Chlorophyll fluorescence induction in anaerobic Scenedesmus obliquus

Fluorescence time curves (Kautsky effect) were studied in anaerobic Scenedesmus obliquus, with an apparatus capable of simultaneous recording of O₂ exchange, and far-red actinic illumination. Results, as interpreted in terms of electron transport reactions, suggest: In the course of becoming anaerobic, fluorescence induction undergoes a series of changes, indicating at least three different effects of the absence of O₂ on electron transport. (1) Immediately on removal of O₂, once the pool of intermediates between the two photo-systems is reduced by light reaction II, electron flow stops, resulting in high fluorescence yield and a cessation of O₂ evolution. O₂ appears to regulate linear electron flow and cyclic feedback of electrons to the intermediate pool. (2) An endogenous reductant formed anaerobically reduces the System II acceptors in the dark. The time course of this reduction is at least biphasic, indicative of inhomogeneity of the primary acceptor pool. Prolonged dark anaerobic treatment induces maximal initial fluorescence which decays rapidly in light and with a System I action spectrum. (3) Anaerobic treatment eventually results in deactivation of the oxidizing side of System II, limiting System II even when the acceptors are oxidized by System I pre-illumination.     

Transfer ribonucleic acid from Scenedesmus obliquus D3: General properties

Transfer RNA has been isolated from the green alga Scenedesmus obliquus by phenol extraction at 60°. The proportions of major bases and some of t     

光、温限制后铜绿微囊藻和斜生栅藻的超补偿生长与竞争效应

研究了铜绿微囊藻(Microcystis aeruginosa)和斜生栅藻(Scenedesmus obliquus)低温和低光照限制后的超补偿效应,以及共培养条件下的竞争效应。结果表明,低温和低光照均显著抑制微藻的生长发育,但低温对铜绿微囊藻的抑制效应更强,而斜生栅藻则对低光胁迫更敏感。经过低光和低温培养后,铜绿微囊藻和斜生栅藻在恢复正常培养时藻细胞密度短期内都表现出超补偿增长效应,但不同藻类超补偿模式不同,斜生栅藻补偿生长时间不超过1周,而铜绿微囊藻的补偿效应可以持续10天;此外,统计结果表明铜绿微囊藻细胞密度对低温限制解除表现出更显著的补偿生长,斜生栅藻则在低光解除后表现出更强的超补偿效应。微藻叶绿素a指标在光恢复条件下都表现出显著的补偿效应,但温度恢复过程中叶绿素a含量与藻密度增长不同步,低温胁迫对恢复正常培养后微藻叶绿素a的形成产生了一定的负效应;铜绿微囊藻产毒株(912)在两种恢复模式下脱氢酶活性显著高于对照,产毒株(912)脱氢酶活性的补偿响应明显高于其它两种材料。共培养实验结果表明斜生栅藻同铜绿微囊藻产毒株(912)相比处于竞争劣势,而在同无毒株(469)的共培实验中,尽管连续正常培养情况下两者竞争能力差异不显著,但在恢复培养条件下斜生栅藻竞争能力显著高于后者。因此产毒型铜绿微囊藻低温和低光后的补偿生长效应以及对斜生栅藻的竞争优势可能是蓝藻爆发的内源性机制之一。     

Harvesting of Scenedesmus obliquus FSP-3 using dispersed ozone flotation

The Scenedesmus obliquus FSP-3, a species with excellent potential for CO₂ capture and lipid production, was harvested using dispersed ozone flotation. While air aeration does not, ozone produces effective solid-liquid separation through flotation. Ozone dose applied for sufficient algal flotation is similar to those used in practical drinking waterworks. The algae removal rate, surface charge, and hydrophobicity of algal cells, and fluorescence characteristics and proteins and polysaccharides contents of algogenic organic matter (AOM) were determined during ozonation. Proteins released from tightly bound AOM are essential to modifying the hydrophobicity of bubble surfaces for easy cell attachment and to forming a top froth layer for collecting floating cells. Humic substances in the suspension scavenge dosed ozone that adversely affects ozone flotation efficiency of algal cells.     

