White light requirements for stimulation of plant growth by malformin

Over a 3-day period, the minimum white fluorescent light intensity required for malformin-induced growth stimulation of etiolated and green cuttings of Phaseolus aureus was approximately 2.6 × 10³ and 0.4 × 10³ ergs/cm² · sec, respectively. High light intensities were unable to inhibit the ability of malformin to stimulate growth. Over 3 days, the minimum photoperiod for malformin-induced growth stimulation using etiolated and green cuttings and a light intensity of 13.5 × 10³ ergs/cm² · sec was 4 hours and 1 hour, respectively. Malformin must be present in the area of growth stimulation during the time of light treatment. Those changes induced by light and required for malformin-induced growth stimulation were estimated to undergo almost complete decay within 1 hour in the dark. By manipulating the experimental technique, it was possible to stimulate the growth of green cuttings with malformin with a 10-min light treatment (13.5 × 10³ ergs/cm² · sec). Although low light intensities and short photoperiods did not allow growth stimulation by malformin using etiolated cuttings, they prevented or alleviated growth inhibition induced by malformin in the dark.     

An investigation of the effects of light on basidiocarp formation of Coprinus domesticus

Coprinus domesticus, grown on a synthetic agar medium, failed to produce primordia and basidiocarps unless exposed to light. Lightdark cycles are not required for maturation of basidiocarps. Short exposure to white light induced primordia, but a longer exposure was necessary for primordia to develop into basidiocarps. The length of exposure to light was related inversely to the length the stipe finally attained. Young basidiocarps were phototropic, growing towards the light. The mycelium of cultures were dark brown following exposure to white and blue light, but the mycelium was light yellow in cultures grown in darkness. The blue end of the visible spectrum at intensities ranging from 1.5–3 × 10⁴ ergs/cm²/sec induced mature basidiocarps, whereas green, red and far red failed to induce basidiocarps and primordia.Department of Biology contribution no. 90     

THE EFFECT OF WAVELENGTH AND INTENSITY OF LIGHT ON THE PROPORTION OF PIGMENTS IN PORPHYRIDIUM CRUENTUM

Brody , Marcia , and Robert Emerson . (U. Illinois, Urbana.) The effect of wavelength and intensity of light on the proportion of pigments in Porphyridium cruentum. Amer. Jour. Bot. 46(6): 433–440. Illus. 1959.—We specify a medium for Porphyridium cruentum made from distilled water and inorganic salts, without addition of earth extract, sea water or other supplements of uncertain composition, which sustains excellent growth through successive transfers. By control of intensity and wavelength of light used for growing the cultures, we have found it possible to vary the concentrations of chlorophyll and phycoerythrin, and also the proportions of these 2 pigments, over a considerable range. The changes in proportion of red and green pigments, in response to culturing in green and in blue light, are opposite to the changes that would be predicted from Engelmann's theory of complementary chromatic adaptation, when the intensities of the blue and green light are of the order of 10⁴ ergs/cm²/sec. (or greater). However, at about 1/100 of this intensity, the changes are in the direction of complementary chromatic adaptation.     

Development of Enzymes Involved in Photosynthetic Carbon Assimilation in Greening Seedlings of Maize (Zea mays)

Upon illumination of dark-grown maize seedlings (5 days old) with incandescent light, there occurred a nearly simultaneous increase, after a certain lag period, in the activities of enzymes engaged in the C₄ pathway and the Calvin-Benson cycle. The light-induced biosynthesis of chlorophyll (a and b) precedes the increase in enzyme activities and proceeds without lag phase. A diphasic feature in the elevation of enzyme activities as a function of the intensities of light provided was observed; the increase in enzyme activities was enhanced by light intensities greater than 10³ ergs per square centimeter per second in comparison with light of lower intensities. Under light intensities greater than 10³ ergs per square centimeter per second, the simultaneous addition of levulinic acid, which inhibited chlorophyll formation, markedly reduced the increase of enzyme activities. However, neither the diphasic light effect nor the inhibitory effect of levulinic acid was observed with ribulose-1,5-bisphosphate carboxylase. The enzyme activities in the dark-grown maize seedlings were enhanced by a brief irradiation with the red light and the red light effect was reversed by the following far red light treatment. The red light-induced increase in the enzyme activities did not accompany chlorophyll synthesis, and was completely inhibited by cycloheximide, indicating that enzyme synthesis rather than activation might be involved. Light may play a dual role in enzyme induction; one is as an energy source through the photosystems at high intensities and the other is presumably as a signal mediated by phytochrome at low intensities.     

Characteristics of mixotrophic growth of Synechocystis sp. in an enclosed photobioreactor

Synechocystis sp. PCC 6803 was grown in a 2.5 l enclosed photobioreactor on medium with or without glucose. The incident light intensities ranged from 1.5 klux to 7 klux. The highest average specific growth rates of mixotrophic culture and photoautotrophic culture were, respectively, 1.3 h^–1 at a light intensity of 7 klux on 3.2 g l^–1 glucose and 0.3 h^–1 at both light intensities of 5 klux and 7 klux. The highest cell density 2.5 g l ^–1 was obtained at both of light intensities 5 klux and 7 klux on 3.2 g glucose l^–1. Glucose consumption decreased with decreasing light intensity. The energy yields of mixotrophic cultures were 4 to 6 times higher than that of photoautotrophic cultures. Light favored mixotrophic growth of Synechocystis sp. PCC 6803, especially at higher light intensities (5–7 klux).     

