光系统Ⅱ反应中心D_1-D_2-Cyt b_(559)复合物低温(77K)荧光发射差异的研究

对菠菜光系统Ⅱ反应中心D_1-D_2-Cytb_(559)复合物进行了系统的低温(77K)荧光发射性质研究。结果表明,D_1-D_2-Cytb_(559)复合物具有681nm和684nm两种波长的低温荧光发射,但两者通常并不是同时存在,而是取决于Ca-680与Ca-670Chla分子的相对含量的。Ca-670Chla含量的增加,会使其低温荧光发射出现在681nm;而Ca-680Chla含量的增加,则会使其低温荧光发射出现在684nm。Ca-670与Ca-680Chla分子的相对含量与不同状态的菠菜叶材料有关。PSⅡ反应中心内周天线CP-47,CP-43多肽的存在是D_1-D_2-Cytb_(591)复合物低温荧光发射红移的原因,而D_1-D_2-Cytb_(559)复合物的不稳定变化则与其蓝移的低温荧光发射有关。     

尖叶拟船叶藓的77K荧光光谱及对强光照的短期适应

报道了东亚特有濒危植物尖叶拟船叶藓(Dolichomitriopsis diversiformis)在不同光质的光照诱导下的低温77K荧光光谱及状态转移的初步研究结果,实验中,尖叶拟船叶藓在77K下出现了3条发射带,分别是F680、F685、F720nm,并没有出现存在于大部分高等植物中的F695nm和F740nm两个峰．经过PSⅡ光诱导后、在77K下出现了F680nm,这个峰在77K下出现是首次报道,而以前的研究认为只在4K下才出现这一条光谱带,这一结果表明尖叶拟船叶藓叶绿体的两个光系统结构与其他高等植物存在着差异。在自然光下,PSⅡ与PSⅠ的总能量比是2.04,经过15min的PSⅡ光(670nm)诱导后,PSⅡ与PSⅠ的总能量比变成了1．28(状态2),当用15min的PSⅠ光(716nm)照射后,PSⅡ与PSⅠ的总能量比从2．04变成了3．4l(状态1)。在自然光下,由尖叶拟船叶藓的光系统的外部LHCⅡ所吸收的激发能是整个光系统激发能的21．19％．这说明尖叶拟船叶藓对光的短期调节能力是21．19％．尖叶拟船叶藓的光系统的外部LHCⅡ有51．7％位于PSⅡ中,48．3％在PSⅠ中.     

华山松叶绿素荧光诱导动力学参数的地理变异及其与树高生长的关系

 利用叶片体内荧光测定技术,检测了南北5个地区华山松种源的荧光诱导动力学参数。结果表明,南方种源具有较强的最大荧光(Fm)和可变荧光(Fv),其光系统Ⅱ(PSⅡ)的潜在活性(Fv／Fo)与原初光能转换效率(Fv／Fm)也明显高于北方种源,并与其树高生长成正相关。另一方面,北方种源的非光化学荧光猝灭系数(QN)比南方种源要大,与它们的树高生长成反相关,而与纬度成正相关。这些实验结果说明,利用荧光诱导动力学技术,可以检测华山松的生长量潜力,在华山松种源选择中有较大的应用前景。     

褐藻77 K荧光特异性研究

测定了裙带菜、叉开网地藻、海带、囊藻、海蒿子、鼠尾藻、萱藻和水云等8种褐藻的77K荧光光谱并同菠菜和红藻条斑紫菜作了比较。结果表明与红藻和高等植物明显不同,褐藻没有作为PSⅠ特征的730 nm荧光峰。按荧光主峰的波长,可以分为二种类型：裙带菜、叉开网地藻、海带和囊藻的荧光主峰位于690 nm,海蒿子、萱藻、水云和鼠尾藻的荧光主峰在705-720 nm。这种77K荧光特异性预示褐藻同高等植物之间在PSⅠ结构上的差异。     

