EFFECTS OF AUXIN ON CELL ENLARGEMENT IN FERN GAMETOPHYTES: DEVELOPMENTAL STAGE OF THE CELL AND RESPONSE TO AUXIN

When gametophytes of Onoclea sensibilis are placed on an auxin-containing medium in darkness, the cells of the plant increase in length and width. Measurements were made to determine the effects of cell age and auxin concentration on the magnitude of the cells’ response. When comparing final size with size before auxin treatment, the youngest cells have the greatest response. However, they also have the greatest endogenous growth. The optimum auxin concentration for increase in length changes with the age of the cells. The greatest length is attained in young cells with an auxin concentration of 10^−-6–10^−-7 m; 10^−-5 m auxin is supraoptimal. The greatest increase in width of the cells is produced by 10^−-5 m auxin, regardless of cell age. The data are believed best to be interpreted as showing an age-dependent change in the capacity of the cells to respond to auxin.     

INFLUENCE OF THE TONIC EFFECT OF GRAVITATION AND AUXIN ON CELL ELONGATION AND POLARITY IN ROOTS

The influence of a longitudinal (tonic) gravitational force and of auxin on the pattern of growth and cell polarity has been studied on intact roots of wheat seedlings. A klinostat technique was used for controlling gravitation. Growth in length was evaluated as cell division activity, rate of cell elongation (μ/h) and duration of elongation (h). Exogenous auxin (1-NAA) increases the rate of cell elongation in all concentrations tested (10⁻⁸ — 3 × 10⁻⁷m ) and shortens the time of elongation with increasing concentration. It promotes rate of cell elongation in roots as it does in shoots. It also accentuates the polar insertion of root hairs and their growth. The tonic effect of gravitation resembles that of an increase in auxin both in light and darkness. The results are discussed in relation to plagiotropic growth of roots, root growth promotions by auxin, and the difference between root and shoot growth.     

REGULATION OF GROWTH AND CELLULAR DIFFERENTIATION IN DEVELOPING AVENA INTERNODES BY GIBBERELLIC ACID AND INDOLE-3-ACETIC ACID

Auxin (IAA) at physiological concentrations causes significant reduction of GA₃-promoted growth in excised Avena stem segments. IAA is thus considered to be a gibberellin antagonist in this system. It was found to act non-competitively in repressing GA₃-augmented growth in these segments. In intercalary meristem cells at the base of the elongating internode, GA₃ blocks cell division activity and causes a marked increase in cell lengthening. IAA substantially promotes lateral expansion in comparable intercalary meristem cells, particularly in the vicinity of vascular bundles underlying the epidermis. It also alters the plane of cell division in differentiating stomata. IAA at high concentrations (10⁻³, 10⁻⁴ m ), in combination with GA₃, overrides the effects of GA₃ on cell lengthening, while with low concentrations of IAA (10⁻⁹, 10⁻¹⁰m ), the effects of GA₃ are clearly dominant. At intermediate concentrations of IAA (10⁻⁶, 10⁻⁷m ), in the presence of GA₃, the effects of this treatment on cell differentiation closely parallel the pattern of differentiation in untreated tissue. It is postulated that a lateral gradient of auxin and gibberellin could control cell expansion in long epidermal cells during intercalary growth of the internode.     

THE COMPARATIVE CELL CYCLE AND METABOLIC EFFECTS OF CHEMICAL TREATMENTS ON ROOT TIP MERISTEMS. I. IOXYNIL

In this study we investigated the cell cycle response of Vicia faba and Pisum sativum root tip meristems to ioxynil treatments at two concentrations, (10^−-4m and 10^−-6m ). After 24 h of treatment at 10^−-4m concentration, O₂ uptake and ATP concentrations were significantly reduced. The mitotic index was reduced and the cell cycle population position was shifted to indicate that previously inhibited cells reformed their nuclei and became tetraploid. Prolonged treatment at this concentration resulted in cell death. Treatment with ioxynil at 10^−-6m reduced the rate of entry into mitosis. Abnormal mitotic figures in all stages were observed, and the ploidy level of mitotically inhibited cells was doubled. These observations indicated that at 10^−-6m concentration ioxynil acts as a preprophase inhibitor, that is, it does not act directly on the mitotic apparatus but does affect processes on which mitosis depends.     

