confirmed by finite element modeling of pairs of CNTs as corrugated beams in contact and highlights the coupled role of elasticity and friction in shrinkage and stability of nanoporous solids. Overall, this study shows that nanoscale porosity can be tailored via the filament density and adhesion at contact points, which is important to the development of lightweight multifunctional materials.

Measuring EGFR Separations on Cells with ∼10 nm Resolution via Fluorophore Localization Imaging with Photobleaching

Detecting receptor dimerisation and other forms of clustering on the cell surface depends on methods capable of determining protein-protein separations with high resolution in the ∼10-50 nm range. However, this distance range poses a significant challenge because it is too large for fluorescence resonance energy transfer and contains distances too small for all other techniques capable of high-resolution in cells. Here we have adapted the technique of fluorophore localisation imaging with photobleaching to measure inter-receptor separations in the cellular environment. Using the epidermal growth factor receptor, a key cancer target molecule, we demonstrate ∼10 nm resolution while continuously covering the range of ∼10-80 nm. By labelling the receptor on cells expressing low receptor numbers with a fluorescent antagonist we have found inter-receptor separations all the way up from 8 nm to 59 nm. Our data are consistent with epidermal growth factor receptors being able to form homo-polymers of at least 10 receptors in the absence of activating ligands. © 2013 Needham et al.