化学交联处理对PSⅡ放氧核心复合物荧光光谱的影响

选用5种交联剂（EDC、DCC、HMDI、EGS和DTSP）,在不同浓度条件下交联处理PSⅡ放氧核心复合物,测定了交联样品的室温荧光发射光谱和荧光激发光谱。结果表明：交联处理对PSⅡ放氧核心复合物叶绿素荧光和蛋白内源荧光都有影响,引起682nm处叶绿素荧光强度的降低、308nm或328nm处蛋白质内源荧光强度的增大或减小,并与处理时所用交联剂的浓度、交联剂的亲疏水性和交联臂长相关。亲水性EDC对PSⅡ的蛋白质中Tyr和Trp残基所处循环境的影响较小;而亲脂性DCC、HMDI、EGS、DTSP对PSⅡ放氧核心复合物蛋白质中Tyr、Trp微环境和682nm处叶绿素荧光影响大,可能它们参与了PSⅡ放氧核心复合物内部的蛋白疏水区域交联。     

低温对水稻类囊体膜蛋白磷酸化及光合机构光能分配的影响

研究了低温胁迫对水稻类囊体膜蛋白磷酸化和光能分配的影响。类囊体膜蛋白组分的SDS-PAGE和免疫印迹分析结果显示,低温弱光条件下光系统Ⅱ(PSⅡ)功能蛋白的稳态水平均有所降低。低温(77K)荧光分析表明,低温处理后类囊体膜光能吸收明显下降,而且FPSⅡ/FPSⅠ的比值均较对照组下降,表明低温弱光条件下有更多的激发能被分配到PSⅠ。低温处理同时还改变了类囊体膜蛋白磷酸化水平,捕光天线LHCⅡ蛋白中lhcb1的磷酸化水平明显降低,lhcb2的磷酸化水平增加,进一步证实lhcb2向PSⅠ移动,改变光能分配。PSⅡ反应中心D1、D2蛋白和核心天线CP43的磷酸化水平增高,有利于PSⅡ二聚体的稳定。     

低温胁迫对2个茶树品种叶片叶绿素荧光特性的影响

以茶树〔Camellia sinensis ( Linn.) O. Ktze.〕品种‘黄金芽’(‘Huangjinya’)和‘迎霜’(‘Yingshuang’)为实验材料,研究了4℃低温胁迫1、2、4和6d对茶树叶片叶绿素荧光特性的影响。结果表明：4℃低温胁迫条件下2个茶树品种叶片的PSⅡ最大光化学效率( Fv/Fm )、PSⅡ潜在活性( Fv/F0)和表观光合电子传递速率( ETR)均显著低于各自的对照(25℃),且总体上随胁迫时间延长逐渐下降;‘黄金芽’叶片的光化学淬灭系数(qP)随低温胁迫时间延长持续下降且低于其对照,而‘迎霜’叶片的qP较其对照的变幅较小,且2个品种的qP总体上与各自的对照无显著差异;随低温胁迫时间延长,2个品种叶片的非光化学淬灭系数( NPQ)均先升高后降低,并在胁迫2 d时达到最高,且总体上高于各自的对照;而2个品种叶片的光合功能相对限制值( LPFD )均随低温胁迫时间延长而增大,且大多高于各自的对照。与各自的对照相比,低温胁迫条件下‘迎霜’叶片的各项叶绿素荧光参数的变幅总体上低于‘黄金芽’。研究结果显示：低温胁迫可直接损伤茶树叶片的PSⅡ反应中心,致使过剩的激发能大量积累于PSⅡ反应中心,最终导致茶树光合作用能力减弱。根据叶绿素荧光参数的比较结果,可以初步判定品种‘迎霜’的耐寒性优于品种‘黄金芽’。     

