Solution NMR Studies of Chlorella Virus DNA Ligase-adenylate

DNA ligases are essential guardians of genome integrity by virtue of their ability to recognize and seal 3′-OH/5′-phosphate nicks in duplex DNA. The substrate binding and three chemical steps of the ligation pathway are coupled to global and local changes in ligase structure, involving both massive protein domain movements and subtle remodeling of atomic contacts in the active site. Here we applied solution NMR spectroscopy to study the conformational dynamics of the Chlorella virus DNA ligase (ChVLig), a minimized eukaryal ATP-dependent ligase consisting of nucleotidyltransferase, OB, and latch domains. Our analysis of backbone ¹⁵N spin relaxation and ¹⁵N,¹H residual dipolar couplings of the covalent ChVLig-AMP intermediate revealed conformational sampling on fast (picosecond to nanosecond) and slow timescales (microsecond to millisecond), indicative of interdomain and intradomain flexibility. We identified local and global changes in ChVLig-AMP structure and dynamics induced by phosphate. In particular, the chemical shift perturbations elicited by phosphate were clustered in the peptide motifs that comprise the active site. We hypothesize that phosphate anion mimics some of the conformational transitions that occur when ligase-adenylate interacts with the nick 5′-phosphate.     

Storage glucan biosynthesis in Cyanidium, Chlorella and Prototheca: evidence for endosymbiosis?

The storage glucans of Chlorella pyrenoidosa and Prototheca zopfii are identical and consist of a linear polyglucan akin to amylose and a branched amylopectin component. The branched glucans of these algae differ markedly from that formed by the hot-springs alga, Cyanidium caldarium. The more highly branched Cyanidium glucan appears to be formed by branching glucosyltransferases which are different from those of the other two algae. The relevance of the data to the possibility of Cyanidium being a Prototheca-like Chlorella that has acquired symbiotic Cyanobacteria as chloroplasts is discussed.     

The effects of detergents DDM and β-OG on the singlet excited state lifetime of the chlorophyll a in cytochrome b6f complex from spinach chloroplasts

The singlet excited state lifetime of the chlorophyll a (Chl a) in cytochrome b ₆ f (Cyt b ₆ f) complex was reported to be shorter than that of free Chl a in methanol, but the value was different for Cyt b ₆ f complexes from different sources (∼200 and ∼600 ps are the two measured results). The present study demonstrated that the singlet excited state lifetime is associated with the detergents n-dodecyl-β-D-maltoside (DDM) and n-octyl-β-D-glucopyranoside (β-OG), but has nothing to do with the different sources of Cyt b ₆ f complexes. Compared with the Cyt b ₆ f dissolved in β-OG, the Cyt b ₆ f in DDM had a lower fluorescence yield, a lower photodegradation rate of Chl a, and a shorter lifetime of Chl a excited state. In short, the singlet excited state lifetime, ∼200 ps, of the Chl a in Cyt b ₆ f complex in DDM is closer to the true in vivo.     

遮荫对3年生东北铁线莲生长特性及品质的影响

采用人工遮荫的方法,研究了遮荫处理(全光照CK、遮荫30%、50%、70%和90%)对东北铁线莲生长特性和品质的影响。结果表明:东北铁线莲株高随着遮荫梯度的增加而增高;30%遮荫处理茎粗最粗,根茎生物最高,与其它处理差异显著;而茎生物量、叶生物量则是30%遮荫处理与CK、70%和90%处理差异显著;东北铁线莲各器官生物量对光照强度的响应不同,表现为根茎生物量比、根冠比随着光照强度的减弱而降低,茎生物量比和叶生物量比则随着光照的减弱而增加,说明在遮荫的条件下,东北铁线莲优先将生物量分配给茎和叶,用来增加叶面积,以便于接受更多的光能,维持自身的生长发育。东北铁线莲光合色素含量随着遮荫梯度的增加而增加,90%遮荫下的东北铁线莲叶片光合色素含量最高,与其他处理差异显著;相反叶绿素a/b的值却最低;尤其是3年生东北铁线莲叶片中叶绿素 b含量随着光照强度的减弱而升高,能够使东北铁线莲充分吸收散射光,保持较强的光合能力。30%遮荫处理东北铁线莲净光合速率为10.86 μmol · m^-2 · s^-1,显著高于其它处理;随着光照强度的减弱东北铁线莲Gs先增加后降低,Ci则是先降低后增加,30%遮荫处理与其它处理间差异显著,但随着遮荫梯度的增加,东北铁线莲的净光合速率降低,光合作用合成产物总量减少,导致70%和90%遮荫条件下生物量较低;遮荫不利于东北铁线莲有效成分齐墩果酸的积累,不同处理,东北铁线莲齐墩果酸含量差异较大,以全光照和30%遮荫处理齐墩果酸含量较高,与其它处理间差异显著。因此,在生产实践上,对其进行适度遮荫,既保证东北铁线莲药材的质量,又提高药材产量;同时,若不以采收种子为目的,建议在8月份以后采收东北铁线莲药材。     