TIME- AND CONCENTRATION-DEPENDENCE OF THE GROWTH-PROMOTING ACTIVITY OF INSULIN AND HISTAMINE IN TETRAHYMENA. APPLICATION OF THE MTT-METHOD FOR THE DETERMINATION OF CELL PROLIFERATION IN A PROTOZOAN MODEL

The unicellular ciliateTetrahymena pyriformiswas treated with different concentrations of insulin or histamine and at different time points the cell density was measured, using a tetrazolium-based semiautomated colorimetric assay (MTT). The assay was suitable to determine the rate of cell proliferation of Tetrahymena. Insulin in each concentration significantly elevated the cell count up to 3h. After that, it was neutral or its effect was insignificant. Histamine at 10^?5m concentration diminished cell count at 3, 5, 7 and 24h. At 10^?6m concentration there was no difference and at 10^?7m concentration it enhanced cell division up to 5h, after that there being no difference. The two hormones have cell division promoting activity for cells of higher animals and the experiments demonstrate this effect already at a unicellular level.     

COBALT EFFECTS ON CELL DIVISION AND CALCIUM UPTAKE IN THE COCCOLITHOPHORID CRICOSPHAERA CARTERAE (HAPTOPHYCEAE)1

Cobalt was found to produce effects on cell division, cell protein, volume, calcium uptake, and ultrastructure of the calcifying alga Cricosphaera carterae (Braarud & Fager.) Braarud. The rate of cell division increased slightly as the added Co concentration of the medium was increased from 0–10 μM. At higher added Co concentrations, the rate of division decreased approximately linearly with concentration until division was blocked almost completely at 100 μM Co. Inhibition of division was reversible after 2 days in Co. Protein content was elevated in Co-treated nondividing cells but the rate of protein synthesis was markedly reduced. Cell volume also increased progressively with Co concentration, and after 72 h in 200 μM Co, the volume was 3.2 times that in culture medium (0.09 μM Co). Co had a dual effect on calcification as indicated by the uptake of ⁴⁵Ca. After 48 h treatment, Ca uptake had increased 53% in 100 μM and decreased 40% in 200 μM Co as compared with cells in 0.09 μM Co. Ultrastructurally, Co caused enlargement of the cell vacuole and the appearance of membrane-bound vacuoles containing electron dense bodies.     

SELENIUM: AN ESSENTIAL ELEMENT FOR GROWTH OF THE COASTAL MARINE DIATOM THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE)1,2

An obligate requirement for selenium is demonstrated in axenic culture of the coastal marine diatom Thalassiosira pseudonana (clone 3H) (Hust.) Hasle and Heimdal grown in artificial seawater medium. Selenium deficiency was characterized by a reduction in growth rate and eventually by a cessation of cell division. The addition of 10⁻¹⁰ M Na₂SeO₃ to nutrient enriched artificial seawater resulted in excellent growth of T. pseudonana and only a slight inhibition of growth occurred at Na₂SeO₃ concentrations of 10⁻³ and 10⁻² M. By contrast, Na₂SeO₄ failed to support growth of T. pseudonana when supplied at concentrations less than 10⁻⁷ M and the growth rate at this concentration was only one quarter of the maximum growth rate. The addition of 10⁻³ and 10⁻² M Na₂SeO₄ to the culture medium was toxic and cell growth was completely inhibited. Eleven trace elements were tested for their ability to replace the selenium requirement by this alga find all were without effect. In selenium-deficient and selenium-starved cultures of T. pseudonana cell volume increased as much as 10-fold as a result of an increase in cell length (along the pervalvar axis) but cell width was constant. It is concluded that selenium is an indispensable element for the growth of T. pseudonana and it should be included as a nutrient enrichment to artificial seawater medium when culturing this alga.     

STUDIES ON CELL METABOLISM AND CELL DIVISION : II. STIMULATION OF CELLULAR OXIDATION AND REVERSIBLE INHIBITION OF CELL DIVISION BY DIHALO AND TRIHALOPHENOLS