表面活性剂对小麦光系统Ⅰ叶绿素结合状态和能量传递的影响

表面活性剂在光合膜色素蛋白复合物的分离纯化和结构功能研究中有十分广泛的应用。为了探讨表面活性剂与光系统Ⅰ（PSⅠ）的相互作用机理,选择了两种具代表性的表面活性剂TritonX-100和十二烷基磺酸钠（SDS）,选取一系列浓度值研究了它们对PSⅠ的相互作用机理,选择了两种具代表性的表面活性剂Triton X-100和十二烷基磺酸钠（SDS）,选取一系列浓度值研究了它们对PSⅠ的影响,结果表明,SDS处理时,PSⅠ在区和蓝区的表观吸收峰值下降,峰位蓝移：TritonX-100使红区吸收峰值下降,峰位蓝称,但却使PSⅠ颗粒蓝区表观吸收升高,四阶导数光谱显示小麦（Triticum aestivumL.)PSⅠ颗粒中较长吸收波长的669nm和683nm状态叶绿素a分子受影响较大,两吸光度值互为消长且变化呈轴对称形式,而649nm组分（叶绿素b)在两作用下变化较小。表面活性剂使PSⅠ颗粒长波长荧光急剧下降,并在680nm左右出现新的荧光发射峰,可见其阻碍了激发能由天线色素向反应中心的传递。以上的变化说明SDS和TritonX-100对PSⅠ载体蛋白的微环境甚至结构产生影响而导致色素与蛋白结合状态发生改变,或从蛋白上脱落下来,最终影响到光能的吸收和能量传递。     

强光高温胁迫对盾叶薯蓣量子产额和77K荧光光谱的影响

以低光强生态型盾叶薯蓣(Dioscorea zingiberensis)为材料,研究强光高温(32℃)对盾叶薯蓣量子产额和77K荧光光谱的影响,以探讨强光高温胁迫与光合作用的关系.结果表明:短期高温强光胁迫引起最大量子产额显著下降,但能迅速恢复,且存在超补偿现象.强光胁迫首先破坏了光系统Ⅱ,表现为胁迫后在77K荧光光谱中出现了F680的荧光峰.670 nm红光诱导后引起部分捕光系统Ⅱ的色素蛋白转移至光系统Ⅰ,导致光系统Ⅰ的荧光发射量增加;716 nm红光诱导后引起捕光系统Ⅱ部分色素蛋白转移到光系统Ⅱ,导致光系统Ⅱ的荧光发射量增加.强光高温胁迫后各个荧光峰的差异程度减少.研究发现,强光高温胁迫引起光系统的能量重新调整,从而出现状态转变;依据F672和F675两个荧光峰的首次出现,预测叶绿体类囊体膜上含叶绿素的多肽或蛋白亚基至少有16个.     

生长期间的光强度对小麦光谱特性的影响

在强光(20 klx)下生长的小麦,其叶片和叶绿体的低温(77K)荧光发射强度明显高于在弱光(2 klx)下生长的小麦。同时,前者的PSⅡ的相对荧光产量与PSⅠ的相对荧光产量的比值也高,表明增加光强度有利于PSⅡ的发育。此外。在强光下生长的小麦具有更高的PSⅡ活性和原初光能转化效率。在等量叶绿素的情况下,生长在弱光下的小麦,其叶绿体在吸收光谱上的两个特异吸收带都有较高的吸收值,表明它们比在强光下生长的小麦叶绿体含有较多的光合膜,从而有更大截获光能的面积。     

Geographical Variation in the Characteristics of Absorption Spectra and Fluorescence Spectra of Pinus armandi

By measuring the derivative absorption spectra of chloroplasts of different populations of Pinus armandi Franch., it was found that southern populations maintained higher absorption at 680 nm than at 670 nm, but some of the northern populations deviated the maximum absorption from 680 nm to 670 nm, which indicated that the activity of PS Ⅱ in some of the northern populations declined. Clear geographical differences also have been found in the positions of emission peaks of PS Ⅰ and PS Ⅱ in the fluorescence emission spectra at 77 K. Analysis of the fluorescence excitation spectra at 77 K revealed geographical changes in the absorption.status of Chl a. Besides, the experimental results indicated that the intact needles of Pinus armandi are not ideal materials to be used in detecting the geographical variation in photochemical reaction process because the presence of thicker coat, resin etc. can conceal the spectral differences in different populations.     

