A changed nitrogenase activity in Rhodospirillum rubrum after substitution of tungsten for molybdenum

The nitrogenase activity in Rhodospirillum rubrum was changed when the cells were made either Mo-deficient or when Mo was replaced by tungsten (W) as trace element in the growth medium: In the absence of N₂, normal Mo cells evolved H₂ (via nitrogenase) from added malate in the light faster than W cells, which in turn evolved H₂ faster than Mo-deficient cells. In the presence of N₂, on the other hand, nitrogen fixation rate in W cells was very close to the low rate found with Mo-deficient cells. Incubation after harvesting of Mo-deficient cells with 2×10^-5 M molybdate or tungstate stimulated the H₂ evolution (similarly with both trace elements) as well as the N₂ fixation (Mo was more effective than W). This indicates that the nitrogenase activity of W cells was truly caused by W and not merely by remaining traces of Mo. The ATP consumption is apparently higher with a W-containing nitrogenase than with the normal Mo-nitrogenase. Further, the affinity to N₂ of the W cells seems to be lower than with the Mo cells.     

Membrane lipid remodeling is required for photosystem II function under low CO2

Membrane lipid remodeling in plants and microalgae has a crucial role in their survival under nutrient-deficient conditions. Aquatic microalgae have low access to CO₂, an essential carbon source for photosynthetic assimilates; however, 70–90 mol% of their membrane lipids are sugar-derived lipids (glycolipids) such as monogalactosyldiacylglycerol (MGDG). In this study, we discovered a new system of membrane lipid remodeling responding to CO₂ in Synechocystis sp. PCC 6803, a unicellular, freshwater cyanobacterium. As compared with higher CO₂ (HC; 1% CO₂), under ambient air (lower CO₂: LC), phosphatidylglycerol (PG) content was increased at the expense of MGDG content. To explore the biological significance of this alteration in content, we generated a transformant of Synechocystis sp. PCC 6803 overexpressing sll0545 gene encoding a putative phosphatidic acid phosphate (oxPAP), which produces diacylglycerol that is used for the synthesis of glycolipids, and examined the effect on membrane lipid remodeling and phototrophic growth responding to LC. Photosystem II (PSII) activity and growth rate were inhibited under LC in oxPAP cells. PG content was substantially reduced, and MGDG and sulfoquinovosyldiacylglycerol contents were increased in oxPAP cells as compared with control cells. These phenotypes in oxPAP cells were recovered under the HC condition or PG supplementation. Increased PG content may be required for proper functioning of PSII under LC conditions.     

Distribution and Redistribution of Molybdenum in Black Gram (Vigna mungo L. Hepper) in Relation to Molybdenum Supply

The effect of seven rates of molybdenum (Mo) supply on the distributionand redistribution of Mo in Vigna mungo (black gram) cv. Reguron a Mo-deficient sandy loam was examined from flower bud appearanceto pod set in one experiment and during pod filling to maturityin another. At the three lowest Mo supply rates, N deficiency symptoms typicalof Mo deficiency appeared, and shoot dry matter and shoot nitrogencontent were depressed. Increasing Mo supply increased Mo concentrationsin all plant parts but the response varied with Mo supply andwith plant part. In leaf blades and petioles, Mo concentrationsincreased slightly when the Mo supply increased from severelydeficient to deficient levels but further increases in Mo supplymarkedly increased the Mo concentrations, particularly in immatureand recently matured leaves. In petioles, Mo concentrationsgenerally exceeded those in the blades which they supportedat all levels of Mo supply. At Mo rates greater than that requiredfor maximum growth, Mo concentrations in basal stem segmentsexceeded those in petioles. Molybdenum concentrations in nodulesexceeded those in above ground plant parts except at the highestlevel of Mo supply where the concentrations in basal stem segmentsexceeded those in nodules. In Mo-adequate plants, Mo contents in the trifoliolate leavesdecreased with time suggesting that Mo was readily remobilized.By contrast, in stem segments at all levels of Mo supply, andin trifoliolate leaves in Mo-deficient plants, Mo contents remainedconstant or increased with time suggesting that Mo was not remobilizedin all plant parts or at all levels of Mo supply. Thus, theresults suggest that in black gram Mo was variably mobile, beingphloem immobile at low Mo supply, but phloem-mobile in all plantparts with the possible exception of stem segments at adequateMo supply. The relevance of these results for the developmentof plant tests for Mo deficiency diagnosis is discussed.Copyright1994, 1999 Academic Press Molybdenum, phloem-mobility, redistribution, black gram, Vigna mungo L. Hepper     