The dihalo and trihalophenols, and phenols containing both halo and nitro substituents in the same molecule, produce, in fertilized eggs of Arbacia punctulata, a rise in rate of oxygen consumption and a reversible block to cell division. To define the conditions which affect the degree of this activity, the following factors have been varied: the arrangement of substituents in the molecule, the concentration of reagent, and the time after fertilization at which the reagent is added. The stimulation of oxygen consumption and reversible block to cell division produced by the dihalophenols are qualitatively the same as those previously produced in fertilized Arbacia eggs by certain dinitrophenols. To yield optimum respiratory effect and maximum division block, it usually requires a higher concentration of dihalo than of the corresponding dinitrophenol. For example, with fertilized Arbacia eggs at 20°C. 2,4-dinitrophenol, in optimum concentration of 3 x 10^–5 molar, raises oxygen consumption to 292 per cent of normal (4). The corresponding values for two dihalo analogues are: 2,4-dichlorophenol, 10^–4 molar and 236 per cent; 2,4-dibromophenol, 6 x 10^–5 molar and 282 per cent. The halophenols differ from the nitrophenols in two interesting respects: (a) The monohalophenols produce little or no oxidative stimulation or division block in fertilized Arbacia eggs; p-nitrophenol is very active in both respects. (b) The symmetrical trihalophenols have an appreciable ability to stimulate oxygen consumption and block division; symmetrical trinitrophenol is inactive in both respects (4). The increases in oxygen consumption produced in fertilized Arbacia eggs by 2,4-dichloro and 2,4-dinitrophenol are larger than the percentage increases given by methylene blue and o-cresol indophenol under the same experimental conditions. The dihalo and dinitrophenols produce a reversible block to the cell division of fertilized marine eggs. The oxidation-reduction indicators, in contrast to the dihalo and dinitrophenols, block cell division irreversibly and fertilized eggs of Arbacia do not recover from optimum respiratory stimulating concentrations of these oxidation-reduction dyes. The present experiments with halophenols are in harmony with and lend considerable support to the hypothesis (4) that nitro and similarly substituted phenols derive their biological activity from the presence and properties of the phenolic OH group, as modified by proper substitution in the phenolic benzene ring.     

THE AFFINITY OF ONION CELL WALLS FOR CALCIUM IONS

Cell walls prepared from onion bulbs were found to exhibit an affinity for Ca⁺⁺. The adsorption of this ion was enhanced by the action of pectin methylesterase. It was confirmed that Ca⁺⁺ reacts with two COO“ groups and the corresponding affinity constant, K, was found to be: log K = 4.25. The action of pectin methylesterase had no effect upon K. The cell walls, as prepared, had 25 % of the total COO⁻ groups occupied by Ca⁺⁺, 14 % by Mg⁺⁺, and 39 % by H⁺. Treatment with acidified ethanol removed all of the metallic cations. K⁺ and Mg⁺⁺ could displace Ca⁺⁺ from the cell walls. At concentrations from 10⁻³ to 3 times 10⁻³ m it required from 4.9 to 13.2 moles of Mg⁺⁺ to displace one mole of Ca⁺⁺. For K⁺ it required 80 moles to displace 1 mole of Ca⁺⁺ at K⁺ concentrations from 0.65 × 10⁻² to 1.6 × 10⁻² M.     

SELENIUM: AN ESSENTIAL ELEMENT FOR GROWTH OF THE COASTAL MARINE DIATOM THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE)1,2

An obligate requirement for selenium is demonstrated in axenic culture of the coastal marine diatom Thalassiosira pseudonana (clone 3H) (Hust.) Hasle and Heimdal grown in artificial seawater medium. Selenium deficiency was characterized by a reduction in growth rate and eventually by a cessation of cell division. The addition of 10⁻¹⁰ M Na₂eO₃ to nutrient enriched artifical seawater resulted in excellent growth of T. pseudonana and only a slight inhibition of growth occurred at Na₂SeO₃ concentrations of 10⁻³ and 10_-2 M. By contrast, Na₂SeO₄ failed to support growth of T. pseudonana when supplied at concentrations less than 10⁻⁷ M and the growth rate at this concentration was only one quarter of the maximum growth rate. The addition of 10⁻³ and 10⁻² M Na₂SeO₄ to the culture medium was toxic and cell growth was completely inhibited. Eleven trace elements were tested for their ability to replace the selenium requirement by this alga and all were without effect. In selenium-deficient and selenium-starved cultures of T. pseudonana cell volume increased as much as 10-fold as a result of an increase in cell length (along the pervalvar axis) but cell width was constant. It is concluded that selenium is an indispensable element for the growth of T. pseudonana and it should be included as a nutrient enrichment to artificial seawater medium when culturing this alga.     