The 520 nm absorbance changes in Scenedesmus obliquus and its relation to Photosystem I

The kinetics (region of seconds) of the light-induced 520 nm absorbance change and its dark reversal have been studied in detail in the wild type and in some pigment and photosynthetic mutants of Scenedesmus obliquus. The following 5 lines of evidence led us to conclude that the signal is entirely due to the photosystem I reaction modified by electron flow from Photosystem II.Gradual blocking of the electron transport with 3(3,4-dichlorophenyl)-1,1-dimethylurea resulted in diminution and ultimate elimination of the biphasic nature of the signal without reducing the extent of the absorbance change or of the dark kinetics. On the contrary, blocking electron flow at the oxidizing side of plastoquinone with 2, $\text{5-}\text{dibromo}\text{-3-}\text{methyl}\text{-6-}\text{isoprophyl}\text{-p-}\text{benzoquinone}$ or inactivating the plastocyanin with KCN, prolonged the dark reversal of the absorbance change apart from abolishing the biphasic nature of the signal.Action spectra clearly indicate that the main signal (I) is due to electron flow in Photosystem I and that its modification (Signal II) is due to the action of Photosystem II.Signal I is pH independent, whereas Signal II demonstrates a strong pH dependence, parallel to the O₂-evolving capacity of the cells.Chloroplast particles isolated from the wild type Scenedesmus cells demonstrated in the absence of any added artificial electron donor or acceptor and also under non-phosphorylation conditions the 520 nm absorbance change with approximately the same magnitude as whole cells. The dark kinetics of the particles were comparatively slower. Removal of plastocyanin and other electron carriers by washing with Triton X-100 slowed down the kinetics of the dark reversal reaction to a greater extent. A similar positive absorbance change at 520 nm and slow dark reversal was also observed in the Photosystem I particles prepared by the Triton method.Mutant C-6E, which contains neither carotenoids nor chlorophyll $\text{b}$ and lacks Photosystem II activity, demonstrates a normal signal I of the 520 nm absorbance change. This latter result contradicts the postulate that carotenoids are the possible cause of the 520 nm absorbance change.     

Transfer ribonucleic acid from Scenedesmus obliquus D3: Purification of the major formylatable methionine-accepting species

The major formylatable methionine tRNA of Scenedesmus obliquus has been purified using two column chromatographic steps: (1) chromatography on     

A comparative approach towards thylakoid membrane proteome analysis of unicellular green alga Scenedesmus obliquus

The chlorophyll (Chl)-containing membrane protein complexes from the green alga Scenedesmus obliquus have been isolated from the thylakoid membranes by solubilization with dodecyl-β-maltoside and fractionation using a sucrose density gradient. The Chl-containing protein fractions were characterized by absorption spectroscopy, tricine SDS PAGE, BN-PAGE, and dynamic light scattering (DLS). BN-PAGE showed the presence of seven protein complexes with molecular weights in the range of 68, 118, 157, 320, 494, 828 and 955 kDa, respectively. Furthermore, light scattering reveals the simultaneous presence of particles of different sizes in the 3-4 nm and 6.0-7.5 nm range, respectively. The smaller size is related to the hydrodynamic radius of the trimer Light Harvesting Complex (LHCII), whereas the larger size is associated with the presence of photosystem I and photosystem II reaction centers. Additionally, functional information regarding protein-protein interactions was deconvoluted using coupling 2-D BN-PAGE, MALDI-TOF MS and a detailed mapping of S. obliquus photosynthetic proteome of the solubilized thylakoid membranes is therefore presented.     

A mutant strain of Scenedesmus obliquus deficient in ribulose diphosphate carboxylase,cytochrome f and Photosystem II activity

The partial reactions of photosynthesis shown by strain F208, a non-photosynthetic mutant strain of Scenedesmus obliquus, have been compared with those performed by other mutant strains which lacked; Photosystem II activity (strains 11 and F131), cytochrome f (strain 50), P-700 and cytochrome f (strain F119), and P-700 (strains F139 and 199). In this respect the properties of strain F208 were those that would be expected if Photosystem II activity and cytochrome f were not present in this strain. Examination of the composition of strain F208 has shown the absence of cytochrome f in both the soluble and the membrane-bound form. The considerably lower level of plastoquinone compared to that found in the wild type is characteristic of the strains which lack Photosystem II activities.Fraction 1 protein could not be detected in extracts of strain F208 by sedimentation velocity experiments in the ultracentrifuge, and only 7% of the wild type ribulose diphosphate carboxylase activity was found after chromatography of these extracts on DEAE-cellulose.The properties of strain F208 are compared with those of the ac-20 and cr-1 strains of Chlamydomonas rheinhardi, both of which have a deficiency of ribulose diphosphate carboxylase which is considered to result from a deficiency of chloroplast ribosomes. Strain F208 resembles these strains in its abnormal chloroplast ultrastructure and its decreased levels of the RNA forms derived from the chloroplast ribosomes when compared with the wild type.Chloroplast fragments isolated from strains of S. obliquus which lacked cytochrome f (strains 50 and F208) were able to use diaminodurene and ascorbate as an electron donor to Photosystem I. Since this reaction was inhibited by mercuric salts it would appear that plastocyanin, but not cytochrome f, was involved in this electron transfer.     

