Measurement of enzymatic and motile activities of Arabidopsis myosins by using Arabidopsis actins

There are two classes of myosin, XI and VIII, in higher plants. Myosin XI moves actin filaments at high speed and its enzyme activity is also very high. In contrast, myosin VIII moves actin filaments very slowly with very low enzyme activity. Because most of these enzymatic and motile activities were measured using animal skeletal muscle α-actin, but not plant actin, they would not accurately reflect the actual activities in plant cells. We thus measured enzymatic and motile activities of the motor domains of two Arabidopsis myosin XI isoforms (MYA2, XI-B), and one Arabidopsis myosin VIII isoform (ATM1), by using three Arabidopsis actin isoforms (ACT1, ACT2, and ACT7). The measured activities were different from those measured by using muscle actin. Moreover, Arabidopsis myosins showed different enzymatic and motile activities when using different Arabidopsis actin isoforms. Our results suggest that plant actin should be used for measuring enzymatic and motile activities of plant myosins and that different actin isoforms in plant cells might function as different tracks along which affinities and velocities of each myosin isoform are modulated.     

Significance of the enzyme complex that synthesizes UMP in ehrlich ascites cells

The de novo biosynthesis of pyrimidine nucleotides is completed by two sequential enzyme activities that convert orotate plus 5-phosphoribosyl-1-pyrophosphate to orotidine-5′-monophosphate (OMP) and PP_i and then decarboxylate OMP to produce 5′-uridylic acid. In mammalian cells the two enzyme activities, orotate phosphoribosyltransferase and orotidine-5′-phosphate decarboxylase, form a normally inseparable enzyme complex. It was previously reported that this complex is able to channel the intermediate product, OMP (Traut, T. W., and Jones, M. E., 1977, J. Biol. Chem.252, 8374–8381). The studies reported here indicate that one advantage of this channeling of OMP is to spare OMP from being degraded to orotidine by a potentially competitive nucleotidase activity. Yeast cells have two separate enzymes instead of an enzyme complex, and lack the ability to channel OMP. The OMP formed in yeast cells is not degraded because these cells lack significant nucleotidase activity. These results suggest that the capability for channeling OMP may have been important in evolving the enzyme complex found in mammalian cells.     

Mutants of the Formyltetrahydrofolate Interconversion Pathway of SACCHAROMYCES CEREVISIAE

Thirteen mutants of Saccharomyces cerevisiae that lack one or more of the three enzyme activities of the pathway for interconversion of tetrahydrofolate coenzymes at the formate level of oxidation have been isolated. They do not require adenine. All fail to complement mutations in the ade3 locus. Mutations that greatly reduce activity for one enzyme also reduce activity for the other two interconversion enzymes. The three enzyme activities cochromatograph on TEAE-cellulose columns. A mutation that eliminates synthetase activity also alters the chromatographic behavior of the remaining cyclohydrolase and dehydrogenase activities. It is suggested that the three activities reside in an enzyme complex encoded by the ade3 locus.     

Expression of Glycogen Phosphorylase Isoforms in Cultured Muscle from Patients with McArdle's Disease Carrying the p.R771PfsX33 PYGM Mutation

Background

Mutations in the PYGM gene encoding skeletal muscle glycogen phosphorylase (GP) cause a metabolic disorder known as McArdle''s disease. Previous studies in muscle biopsies and cultured muscle cells from McArdle patients have shown that PYGM mutations abolish GP activity in skeletal muscle, but that the enzyme activity reappears when muscle cells are in culture. The identification of the GP isoenzyme that accounts for this activity remains controversial.

Methodology/Principal Findings

In this study we present two related patients harbouring a novel PYGM mutation, p.R771PfsX33. In the patients'' skeletal muscle biopsies, PYGM mRNA levels were ∼60% lower than those observed in two matched healthy controls; biochemical analysis of a patient muscle biopsy resulted in undetectable GP protein and GP activity. A strong reduction of the PYGM mRNA was observed in cultured muscle cells from patients and controls, as compared to the levels observed in muscle tissue. In cultured cells, PYGM mRNA levels were negligible regardless of the differentiation stage. After a 12 day period of differentiation similar expression of the brain and liver isoforms were observed at the mRNA level in cells from patients and controls. Total GP activity (measured with AMP) was not different either; however, the active GP activity and immunoreactive GP protein levels were lower in patients'' cell cultures. GP immunoreactivity was mainly due to brain and liver GP but muscle GP seemed to be responsible for the differences.