Lipid and hydrocarbon production byBotryococcus spp. under nitrogen limitation and anaerobiosis

The production of lipids and hydrocarbons in batch cultures of the algaeBotryococcus braunii andB. protuberans has been studied with respect to nitrogen limitation under aerobic and anaerobic conditions. Nitrogen deficiency significantly decreased the dry weight, chlorophylla and protein contents but the amounts of carotenoids, carbohydrates and lipids increased in both the species. Nitrogen starvation gave a 1.6-fold increase in lipid content. Anaerobiosis under nitrogen deficient conditions gave greater lipid production than anaerobiosis in nitrogen supplemented medium. Under nitrogen deficiency, the hydrocarbon fraction increased and the polar lipids decreased. Anaerobiosis induced hydrocarbon synthesis more significantly than nitrogen deficiency but decreased other non-polar and polar lipids.     

The effects of nitrogen deficiency on pigments and lipids of cyanobacteria

In contrast to what happens in higher plants and eukaryotic algae, a nitrogen deficiency during growth causes a change in pigment composition but no significant changes in whole cell lipid and fatty acid composition of the two Cyanobacteria, Pseudanabaena sp. (strain M2) and Oscillatoria splendida (strain L3). Nitrogen deficiency does not affect the cellular content in chlorophyll a, but it causes a selective loss in phycobiliproteins; carotenoid content increases with phycocyanin depletion. The major cellular lipids in both Cyanobacteria studied are monogalactosyl diacylglycerol, digalactosyl diacylglycerol, sulfoquinovosyl diacylglycerol, and phosphatidylglycerol. The fatty acid composition is particularly interesting as both these filamentous Oscillatoriaceae show important contents in α- and γ-linolenic (18:3) and parinaric (18:4) acids. This seems to be very unusual in Cyanobacteria.     

Cold-induced changes in brown adipose tissue thermogenic capacity of immunocompetent and immunodeficient hairless mice

Mild cold acclimation (22°C, 3 weeks) of hairless mice was shown to increase 5-fold the brown adipose tissue uncoupling protein content in immunodeficient BALB/c nu/nu mice, but by only 2.3-fold in immunocompetent BFU mice. The difference in activation of brown adipose tissue thermogenic capacity was due to a 2-fold increase in the content of brown adipose tissue in nu/nu mice only, which was paralleled by an increase in brown adipose tissue protein but not DNA content. Likewise, only in nu/nu mice the cold acclimation increased the reaction of natural killer cells in blood and peritoneal exudate with a shift from spleen to lymph nodes and increased the phagocytic index. The results indicate that the immune system may influence the defence against cold at the level of brown adipose tissue thermogenesis.Abbreviations AU arbitrary unit(s) - bw body weight - HEMA 2-hydromethyl-metacrylate copolymer - BAT brown adipose tissue - UCP uncoupling protein - ATPase mitochondrial F_oF₁-ATPsynthase - IL-1 interleukin 1 - TNF tumour necrosis factor - NK cells natural killer cells - T _a ambient temperature     