Properties and subcellular distribution of ribonuclease activity in Chlorella

RNase activity from Chlorella was partially purified. Two RNase activities were demonstrated, one soluble and the other ribosomal. The effects on ribonuclease activity of variations in pH and temperature, and of Mg²⁺, Na⁺, and mononucleotides were examined. The RNase activities (phosphodiesterases EC 3.1.4.23) were both endonucleolytic, releasing oligonucleotides, and cyclic nucleotide intermediates, but exhibited different specificities in releasing mononucleotides from RNA. The ribosomal activity released 3′-GMP, and after prolonged incubation 3′-UMP, but the soluble activity released 3′-GMP, 3′-AMP and 3′-UMP. Neither ofthe RNase preparations hydrolysed DNA, nor released 5′-nucleotides from RNA. Increased ribosomal RNase activity was related to dissociation of ribosomes, and latency of ribosomal RNase activity was demonstrated. The possible in vivo distribution of RNases is discussed.     

Isolation of a fast decay in submillisecond Chlorella luminescence

Using a simple He-Ne (632.8-nm) laser phosphoroscope steady-state luminescence from Chlorella pyrenoidosa was studied from 50 μs to 1.1 ms between 1 ms long exciting flashes. The following results were obtained: (1) prior freezing or ultraviolet irradiation changed the time course of the luminescence to a rapid decay with a half-time of about 110 μs; (2) 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) suppressed the 110-μs luminescence; (3) spectrally, all observed luminescence was, within possible error, identical to fluorescence; (4) no effect on the luminescence intensity from pulsed magnetic fields up to 30 kgauss was observed; (5) the relative fluorescence yield, measured simultaneously with luminescence, was found to be constant.Our principal conclusions, supported mainly by experiments with DCMU, are: (1) the 110-μs decay is a distinct component of the total steady-state luminescence; (2) prior freezing or ultraviolet irradiation isolates this component of the luminescence by suppressing all other components; (3) the half-time and intensity of this component are temperature independent in the interval 0–22 °C.     

Characterization of Chlorella phosphorylase: The glucan specificity and effect of temperature

The glucan specificity of α-glucan phosphorylase partially purified from Chlorella vulgaris was characterized in both directions of glucan phosph     

遮荫对连翘光合特性和叶绿素荧光参数的影响

本文系统研究了不同程度的遮荫(0%、43%、70%、97%)处理对连翘叶片光合特性和叶绿素a荧光参数的影响。结果表明：随着遮荫程度增加,最大净光合速率、光补偿点、光饱和点、暗呼吸速率降低;净光合速率日变化均呈单峰型,峰值和光能利用率增加;叶绿素a b、叶绿素a、叶绿素b含量增加,叶绿素a/b降低;叶绿素a荧光参数Fv/Fm和Fv/Fo日变化呈单谷曲线,值均高于全光照的,且随着遮荫程度的提高其值均依次增加。这说明,连翘是一种耐荫性很强的植物,遮荫可使其降低光补偿点、光饱和点、净光合速率、暗呼吸速率以及叶绿素a/b,增加总叶绿素、叶绿素a、叶绿素b含量、光能利用率以及PSⅡ原初光能转化效率和潜在活性,从而增强其在弱光条件下的生长发育能力。     

Postingestive sorting of living and heat-killed Chlorella within the sea scallop, Placopecten magellanicus (Gmelin)

Suspension-feeding bivalves may enhance the energy value of their food supply by sorting particles both before and after ingestion. Previous research has indicated that the sea scallop (Placopecten magellanicus (Gmelin) (Mollusca: Bivalvia)) is capable of sorting particles within the gut both on the basis of physical properties (particle size and density) as well as chemical properties. In this study, the ability of the sea scallop to sort living from dead material solely on the basis of chemical properties was tested. The microalga Chlorella (Chlorophyta: Chlorophyceae) was chosen as the test particle because its thick cell wall remains physically intact following heat treatment, while its carbon, nitrogen, and chlorophyll a content declines. Scallops were fed a mixture of radiolabelled live and heat-killed Chlorella. We demonstrate that P. magellanicus can distinguish between living and dead algae, retaining live Chlorella cells longer than heat-killed cells. This ability to detect the subtle chemical differences between living algal material and detrital material would enhance the digestive efficiency of this species by reducing the amount of energy expended, digesting poor-quality materials. This paper presents the first study of the ability of a bivalve to distinguish between two physically identical but nutritionally different forms of the same species of microalgae.     

