[12] Isolation of mutants from Euglena gracilis

Schiff, Jerome A.; Lyman, Harvard; Russell, George K.

doi:10.1016/s0076-6879(71)23088-3

Cited by 79 publications

(49 citation statements)

References 36 publications

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“…Photosynthetic carbon dioxide fixation was assayed as described previously (Diamond & Schiff,I 970). Viability and green-colony forming ability were determined by dilution and plating on pH 3.5 medium (Schiff, Lyman & Russell, 1971) since plating on a low pH medium prevented further streptomycin entry. Methods for fluorescence microscopy have been described (Klein,Schiff & Holowinsky,I 972).…”

Section: Introductionmentioning

confidence: 99%

Events Surrounding the Early Development of Euglena Chloroplasts: Experiments with Streptomycin in Non-dividing Cells

Bovarnick

Chang

Schiff

et al. 1974

Journal of General Microbiology

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The concentration of streptomycin (Sm) which selectively inhibits light-induced chloroplast development in non-dividing Euglena is the same as that which induces the loss of green-colony forming ability in dividing organisms. This concentration of Sm has no effect on division or viability. Chlorophyll synthesis is insensitive to streptomycin for the first 1 2 h of development but is strongly inhibited after this time. Between 72 and 96 h after the beginning of chloroplast development, Sm-treated organisms contain 10 yo of the chlorophyll and 24 of the carotenoids of algae developing in the absence of the antibiotic. The chlorophyll-to-carotenoid ratio in treated organisms at 72 to 96 h is 0.9, the same as is found at 1 2 h for organisms developing in the absence of Sm. In the presence of streptomycin, Euglena never develops the ability to fix CO, photosynthetically, although CO, fixation after I 2 h of development in the absence of the antibiotic can be readily detected. At I 2 h of chloroplast development the following parameters are at comparable levels in Sm-treated and untreated organisms: the bound forms of chlorophyll, concentration of cytochrome 552, the activities of ribulose diphosphate carboxylase, NADP-triose phosphate dehydrogenase, the enzymes converting ribose-yphosphate to ribulose diphosphate, and photosystem I1 activity measured as dye reduction.

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Section: Introductionmentioning

confidence: 99%

Events Surrounding the Early Development of Euglena Chloroplasts: Experiments with Streptomycin in Non-dividing Cells

Bovarnick

Chang

Schiff

et al. 1974

Journal of General Microbiology

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“…Older colonies of the organism are golden in color, and the cells contain low levels of chlorophyll and are capable of photosynthesis. Although the several lines of evidence that indicate that the mutants used in these studies are probably chloroplastic have been discussed in some detail (4,5), it seems appropriate to review them very briefly here. Succinctly stated these are: (a) Ultraviolet and x-ray killing curves show a multiplicity of about 8, indicating that the nuclear chromosomal complement is octaploid.…”

Section: Methodsmentioning

confidence: 99%

A Cytoplasmic Regulatory Mutant of Euglena: Constitutivity for the Light-Inducible Chloroplast Transfer RNAs

Goins

Reynolds²,

Schiff³

et al. 1973

Proc. Natl. Acad. Sci. U.S.A.

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An ultraviolet-induced cytoplasmic mutant (GIBU) In Euglena, the chloroplastic species of tRNA and of aminoacyl-tRNA synthetases appear to be quantitatively regulated by light; i.e., exposure of dark-grown wild-type cells to light results in a striking increase in the quantity of these organelle translational macromolecules (1-3). The present report represents an initial attempt to elucidate the control mechanism(s) involved in the light induction of chloroplast tRNAs, through the use of chloroplhstic mutants (4-6) ofEuglena. In these studies we examined mutants of Euglena gracilis Klebs var. bacillaris Pringsheim for the loss of normal control of chloroplast tRNA biosynthesis. In particular, we describe one mutant (GIBU), which was induced by ultraviolet irradiation, but appears to be a regulatory "constitutive" mutant in the sense that, in contrast to wild type, certain chloroplast tRNAs are present in repressed (or darkgrown) mutant cells at approximately the same level as that found in induced (or light-grown) cells.MATERIALS AND METHODS tRNA Was Prepared from Euglena cells in the logarithmic growth phase as described (3).tRNA Chromatography. The reversed-phase system described by Weiss and Kelmers (7) was used -with columns 225 X 1.5 cm. For chromatographic comparisons, approximately equal amounts (in cpm) of 14C-and 3H-labeled aminoacyl-tRNAs were applied to the columns. The chromatography is described in the legend to tRNAs from cells grown in the light and the dark were acylated with a synthetase preparation (1) from cells grown in the light and chromatographed as described in (A) except that the NaCl gradient was from 0.40-0.55 M. 1749I To whom reprint requests should be addressed.