INFLUENCE OF CERTAIN PLANT GROWTH REGULATORS AND CROWN-GALL RELATED SUBSTANCES ON BUD FORMATION AND GAMETOPHORE DEVELOPMENT OF THE MOSS PYLAISIELLA SELWYNII

Bud formation and gametophore development were studied in the moss Pylaisiella selwynii (Kindb.) Steere and Anderson grown from spores in a liquid medium consisting of inorganic salts. Indoleacetic acid and ethrel increased bud formation within a narrow concentration range. Copious bud formation was obtained with the five cytokinins tested at concentrations varying from 5 X 10⁻⁶ to 5 X 10⁻¹⁴ M. Except for about 10 % of the buds obtained with 6-γ, γ-dimethylallylaminopurine at 5 times 10⁻¹⁴ m, the cytokinin-induced buds failed to develop into normal gametophores. Octopine, lysopine, and octopinic acid, substances obtained from crown-gall tumors, increased bud formation at 10⁻³ m. On lysopine-treated plants these buds developed into typical gametophores. Gemma-like structures were obtained with octopine but no gametophores. l -arginine and l -lysine, the amino acids which respectively occur in octopine and lysopine, failed to induce gametophore formation although buds were obtained with 10⁻³ m lysine. γ-Guanidinobutyric acid induced bud formation at 10⁻³ m, but these buds developed into highly abnormal gametophores. The failure of buds obtained with many of these treatments to develop into gametophores appeared to result from the formation of new cell walls in other than the normal geometrical relationship during initial divisions of the pro-bud. The relevance of the findings to the crown-gall problem is discussed.     

THE RESPONSE OF MALE GAMETES OF ALLOMYCES TO THE SEXUAL HORMONE SIRENIN

Attraction of male gametes of Allomyces to sirenin concentrations as low as 10^−-10 m has been demonstrated. Increasingly strong responses are observed to concentrations up to 10^−-6 m. The response is slightly weaker at 10^−-5 m and very weak at 10^−-4 m. Microscopic studies suggest that the attraction comes into the category of tropotaxis, a direct approach to a sirenin source occurring. Male gametes have been shown to inactivate sirenin. The possibility that this inactivation is an adaptation process enabling the gametes to maintain sirenin sensitivity over a 100,000-fold concentration range is discussed.     

LIGHT/DARK-PHASED CELL DIVISION IN EUGLENA GRACILIS (Z) (EUGLENOPHYCEAE) IN PO4-LIMITED CONTINUOUS CULTURE1

Eugene gracilis Klebs (Z) was grown in a cyclostat (continuous culture on a light/dark cycle) at growth limiting levels of phosphate. Cell division was restricted to the dark period regardless of the proportion of the cells dividing during each 24 h period. Growth rate, as reflected by the amplitude of the cell density oscillation, was correlated with dilution rate. The width of the division gate was analyzed using a phasing index and found to be narrowest at dilution rates where the mean generation time of the cell population was an even multiple of 24 h. The effect was attributed to enhanced phasing of the cell division process by the biological clock of Euglena. Residual phosphate levels in the cyclostat were less than 0.3 μM PO₄ at all submaximal growth rates. Cellular phosphorus concentration increased with dilution rate as described by a hyperbola saturating at D_max= 0.74 day⁻¹ with 8 × 10⁻⁸μM P/cell as the minimum intracellular phosphorus concentration for growth. The results are discussed, in terms of the inherent similarities and differences between a cyclostat and a steady state chemostat, and the advantages of the cyclostat for studies in phytoplankton ecology.     

THE EFFECTS OF KINETIN AND NAPHTHALENEACETIC ACID ON IN VITRO SHOOT MULTIPLICATION AND ROOTING IN THE FISHTAIL FERN

This study is concerned with the determination of the shoot and root inducing effects of kinetin (K) and naphthaleneacetic acid (NAA) on shoots ofthe fishtail fern (Nephrolepisfalcata formafurcans) in sterile tissue culture. The data shows that K is the major factor involved in maximal shoot production. The NAA is not essential. Specific concentrations ofNAA must be present with specific concentrations of K for maximal root production. At the same time, the data also demonstrate that shoots on media containing no NAA but with K concentrations of 5 × 10⁻⁷ and 10⁻⁶ M produced as many roots as with any other NAA concentration. The data can be used as a guide to rapid commercial propagation of fishtail fern, and demonstrate that media available commercially for Boston fern multiplication will induce maximal shoot production in fishtail fern.     