铜绿微囊藻和斜生栅藻非稳态营养盐限制条件下的生长竞争特性

采用“脉冲”添加方法进行了非稳态条件下铜绿微囊藻（M.）和斜生栅藻（S.）分别在氮磷单营养盐和双营养盐限制时的共培养试验。试验结果显示：当添加频率为1d时,无论何种营养盐限制,M.均成为优势藻种。氮限制条件下,氮时均浓度范围在0.3—2.4 mg/L时,M.始终具有竞争优势。磷限制条件下,磷浓度范围在0.018—0.035 mg/L时,S.只在生长初期阶段占优。氮磷双营养盐限制条件下,添加液的氮磷质量比为35:1（设定为最优比）,添加频率为8 d时,两种藻表现出共生特征;而偏离最优比时（N:P=70:1,17:1）,在不同的添加频率下均未出现共生现象,且氮的时均浓度为0.6—4.8 mg/L时（70:1）,M.具有竞争优势,而降低为0.15—0.3 mg/L时（17:1）,S.占优。随着添加频率的变化,两种藻的细胞大小也会随之改变,S.随着营养盐浓度的降低而增大,且在双营养盐限制条件下变化更显著。上述试验结果分析表明：两种藻竞争能力与添加频率相关,在藻种浓度的变化上,按照‘拾遗－机会’交替竞争理论,M.表现出机会主义者特征,而S.则表现出拾遗者的特征,两者的共生特征也符合‘中度干扰’假说。藻细胞大小变化表明,两种藻均可以改变大小实现最大限度争夺受限营养盐。在低浓度时,S.细胞大小的变化同样也变现出了“拾遗者”的特征。     

PSII-efficiency,polysaccharide production,and phenotypic plasticity of Scenedesmus obliquus in response to changes in metabolic carbon flux

To investigate the response of Scenedesmus obliquus to changes in metabolic carbon flux, S. obliquus was cultured in medium with different concentrations of glyoxylate over 9 days. Results showed that growth rates were not affected in the lower concentration glyoxylate (0.25 and 0.5 mM). However, growth rate of S. obliquus was inhibited in the higher concentration glyoxylate (0.85 and 1.25 mM) during the early phase before recovering at higher densities. Changes in growth rates in different glyoxylate concentrations were in line with changes in F_v/F_m and Φ_PSII. Colony formation was observed in S. obliquus in the four glyoxylate treatments. As a consequence, the mean number of cells per particle of S. obliquus in the glyoxylate treatments were significantly higher than those in the control. The total polysaccharide content of S. obliquus cells increased with increased glyoxylate concentrations. The increased glyoxylate-stimulated polysaccharide levels were directly correlated with colony size of S. obliquus.     

NMR-based metabonomic analysis on effect of light on production of antioxidant phenolic compounds in submerged cultures of Inonotus obliquus

This study was designed to investigate the light effect on biosynthesis of antioxidant phenolic compounds by Inonotus obliquus grown in submerged cultures using ¹H NMR spectroscopy combining multivariate pattern recognition strategies. I. obliquus were exposed to a range of light conditions and resultant data were compared to those from field-grown sclerotia and the mycelia grown in daylight. Daylight illumination inhibited biosynthesis of davallialactone and phelligridins and other hispidin analogs. Continuous darkness enhanced the formation of phelligridins, davallialactone and inoscavins. Phelligridins and davallialactone also occurred in the mycelia grown in blue and red light with levels lower than those found in darkness. In addition, polyphenols synthesized under daylight conditions showed less potential antioxidant activity than those determined with other light regimes. These findings demonstrate that light regulates biosynthesis of polyphenols in I. obliquus and their subsequent antioxidant activities, and ¹H NMR-based metabolic profiling is a cost-effective approach for evaluating light effects on fungal metabolisms.     

