磷饥饿下硅藻光系统Ⅱ光化学反应及其对高光强的响应

海洋暖化导致海水上部混合层变浅,使营养盐限制情况加剧,也导致硅藻接收到的阳光辐射量增加。高光及营养盐限制的双重胁迫会影响硅藻的光合产量。该研究主要探索硅藻应对磷饥饿及高光胁迫的光合生理调控,以进一步了解海洋环境变化对硅藻光合作用的影响。将体积不同的假微型海链藻(Thalassiosira pseudonana)和威氏海链藻(T.weissflogii)在磷饥饿条件下培养,监测其光系统Ⅱ (PSII)功能变化,并将其置于高光强下,研究其光合生理响应。结果表明:磷饥饿条件下,体积较小的假微型海链藻PSII活性逐渐下降,从与D2蛋白结合的质体醌Q_A^–到与D1蛋白结合的质体醌Q_B的电子传递效率降低,单位反应中心捕获的用于电子传递的能量下降,并诱导产生非光化学淬灭,而体积较大的威氏海链藻则能够维持较长时间的PSII活性;假微型海链藻在磷充足条件下相比威氏海链藻具有较高的PSII光失活功能截面(σ_i)值,易发生光抑制,但同时也具有高的PSII修复速率,磷饥饿对其光抑制敏感度无显著影响,而威氏海链藻在磷饥饿条件下...     

光强调控三角褐指藻对海洋酸化的生理学响应

光和海洋酸化(CO₂浓度升高)分别对海洋硅藻的光合能力具有不同程度的影响, 但两者的耦合响应被较少关注。研究以三角褐指藻作为实验材料, 测定了不同光强下CO₂浓度升高对三角褐指藻的生长、净光合速率、生化组分、胞外碳酸酐酶(eCA)活性和核酮糖-1,5-二磷酸羧化/氧化酶(Rubisco)活性的影响。结果显示在低光下, CO₂浓度对三角褐指藻的生长和净光合速率(P_n)并没有显著影响, 而在高光下, 具有明显的影响。无论是在高光或是低光下, eCA活性、叶绿素和可溶性蛋白的含量都随着CO₂浓度的升高而降低。在低光下, 高浓度CO₂ (HC)培养下的Rubisco活性分别是低浓度CO₂ (LC)和中浓度CO₂ (MC)的2.42和1.39倍, 而在高光下, HC培养下的Rubisco活性分别是LC和MC的6.72和3.45倍。以上结果表明硅藻能够通过调节光合生理特征和CCM运行中能量的分配来适应环境中光强和CO₂浓度的变化。     

氮限制对硅藻三角褐指藻光系统Ⅱ光化学反应的影响

本文以硅藻三角褐指藻为实验材料,设置氮充足(882.35μmol·L~(-1))和氮限制(40μmol·L~(-1))两种营养盐供应水平进行培养,以探讨氮限制条件下三角褐指藻光系统Ⅱ(PSII)光化学反应及其对高光强的耐受性变化。结果表明:氮限制抑制三角褐指藻的生长,降低其叶绿素a含量,并加剧其受光抑制程度;通过分析快速叶绿素荧光诱导动力学曲线发现,氮限制使得电子从Q_A~-向QB的传递受阻,且高光强作用后,Q_A~-的瞬时积累进一步增加;此外,氮限制条件下,活性反应中心部分关闭,藻细胞单位反应中心吸收的能量(ABS/RC)、捕获的用于还原Q_A的能量(TRo/RC)以及耗散的能量(DI_o/RC)均明显升高;而高光的耦合作用使得PSII反应中心进一步失活;氮限制影响三角褐指藻PSII光化学反应,进而降低其对高光的耐受能力。     