APICAL DOMINANCE: THE EFFECTS OF GROWTH REGULATORS ON THE GAMETOPHYTE OF ANEMIA PHYLLITIDIS

Gametophytes of the fern, Anemia phyllitidis (L.) Sw. were aseptically cultured until nearly mature, then collected and surgically divided into meristematic and nonmeristematic halves. Indoleacetic acid, benzyladenine, abscisic acid, and the auxin inhibitor, triiodobenzoic acid, were employed to elucidate the role of growth regulators during regeneration of adventitious gametophytes. Explants possessing actively growing meristems failed to produce adventitious thalli regardless of treatment. Untreated nonmeristematic halves initiated numerous filamentous outgrowths within two weeks of excision. These outgrowths eventually developed into the cordate prothalli typical of this species. Nontoxic levels of IAA (10⁻⁴ g/1 and 10⁻⁵ g/1) promoted early regeneration by basal explants. TIBA, at 10⁻⁵ m, completely suppressed regeneration. Benzyladenine, at 10⁻² g/1, exerted its most striking effect by greatly increasing the number of adventitious outgrowths formed by treated basal explants. ABA at all concentrations delayed regeneration and reduced the number of basal halves that displayed renewed mitotic activity. This evidence suggests that auxin acts as an essential promoter of regeneration, and that the maintenance of endogenous hormonal interactions is necessary to preserve the form of an intact gametophyte.     

CALCIUM DELAYS THE ONSET OF CELL DIVISION AND RHIZOID ELONGATION IN GEMMAE OF THE CALCIFUGE FERN,VITTARIA GRAMINIFOLIA

Vittaria graminifolia gametophytes reproduce asexually by linear six-celled propagules known as gemmae. It has been shown previously that asymmetric cell division and rhizoid elongation in gemmae are inhibited by millimolar concentrations of Ca^{+ +}. The present paper shows that millimolar Ca^{+ +} delays the onset of cell division at a point prior to mitosis, without affecting the maximal rate of cell division. Ca^{+ +} is most effective in delaying cell division when it is present during the first 24 hours of culture, 2 or 3 days before the initiation of cell division. Millimolar Ca^{+ +} inhibits rhizoid elongation by delaying its onset, while also reducing its maximal rate. Ca^{+ +} is also most effective in delaying rhizoid elongation during the first 24 hours of culture. Culture of gemmae on Ethylglycol-bis-(aminoethyl ether)-N,N,N‘,N‘-tetraacetic acid-buffered media shows that the maximum frequency of cell division occurs at pCa 7, while for the initiation of rhizoid elongation, it occurs at pCa 5.     

AUXIN-INDUCED HYPONASTY OF THE LEAF BLADE OF PHASEOLUS VULGARIS

The lateral margins of immature primary leaf blades of Phaseolus vulgaris L. cv. ‘Pinto’ curve up and in toward the midrib when auxin is applied to the leaf. The leaves are most sensitive to auxin shortly after they first unfold and leaves which have grown to about 60 % or more of their ultimate area no longer give this hyponastic response. The response is specific for auxins and is inhibited by the anti-auxins, trans-cinnamic acid and para-chlorophenoxyisobutyric acid. Ethylene and ethylene-generating compounds failed to induce hyponasty, suggesting the response is due to a positive growth promotion by auxin. Measurements of the distance between the lateral margins of the leaf at its maximum width were used to provide quantitative estimates of the degree of hyponasty. Between 2 and 4 hr after auxin application a direct proportionality was found between the amount of curvature and the logarithm of the indoleacetic acid concentration over the range of 10⁻⁶ to 10⁻³ m. The relative sensitivity of the leaves to different auxins was qualitatively similar to that observed in many straight-growth bioassays. Similar responses were obtained when auxin was applied by a carborundum wounding procedure. Potential applications of this auxin bioassay for investigations of the role of auxin in the normal plagiotropic growth behavior of leaf lamina and of the role of auxin in the initiation of various plant diseases are suggested.     

THE EFFECT OF SELENITE ON THE PHYSIOLOGICAL AND MORPHOLOGICAL PROPERTIES OF THE BLUE-GREEN ALGA PHORMIDIUM LURIDUM VAR. OLIVACEA1

Phormidium luridum cultures were treated with sodium selenite in concentrations ranging from 10^?6 M to 10^?2 M. In contrast to the increasing culture turbidity of control and 10^?6 M selenite cultures, the turbidity of the other selenite cultures declined in proportion to time and selenite concentration. Chlorophyll extraction revealed similar results. Photosynthetic activity was inhibited within 6 hr in all cultures except control and 10^?6 M selenite. Phormidium at concentrations greater than 10^?6 M selenite showed a gradual loss of the bright green color and turned semitransparent. Cell-associated granules of reduced selenium were observed at higher selenite concentrations. Other structural changes observed were the presence of intracellular and intercellular spaces, spheroplast formation, and gradual cell lysis. Protein analyses of total cell samples and supernatant fractions confirmed cellular breakdown of selenite-treated algal cells.     

