Diurnal Changes in Asparaginase Activity in Pea Leaves: II. REGULATION OF ACTIVITY

Pea leaf asparaginase is stabilized by asparagine and aspartateduring incubation. In crude extracts this effect was enhancedby products of the light reaction (NADPH, NADH, or reduced ferredoxin),but these compounds were ineffective on the purified enzyme,or in the absence of asparagine. MgATP, MgADP and oxidized ferredoxinreduced asparaginase activity in purified preparation reducedor oxidized glutathione had no effect. Asparaginase activitydoes not appear to be modulated via phosphorylation/dephosphorylation.The presence of calcium during extraction increased asparaginaseactivity more than 2-fold, but addition of calcium to extractsprepared in its absence had no effect; calmodulin had no effecton activity. Co-extraction of light- and dark-treated tissueshowed that soluble factors are not responsible for the diurnalvariation in asparaginase activity. Association of asparaginasewith membranes did not account for changes in extractable activity.Use of the protein synthesis inhibitors cycloheximide, puromycin,emetine, actinomycin D and cordycepin and the thiol proteaseinhibitor leupeptin suggested that mRNA and protein synthesisare required for the increase of asparaginase activity duringthe light period and that proteolytic degradation accounts forthe decrease during the dark. Key words: Pisum sativum, asparaginase, protein synthesis, proteolysis.     

Diurnal Changes in Asparaginase Activity in Pea Leaves: I. THE REQUIREMENT FOR LIGHT FOR INCREASED ACTIVITY

During the growth of leaves of Pisum sativum L., levels of asparaginase(E.C. 3.5.1.1 [EC] ) showed a diurnal variation during a 3 d periodof leaf expansion, increasing in the light and decreasing inthe dark period; the greatest diurnal variation being foundin half-expanded leaves. Asparaginase activity in half-expandedleaves reached a maximum after 4 h exposure to light and thisactivity was maintained over the rest of the light period. Changesin asparaginase activity were not influenced by diurnal temperaturechanges. The increase in asparaginase activity during the lightperiod was directly proportional to the photon flux densityover the range 0–285 µmol m^-2 s^-1 PAR. The increaseof asparaginase activity during illumination of detached leaveswas inhibited by the photosynthetic electron transport inhibitors3-(3', 4'-dichlorophenyl)-1, 1-dimethylurea (DCMU) and atrazine.These observations indicate that the increase in asparaginaseactivity in half-expanded leaves is dependent upon non-cyclicelectron transport. Key words: Pisum sativum, asparaginase, photosynthetic electron transport     

A New Species of Kudoa (Myxosporidea: Chloromyxidae) from the Spot,Leiostomus xanthurus Lacépède,in Clear Lake,Texas*

SYNOPSIS. Kudoa branchiata sp. n. (Myxosporidea: Chloromyxidae) is described from the gills of the marine sciaenid fish, Leiostomus xanthurus Lacépède, from Clear Lake, Texas. Twelve of the 429 hosts examined from 10 September 1970 to 28 April 1971, were infected. Eight of the 12 infected fish were collected in February and March 1971—a period during which only 13% of the total host sample was taken. The mean total length of infected hosts was 149 mm, with a range of 112–185 mm. The mean number of myxosporidean cysts per infected host was 3.5, with a range of 1–11.     

The causes of colour and colour change in caterpillars of the poplar and eyed hawkmoths (Laothoe populi and Smerinthus ocellata)

Caterpillars of the poplar and eyed hawkmoths (Laothoe populi and Smerinthus ocellata respectively) were reared under different conditions in order to determine why final instar caterpillars vary in colour. Poplar hawkmoth caterpillars normally rest on the undersides of leaves. Dull green and redspotted caterpillars are genetically determined polymorphisms. Caterpillars that are not dull green, however, can become white when fed on Populus alba or yellow-green when fed on Salix fragilis. Experiments showed that it is the reflective qualities of the leaves that determines which colour the caterpillar develops: if the young larva sees white then it becomes white, but if it sees green, grey or black then it becomes yellow-green. Young eyed hawkmoth larvae always developed into grey-green final instar caterpillars under our rather poorly-illuminated rearing conditions, but when reared on wild plants in white muslin sleeves they became whitish-green. In this species also it appears that colour of the final instar is determined by the reflectance of the substrate perceived by the young caterpillar.     