倪氏拟多甲藻叶绿素荧光活性对环境因子的响应

采用叶绿素荧光分析技术探讨了温度、pH、光强对水华优势种倪氏拟多甲藻光合活性的影响。结果表明, 倪氏拟多甲藻光系统Ⅱ的量子产量Y(Ⅱ)和最大光化学效率(Fv/Fm)随温度(7.5-20.0℃)升高而显著增加(P0.05), 在低温下电子传递速率未受阻, 细胞在7.5-20.0℃内均有高光合活性; 10.0 ℃下的光合活性随pH增大先升高后降低, 峰值出现在pH 7.3时, 光合活性顺序为: 弱碱性 中性 酸性; 快速叶绿素荧光动力学曲线分析显示pH 7.3下的光合活性为典型OJIP曲线, 其他pH下PSⅡ反应中心、电子受体库受损, 显示该藻适应较窄的pH范围, pH7.0-8.0内是其适宜的条件; 快速光响应曲线显示其半饱和光强Ek为385.52 mol photons/(m2s), 表明其具有高光饱和点, 耐受高光强。研究表明藻细胞光合活性对温度和光强变化有较强适应性, 对pH的变化敏感, 弱碱性条件是其光合作用的适宜条件; 低温时细胞通过环式电子链提高光化学效率, 降低高光强可能带来的光损伤; 弱酸性(pH 5.0)会同时损伤其光系统Ⅰ和光系统Ⅱ, 造成其光化学效率的显著下降;倪氏拟多甲藻在低温和高光强下的独特光合特性使其在春季淡水水体中占据竞争优势, 是其形成水华的内在原因。     

氮磷营养限制影响三角褐指藻光合无机碳利用和碳酸酐酶活性

以海洋硅藻三角褐指藻为实验材料, 研究了不同氮磷比培养对其光合无机碳利用和碳酸酐酶活性的影响, 结果显示三角褐指藻生长速率在N:P=16:1时最大, 高于或低于16:1时明显下降, 表明其最适生长受到氮磷的限制。氮限制(N:P=4:1或1:1)导致叶绿素a含量分别下降30.1% 和47.6%, 磷限制(N:P=64:1或256:1)下降39.1%和52.4%, 但氮或磷限制对叶绿素c含量并没有明显影响。不同营养水平培养对光饱和光合速率具有明显的影响, 与营养充足培养相比, 在严重氮磷限制(N:P=1:1或256:1)培养下光饱和光合速率分别下降39.7%和48.0%, 光合效率与暗呼吸速率也明显下降。在氮磷限制培养下藻细胞pH补偿点明显下降; K0.5CO2值在磷限制下降低30%, 表明磷限制有助于提高细胞对CO2的亲和力, 但氮限制并没有明显影响。在氮磷限制培养的细胞反应液中Fe (CN)63-浓度下降速率较慢, 表明在氮磷限制环境中生长的细胞质膜氧化还原能力明显低于营养充足条件下生长的细胞。氮磷限制也导致胞内、外碳酸酐酶活性明显下降, 其中在氮限制下胞外碳酸酐酶活性分别下降50%和37.5%, 在磷限制下下降22.3%和42.1%。严重的氮(N:P=1:1)或磷(N:P=256:1)限制导致胞内碳酸酐酶活性下降36.5%和42.9%。研究结果表明, 三角褐指藻细胞在氮磷营养限制的环境中, 可以通过调节叶绿素含量、无机碳的利用方式和碳酸酐酶的活性以维持适度的生长。       

光发酵强化三角褐指藻生产岩藻黄素

本研究旨在建立光发酵培养三角褐指藻高效生产岩藻黄素的技术体系。在5 L光发酵罐中,系统研究了兼养条件下初始光强、氮源种类和浓度以及光质对于三角褐指藻生物量浓度和岩藻黄素积累的效果。结果表明,在初始光强为100μmol/(m²·s)红蓝(R:B=6:1)混合光、含氮量为0.02 mol/L的胰蛋白胨和尿素混合氮源(1:1, N mol/N mol)优化条件下,三角褐指藻生物量浓度、岩藻黄素含量和产率分别达到了最大值3.80 g/L、13.44 mg/g和4.70 mg/(L·d),比优化前分别提高了1.41、1.33和2.05倍。本研究开发了强化三角褐指藻光发酵生产岩藻黄素的关键技术,促进了海洋天然产物开发。     

