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 organization in dark-grown and light-grown pine (Pinus brutia) and barley (Hordeum vulgare)

A study was conducted comparing the organization of chlorophyll during development of the photosynthetic apparatus in dark-grown and light-grown pine and barley. The rationale was that gymnosperms, but not angiosperms, have a capacity to synthesize chlorophyll in darkness. Seedlings of Pinus brutia were germinated and grown in darkness or under photoperiodic (day/night) conditions. The low-temperature (77 K) fluorescence spectra of newly-emerging dark-grown seedlings exhibited a single fluorescence band peaking at 678–679 nm, which decayed primarily with a ∼5.5 ns lifetime. Over the first few days of growth, the emission shifted to longer wavelengths and a subnanosecond lifetime component became prevalent. After several days of dark growth the emission spectrum and lifetime profile of the low temperature fluorescence came to resemble those of light-grown pine and barley. At room temperature, dark-grown pine showed little variable fluorescence, though addition of DCMU caused a substantial fluorescence rise. Illumination with moderate light for a few hours was sufficient to 'photoinduce' the appearance of normal variable fluorescence. At 77 K, DCMU-treated samples clearly showed a very long-lived (∼40 ns) fluorescence lifetime component in light-grown pine and barley. This component was undetectable in dark-grown pine. If, however, dark-grown samples were illuminated either before or after DCMU addition and then frozen to 77 K, the ∼40 ns lifetime component appeared at a fluorescence intensity similar to that in light-grown samples. These results are explained primarily in terms of photoactivation of the photosystem II (PSII) donor side. The temporary maintenance of PSII in an inactive, highly-quenched state is suggested to provide an available, yet protected precursor for active PSII.     

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.     

Effect of Different Light Quality on Photosynthetic Characteristics of Cucumber Leaves

Cucumber (Cucumis sativus L. cultivar "Changchun Mici") seedlings were cultured in Hoagland solution under irradiation with different light spectra (8 h per day) for 20 days. The red light (λmax 658 nm, λ1/2 25 nm), blue light (λmax 450 nm, λ1/2 43 nm) and white fluorescent light possessed the same fluent rate (20 μmol· m-2·s-1 ). The experimental results showed that chlorophyll content of the leaves grown under white light was 7 % and 22.4% higher than those in red and blue light, respectively. Compared with white and blue light, red light induced a lower Chl a/b ratio and a higher level of Chl b in the cucumber leaves. Measurements of the low temperature (77 K) fluorescence emission spectra and kinetics of Chl a fluorescence induction of the leaves proved that the leaves grown under red light expressed the highest PSⅡ and the lowest PSⅠactivities while the leaves under blue light had the lowest PSⅡand the highest PSⅠ activities. The O2 evolution rate of red light-grown leaves was 44.9% higher than that of the white light-grown leaves, while blue light effect was similar to that of white in respect of O2 evolution. It is concluded that light quality is an important factor in regulating the development and activities of PSⅡ and PSⅡand the O2 evolution of photosynthesis in cucumber leaves.     

Fe^3＋对雨生红球藻（Haematococcus pluvialis）生长及荧光特性的影响

为探求适宜雨生红球藻CG-06株生长的Fe^3＋浓度和Fe^3＋对藻细胞荧光特性的影响,以BBM为基础培养基,选用EDTA—FeNa（Ⅲ）为Fe^3＋源,设置0、1．79、8．95、17．9、35．8、71．6μmol·L^-1 6种Fe^3＋浓度梯度,实验测定藻的生长并分析不同Fe^3＋对藻细胞叶绿素荧光和77K低温荧光等的影响。结果表明：适合雨生红球藻CG-06株生长的Fe^3＋浓度为8．95μmol·L^-1,Fe^3＋浓度较高时,雨生红球藻的生长受到抑制,Fe^3＋浓度低于1．79μmol·L^-1将产生低铁限制。Walz—PAM测定数据显示,在铁不足或高铁抑制条件下,雨生红球藻光系统Ⅱ活性明显下降,开放态的反应中心数目减少,光合作用受到抑制。77K低温荧光光谱显示,在铁不足或高铁抑制条件下,710nm荧光峰降低,684nm和694nm荧光峰相对增强,说明能量在两个光系统分配上发生变化,能量更多的分配给光系统Ⅱ,限制了光系统Ⅰ的活性。在高铁抑制条件下,CP47蛋白荧光峰降低,CP43蛋白荧光峰增强,推测存在D1蛋白降解。     

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.     