颗石藻Pleurochrysis carterae抗捕食特征

颗石藻Pleurochrysis carterae是沿海水域中常见钙化微藻,易形成高密度水华,也是养殖环境致害种之一。抗捕食防御能力可能是其种群增殖优势的一个重要原因。以卤虫作为捕食者,分析了颗石藻P.carterae抗捕食现象,以及在捕食压力下的重要生理生化响应特征,以期为颗石藻P.carterea抗捕食机制研究及其高密度增殖机理提供参考。研究结果显示:(1)当颗石藻P.carterae比例增加时,卤虫对微藻的摄食率显著降低,且存活率显著下降,显示该藻具抗捕食能力。(2)以卤虫饵料微藻球等鞭金藻(Isochrysis galbana)为对照,比较研究发现,相同的捕食压力下,饵料金藻的叶绿素荧光参数(电子传递速率ETR和最大量子产率Fv/Fm)显著降低,但颗石藻P.carterae的ETR和Fv/Fm没有显著变化,显示颗石藻P.carterae对卤虫抗捕食作用。(3)相对于没有捕食压力的对照组,捕食压力下,饵料金藻I.galbana的脂类组成没有显著差异。但是,颗石藻P.carterae的脂类组成则发生了显著变化,主要表现在对细胞叶绿体有重要作用的单半乳糖甘油二酯(MGDG),双半乳糖甘油二酯(DGDG),磷脂酰甘油二酯(PG)含量上升,与促细胞分裂相关的二酰甘油(DAG)和磷脂酰肌醇(PI)也上升。这些脂类代谢物的变化可能在其种群水平上抵抗捕食并实现种群增殖中发挥作用。(4)培养介质中磷的状态对颗石藻P.carterae细胞二甲基巯基丙酸(Dimethyl sulfonio propionate,DMSP)含量有显著影响,且影响颗石藻P.carterae对卤虫的致害效应:缺磷条件下生长的颗石藻P.carterae首先使卤虫受害。当培养液中仅以ATP为磷源时,颗石藻P.carterae的卤虫致害效应则降低。研究证明,颗石藻P.carterae具有抗捕食能力,细胞的脂类代谢物质以及DMSP可能在抗捕食防御中发挥作用。     

Response of Chlorella to Scandium and Potassium at Various pH

The response of Chlorella pyrenoidosa cells to moderate potassium deficiency and to the addition of various scandium amounts in the nutrient solution were studied at different pH. The effects on growth, chlorophyll content, oxygen consumption, and the ¹⁴CO₂ incorporation in photosynthesizing cells were measured. Considerable metabolic changes inside the cells were registered under the given nutrient conditions. The separated ¹⁴C-labelled compounds showed, at acid and neutral pH in complete nutrient solution, an increase in the relative content of sugar phosphates and a decrease in the relative content of sucrose, caused by increasing scandium concentrations. Moderate potassium deficiency caused a diminution of the relative content of sugar phosphates. The relative content of amino acids was increased by scandium added to complete nutrient solution, but it decreased under potassium deficiency.     

After X-irradiation a transient arrest of L929 cells inG 2-phase coincides with a rapid elevation of the level of O6-alkylguanine-DNA alkyltransferase

Summary Following X-irradiation of exponentially growing L929 cells two major phenomena have been observed. First, there was a delay in cell division which can be ascribed to the arrest of cells in theG ₂-phase (G ₂-block), and, second, the cellular content of the O⁶-alkylguanine-DNA alkyltransferase (AGT) was markedly increased. Flow cytometrical DNA-measurements revealed that cells began to accumulate in theG ₂-phase 4 h after irradiation (p.r.) irrespective of the X-ray dose, while both the fraction of cells blocked inG ₂ and the time period the cells persisted inG ₂ increased with the radiation dose. About 24 h past release from theG ₂-block the distribution of cells in the cell cycle was similar to that of untreated control cells. In comparison with control cells the AGT content in irradiated cells (4 Gy) was highest at about 48 h p.r. (3.4-fold increase). The highest ratio of increase in AGT was, however, observed to occur between about 4 and 13 h p.r. (2.6-fold increase). As shown by flow cytometrical measurements using a BrdUrd/DNA double labeling technique, this rapid primary increase in AGT coincides very well with the entrance of cells into theG ₂-phase. This indicates that the cellular AGT content in X-irradiated (parental) cells started to exceed the basal level at the beginning of theG ₂-phase, but not before or during theS-phase. Once the AGT level was elevated it continued to increase for 2 to 3 cell doubling times.Paper given at the Workshop Molecular Radiation Biology. German Section of the DNA Repair Network, München-Neuherberg, 21.-23.3.1990     

Lipid Peroxidation in Choline-Methionine Deficiency

A deficiency of choline and methionine is hepatocarcinogenic and is associated with an apparent increase in lipid peroxidation. In this study the susceptibility of microsomes and nuclei to ferritin-dependent lipid peroxidation is examined together with the status of the peroxidation- protective systems. Choline-methionine deficiency caused an increase in Se-independent GSH peroxidases (GSH transferase subunit 2) and membrane vitamin E but a decrease in Se-dependent GSH peroxidase and microsomal GSH peroxidase activity. Choline-methionine deficient microsomes and nuclei were 4-fold more susceptible to lipid peroxidation induced in vitro by physiological concentrations of ferritin/ascorbate/ADP; and the peroxidation was less effectively inhibited by GSH and soluble GSH peroxidases than controls. The results indicate that a decreased level of Se-dependent and membrane GSH peroxidases is involved in the increase in lipid peroxidation observed in choline-methionine deficiency.     