濒危植物长序榆(Ulmus elongata)幼苗叶绿素荧光特性的初步研究

用Li-6400XT便携式光合作用仪对濒危植物长序榆幼苗的各叶绿素荧光参数的日变化和快速光响应曲线进行了测定。结果发现,光系统Ⅱ(PSⅡ)的实际光化学效率(ΦPSⅡ)、电子传递速率(ETR)在整个白天阶段较稳定,下午18:00显著下降。光化学淬灭(qP)先增大后减小。非光化学淬灭(NPQ)呈现出与光化学淬灭(qP)相反的变化趋势,中午最低,说明长序榆幼苗光能利用率较高。快速光曲线表明实际光化学效率(ΦPSⅡ)和光化学淬灭(qP)随着光合有效辐射(PAR)的增大而减小,电子传递速率(ETR)和非光化学淬灭(NPQ)随着光合有效辐射(PAR)的增大而增大。使用幂函数能够很好的拟合实际光化学效率(ΦPSⅡ)和电子传递速率(ETR)随光强的变化,而对数函数能较好的拟合实际光化学淬灭(qP)和非光化学淬灭(NPQ)随光强的变化。     

经济海藻红毛菜原位光合作用日变化

利用光合气体交换和叶绿素a荧光技术测定了原位沉水和干出条件下红毛菜光合作用的日变化,结果显示与沉水藻体相比,中午干出藻体光合速率、光系统II最大光化学效率(Fv/Fm)、实际光化学效率(ΦPSⅡ)、光化学淬灭系数(qP),光响应曲线初始斜率(ɑETR)和相对电子传递速率(rETR)值下降更明显。干出藻体重新入水后,其叶绿素a荧光参数在两个小时内可以完全恢复到沉水藻体的水平。红毛菜叶绿素a、类胡萝卜素、藻红蛋白的含量在一天中并没有出现明显变化。以上结果表明红毛菜日生长在中午要经历光抑制过程,干出状态下光抑制更严重;干出和沉水藻体光合速率都可以在傍晚得以恢复;红毛菜光系统II反应中心可以通过增加热耗散和降低光合电子传递速率等策略来应对光抑制。     

大气CO_2增加对不同生长光强下龙须菜光合生理特性的影响

为了探讨未来大气CO2升高对不同生长光强下大型海藻的影响,选取经济红藻龙须菜为实验材料,研究了其生长速率、光合作用、呼吸作用、叶绿素荧光参数以及光合色素对CO2和光强的响应。实验设置两个CO2浓度,正常空气水平CO2浓度(390μL/L)和高CO2浓度(1000μL/L);两个光强梯度,高光(300μmol m-2s-1)和低光(100μmol m-2s-1)。结果表明,CO2和光强对龙须菜的生长和光合作用有明显的交互作用。大气CO2升高并没有显著影响龙须菜的生长速率,但在不同CO2处理下,龙须菜对光强的响应不同。在空气水平下,光强的变化对其生长速率影响不显著。而在高CO2作用下,高光处理下的藻体有更高的生长速率。CO2显著促进高光生长下龙须菜的呼吸作用速率,但是在低光下作用不明显。而对于光合作用速率来说,低光培养下的藻体CO2表现为负面效应,但对高光下生长的藻体作用不明显。CO2增加没有改变龙须菜生长状态下的电子传递速率,但在高光下,CO2表现为一定的抑制作用。CO2显著降低了龙须菜天线色素藻红蛋白和叶绿素a的含量。这些CO2与光强的结合效应表明,大气CO2的升高对龙须菜光合生理特性的影响随着光强的变化而呈现不同的效应,在未来评估CO2的增加对大型海藻的影响时,要充分考虑其他环境因子的耦合效应。     

高温胁迫下苋菜的叶绿素荧光特性

为了探明高温胁迫对苋菜(Amaranthus tricolor L.)光合过程的影响,用不同温度(25、30、35、40、45℃)处理苋菜植株1h后,随即测定了其叶绿素荧光动力学参数和快速光响应曲线特征参数的变化.结果表明:40℃以上高温胁迫下,苋菜叶片的光系统Ⅱ(PSⅡ)潜在光化学效率(Fv/Fo)、最大光化学效率(Fv/Fm)下降;最大荧光(Fm)、光合电子传递速率(ETR)、PSⅡ实际光化学效率(Yield)、光化学淬灭系数(qP)也均有所下降;而初始荧光(F.)和非光化学淬灭系数(NPQ)在40℃以上高温胁迫下显著上升.叶绿素荧光快速光响应曲线测定结果表明,初始斜率α、最大相对电子传递速率ETRmax和半饱和光强Ik在40℃以上高温胁迫下有所下降.研究表明,40℃以上高温胁迫对苋菜的光能的吸收、转换、光合电子传递和强光耐受能力等均有一定的影响.     