The Chlorophyll-Protein Complexes Resolved from Ultrasonically Broken Thylakoid Memb-ranes of Blue-Green Algae

When the thylakoid membranes of blue-green algae were broken by ultrasonic vibrations and subjected to polyacrylamide gel electrophoresis at 4℃, six green zones were resolved. They were designated as CPIa, CPlb, CPI; CPal, CPa2, and FC. The absorption spectrum of CPI had a red maximum at 674 nm and a peak in the blue at 435 nm. It was identified as PS chlorophyll a-protein Complex, but was contaminated with minor PSⅡ which was implied by the appearance of fluorescence emission peak at 680 nm besides the main one at 725 nm at 77 K. The spectral properties of CPIa and CPlb were similar to that of CPl. The absorption spectra of CPa1 and CPa2 were similar, both having red maxima at 667 nm and peaks in the blue at 431.5 nm. Their fluorescence emission had the same peaks at 684 nm at 77 K indicating that they belonged to PSⅡ. It was recognized that CPal of 47 kD is the reaction center complex of photosystem Ⅱ and CPa2 of 40 kD is the internal antenna complex of photosystem Ⅱ. The spectral characteristics of the chlorophyll-protein complexes resolved by ultrasonic method were similar to those of the same complexes resolved by SDS solubilization, except the absorbance positions of CPa1 and CPa2 in the blue peak and the red one which shifted to blue about 3–5 nm. It was calculated that in thylakoid membranes of blue-green algae 40.93% chlorophyll was in PSⅠ, while 38.78% of chlorophyll in PSⅡ. The difference of chlorophyll contents between PSⅠ and PSⅡ was only 2.15%. Concerning the fact that minor PSⅡ compound remained in the part of PSⅠ zones, it might be concluded that the distribution of chlorophyll between PSⅠ and PSⅡ in blue-green algae was equal. This result was in agreement with the hypothesis that PSⅠ and PSⅡ operates in series in photosynthetic electron transport.     

Chlorophyll-Protein Complexes of Blue-Green Algae Isolated by a New Gel Electrophoresis System with High Resolving Power

At least 13 chlorophyll bands from the thylakoid membranes of blue-green algae could be clearly resolved by SDS-PAGE employing a new improved procedure. They were designated as CPIa, CPIb, CPIc, CPId, CPIe, CPIf, CPIg, CHIh, CPal, CPa2, CPa3, CPa4 and FC. 8 chlorophyll-protein complexes, CPIa-CPIh, had the same absorption spectrum at 676 nm in the red and 436 nm in the blue region. They belonged to the chlorophyll-protein complexes of PS Ⅰ. 4 chlorophyll-protein complexes, CPal-CPa4, had a red absorption peak at 670­672 nm and a blue one at 436 nm. Their fluorescence emission peak at 77K was at 685 nm. They were chlorophyll-protein complexes of PS Ⅱ.     

Structure and composition of the photochemical apparatus of the antarctic green alga,Chlamydomonas subcaudata

The green alga, Chlamydomonas subcaudata, collected from a perennially ice-covered Antarctic lake, was able to grow at temperatures of 16°C or lower, but not at temperatures of 20°C or higher, which confirmed its psychrophilic nature. Low temperature (77 K) Chl a fluorescence emission spectra of whole cells of the mesophile, C. reinhardtii, indicated the presence of major emission bands at 681 and 709 nm associated with PS II and PS I, respectively. In contrast, emission spectra of whole cells of C. subcaudata exhibited major emission bands at 681 and 692 nm associated with PS II, but the absence of a major PS I emission band at 709 nm. These results for C. subcaudata were consistent with: (1) low ratio of Chl a/b (1.80); (2) low levels of PsaA/PsaB heterodimer as well as specific Lhca polypeptides as determined by immunoblotting, (3) decreased levels of the Chl-protein complexes CP1 and LHC I associated with PS I; and (4) an increased stability of the oligomeric form of LHC II as assessed by non-denaturing gel electrophoresis in the psychrophile compared to the mesophile. Furthermore, immunoblotting indicated that the stoichiometry of PS II:PS I:CF₁ is significantly altered in C. subcaudata compared to the mesophile. Even though the psychrophile is adapted to growth at low irradiance, it retained the capacity to adjust the total xanthophyll cycle pool size as well as the epoxidation state of the xanthophyll cycle. Despite these differences, the psychrophile and mesophile exhibited comparable photosynthetic efficiency for O₂ evolution regardless of growth conditions. P^max for both Chlamydomonas species was similar only when grown under identical conditions. We suggest that these photosynthetic characteristics of the Antarctic psychrophile reflect the unusual light and low temperature regime to which it is adapted.     