Protochlorophyll(ide) forms in hypocotyls of dark-grown bean (Phaseolus vulgaris)

The protochlorophyll(ide) forms and plastid ultrastructure were investigated in hypocotyls of dark-grown seedlings of kidney bean ( Phaseolus vulgaris L. cv. Brede zonder draad). By deconvolution of the fluorescence emission spectra into Gaussian components three protochlorophyll(ide) forms were found with maxima at 633, 642 and 657 nm, respectively. The ratio of protochlorophyll(ide) emitting at 657 nm to protochlorophyll(ide) emitting at 633 nm decreased downwards the hypocotyl. The gradient was established already after 4 days in dark-grown Phaseolus and was also seen in hypocotyls of 7-day-old dark-grown plants of 8 other species. Ultrastructural observations revealed a plastid developmental sequence along the hypocotyl. Plastids in the upper parts of the hypocotyl contained prolamellar bodies typical of etiolated leaves while those in the lower parts contained only stroma lamellae. Immunological detection of NADPH-protochlorophyllide oxidoreductase (EC 1.3.1.33) on nitrocellulose membranes after sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDSPAGE) indicated the occurrence of the enzyme in upper, middle and lower sections of hypocotyls and in the root tips.     

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。     

The development and ageing of membranes from etioplasts of Avena sativa

A growth period from day 4 to day 11 after sowing of dark-grown oat seedlings ( Avena sativa L. cv. Flämingskrone) was studied. Etioplasts were prepared from various stages. By means of one and two dimensional gel eletrophoresis developmental changes in the polypeptide pattern of the prolamellar body-prothylakoid complex of etioplasts were studied. Polypeptides were described by their molecular weights and pI values after isoelectric focusing. The polypeptide map changes continuously with age, with the onset of effects of senescence around day 7. Concomitantly, proto-chlorophyllide and proteins are degraded, with a higher degradation rate for proto-chlorophyllide. An analysis of protochlorophyllide fluorescence displayed a constant ratio (fluorescence emission at 650 nm/fluorescence emission at 630 nm) from day 6 to day 11. It is concluded, that the best stages for greening studies of dark-grown oat seedlings occur between day 4 and day 6 after sowing.     

Effects of Cucumber Mosaic Virus Infection on Photosynthetic Activities of Tobacco Leaves and Chloroplasts

Changes in photosynthetic activities were studied with tobacco (Nicotiana tabacum L.) leaves and chloroplasts infected by cucumber mosaic virus (CMV) at the top, middle and bottom located leaves. Net photosynthetic rate was reduced at all three positioned leaves, with the maximum reduction occurring at the top leaves (31.9% of control). The infected chloroplasts showed a reduction in electron transport rates of the whole chain electron transport, photosystem Ⅱ (PSⅡ) and photosystem Ⅰ (PSⅠ). Since the decline in the whole chain electron transport (15.6% of control, H2O→MV) closely paralleled the decline in PSⅡ activity (20.9% of control, H2O→PBQ), the inhibition of the latter was probably responsible for the overall decrease. Chlorophyll a fluorescence measurements showed a variable reduced fluorescence yield (Fv/Fo) which indicated that PSⅡ was impaired and the CO2 assimilation was disturbed by CMV infection. Fluorescence emission spectra at 77 K indicated that energy distribution between PSⅡ and PSⅠ was affected. F686/F734 of infected leaves and chloroplasts increased and the greatest increase (331.1% of control ) was found in the top leaves. These data may conclude that the infection inhibited mainly the PSⅡ activity.     