CO2 fixation kinetics of Halothiobacillus neapolitanus mutant carboxysomes lacking carbonic anhydrase suggest the shell acts as a diffusional barrier for CO2

The widely accepted models for the role of carboxysomes in the carbon-concentrating mechanism of autotrophic bacteria predict the carboxysomal carbonic anhydrase to be a crucial component. The enzyme is thought to dehydrate abundant cytosolic bicarbonate and provide ribulose 1.5-bisphosphate carboxylase/oxygenase (RubisCO) sequestered within the carboxysome with sufficiently high concentrations of its substrate, CO(2), to permit its efficient fixation onto ribulose 1,5-bisphosphate. In this study, structure and function of carboxysomes purified from wild type Halothiobacillus neapolitanus and from a high CO(2)-requiring mutant that is devoid of carboxysomal carbonic anhydrase were compared. The kinetic constants for the carbon fixation reaction confirmed the importance of a functional carboxysomal carbonic anhydrase for efficient catalysis by RubisCO. Furthermore, comparisons of the reaction in intact and broken microcompartments and by purified carboxysomal RubisCO implicated the protein shell of the microcompartment as impeding diffusion of CO(2) into and out of the carboxysome interior.     

Effect of inorganic carbon supply on the photosynthetic physiology of Gracilaria tenuistipitata

Gracilaria tenuistipitata Zhang et Xia was cultured for 15 d at low, normal and high inorganic carbon concentrations under constant light, temperature and nutrient conditons. Carbonic anhydrase (CA; EC 4.2.1.1.) activity, ribulose-1,5-bisphosphate carboxylase/ oxygenase (Rubisco; EC 4.1.1.39) content, pigment content and C/N ratio were measured, and also the photosynthesis and growth rates. Both Rubisco content and CA activity increased under conditions of low inorganic carbon (C_i) but decreased at high C_i with respect to the control. The amount of pigments declined considerably at high C_i and was slightly higher at low C_i. The maximum rate of photosynthesis and the photosynthetic efficiency increased in low C_i and the opposite was found at high C_i concentration. The effects of C_i concentration on maximum rate of photosynthesis and photosynthetic efficiency are discussed in relation to the variation in pigment and Rubisco contents and CA activity. The data indicate that C_i may be an important factor controlling the photosynthetic physiology of G. tenuistipitata with regard, not only to the enzymes of C_i metabolism, but also to the pigment content.Abbreviations APS_max maximum apparent photosynthetic rate - CA carbonic anhydrase - Chl chlorophyll - C_i inorganic carbon - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase This work has been supported by grants No. PB91-0962 and No. MAR90-0365 from Spanish Direction for Science and Technology (DIGICYT). M.J. G-S holds a fellowship from the DIGICYT.     

Physiological and Biochemical Response of the Photosynthetic Apparatus of Two Marine Diatoms to Fe Stress

Flavodoxin is a small electron-transfer protein capable of replacing ferredoxin during periods of Fe deficiency. When evaluating the suitability of flavodoxin as a diagnostic indicator for Fe limitation of phytoplankton growth, we examined its expression in two marine diatoms we cultured using trace-metal-buffered medium. Thalassio-sira weissflogii and Phaeodactylum tricornutum were cultured in ethylenediaminetetraacetic acid-buffered Sargasso Sea water containing from 10 to 1000 nM added Fe. Trace-metal-buffered cultures of each diatom maintained high growth rates across the entire range of Fe additions. Similarly, declines in chlorophyll/cell and in the ratio of photosystem II variable-to-maximum fluorescence were negligible (P. tricornutum) to moderate (T. weissflogii; 54% decline in chlorophyll/cell and 22% decrease in variable-to-maximum fluorescence). Moreover, only minor variations in photosynthetic parameters were observed across the range of additions. In contrast, flavodoxin was expressed to high levels in low-Fe cultures. Despite the inverse relationship between flavodoxin expression and Fe content of the medium, its expression was seemingly independent of any of the indicators of cell physiology that were assayed. It appears that flavodoxin is expressed as an early-stage response to Fe stress and that its accumulation need not be intimately connected to limitations imposed by Fe on the growth rate of these diatoms.     