Conclusions/Significance

These results indicate that in both patients'' and controls'' cell cultures, unlike in skeletal muscle tissue, most of the protein and GP activities result from the expression of brain GP and liver GP genes, although there is still some activity resulting from the expression of the muscle GP gene. More research is necessary to clarify the differential mechanisms of metabolic adaptations that McArdle cultures undergo in vitro.     

Heteromultimeric structure of the nitrate reductase complex of Chlamydomonas reinhardii

The NAD(P)H-nitrate reductase complex (overall-NR) of Chlamydomonas reinhardii exhibits two partial activities: NAD(P)H-cytochrome c reductase (diaphorase) and reduced benzyl viologen-NR (terminal-NR). Mild tryptic digestion of the enzyme complex resulted in the loss of both overall and terminal-NR activities, whereas diaphorase activity remained unaltered. The diaphorase activity of mutant 104 and the terminal-NR activity of mutant 305 of C. reinhardii, which are the sole activities related to NR present in these mutants, responded to tryptic treatment to the same extent as the corresponding activities of the wild enzyme complex. Trypsin disassembled the 220-kd NR native complex by destroying the aggregation capability of the diaphorase subunits without affecting their activity nor molecular size (45 kd). A 67-kd thermostable protein, containing molybdenum co-factor, was also released from trypsin-treated NR. This protein lacked diaphorase and NR activities but was able to reconstitute the overall-NR complex by complementation with untreated diaphorase subunit of mutant 104. Our results support a tetrameric structure for the C. reinhardii NR complex, containing two kinds of subunits.     

Polarographic evaluation of mitochondrial enzymes activity in isolated mitochondria and in permeabilized human muscle cells with inherited mitochondrial defects

Inherited disturbances of the mitochondrial energy generating system represent a heterogeneous group of disorders associated with a broad spectrum of metabolic abnormalities and clinical symptoms. We used the polarographic and spectrophotometric method for detection of mitochondrial disorders, because these two techniques provide a different insight into mitochondrial function. In six patients suspected of mitochondrial disease we found defects of complex I (two patients), complex III (one patient), complex IV (two patients) and a combination of defect of complex III and IV (one patient). Citrate synthase activity, used as the reference enzyme, was not changed. A comparison of the two methods showed several differences in evaluation of mitochondrial enzymes activity due to the fact that both methods used different conditions for enzyme activity measurements. In contrast to oxygen consumption measurements, where the function of the whole-integrated respiratory chain is characterized, spectrophotometric measurements characterize activities of isolated complexes in disintegrated membranes. However, it may be concluded from our experiments that both methods provide useful and complementary data about mitochondrial energetic functions. Whereas spectrophotometric data are suitable for evaluation of maximal enzyme activities of mitochondrial enzyme complexes, polarographic data provide better information about enzyme activities in cells with mitochondrial defects under in situ conditions.     

Impact of the Mitochondrial Genetic Background in Complex III Deficiency

Background

In recent years clinical evidence has emphasized the importance of the mtDNA genetic background that hosts a primary pathogenic mutation in the clinical expression of mitochondrial disorders, but little experimental confirmation has been provided. We have analyzed the pathogenic role of a novel homoplasmic mutation (m.15533 A>G) in the cytochrome b (MT-CYB) gene in a patient presenting with lactic acidosis, seizures, mild mental delay, and behaviour abnormalities.

Methodology

Spectrophotometric analyses of the respiratory chain enzyme activities were performed in different tissues, the whole muscle mitochondrial DNA of the patient was sequenced, and the novel mutation was confirmed by PCR-RFLP. Transmitochondrial cybrids were constructed to confirm the pathogenicity of the mutation, and assembly/stability studies were carried out in fibroblasts and cybrids by means of mitochondrial translation inhibition in combination with blue native gel electrophoresis.