Fe3+对雨生红球藻(Haematococcus pluvialis)生长及荧光特性的影响

为探求适宜雨生红球藻CG-06株生长的Fe³⁺浓度和Fe³⁺对藻细胞荧光特性的影响,以BBM为基础培养基,选用EDTA-FeNa(Ⅲ)为Fe³⁺源,设置0、1.79、8.95、17.9、35.8、71.6μmol·L^-16种Fe³⁺浓度梯度,实验测定藻的生长并分析不同Fe³⁺对藻细胞叶绿素荧光和77 K低温荧光等的影响。结果表明:适合雨生红球藻CG-06株生长的Fe³⁺浓度为8.95μmol·L^-1,Fe³⁺浓度较高时,雨生红球藻的生长受到抑制,Fe³⁺浓度低于1·79μmol.L^-1将产生低铁限制。W alz-PAM测定数据显示,在铁不足或高铁抑制条件下,雨生红球藻光系统Ⅱ活性明显下降,开放态的反应中心数目减少,光合作用受到抑制。77 K低温荧光光谱显示,在铁不足或高铁抑制条件下,710 nm荧光峰降低,684 nm和694 nm荧光峰相对增强,说明能量在两个光系统分配上发生变化,能量更多的分配给光系统Ⅱ,限制了光系统Ⅰ的活性。在高铁抑制条件下,CP₄₇蛋白荧光峰降低,CP₄₃蛋白荧光峰增强,推测存在D₁蛋白降解。     

褐藻光合作用色素—蛋白质复合物——研究进展和问题

本文综述了近二十年来褐藻色素-蛋白质复合物的研究进展,包括色素—蛋白质复合物分离技术、褐藻的光系统Ⅰ、光系统Ⅱ及捕光色素—蛋白质复合物研究进展。并就褐藻色素—蛋白质复合物分离技术中存在的问题、褐藻的特点、褐藻与其它光合生物的色素—蛋白质复合物的同源性以及褐藻PSI复合物77 K荧光发射的特点等进行了讨论。     

Effects of Glycerol on the Fluorescence Spectra and Chloroplast Ultrastructure of Phaeodactylum tricornutum (Bacillariophyta)

Responses of the photosynthetic activity of Phaeodactylum tricornutum (Bacillariophyta) to organic carbon glycerol were investigated. The growth rate, photosynthetic pigments, 77 K fluorescence spectra, and chloroplast ultrastructure of P. tricornutum were examined under photoautotrophic, mixotrophic, and photoheterotrophic conditions. The results showed that the specific growth rate was the fastest under mixotrophic conditions. The cell photosynthetic pigment content and values of Chl a/Chl c were reduced under mixotrophic and photoheterotrophic conditions. The value of carotenoid/Chl a was enhanced under mixotrophic conditions, but was decreased under photoheterotrophic conditions. In comparison with photoautotrophic conditions, the fluorescence emission peaks and fluorescence excitation peaks were not shifted. The relative fluorescence of photosystem (PS) Ⅰ and PS Ⅱ and the values of F6851F710 and F685/F738 were decreased. Chloroplast thylakoid pairs were less packed under mixotrophic and photoheterotrophic conditions. There was a strong correlation between degree of chloroplast thylakoid packing and the excitation energy kept in PS Ⅱ. These results suggested that the PS Ⅱ activity was reduced by glycerol under mixotrophic conditions, thereby leading to repression of the photosynthetic activity.     