Study of the relationship between thyroid hormones and lipid metabolism during KClO4-induced metamorphosis of landlocked lamprey,Petromyzon marinus

This study examines the role of thyroid hormones (TH) (thyroxine and triiodothyronine) in regulating lipid metabolism of landlocked larval sea lampreys, Petromyzon marinus. Larvae were treated with either thyroxine (0.5 or 1 mg l⁻¹ water) or triiodothyronine (0.25 or 1 mg l⁻¹ water) in the presence or absence of the goitrogen, potassium perchlorate (KClO₄) (0.05% w/v), for 4, 8, and 16 weeks. Treatment with KClO₄ alone, which induced metamorphosis after 8 weeks and lowered plasma TH levels, reduced hepatic and renal total lipid content after 8 weeks of treatment. KClO₄-induced lipid depletion after the 8-week treatment was supported by an increased rate of hepatic lipolysis, as indicated by increased triacylglycerol lipase activity. Furthermore, reduced lipogenesis in the liver was indicated by decreased hepatic acetyl-CoA carboxylase and diacylglycerol acyltransferase (DGAT) activities, and by decreased renal DGAT activity following 8 weeks of KClO₄ treatment. Treatment of larvae for 4 weeks with TH alone resulted in either no change or a slight increase of lipid in the liver and kidney. TH treatments in combination with KClO₄ failed to induce metamorphosis and, after up to 8 weeks, several TH treatments blocked changes in lipid content and enzyme activity associated with KClO₄-induced metamorphosis. These experimental results suggest that TH deficiency during metamorphosis may promote lipid catabolism, while the presence of TH tends to protect/promote lipid reserves, perhaps favoring the larval condition. The actions of TH and KClO₄ on metamorphosis-associated lipid metabolism in sea lampreys may be direct, permissive, and/or indirect via other factors.     

Changes of phenolic metabolism and oxidative status in nitrogen-deficient Matricaria chamomilla plants

The effects of nitrogen deficiency on selected physiological attributes, phenylalanine ammonia-lyase (PAL, EC. 4.3.1.5) activity, phenolic contents, peroxidase (EC. 1.11.1.7) and catalase (EC. 1.11.1.6) activities, lipid peroxidation status and H₂O₂ accumulation were studied in N-deficient Matricaria chamomilla (L.) over 12 days. N deficiency enhanced root growth and inhibited shoot growth. Chlorophyll composition and F _v/F _m were not affected by N stress, but nitrogen and soluble proteins decreased in both the rosettes and the roots. PAL activity, expressed per mg protein, was enhanced in N-deficient rosettes and tended to decrease by the end of the experiment, while in the roots PAL activity was maintained. Total phenolic contents increased in both rosettes and roots. Peroxidase and catalase activities in N-deficient rosettes tended to decrease by the end of the experiment, while in the roots they increased on the 12th day of deficiency. Furthermore, lipid peroxidation status increased in N-deficient roots on the 12th day, indicating that antioxidative protection was insufficient to scavenge reactive oxygen species being generated. Surprisingly, H₂O₂ content was even lower in N-deficient roots by the end of the experiment, while in the leaves increased. This observation in correlation to lipid peroxidation and H₂O₂ degradation is discussed. The importance of PAL activity and phenolic metabolites in combination with antioxidative enzymes for plant protection against oxidative stress and the significance of PAL activity for the mobilization of N availability in N-deficient tissue are also discussed in view of existing information.     