Principal Findings

Biochemical analyses revealed a decrease in respiratory chain complex III activity in patient''s skeletal muscle, and a combined enzyme defect of complexes III and IV in fibroblasts. Mutant transmitochondrial cybrids restored normal enzyme activities and steady-state protein levels, the mutation was mildly conserved along evolution, and the proband''s mother and maternal aunt, both clinically unaffected, also harboured the homoplasmic mutation. These data suggested a nuclear genetic origin of the disease. However, by forcing the de novo functioning of the OXPHOS system, a severe delay in the biogenesis of the respiratory chain complexes was observed in the mutants, which demonstrated a direct functional effect of the mitochondrial genetic background.

Conclusions

Our results point to possible pitfalls in the detection of pathogenic mitochondrial mutations, and highlight the role of the genetic mtDNA background in the development of mitochondrial disorders.     

Studies on the subunits of the anthranilate synthetase-phosphoribosyltransferase enzyme complex from Salmonella typhimurium.

Both uncomplexed subunits of the anthranilate synthetase-phosphoribosyltransferase enzyme complex from Salmonella typhimurium have an absolute requirement for divalent metal ions which can be satisfied by Mg²⁺, Mn²⁺, or Co²⁺. The metal ion kinetics for uncomplexed anthranilate synthetase give biphasic double-reciprocal plots and higher apparent K_m values than those for anthranilate synthetase in the enzyme complex. In contrast, the apparent K_m values for phosphoribosyltransferase are the same whether the enzyme is uncomplexed or complexed with anthranilate synthetase. This suggests that the metal ion sites on anthranilate synthetase, but not those on phosphoribosyltransferase, are altered upon formation of the enzyme complex. These results and the results of studies reported by others, suggest that complex formation between anthranilate synthetase and phosphoribosyltransferase leads to marked alterations at the active site of the former, but not the latter enzyme. Uncomplexed anthranilate synthetase can be stoichiometrically labeled with Co(III) under conditions which lead to inactivation of 75% of its activity. A comparison of the effects of anthranilate and tryptophan on phosphoribosyltransferase activity in the uncomplexed and complexed forms shows that anthranilate, but not tryptophan, inhibits the uncomplexed enzyme. The complexed phosphoribosyltransferase shows substrate inhibition by anthranilate binding to the phosphoribosyltransferase subunits. In contrast, in a tryptophan-hypersensitive variant complex, anthranilate inhibits phosphoribosyltransferase activity by acting on the anthranilate synthetase subunits. The data are interpreted to mean that there are two classes of binding sites for anthranilate, one on each type of subunit, which may participate in the regulation of anthranilate synthetase and phosphoribosyltransferase under different conditions.     

Improvement of cellulase activity in Trichoderma reesei by heterologous expression of a beta-glucosidase gene from Penicillium decumbens

Trichoderma reesei is a well-known cellulase producer and widely applied in enzyme industry. To increase its ability to efficiently decompose cellulose, the beta-glucosidase activity of its enzyme cocktail needs to be enhanced. In this study, a beta-glucosidase I coding sequence from Penicillium decumbens was ligated with the cellobiohydrolase I (cbh1) promoter of T. reesei and introduced into the genome of T. reesei strain Rut-C30 by Agrobacterium-mediated transformation. In comparison to that from the parent strain, the beta-glucosidase activity of the enzyme complexes from two selected transformants increased 6- to 8-fold and their filter paper activity (FPAs) was enhanced by 30% on average. The transformant's saccharifying ability towards pretreated cornstalk was also significantly enhanced. To further confirm the effect of heterologous beta-glucosidase on the cellulase activity of T. reesei, the heterologously expressed pBGL1 was purified and added to the enzyme complex produced by T. reesei Rut-C30. Supplementation of the Rut-C30 enzyme complex with pBGL1 brought about 80% increase of glucose yield during the saccharification of pretreated cornstalk. Our results indicated that the heterologous expression of a beta-glucosidase gene in T. reesei might produce balanced cellulase preparation.     