THE ESTIMATION OF SULPHATE-REDUCING BACTERIA (D. DESULPHURICANS)

SUMMARY: Sodium sulphide or cysteine stimulated the growth of sulphate-reducing bacteria; small populations often did not grow without such supplements. Ascorbic acid, glutathione or thiolacetic acid had similar properties but thiolacetic acid was sometimes inhibitory, Dilution counts in liquid media or colony counts in agar media did not bear any regular relation to the total count unless one of these supplements was present. With suitable precautions colony counts reaching 50 to 60% of the total count were obtained in media incorporating cysteine and a ferrous salt (as an indicator of sulphide formation).
Samples of natural origin containing sulphate-reducing bacteria gave greater viable counts in cysteine-iron media than in unsupplemented media. Blackend culture tubes with natural populations were sometimes due to cysteine-decomposing organisms; further examination of positive tubes was therefore necessary.     

Flow Cytometric Analysis of Immunoglobulin and Complement Component C3 on the Surface of Trypanosoma lewisi1

ABSTRACT. We have reinvestigated whether surface immunoglobulin (sIg) on Trypanosoma lewisi is antibody directed toward parasite antigen by using flow cytometry to analyze parasites stained with fluoresceinated F(ab′)₂ fragments of antibodies to rat IgG and IgM. We have confirmed that IgG antibody to the parasites is present both in the serum of rats and on the surface of parasites between the fourth and twentieth days of infection, that the amount of sIg per cell increases as the infection progresses, and that considerably more IgG is present on parasites harvested from intact rats than on those from rats that had been immunosuppressed by whole body γ-irradiation. In addition sIgM was detected on trypanosomes from intact, but not on parasites from irradiated rats. We have also made two observations suggesting that not all sIg is specific antibody made in response to T. lewisi. First, a low but significant amount of sIgG was detected on parasites throughout infection in irradiated rats; no sIgM was detected on these parasites. Second, when parasites harvested from immunosuppressed rats were incubated in normal rat serum, the amount of both sIgG and sIgM detected by flow immunofluorescence increased. Parasites harvested from intact animals bound IgM but not IgG from normal rat serum. These results suggest either that natural antibody to the trypanosomes is present in the serum of uninfected rats or that some rat immunoglobulins bind to structures on the trypanosome surface in ways that do not depend on usual antigen-antibody interactions. Finally, flow immunofluorescence was also used to detect complement component C3 on the surface of both intact and trypsinized bloodstream forms harvested from intact or immunosuppressed rats. The amount of sC3 per cell did not increase until late in the infection and consequently did not correlate with the increase of sIgG. Therefore, T. lewisi avoids destruction by the immune system although immune effector molecules, IgG, IgM, and C3, are on its surface.     

A New Species of Henneguya (Myxosporidea: Myxobolidae) from the Black Drum Pogonias cromis in Clear Lake, Texas

SYNOPSIS. Henneguya texana n. sp. (Myxosporidea: Myxobolidae) is described from the gill filaments of 2 out of 47 specimens of marine sciaenid fish Pogonias cromis collected from Clear Lake, Texas. The spore body is 7–9 by 6–7 μ (mean 8.1 by 6.7 μ) with a thickness of 4–5 μ (mean 4.7 μ). The 2 fused, tapering caudal processes are 36–59 μ (mean 48.65 μ) long. The parasitized fish were collected in December 1970 and January 1971 were 168 and 482 mm in total length, respectively.