Overexpression of a diacylglycerol acyltransferase gene in Phaeodactylum tricornutum directs carbon towards lipid biosynthesis

Under nutrient deplete conditions, diatoms accumulate between 15% to 25% of their dry weight as lipids, primarily as triacylglycerols (TAGs). As in most eukaryotes, these organisms produce TAGs via the acyl‐CoA dependent Kennedy pathway. The last step in this pathway is catalyzed by diacylglycerol acyltransferase (DGAT) that acylates diacylglycerol (DAG) to produce TAG. To test our hypothesis that DGAT plays a major role in controlling the flux of carbon towards lipids, we overexpressed a specific type II DGAT gene, DGAT2D, in the model diatom Phaeodactylum tricornutum. The transformants had 50‐ to 100‐fold higher DGAT2D mRNA levels and the abundance of the enzyme increased 30‐ to 50‐fold. More important, these cells had a 2‐fold higher total lipid content and incorporated carbon into lipids more efficiently than the wild type (WT) while growing only 15% slower at light saturation. Based on a flux analysis using ¹³C as a tracer, we found that the increase in lipids was achieved via increased fluxes through pyruvate and acetyl‐CoA. Our results reveal overexpression of DAGT2D increases the flux of photosynthetically fixed carbon towards lipids, and leads to a higher lipid content than exponentially grown WT cells.     

Changes in Thermostability of Photosystem Ⅱ and Leaf Lipid Composition of Rice Mutant with Deficiency of Light-harvesting Chlorophyll a/b Protein Complexes

We studied the difference in thermostability of photosystem Ⅱ （PSII） and leaf lipid composition between a T-DNA insertion mutant rice （Oryza sativa L.） VG28 and its wild type Zhonghuau. Native green gel and SDS-PAGE electrophoreses revealed that the mutant VG28 lacked all light-harvesting chlorophyll a/b protein complexes. Both the mutant and wild type were sensitive to high temperatures, and the maximal efficiency of PSII photochemistry （FJ Fm） and oxygen-evolving activity of PSII in leaves significantly decreased with increasing temperature. However, the PSII activity of the mutant was markedly more sensitive to high temperatures than that of the wild type. Lipid composition analysis showed that the mutant had less phosphatidylglycerol and sulfoquinovosyl diacylglycerol compared with the wild type. Fatty acid analysis revealed that the mutant had an obvious decrease in the content of 16：1t and a marked increase in the content of 18：3 compared with the wild type. The effects of lipid composition and unsaturation of membrane lipids on the thermostability of PSII are discussed.     

Alternation in the Glycolipid Transfer Protein Expression Causes Changes in the Cellular Lipidome

The glycolipid transfer protein (GLTP) catalyzes the binding and transport of glycolipids, but not phospholipids or neutral lipids. With its all-alpha helical fold, it is the founding member for a new superfamily, however its biological role still remains unclear. We have analyzed changes in the HeLa cell lipidome in response to down- and up-regulation of GLTP expression. We used metabolic labeling and thin layer chromatography analysis, complemented with a lipidomics mass spectroscopic approach. HeLa cells were treated with GLTP siRNA or were transiently overexpressing the GLTP gene. We identified eight different lipid classes that changed as a result of the GLTP down- or up-regulation treatments; glucosylceramide, lactosylceramide, globotriaosylceramide, ceramide, sphingomyelin, cholesterol-esters, diacylglycerol and phosphatidylserine. We discovered that the amount of globotriaosylceramide (Gb₃) was extensively lowered after down-regulation of GLTP. Further, an up-regulation of GLTP caused a substantial increase in both the Gb₃ and glucosylceramide levels compared to the controls. Total galactosylceramide levels remained unchanged. Both lactosylceramide and ceramide showed small changes, an increase with increasing GLTP and a decrease in the HeLa cell GLTP knockdowns. The cholesterol-esters and diacylglycerol masses increased in cells that had upregulated GLTP protein levels, wheras down-regulation did not affect their amounts. For the glycerophospholipids, phosphatidylserine was the only species that was lower in GLTP overexpressing cells. Phosphatidylethanolamine, phosphatidylglyerol and phosphatidylinositol remained unaltered. A total of 142 lipid species were profiled and quantified using shotgun lipidomics analyses. This work provides for the first time insights into how alternations in the levels of a protein that binds and transfers glycolipids affects the cellular lipid metabolism. We discuss the observed changes in the lipidome and how these relate to GLTP. We suggest, that GLTP not only could be a significant player in cellular sphingolipid metabolism, but also could have a much broader role in the overall lipid metabolism.     