Regulation of acetyl-CoA carboxylase in rat mammary gland. Effects of starvation and of insulin and prolactin deficiency on the fraction of the enzyme in the active form in vivo

The `initial' (I), endogenous phosphatase-activated (A) and citrate-activated (C) activities of acetyl-CoA carboxylase were measured in mammary-gland extracts of pregnant and lactating rats. There was a 10-fold increase in the A and C enzyme activities in the transition from early to peak lactation [cf. data of Mackall & Lane (1977) Biochem. J. 162, 635–642], but there was no significant increase in the ratio of the initial activity to the A and C activities of the enzyme. Starvation (24h) or short-term (3h) streptozotocin-induced diabetes both resulted in a 40% decrease in I/A and I/C activity ratios. In starvation this was accompanied by a decrease in the absolute values of the A and C activities such that the initial activity in mammary glands of starved animals was 45% that in glands from fed animals. Insulin treatment of starved or diabetic animals 60min before killing increased the I activity without affecting the A or C enzyme activities. Removal of the pups for 24h from animals in peak lactation (weaning) resulted in a marked but similar decrease in all three activities such that, although the initial activity was only 10% of that in suckled animals, the I/A and I/C activity ratios remained high and unaltered. Inhibition of prolactin secretion by injection of 2-bromo-α-ergocryptine gave qualitatively similar results to those during weaning. Simultaneous administration of ovine prolactin completely prevented the effects of bromoergocryptine. It is suggested that the initial activity of acetyl-CoA carboxylase in rat mammary gland is regulated by at least two parallel mechanisms: (i) an acute regulation of the proportion of the enzyme in the active state and (ii) a longer-term modulation of enzyme concentration in the gland. Insulin appeared to mediate its acute effects through mechanism (i), whereas prolactin had longer-term effects on enzyme concentration in the gland. A comparison of initial enzyme activities (I) obtained in the present study with rates of lipogenesis measured in vivo [Agius & Williamson (1980) Biochem. J. 192, 361–364; Munday & Williamson (1981) Biochem. J. 196, 831–837] gave good agreement between the two sets of data for all conditions studied except for 24h-starved and streptozotocin-diabetic animals. It is suggested that acetyl-CoA carboxylase activity is rate-limiting for lipogenesis in the mammary gland in normal, fed, suckled or weaned animals but that in starved and short-term diabetic animals changes in the activity of the enzyme by covalent modification alone may not be sufficient to maintain the enzyme in its rate-limiting role.     

The effect of exercise training and water extract from propolis intake on the antioxidant enzymes activity of skeletal muscle and liver in rat

[Purpose]

In this study, the authors have intended to investigate the effects that the exercise training and the intake of the water extract from propolis have on the activity of antioxidant enzymes.

[Methods]

For this purpose, the exercise training (70% VO₂max treadmill running exercise for 60min)of 5 times per week for six weeks and the intake (50mg/kg/day) of the water extract from propolis were performed by separating the experimental animals (SD rats, n=32) into CON(n=8) group, CON+Ex(n=8), PA(n=8), and PA+Ex(n=8).

[Results]

As a result, the following conclusions were obtained: The concentration of the blood glucose and insulin of the CON+Ex group and PA+Ex group which are the exercise parallel group were significantly decreased in comparison with the control group, whereas if comparing the glycogen concentration in skeletal muscle and liver tissue between the exercise parallel group and the CON group, the former showed significantly high value in comparison with the latter (p < .05). In the case of the activity of the antioxidant enzyme in the skeletal muscle and the liver tissue, the activities of SOD, GPX and CAT in the gastrocnemius muscle tissue of the experimental animals showed significantly high value in PA+Ex group in comparison with other experimental groups (p < .05). In addition, the SOD activity in the liver tissue showed that only PA+Ex group was significantly increased, whereas GDX activity showed significantly higher value in CON+Ex group and PA group than CON group (p < .05). However, the activity of CAT in the liver tissue showed that there is no difference between the experimental groups. As a result that measured the concentration of MDA in order to evaluate the damage level of the tissue by oxygen free radicals, the difference between the groups in the liver tissue was not shown, while it was shown that only PA+Ex group in the skeletal muscle tissue was significantly decreased in comparison with other experimental groups (p < .05).