A backflow of electrons around Photosystem II in Chlorella cells

A study was made with a modulated oxygen electrode of the effect of variations of oxygen concentration on photosynthetic oxygen evolution from algal cells. When Chlorella vulgaris is examined with a modulated 650 nm light at 22°C, both the oxygen yield and the phase lag between the modulated oxygen signal and the light modulations have virtually constant values between 800 and 120 ergs · cm^?1 · s^?1 if the bathing medium is in equilibrium with the air. Similar results are obtained at 32°C between 1600 and 120 ergs · cm^?2 · s^?1. Under anerobic conditions both the oxygen yield and the phase lag decrease if the light intensity is lowered below about 500 ergs · cm^?2 · s^?1 at 22°C or about 1000 ergs · cm^?2 · s^?1 at 32°C. A modulated 706 nm beam also gives rise to these phenomena but only at significantly lower rates of oxygen evolution. The cells of Anacystis nidulans and Porphyridium cruentum appear to react in the same way to anaerobic conditions as C. vulgaris. An examination of possible mechanisms to explain these results was performed using a computer simulation of photosynthetic electron transport. The simulation suggests that a backflow of electrons from a redox pool between the Photosystems to the rate-limiting reaction between Photosystem II and the water-splitting act can cause a decrease in oxygen yield and phase lag. If the pool between the Photosystems is in a very reduced state a significant cyclic flow is expected, whereas if the pool is largely oxidized little or no cyclic flow should occur. It is shown that the effects of 706 nm illumination and removal of oxygen can be interpreted in accordance with these proposals. Since a partial inhibition of oxygen evolution by 3-(3.4-dichlorophenyl)-1,1-dimethylurea (10^?8 M) magnifies the decreases in oxygen yield and phase lag, it is proposed that the pool which cycles back electrons is in front of the site of 3-(3,4-dichlorophenyl)-1,1-dimethylurea inhibition and is probably the initial electron acceptor pool after Photosystem II.     

重要理化因子对小球藻生长和油脂产量的影响

本文采用通气培养的方法研究了N、P、Fe3 、盐度、光照强度、温度对小球藻(Chlorella sp. XQ-200419)生长速率、生物量和油脂产量的影响。主要结果如下：N浓度对小球藻的生长和油脂产量均有显著的影响,在KNO3浓度0.05—0.3g/L范围内,小球藻生长速率随N浓度的增加而提高,并积累更多的生物量,而油脂含量随之递减,KNO3浓度为0.3g/L时,油脂产量最高。小球藻对P浓度变化的适应范围很大,K2HPO4浓度在10—160mg/L范围内,对小球藻的生长和油脂产量都没有显著影响。在小球藻培养后期补加不同浓度Fe3 对其生长速率没有显著影响,总脂含量随着Fe3 浓度升高呈现上升的趋势,均比对照有极显著提高,Fe3 浓度为0.75mmol/L时油脂产量最高。盐度对小球藻的生长有一定的抑制作用;油脂含量先随着盐度的增大而提高,当NaCl浓度达到0.6mol/L, 油脂含量又显著降低;油脂含量和油脂产量均在盐度为0.2mol/L时最高。光照强度对处于生长后期的小球藻的生长影响不大,但影响其油脂积累,小球藻的油脂含量和产量随光照强度的增大而显著提高,当光照强度增至280μmolm-2s-1时,油脂含量和油脂产量最高。温度对小球藻的生长速率、生物量、油脂含量和油脂产量都有显著的影响,在15-40℃范围内,随着培养温度的升高,生长速率、生物量、油脂含量和油脂产量都经历了一个先上升然后下降的过程,适合小球藻生长、积累油脂的温度范围是20-35℃,30-35℃时油脂产量最高,40℃时生物量、油脂含量和产量都最低。理化因子对生长和油脂含量的影响分为两种情况：1. 温度、光强、铁浓度和盐度的影响表现为在适宜生长的条件下提高油脂含量,这种模式可以称为“适宜模式”;2. 氮浓度的影响表现为在不利于生长的条件下提高油脂含量,这种模式可以称为“胁迫模式”。两种模式都可以提高油脂含量,但是,只有适宜模式才可以提高油脂产量。在筛选小球藻优良产油藻种时要注意,只有在适宜的培养条件下油脂含量高的藻种才具有高产油潜力。     

Electron and proton transfers from P-430 to ferredoxin-NADP-reductase in Chlorella cells

After blocking Photosystem II on whole Chlorella cells, we measured the absorption changes between 0°C and ?10°C.The absorption changes measured 2 μs after the beginning of a Xenon Flash are the sum of changes due to P⁺-700 and changes due to P^?-430 (after the subtraction of the carotenoid triplet change and of the electrochromic effect).The reduction of P^?-430 is not resolved by our technique. Its reoxidation presents a half-time around 1 μs at 0°C and around 2 μs at ?10°C.The reduction and protonation of ferredoxin-NADP-reductase to its neutral semi-quinoid form FNRH° present a half-time of about 3 μs at 0°C and 6 μs at ?10°C.The presence of only one photoreducible ferredoxin-NADP-reductase per Photosystem I center is confirmed. The acceptor preceding ferredoxin-NADP-reductase is not ferredoxin, but is an acceptor X' the differential extinction coefficients of which are weak or null from 420 nm to 480 nm.Tentative explanations which would reconcile these results with what was already known about ferredoxin are proposed.     