桦褐孔菌三萜类物质的提取与含量测定

以桦褐孔菌发酵菌丝体为材料,通过对溶剂乙醇(95%)、甲醇、乙酸乙酯、丙酮、异丙醇、正己烷和氯仿的提取效果比较表明,提取三萜类化合物的最佳溶剂为异丙醇,提取时间为24h,每个样品所需溶剂量(6mL)和菌丝体样品量(100mg)较少,并可同时对大量样品进行有效提取。以白桦脂醇为标准品,对香草醛-冰醋酸-高氯酸分光光度法进行评价,证明该方法简单快速、准确度高、试验误差小、重复性好。利用此方法对桦褐孔菌的野生菌核和发酵菌丝体内三萜类化合物含量进行测定,结果表明野生菌核(59.86mg/g)和发酵菌丝体(94.92mg/g)中都含有很高的三萜类化合物,且发酵菌丝体中三萜类化合物含量高于野生菌核。因此在桦褐孔菌产品开发中,发酵菌丝体可代替野生菌核进行大工业化生产。     

桦褐孔菌三萜类物质的提取与含量测定

以桦褐孔菌发酵菌丝体为材料,通过对溶剂乙醇（95%）、甲醇、乙酸乙酯、丙酮、异丙醇、正己烷和氯仿的提取效果比较表明,提取三萜类化合物的最佳溶剂为异丙醇,提取时间为24h,每个样品所需溶剂量（6mL）和菌丝体样品量（100mg）较少,并可同时对大量样品进行有效提取。以白桦脂醇为标准品,对香草醛-冰醋酸-高氯酸分光光度法进行评价,证明该方法简单快速、准确度高、试验误差小、重复性好。利用此方法对桦褐孔菌的野生菌核和发酵菌丝体内三萜类化合物含量进行测定,结果表明野生菌核（59.86mg/g）和发酵菌丝体（94.92mg/g）中都含有很高的三萜类化合物,且发酵菌丝体中三萜类化合物含量高于野生菌核。因此在桦褐孔菌产品开发中,发酵菌丝体可代替野生菌核进行大工业化生产。     

桦褐孔菌提取物抗氧化活性研究

以乙酸乙酯和甲醇为提取剂,采用索氏提取法,剩余残渣采用热水浸提,最终得到桦褐孔菌不同极性提取物,对其DPPH自由基、羟基自由基及超氧阴离子自由基清除活性作用进行了研究,确定桦褐孔菌的抗氧化能力,为深入研究和开发桦褐孔菌功能性食品奠定理论基础。实验结果表明桦褐孔菌具有较好的抗氧化活性,其中乙酸乙酯提取物的DPPH自由基清除率、羟基自由基清除率和超氧阴离子自由基清除率均高于其他两组分及BHT,桦褐孔菌提取物有望成为功能性食品组分中合成抗氧化剂的天然替代品。     

Time course of photosynthetic response to changes in incident light energy

The response of whole leaf photosynthetic rate in Fragaria virginiana to sudden changes in photosynthetically active radiation (PAR) is described. Two components of the response, consisting of a time lag and time constant, are estimated under varying PAR changes for plants grown under two different light regimes. Both the time lag and time constant are found to vary with PAR but not with growth light regime. A model of Thornley for leaf photosynthetic response is refuted and an alternative form is discussed.     

Novel fluorescent protein from Hydnophora rigida possesses green emission

Fluorescent proteins are a family of proteins capable of producing fluorescence at various specific wavelengths of ultra violet light. We have previously reported the identification and characterization of a novel cyan fluorescent protein (HriCFP) from a reef coral species, Hydnophora rigida. In search of new members of the diverse family of fluorescent proteins, here we report a new green fluorescent protein (HriGFP) from H. rigida. HriGFP was identified, cloned, expressed in Escherichia coli and purified to homogeneity by metal affinity and size exclusion chromatography. The dynamic light scattering and gel filtration experiments suggested the presence of monomers in solution. The peptide mass fingerprint on the purified protein established the identity of HriGFP. HriGFP had excitation peak at 507 nm and emission peak at 527 nm. HriGFP was similar to HriCFP except the last 16 amino acid sequence at the C-terminal; however, they have shown least similarity with other known fluorescent proteins. Moreover the computational model suggests that HriGFP is a globular protein which consists of 6 α-helices and 3 β-sheets. Taken together our results suggested that HriGFP is a novel naturally occurring fluorescent protein that exists as a monomer in solution.     

Novel fluorescent protein from Hydnophora rigida possess cyano emission

Currently, a broad diversity of fluorescent proteins among marine organisms range from cyano-red emissions. Fluorescent proteins differ in their DNA sequences from green fluorescent protein (GFP). We identified cDNA encoding the gene of a new protein from the reef coral Hydnophora rigida of the Merulinidae family. Both the spectral properties and putative primary sequence of the protein has been determined. The cloned cDNA encode peptide we call HriCFP is comprised of 134 amino acids. It has characteristics of a cyano fluorescent protein (HriCFP) and its sequence is markedly different from known GFP from the hydroid jellyfish Aequorea victoria. HriCFP was cloned, expressed, purified and exist as monomer. The peptide mass finger print on the purified protein confirmed identity of HriCFP.