[Conclusion]

Taken together the above findings, it is considered that the parallel treatment of the exercise training and the water extract from propolis can not only increase the use of glycogen of the skeletal muscle and liver tissue, but also it can give the effect to suppress the creation of active oxygen by inducing the activity of the antioxidant enzyme in the body, and in the future, the possibility as the exercise supplements and the antioxidant of the water-soluble propolis are expected.     

The influence of diet and gastrointestinal fermentation on key enzymes of substrate utilization in marine teleost fishes

Three closely related marine teleosts with similar size, swimming mode, and habitat preference were compared to test the hypothesis that energy metabolism is linked to diet choice in the wild. Key substrate-utilization enzyme activities were assayed from white locomotory muscle and liver in a carnivore (Scorpis violaceus), an omnivore (Girella tricuspidata), and a herbivore (Kyphosus sydneyanus) collected from their natural reef habitat in northeastern New Zealand. The similar energy requirements of the study fishes were reflected in specific enzyme activities of white muscle and suggested high dependence on endogenous fuel that is independent of dietary carbohydrate intake. Clear differences were found in enzymes of hepatic carbohydrate, fat, and ketone body metabolism that appear linked to diet choice and levels of gastrointestinal fermentation. Hepatic metabolism of fat and ketone bodies was also examined in New Zealand samples of the omnivorous Girella cyanea and the herbivorous Kyphosus bigibbus, and the tropical herbivorous species Kyphosus vaigiensis and Kyphosus cinerascens collected from the Great Barrier Reef, Australia. Overall, the results suggest that, like ruminants, herbivorous fishes such as Kyphosus species that rely upon gastrointestinal fermentation preferentially use lipids as major metabolic substrates by an increased capacity for lipid metabolism, and a lower capacity for glycolysis.     

Feasibility test of utilizing Saccharophagus degradans 2-40T as the source of crude enzyme for the saccharification of lignocellulose

In the conversion of lignocellulose into high-value products, including fuels and chemicals, the production of cellulase and the enzymatic hydrolysis for producing fermentable sugar are the largest contributors to the cost of production of the final products. The marine bacterium Saccharophagus degradans 2-40^T can degrade more than ten different complex polysaccharides found in the ocean, including cellulose and xylan. Accordingly, S. degradans has been actively considered as a practical source of crude enzymes needed for the saccharification of lignocellulose to produce ethanol by others including a leading commercial company. However, the overall enzyme system of S. degradans for hydrolyzing cellulose and hemicellulose has not been quantitatively evaluated yet in comparison with commercial enzymes. In this study, the inductions and activities of cellulase and xylanase of cell-free lysate of S. degradans were investigated. The growth of S. degradans cells and the activities of cellulase and xylanase were promoted by adding 2 % of cellulose and xylan mixture (cellulose:xylan = 4:3 in mass ratio) to the aquarium salt medium supplemented with 0.2 % glucose. The specific cellulase activity of the cell-free lysate of S. degradans, as determined by the filter paper activity assay, was approximately 70 times lower than those of commercial cellulases, including Celluclast 1.5 L and Accellerase 1000. These results imply that significant improvement in the cellulase activity of S. degradans is needed for the industrial uses of S. degradans as the enzyme source.     