Effect of growth conditions on carboxylating enzymes of Zea mays plants

Phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31) and ribulose-1,5-bisphospate (RuBP) carboxylase (EC 4.1.1.39) activities in leaves of different maize hybrids grown under field conditions (high light intensity) and in a growth chamber (low light intensity) were determined. Light intensity and leaf age affected PEP carboxylase activity, whereas RuBP carboxylase was affected by leaf age only at low light intensity. PEP carboxylase/RuBP carboxylase activity ratio decreased according to light intensity and leaf age. Results demonstrate that Zea mays grown under field conditions is a typical C₄ species in all leaves independently from their position on the stem, whereas it may be a C₃ plant when it is grown in a growth chamber at low light intensityAbbreviations PEP phosphoenolpyruvate - RuBP ribulose-1,5-bisphosphate     

Characterization of the carboxysomal carbonic anhydrase CsoSCA from Halothiobacillus neapolitanus

In cyanobacteria and many chemolithotrophic bacteria, the CO(2)-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) is sequestered into polyhedral protein bodies called carboxysomes. The carboxysome is believed to function as a microcompartment that enhances the catalytic efficacy of RubisCO by providing the enzyme with its substrate, CO(2), through the action of the shell protein CsoSCA, which is a novel carbonic anhydrase. In the work reported here, the biochemical properties of purified, recombinant CsoSCA were studied, and the catalytic characteristics of the carbonic anhydrase for the CO(2) hydration and bicarbonate dehydration reactions were compared with those of intact and ruptured carboxysomes. The low apparent catalytic rates measured for CsoSCA in intact carboxysomes suggest that the protein shell acts as a barrier for the CO(2) that has been produced by CsoSCA through directional dehydration of cytoplasmic bicarbonate. This CO(2) trap provides the sequestered RubisCO with ample substrate for efficient fixation and constitutes a means by which microcompartmentalization enhances the catalytic efficiency of this enzyme.     

Carboxylating enzymes and carbonic anhydrase functions were suppressed by zinc deficiency in maize and chickpea plants

Maize and chickpea plants were grown in a controlled environment with 0.5 M Zn or without Zn and various photosynthetic reactions were studied. The chlorophyll level, the rate of photosynthesis and photosystem II activity, the activity of carboxylating enzymes and that of carbonic anhydrase were suppressed by Zn deficiency in both plant species. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was quantified using polyacrylamide gel electrophoresis. Growing plants in a medium without Zn caused a decrease in the total protein level and in the levels of large and small subunits of Rubisco.     

Structural characterization of beta-D-(1-->3)-glucans from different growth phases of the marine diatoms Chaetoceros mülleri and Thalassiosira weissflogii

We have investigated the content and structure of the chrysolaminarans isolated from the two marine diatoms Chaetoceros mülleri and Thalassiosira weissflogii. Samples were taken from different phases of growth, and the structure of the chrysolaminaran was seen in relation to the specific growth rate of the diatoms. The structure determined for the glucan from C. mülleri was found not to vary with different specific growth rates. T. weissflogii showed some variance in the structure, both throughout the different stages of growth and between samples taken from the stationary phase. C. mülleri was found to have a chrysolaminaran with a degree of polymerization (DP) of 22-24 and a degree of beta-(1-->6) branching of 0.006-0.009. These results corresponded well with previous results obtained in our laboratories. The chrysolaminaran isolated from T. weissflogii was found to have a DP of 5-13 and no beta-(1-->6) branching. This is to our knowledge the first characterization of the chrysolaminaran from T. weissflogii.     

Comparative studies on photosynthetic enzymes of the symbiotic Chlorella from Paramecium bursaria and other symbiotic and non-symbiotic Chlorella strains

The activities of ribulose 1,5-bisphosphate carboxylase and of carbonic anhydrase were studied in cell-free extracts of two symbiotic Chlorella strains isolated from Paramecium bursaria and from Spongilla sp., and of two nonsymbiotic strains of Chlorella (Chlorella fusca and Chlorella vulgaris) cultivated at varied CO₂-concentrations. The symbiotic Chlorella of Paramecium bursaria differs distinctly from the other Chlorella strains by a higher activity of ribulose 1,5-bisphosphate carboxylase, which is independent of the actual CO₂-concentration, and by a lack of carbonic anhydrase activity. These properties are discussed with respect to their ecological significance.Abbreviations CA carbonic anhydrase - Pbi Paramecium bursaria isolate - RuBP ribulose 1,5-bisphosphate Dedicated to Prof. Dr. André Pirson on the occasion of his 70th birthday