Effects of chlorophyll availability on phycobilisomes in Synechocystis sp. PCC 6803

Inactivation of the chlL gene in Synechocystis sp. PCC 6803 resulted in negligible chlorophyll content when the mutant was grown in darkness. Upon phycocyanin excitation at 580 nm, the 77K fluorescence spectrum of dark-grown cells showed three peaks at 648 nm, 665 nm, and 685 nm, this last being the largest. This reflects the functional presence of major components of phycobilisomes, including phycocyanin, allophycocyanin, and the terminal emitter, and efficient energy transfer between these components. As expected, no fluorescence emission peaks corresponding to chlorophyll in the photosystems were observed. Intact phycobilisomes could be isolated from the dark-grown chlL-deletion mutant. However, the phycobilisomes had a lower efficiency of energy transfer than did those isolated from the light-grown mutant, probably because of a decreased phycobilisome stability in the absence of chlorophyll. Exposing the dark-grown chlL-deletion mutant to light triggered the biosynthesis of chlorophyll. For the first 6 h in the light, upon phycocyanin excitation at 580 nm, the 77K fluorescence emission spectrum of greening cells was identical to that of dark-grown cells that lacked significant amounts of chlorophyll. With increased chlorophyll synthesis, gradual energy transfer from phycobilisomes to the two photosystems can be demonstrated.     

The Influence of Surfactants on Chlorophyll Binding Status and Excitation Energy Transfer in Photosystem ￠? of Wheat (in English)

Surfactants are widely used in the purification and research of structure and function of the protein complexes in photosynthetic membrane. To elucidate the mechanism of interaction between surfactants and photosystem Ⅰ (PSⅠ), effects of two typical surfactants, Triton X-100 and sodium dodecyl sulfate (SDS) on PSⅠ, were studied at different concentrations. The results were: SDS led to the reduction of apparent absorption intensity and blue shift of absorption peaks; while Triton X-100 led to the decrease of apparent absorption intensity in red region and blue shift of the peak, but to the increase of apparent absorption intensity in blue region. The fourth derivative spectra show that the longwavelength (669 nm and 683 nm) absorbing chlorophyll a was affected greatly and their relative changes of absorbance were axially symmetrical. The presence of surfactant could make the long wavelength fluorescence emission decrease greatly and a new fluorescence peak appeared around 680 nm, it was obvious that the surfactant interceded the transfer of excitation energy from antenna pigments to reaction center. The surfactants might affect the microenvironment of proteins, even the structure of PSⅠ protein subunits and hence changed the binding status of pigments with protein subunits, or the pigments might be released from the subunits. All of these might affect the absorption and the transfer of excitation energy.     

Photoinhibition in Shaded Cotton Leaves After Exposing to High Light and the Time Course of Its Restoration (in English)

Chlorophyll fluorescence emission, pigment composition and photosynthetic rate of shade-grown cotton (Gossypium hirsutum L.) plants were measured immediately after suddenly exposing to full sunlight and at regular intervals there after within 15 d. Photoinhibition occurred in shade-grown cotton leaves immediately after exposed to full sunlight. The chlorophyll fluorescence parameter Fv / Fm and ΦPSⅡ , which reflect the efficiency of PSⅡ, obviously decreased in shade-grown leaves, much lower than that of the full sunlight-grown leaves. On the contrary, Fo value was sharply increased. Neither of these parameters could completely recover till next morning. The photoinhibition was chronic and continued for about 4 d, while the Fv / Fm and the net photosynthetic rate ( Pn ) continued to decline, then began to increase gradually 6 d later and turned stable after 10-12 d, appearing as an acclimation phenomenon. However, the final value of Fv / Fm and Pn did not reach the level as in those leaves grown in the full sunlight ever before. The final Pn was higher by 60% than that before exposure, but lower for more than 40% than that of the full sunlight-grown leaves. The most notable response of chloroplast pigment composition was a pronounced increase in the pool size of carotenoids in xanthophyll cycle over a period of 3 d. The results indicated that when shade-grown cotton seedlings were suddenly transferred to the full sunlight, the decline of Fv / Fm and Pn might associate with the damage of the PSⅡ reaction center. During the light acclimation, photoprotective mechanisms such as the xanthophyll cycle-dependent energy dissipation were increased, so that photodamage in leaves transferred from low to high light might be reduced.     