Effect of fasting on the composition of the fat body lipid of Dipetalogaster maximus, Triatoma infestans and Panstrongylus megistus (Hemiptera: Reduviidae)

Modifications in content and lipid composition induced by fasting were examined in fat bodies from adults of Triatominae, Dipetalogaster maximus, Triatoma infestans and Panstrongylus megistus. With fasting, total lipid stores dropped approximately 50% for T. infestans and more than 70% for P. megistus. Total lipids analyzed by thin layer chromatography and fractionated by column chromatography on Unisil showed triacylglycerols as the main component in the three species, although P. megistus showed high levels of diacylglycerols (31–46%). Cholesterol amounted to 8–15%. In diacylglycerol fractions, C_16:0, C_18:1 and C_18:0 fatty acids were detected; their ratio varied with species but it was not dependent on nutritional status. In triacylglycerol fractions C_18:1 fatty acid was the major component at different times (48–68%); the ratio of monounsaturated to saturated in this fraction was 1.3, 2.6 and 1.2 for D. maximus, T. infestans and P. megistus respectively. The remarkable drop in lipid stores without noticeable changes in their relative composition would suggest that all types of lipid are used at similar rates. The higher content of diacylglycerols in P. megistus may be associated with the better flight performance of this species. Accepted: 4 August 1998     

Kontrolle der Eiablage der Brachfliege (Leptohylemyia coarctata Fallén) als Voraussetzung für die Befallsprognose und Bekämpfung

Study of bean plants infected by tabacco mosaic virus and potato X virus was performed. Virus infection of bean plants induced the injury of structural state of photosynthetic apparatus, viz., the a/b chlorophyll ratio decreased and the main photosynthetic membrane lipid monogalactosyl diacylglycerol degradation took place with parallel accumulation of intermediate compound suggested to perform a signal function. Phospholipid and free sterol content increase pointed out on the changes of the membrane fluidity and permeability. Accumulation of sulphoquinovosyl diacylglycerol in infected plants evidences the probability of latter to play important roles in adaptive reactions of photosynthetic apparatus.     

Decreased myo-lnositol Uptake Is Associated with Reduced Bradykinin-Stimulated Phosphatidylinositol Synthesis and Diacylglycerol Content in Cultured Neuroblastoma Cells Exposed to L-Fucose

Abstract: L-Fucose is a potent, competitive inhibitor of myo-inositol transport by cultured mammalian cells. Chronic exposure of neuroblastoma cells to L-fucose causes a concentration-dependent decrease in myo-inositol content, accumulation, and incorporation into phosphoinositides. In these studies, L-fucose supplementation of culture medium was used to assess the effect of decreased myo-inositol metabolism and content on bradykinin-stimulated phosphatidylinositol synthesis and diacylglycerol production. Chronic exposure of cells to 30 mML-fucose caused a sustained decrease in bradykinin-stimulated, but not basal, ³H-inositol phosphate release and ³²P incorporation into phosphatidylinositol in cells incubated in serum-free, unsupplemented medium. In addition, ³²P incorporation into phosphatidylinositol 4-phosphate and phosphatidylinositol 4, 5-bisphosphate was not altered in L-fucose-conditioned cells. Acute exposure of cells to serum-free medium containing 30 mM L-fucose did not affect either basal or bradykinin-stimulated ³²P incorporation into phosphatidylinositol. Basal diacylglycerol content was decreased by 20% in cells chronically exposed to 30 mM L-fucose, although analysis of the molecular species profile revealed no compositional change. Bradykinin stimulated diacylglycerol production in neuroblastoma cells by increasing the hydrolysis of both phosphoinositides and phosphatidylcholine. Bradykinin-stimulated production of total diacylglycerol was similar for control and L-fucose-conditioned cells. However, there was a decrease in the bradykinin-induced generation of the 1 -stearoyl-2-arachidonoyl diacylglycerol molecular species in the cells chronically exposed to 30 mM L-fucose. This molecular species accounts for about 70% of the composition of phosphoinositides, but only 10% of phosphatidylcholine. The results suggest that a decrease in myo-inositol uptake results in diminished agonist-induced phosphatidylinositol synthesis and phosphoinositide hydrolysis in cultured neuroblastoma cells grown in L-fucose-containing medium.