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“…Thylakoids from wild-type (WT) cells, solubilized in these detergents and subjected to polyacrylamide gel electrophoresis at 0°C, yield the following CPs, in order of relative molecular weight, containing the pigments shown in parentheses with their respective molar ratios where determined: CP Ia (Chi a, diadinoxanthin and 0-carotene; 100:12:5); CP I (Chi a and 0-carotene; 100:6-12); CPx (Chi and carotenoids); LHCP2 (light-harvesting CP oligomer) (Chl a, Chl b, diadinoxanthin and neoxanthin; 12:4:3:1); CPy (Chl a, diadinoxanthin and 0-carotene; 100:14:8); CPa (Chi a and 0-carotene; 100:18-25) and LHCP (monomer) (Chl a, Chl b, diadinoxanthin and neoxanthin; 12:6:4:1). The [30] for nomenclature) were cultured in I L batches of Hutner's pH 3.5 medium (15) at 26°C. In most cases, the 2 L Erlenmeyer flasks containing the cultures were placed on a rotary platform shaker adjusted to 110 cycles/ min and were illuminated from above at a fluence rate of 4.0 W/m2 provided by alternating red and cool-white fluorescent lamps (Sylvania) screened with a plastic mesh to reduce the light intensity.…”

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confidence: 99%

“…In addition, cells of Euglena contain only one-third to one-half of the amount of Chl b (relative to Chl a) characteristic of higher plants. Several [30] for nomenclature) were cultured in I L batches of Hutner's pH 3.5 medium (15) at 26°C. In most cases, the 2 L Erlenmeyer flasks containing the cultures were placed on a rotary platform shaker adjusted to 110 cycles/ min and were illuminated from above at a fluence rate of 4 (v:v) was pumped (Lab Data Control Constametric III HPLC pump) through the column at a flow rate of 1.00 ml/min; the corresponding back pressure was about 5.5 x 106 N/M2 (800 p.s.i.).…”

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confidence: 99%

Chlorophyll-Protein Complexes from Euglena gracilis and Mutants Deficient in Chlorophyll b

Cunningham¹,

Schiff²

1986

Plant Physiol.

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The use of n-octyl-ft-n-glucopyranoside along with sodium dodecyl sulfate improves the retention of chlorophyll (Chl) by chlorophyll-protein complexes (CPs) prepared from thylakoids of Euglena grailis Klebs var bacillaris Cori and yields several additional complexes. Thylakoids from wild-type (WT) cells, solubilized in these detergents and subjected to polyacrylamide gel electrophoresis at 0°C, yield the following CPs, in order of relative molecular weight, containing the pigments shown in parentheses with their respective molar ratios where determined: CP Ia (Chi a, diadinoxanthin and 0-carotene; 100:12:5); CP I (Chi a and 0-carotene; 100:6-12); CPx (Chi and carotenoids); LHCP2 (light-harvesting CP oligomer) (Chl a, Chl b, diadinoxanthin and neoxanthin; 12:4:3:1); CPy (Chl a, diadinoxanthin and 0-carotene; 100:14:8); CPa (Chi a and 0-carotene; 100:18-25) and LHCP (monomer) (Chl a, Chl b, diadinoxanthin and neoxanthin; 12:6:4:1). The [30] for nomenclature) were cultured in I L batches of Hutner's pH 3.5 medium (15) at 26°C. In most cases, the 2 L Erlenmeyer flasks containing the cultures were placed on a rotary platform shaker adjusted to 110 cycles/ min and were illuminated from above at a fluence rate of 4.0 W/m2 provided by alternating red and cool-white fluorescent lamps (Sylvania) screened with a plastic mesh to reduce the light intensity. For some experiments, cultures were grown without shaking (standing) on a glass shelf and were illuminated from above and below at a fluence rate of 14.0 W/m2 with the unscreened fluorescent lamps described above. All cultures were harvested 2 d after cell division had ceased; at this point the cellular Chl contents had reached stable values. Cell density was determined with a Coulter model Zb cell counter (100 AM aperture).Preparation of Thylakoids. Cultures (usually 10-20 L) of Euglena were filtered through four layers of cheesecloth and the cells were pelleted by centrifugation at 100g for 2 min at 4°C.All subsequent procedures were carried out at 4°C or on ice. The cell pellets were combined, washed with ice-cold distilled H20, and resuspended in a BS containing 0.3 M sucrose and 50 mm Tris-HCl at pH 7.6. Cells were pelleted by centrifugation at 1 640g for 5 min in preweighed centrifuge bottles, the supernatant fluid was removed by aspiration, and the weights of the pellets were determined. BS containing 1.0 mM PMSF (Sigma) was 223 www.plantphysiol.org on May 10, 2018 -Published by Downloaded from

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[12] Isolation of mutants from Euglena gracilis

Cited by 79 publications

References 36 publications

Events Surrounding the Early Development of Euglena Chloroplasts: Experiments with Streptomycin in Non-dividing Cells

Events Surrounding the Early Development of Euglena Chloroplasts: Experiments with Streptomycin in Non-dividing Cells

A Cytoplasmic Regulatory Mutant of Euglena: Constitutivity for the Light-Inducible Chloroplast Transfer RNAs

Chlorophyll-Protein Complexes from Euglena gracilis and Mutants Deficient in Chlorophyll b

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