The development of antenna complexes of barley (Hordeum vulgare cv. Akcent) under different light conditions as judged from the analysis of 77 K chlorophyll a fluorescence spectra

Using 77 K chlorophyll a (Chl a) fluorescence spectra in vivo, the development was studied of Photosystems II (PS II) and I (PS I) during greening of barley under intermittent light followed by continuous light at low (LI, 50 μmol m⁻² s⁻¹) and high (HI, 1000 μmol m⁻² s⁻¹) irradiances. The greening at HI intermittent light was accompanied with significantly reduced fluorescence intensity from Chl b excitation for both PS II (F685) and PS I (F743), in comparison with LI plants, indicating that assembly of light-harvesting complexes (LHC) of both photosystems was affected to a similar degree. During greening at continuous HI, a slower increase of emission from Chl b excitation in PS II as compared with PS I was observed, indicating a preferred reduction in the accumulation of LHC II. The following characteristics of 77 K Chl a fluorescence spectra documented the photoprotective function of an elevated content of carotenoids in HI leaves: (1) a pronounced suppression of Soret region of excitation spectra (410–450 nm) in comparison with the red region (670–690 nm) during the early stage of greening indicated a strongly reduced excitation energy transfer from carotenoids to the Chl a fluorescing forms within PS I and PS II; (2) changes in the shape of the excitation band of Chl b and carotenoids (460–490 nm) during greening under continuous light confirmed that the energy transfer from carotenoids to Chl a within PS II remained lower as compared with the LI plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

毕氏海蓬子光系统Ⅱ颗粒的分离与鉴定

采用去污剂TritonX-100增溶类囊体膜和高速离心的方法,首次分离和纯化了毕氏海蓬子的光系统Ⅱ（photosystemⅡ,PSⅡ）颗粒,通过光谱学和SDS-PAGE对其进行鉴定并与类囊体膜进行比较。室温吸收光谱结果表明,PSⅡ颗粒在蓝区的叶绿素（chlorophyll,ChOb和胡萝卜素类吸收峰为485nm,在红区的Ch1b吸收峰为655nm,这两个峰值均低于类囊体膜中的。77K荧光发射光谱结果表明,提取的PSⅡ颗粒基本不含光系统Ⅰ（photosystemⅠ,PSI）的低温荧光反射峰737nm。77K荧光激发光谱结果显示,海蓬子PSⅡ颗粒在470-485am之间的Ch1b 和胡萝卜素类的荧光发射峰明显低于类囊体膜的。这说明在PSⅡ中大部分的PSI已被除去。电泳结果显示,海蓬子PSⅡ颗粒缺少PSI反应中心蛋白质亚基PsaA和PsaB,这说明提取到的PSⅡ纯度较高,这为进一步研究毕氏海蓬子PSⅡ的结构与功能奠定基础。     

Excitation spectra of chlorophyll fluorescence in spinach and barley chloroplasts at 4 K

Excitation spectra of chlorophyll a fluorescence in chloroplasts from spinach and barley were measured at 4.2 K. The spectra showed about the same resolution as the corresponding absorption spectra. Excitation spectra for long-wave chlorophyll a emission (738 or 733 nm) indicate that the main absorption maximum of the photosystem (PS) I complex is at 680 nm, with minor bands at longer wavelengths. From the corresponding excitation spectra it was concluded that the emission bands at 686 and 695 nm both originate from the PS II complex. The main absorption bands of this complex were at 676 and 684 nm. The PS I and PS II excitation spectra both showed a contribution by the light-harvesting chlorophyll $\text{a}\text{b}$ protein(s), but direct energy transfer from PS II to PS I was not observed at 4 K. Omission of Mg²⁺ from the suspension favored energy transfer from the light-harvesting protein to PS I. Excitation spectra of a chlorophyll b-less mutant of barley showed an average efficiency of 50–60% for energy transfer from β-carotene to chlorophyll a in the PS I and in the PS II complexes.