Multiple proteins with single activities or a single protein with multiple activities: The conundrum of cell surface NADH oxidoreductases

Reduction of the cell-impermeable tetrazolium salt WST-1 has been used to characterise two plasma membrane NADH oxidoreductase activities in human cells. The trans activity, measured with WST-1 and the intermediate electron acceptor mPMS, utilises reducing equivalents from intracellular sources, while the surface activity, measured with WST-1 and extracellular NADH, is independent of intracellular metabolism. Whether these two activities involve distinct proteins or are inherent to a single protein is unclear. In this work, we have attempted to address this question by examining the relationship between the trans and surface WST-1-reducing activities and a third well-characterised family of cell surface oxidases, the ECTO-NOX proteins. Using blue native-polyacrylamide gel electrophoresis, we have identified a complex in the plasma membranes of human 143B osteosarcoma cells responsible for the NADH-dependent reduction of WST-1. The dye-reducing activity of the 300 kDa complex was attributed to a 70 kDa NADH oxidoreductase activity that cross-reacted with antisera against the ECTO-NOX protein CNOX. Differences in enzyme activities and inhibitor profiles between the WST-1-reducing NADH oxidoreductase enzyme in the presence of NADH or mPMS and the ECTO-NOX family are reconciled in terms of the different purification methods and assay systems used to study these proteins.     

Catalytic properties of lactate dehydrogenase in Homarus americanus.

Lobster tail and leg lactate dehydrogenases (LDH) have been characterized kinetically. The four binding sites for reduced coenzyme have been shown to be equivalent for the enzyme purified from lobster tail muscle. For the reduced form of 3-acetyl pyridineadenine dinucleotide, the K_a = 1.4 × 10⁷ M^?1 S^?1. The activity of the enzyme purified from the tail muscle is severely inhibited (90%) by high levels of pyruvate (10 mm) when assayed for pyruvate reductase activity at 11 °C; the reductase activity measured using the enzyme from the walking leg muscle was not inhibited by these high levels of pyruvate. Evidence is presented which indicates that the LDH from the tail muscle of the East Coast lobster forms an abortive ternary complex (enzyme-NAD⁺-pyruvate) which accounts for these inhibitory kinetics. The data suggest that the LDH from the tail muscles of the invertebrate lobster represents a “kinetic” heart-type l-specific LDH and that from the walking legs, a “kinetic” muscle-type l-specific LDH.     

On the lack of inotropy of cardiac glycosides on skeletal muscle: a comparison of Na+, K+-ATPases from skeletal and cardiac muscle.

Comparison of Na,K-ATPase from skeletal and cardiac muscle revealed that, although the skeletal muscle enzyme was only slightly less sensitive to inhibition by ouabain, the rates of [³H]ouabain binding to, and dissociation from, the skeletal enzyme were much faster than the corresponding rates for the cardiac enzyme. The skeletal muscle enzyme required higher concentrations of potassium to stabilize the ouabainenzyme complex and to stimulate the K⁺-phosphatase activity. The K⁺-phosphatase activity was only 8% of the Na,K-ATPase activity of the skeletal muscle enzyme, compared to 22% for the cardiac preparation. The glycoprotein subunit found in Na,K-ATPases from cardiac and many other tissues appeared to be absent in the enzyme from skeletal muscle. The differences in binding and dissociation rates for ouabain suggest that there may be significant differences in the structure of the digitalis receptor in the two enzymes. The I₅₀ for ouabain inhibition of the skeletal muscle Na,K-ATPase was, however, only slightly higher than for the cardiac enzyme, suggesting that the lack of an inotropic effect of cardiac glycosides on skeletal muscle could not be due to failure of the digitalis drugs to bind to and inhibit the membrane-linked sodium pump.     

Oxidase reactions of tomato anionic peroxidase

Tomato (Lycopersicon esculentum Mill) anionic peroxidase was found to catalyze oxidase reactions with NADH, glutathione, dithiothreitol, oxaloacetate, and hydroquinone as substrates with a mean activity 30% that of horseradish peroxidase; this is in contrast to the negligible activity of the tomato enzyme as compared to the horseradish enzyme in catalyzing an indoleacetic acid-oxidase reaction with only Mn²⁺ and a phenol as cofactors. Substitution of Ce³⁺ for Mn²⁺ produced an 18-fold larger response with the tomato enzyme than with the horseradish enzyme, suggesting a significant difference in the autocatalytic indoleacetic acid-oxidase reactions with these two enzymes. In attempting to compare enzyme activities with 2,4-dichlorophenol as a cofactor, it was found that reaction rates increased exponentially with both increasing cofactor concentration and increasing enzyme concentration. While the former response may be analogous to allosteric control of enzyme activity, the latter response is contrary to the principle that reaction rate is proportional to enzyme concentration, and additionally makes any comparison of enzyme activity difficult.     