贝壳砂生境酸枣叶片光合生理参数的水分响应特征

以黄河三角洲贝壳堤岛优势灌木酸枣(Ziziphus jujuba var. spinosus)为试验材料,模拟贝壳砂生境系列水分条件,测定分析酸枣叶片在系列水分梯度下的光合参数光响应及叶绿素荧光参数,阐明酸枣主要光合生理参数的水分响应特征。结果表明:(1) 酸枣叶片净光合速率(Pn)、蒸腾速率(Tr)和水分利用效率(WUE)随土壤水分的增多先增大后减小, Tr对土壤相对含水量(Wr)的敏感程度大于Pn,因而WUE维持在较高水平。(2) 酸枣叶片Pn下降的原因在Wr > 25%时以气孔限制为主;Wr < 25%时以非气孔限制为主,光合机构受到不易逆转的破坏。(3) 酸枣叶片最大净光合速率、暗呼吸速率、表观量子效率(AQY)和光饱和点随土壤水分的增多先增大后减小,光补偿点、光抑制项(β)和光饱和项则先减小后增大。(4)在Wr为80%时,酸枣叶片PSⅡ反应中心的光化学转化效率最高。当Wr < 30%时,AQY和潜在光化学效率迅速减小,β迅速增大,酸枣光抑制明显。当Wr < 25%时,非光化学淬灭系数迅速减小,初始荧光迅速增大,酸枣PSⅡ受到不可逆的破坏。(5) Wr在11%-25%内为低产低效水,Wr在25%-58%内为中产中效水,Wr在58%-80%内为高产高效水,Wr在80%-95%内为中产低效水。贝壳砂生境酸枣叶片的光合作用对水分逆境具有较强的生理适应性和可塑性,在Wr为58%-80%内,酸枣光合生理活性较高,利于酸枣苗木生长。     

短期环割对骆驼刺气孔导度及叶绿素荧光的影响

为研究骆驼刺对不同环割的生理响应,以塔克拉玛干沙漠南缘策勒绿洲外围骆驼刺为试验材料,设置了对照(CK),韧皮部半割(PS),韧皮部全割(PF),木质部半割(XS)4种处理,研究了骆驼刺的气孔导度、光合色素含量、Chl a/b、叶绿素荧光参数和曲线在不同环割处理下的变化特征。结果表明:短期韧皮部半割对骆驼刺各项生理指标影响不明显;韧皮部全割下,骆驼刺气孔导度、光合色素含量、叶绿素荧光参数有了不同程度的下降;木质部半割显著降低了骆驼刺气孔导度但光合色素含量和大部分荧光参数没有发生变化。处理后第5天和第10天各项参数相差不大,短期内环割对骆驼刺的伤害程度随时间推移变化较小。总体来看,短期环割处理对骆驼刺伤害程度是PFXSPSCK。在荒漠化防治过程中,应尽量避免骆驼刺的韧皮部被全部剥蚀。     

Turnover kinetics of Photosystem I measured by the electrochromic effect in Chlorella

The rise kinetics of the absorption changes induced at 515 nm and 480 nm by a flash were studied using two types of xenon flashes of different durations. The ‘slow’ rise of the absorption change ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= 15–20 μ}\text{s}$) observed by Cox and Delosme (1978 C.R. Acad. Sci. (Paris) Sér. D 282, 775–778) and Joliot P., Delosme, R. and Joliot, A. ((1977) Biochim. Biophys. Acta 459, 47–57) was found to be due to double hits occurring in the reaction centers of System I during the flash.The turnover kinetics of the reaction centers of System I after a short flash were studied by a double flash method. They are in agreement with a second order reaction between P⁺-700 and its electron donor.     

Cytochrome cs from Rhodymenia palmata and Porphyra umbilicalis and the amino acid sequences of their N-terminal regions

Basic c-type cytochromes homologous with plant and animal mitochondrial cytochrome c have been isolated and purified from Rhodymenia palmata and Porphyra umbilicalis. The N-terminal regions have been analysed using a Beckman 890C automatic sequencer. When compared to animal cytochrome c, the Rhodymenia cytochrome c has an unblocked N-terminal tail of 10 amino acids, whereas Porphyra has an unblocked N-terminal tail of only a single amino acid.