THE EFFECT OF AUXINS ON PROTOPLASMIC STREAMING. III

1. A new method is described which gives a continuous record of the absolute rate of protoplasmic streaming in epidermal cells of the Avena coleoptile. 2. With this method a study was made of the influence of malate and iodoacetate on streaming velocity, in order to make correlations with the previously established effects of these substances on growth and respiration. 3. In the presence of optimum concentrations of indole-3-acetic acid in freshly cut sections, malate had no effect on streaming. In the presence of very low concentrations of the auxin, however, malate increased the range of response, so that the threshold of auxin sensitivity was lowered some ten times by the malate. Malate alone had no effect on streaming. 4. In coleoptile sections, soaked overnight in sugar solution or in water, the acceleration of streaming normally caused by auxin almost disappears, but the presence of malate causes large accelerations of streaming by the auxin. 5. Similarly, in sections from old coleoptiles, which no longer show acceleration of streaming by auxin, the acceleration is restored when malate is added together with the auxin. 6. Malate does not enter the cell as rapidly as does auxin, but easily detectable amounts penetrate within 30 minutes. 7. Iodoacetate in the concentration which inhibits growth (5 x 10^–5 M) completely inhibits the acceleration of streaming by auxin. In still lower concentrations iodoacetate slightly accelerates streaming. Higher concentrations, up to 2 x 10^–4 M, did not reduce the rate of streaming below that of controls without auxin. The effect of iodoacetate is therefore to inhibit the acceleration caused by auxin and not to affect the basal streaming rate. 8. It is concluded that, just as for growth and respiration, malate is necessary for the response to auxin shown by acceleration of streaming. This further strengthens the triple parallel between the effects of auxin on streaming, growth, and respiration, all of which are apparently mediated by the 4-carbon acid system.     

STUDIES ON CELL METABOLISM AND CELL DIVISION : III. OXYGEN CONSUMPTION AND CELL DIVISION OF FERTILIZED SEA URCHIN EGGS IN THE PRESENCE OF RESPIRATORY INHIBITORS

1. The effects of a number of respiratory inhibiting agents on the cell division of fertilized eggs of Arbacia punctulata have been determined. For eggs initially exposed to the reagents at 30 minutes after fertilization at 20°C., the levels of oxygen consumption prevailing in the minimum concentrations of reagents which produced complete cleavage block were (as percentages of the control): In 0.4 per cent O₂-99.6 per cent N₂, 32; in 0.7 per cent O₂-99.3 per cent CO, 32; in 1.6 x 10^–4 M potassium cyanide, 34; in 1 x 10^–3 M phenylurethane, 70; in 4 x 10^–3 M 5-isoamyl-5-ethyl barbituric acid, 20; in 3 x 10^–4 M iodoacetic acid, 53. 2. The carbon monoxide inhibition of oxygen consumption and cell division was reversed by light. The percentage inhibition of oxygen consumption by carbon monoxide in the dark is described by the usual mass action equation with K, the inhibition constant, equal to approximately 60, as compared to values of 5 to 10 for yeast and muscle. In 20 per cent O₂-80 per cent CO in the dark there was a slight stimulation of oxygen consumption, averaging 20 per cent. 3. Spectroscopic examination of fertilized and unfertilized Arbacia eggs reduced by hydrosulfite revealed no cytochrome bands. The thickness and density of the egg suspension was such as to indicate that, if cytochrome is present at all, the amount in Arbacia eggs is extremely small as compared to that in other tissues having a comparable rate of oxygen consumption. 4. Three reagents poisoning copper catalyses, potassium dithio-oxalate (10^–2 M), diphenylthiocarbazone (10^–4 M), and isonitrosoacetophenone (2 x 10^–3 M) produced no inhibition of division of fertilized Arbacia eggs. 5. These results indicate that the respiratory processes required to support division in the Arbacia egg may perhaps differ in certain essential steps from the principal respiratory processes in yeast and muscle.