超高产杂交稻‘华安3号’冠层不同衰老程度叶片的光合功能

 较系统地研究了抽穗期超高产杂交稻‘华安3号’（`X075’×`紫恢100’）冠层顶部5片叶片的光合功能。结果表明,‘华安3号’剑叶的光系统Ⅱ（PSＩＩ）光化学最大效率(Fv/Fm)、开放的PSⅡ反应中心捕获激发能效率（Fv′/Fm′）、PSⅡ电子传递量子效率（ΦPSⅡ）、光化学猝灭系数（qP）、表观电子传递效率（ETR）、光合色素尤其是叶绿素（Chl）和类胡萝卜素（Car）中的新黄素、黄体素和β-胡萝卜素（β-Car）的含量等均优于其下的各叶,而PSⅡ的激发压力（1-qP）低于其它叶片。经对叶片低温（77K）荧光发射光谱的Gaussian解析,与其它各叶片相比,剑叶PSⅡ核心天线复合物CP47和光系统Ⅰ（PSⅠ）的含量较高,而非活性的PSⅡ捕光色素蛋白复合体（LHCⅡ）聚集态含量较少。研究证明：1）水稻在决定籽粒产量的生育后期,其干物质的积累主要是由冠层最上面的3片叶的光合作用所提供;2）在叶片衰老过程中,光合反应中心的衰老早于天线系统;3）杂交稻的光保护途径之一,可能在于光抑制条件下通过增加PSⅠ含量及其对光能的吸收并刺激环式电子传递高速运转,从而对光合器起保护作用;4）水稻叶片在衰老过程中,可能通过部分Chl b还原为Chl a,以降低LHCⅡ的含量,从而减少对光能的捕获,达到降低光抑制的伤害。     

毕氏海蓬子类囊体膜与卵磷脂重组脂质体的耐热性研究

采用卵磷脂(PC)构建脂质体,然后将毕氏海蓬子类囊体膜蛋白复合物重组到脂质体中.分析不同温度(25℃、35℃、45℃和55℃)处理后蛋白脂质体的电子传递活性、吸收光谱和荧光光谱的变化,以探讨膜脂与膜蛋白在高温胁迫下的交互作用.结果显示:蛋白脂质体光系统Ⅱ(PSⅡ)的放氧活性和光系统Ⅰ(PSⅠ)的耗氧活性随着PC比例的提高而增加,在PC与类囊体膜比例为4∶1(Lipid∶Chl,w/w)时达到最高,同时蛋白脂质体的吸收光谱和荧光光谱也呈上升趋势;在PC与类囊体膜重组比例为4∶1条件下,高温处理后的蛋白脂质体的PSⅡ放氧活性和PSⅠ耗氧活性显著大于未经重组的,其吸收光谱和荧光光谱峰值下降幅度低于未经重组的,且峰位基本没有变化.研究表明,PC可能通过增加结合天线的大小来促进蛋白脂质体对光能的吸收和能量从外周天线到PSⅡ和PSⅠ核心复合物的传递;在脂质体中,PC与类囊体膜的交互作用提高了PSⅡ和PSⅠ在高温胁迫下的光化学效率,增强了PSⅡ和PSⅠ的耐热性.     

Mg2+对生长在不同光强下的小麦叶绿体光合功能的影响

研究结果表明,Mg~(2+)对生长在不同光强度下的小麦叶绿体光合功能有不同影响。与生长在低光强(2×10~8勒克斯)下的小麦叶绿体相比,Mg~(2+)更加明显地降低从生长在高光强(2×10~4勒克斯)下的小麦所分离的叶绿体的吸收光谱在红区和蓝区的吸收峰值;但它更大幅度地提高后者在低温(77K)下的PSⅡ相对荧光产量(F_(687))与PSⅠ相对荧光产量(F_(742))的比值,PSⅡ活性和PSⅡ原初光能转化效率。实验结果证明,更高的光强度可能有利于叶绿体形成更多可流动的LHC-Ⅱ和LHC-Ⅰ。