西宁南山4种灌木根际和非根际土壤微生物、酶活性和养分特征

西宁南山区植被退化情况严重,人工造林植被恢复被看作是最有效的恢复手段,其中选择合适造林树种尤为关键。选择人工种植的唐古特白刺Nitraria tangutorum、柠条Caragana korshinskii、西北小蘗Berberis vernae和短叶锦鸡儿Caragana brevifolia共4种灌木树种造林试验区为研究对象,通过测定根际和非根际土壤微生物数量、酶活性及养分含量,综合比较种植4种灌木树种根际和非根际土壤肥力差异,科学评价其对土壤的改善效果。研究表明:(1)土壤微生物数量和酶活性总体呈现出根际高于非根际的规律,仅放线菌数量和脲酶活性出现了根际低于非根际现象。(2)土壤养分方面,4种灌木根际土壤和非根际土壤p H值、全N、全P、全K含量差异不显著,有机质、有效P、速效K含量均呈现出根际非根际,而碱解N则是根际非根际。(3)土壤酶活性与土壤微生物数量相关性不显著,土壤有机质含量与土壤细菌、真菌数量呈极显著正相关,有效P含量与土壤细菌、真菌和放线菌数量呈极显著正相关,速效K含量与过氧化氢酶、酸性磷酸酶活性呈显著正相关,全N、碱解N含量均与脲酶活性呈显著正相关。(4)从土壤肥力综合水平来看,根际非根际,其中根际土壤中西北小蘗柠条短叶锦鸡儿唐古特白刺,研究结果表明西北小蘗和柠条能大幅提高土壤肥力,改良土壤效果较好。     

Characterization of an Autosomal Rudimentary-Shaped Wing Mutation in DROSOPHILA MELANOGASTER That Affects Pyrimidine Synthesis

A new autosomal mutation, rudimental (ral), which causes rudimentary-shaped wings in Drosophila melanogaster, has been isolated following ethyl methanesulfonate (EMS) mutagenesis. The wing phenotype of rudimental is identical to that of the X-linked rudimentary (r) mutation, which affects the first three enzymes in the pyrimidine biosynthetic pathway. The autosomal mutant maps very close to ebony (3–70.7) at 70.42 on the right arm of chromosome 3. Analysis of the enzyme activities of orotate phosphoribosyltransferase (OPRTase) and orotidylate decarboxylase (ODCase) indicates that the ral^a26a allele has less than wild-type activity for both enzymes. This result is discussed in light of the fact that the OPRTase and ODCase activities are part of an enzyme complex, as are the carbamyl phosphate synthetase (CPSase), aspartate transcarbamylase (ATCase) and dihydroorotase (DHOase) activities, which are encoded by the complex rudimentary locus. We suggest that rudimental is also a complex locus.     

Development of catabolic pathways in insect flight muscles. A comparative study

Activities of enzymes representative of glycolytic and β-oxidative pathways and citric acid and glycerophosphate cycles were measured in the developing flight muscles of three species: Calliphora erythrocephala, Locusta migratoria, and Philosamia cynthia. The activities were measured in vitro under optimal conditions.The enzyme pattern of young flight muscles is quite different from the adult pattern. In the second half of the developmental period final differentiation towards the adult metabolic pattern takes place, in Calliphora leading to exclusively carbohydrate-oxidizing capacities, in Locusta to properties enabling both aerobic glycolytic and β-oxidative processes, whereas Philosamia becomes oriented to fatty acid oxidation. This differentiation starts after a temporary rise of lactate dehydrogenase activity, a phenomenon that seems to be connected with invagination of tracheoblasts into the muscle fibres. This tracheolization might be necessary for differentiation towards the species specific metabolic properties of the adult flight muscle.Theoretical aspects of the enzyme activities, as they were measured in the in vitro assays, are discussed and related to the physiological qualities of the flight muscles of the three species investigated.