COUNTERACTING THE RETARDING AND INHIBITORY EFFECTS OF STRONG ULTRAVIOLET ON FUCUS EGGS BY WHITE LIGHT

1. Strong dosages (20,000–50,000 ergs per mm.²) of ultraviolet light, predominantly of the wave-length 2537 Å, greatly retard and inhibit the development of rhizoids in Fucus eggs irradiated at about 8 hours after fertilization. 2. If white light shines on the eggs after the irradiation by ultraviolet is terminated, the white light causes a considerable degree of recovery from the retarding and inhibiting effects. 3. If strong white light shines on the eggs during the ultraviolet irradiation, its effect is even more marked in protecting the cells from the damaging effects of the ultraviolet.     

Nitrogen Fixation by Thermophilic Blue-Green Algae (Cyanobacteria): Temperature Characteristics and Potential Use in Biophotolysis

Thermophilic, nitrogen-fixing, blue-green algae (cyanobacteria) were investigated for use in biophotolysis. Three strains of Mastigocladus laminosus were tested and were found to be equally effective in biophotolysis as judged by nitrogenase activity. The alga, M. laminosus NZ-86-m, which was chosen for further study, grew well in the temperature range from 35 to 50°C, with optimum growth at 45°C, at which temperature acetylene reduction activity was also greatest. The maximum tolerable temperature was 55°C. Acetylene reduction activity was saturated at a light intensity of 1 × 10⁴ ergs cm⁻² s⁻¹. Atmospheric oxygen tension was found to be slightly inhibitory to acetylene reduction of both slowly growing and exponentially growing cultures. Nonsterile continuous cultures, which were conducted to test problems of culture maintenance, could be operated for 2 months without any significant decrease in nitrogenase activity or contamination by other algae. Nitrogen-starved cultures of M. laminosus NZ-86-m produced hydrogen at comparable rates to Anabaena cylindrica. The conversion efficiency of light to hydrogen energy at maximum rates of hydrogen production was 2.7%.     

Effect of light intensity on macromolecular synthesis in cyanobacteria

The light-dependent incorporation of NaH¹⁴CO₃ into low molecular weight compounds, polysaccharide, or protein was determined in cultures of the cyanobacteriumMerismopedia tenuissima incubated at a series of light intensities. There was an inverse relationship between incorporation into polysaccharide and protein. At light intensities of 90 E/m²/sec or above, relative incorporation of radioisotope into polysaccharide was greatest and relative incorporation into protein was lowest. Optimal relative protein accumulation occurred in samples incubated at 20 E/m²/sec. A broader optimum of light intensity for maximal protein accumulation was found if ammonia rather than nitrate was the nitrogen source. Physiological adaptation of cultures to growth at a particular light intensity did not alter the pattern of macromolecular incorporation when those cultures were tested over the series of light intensities. The response of cultures ofOscillatoria rubescens to light intensity was similar to that ofM. tenuissima, although incorporation into low molecular weight compounds was significantly greater.The effect of light intensity on macromolecular synthesis in a natural population ofOscillatoria rubescens was also determined. A pattern similar to that observed in batch cultures ofO. rubescens was occasionally found, but in other experiments there was no increase in relative protein incorporation when light intensity was decreased.     

THE ABSORPTION OF OXYGEN FROM LIQUID CULTURES OF BLUE-GREEN ALGAE BY ALKALINE PYROGALLOL

SUMMARY

A mixture of pyrogallol with sodium hydroxide and sodium carbonate (2:1:1) in a separate container absorbed up to 75% of the oxygen in a bicarbonate-carbonate buffered medium with more or less no change in the pH of the medium. A system in which two test tubes were added to the culture vessel, one containing saturated sodium bicarbonate and the other pyrogallol plus sodium hydroxide gave similar results, but were not investigated further because of difficulties in handling. Optimal conditions for absorption of oxygen from 100 ml of the growth medium was 1 mMole pyrogallol +0,5 mMole sodium hydroxide + 0,5 mMole sodium carbonate. The addition of this pyrogallol to alkali ratio to cultures of two Microcystis and one Synechococcus isolate in rubber stoppered flasks gave stoichiometric increases in yield which was not due to carbonate enrichment but to a lowering of oxygen tension. The data may mean that even under relatively low light intensities (1 - 4 × 10³ ergs cm^?2 sec^?1) photooxidation occurs.     

Responses of Heterotrophic Cultures of Chlorella vulgaris Beyerinck to Darkness and Light. II. Action Spectrum for and Mechanism of the Light Requirement for Heterotrophic Growth

Chlorella vulgaris Beyerinck (Emerson's strain), fails to grow in the dark even when sugars are provided. This phenomenon was clearly demonstrated in the alga, C. vulgaris, for which the growth rate in darkness on a glucose medium remained constant for 2 days and then declined to approach zero. Pigment concentrations also declined in darkness. Changes in flow rate of 1% CO₂-in-air from zero to 7 ml per minute caused a progressive increase in the dark growth rate over a 5-day period, but did not maintain growth in the dark. Rates above 7 ml per minute produced no changes in growth rates.

White light intensities below the compensation point of the alga maintained heterotrophic growth. The saturation value for this response was 0.8 μw/cm². White light also initiated growth in nongrowing cultures transferred from darkness to light.

The action spectrum for heterotrophic growth indicated a porphyrin as the active pigment. Light in the 425 mμ region was 4 times as effective as white light in stimulating heterotrophic growth. A secondary peak of growth stimulation occurred in the 575 mμ region.

The respiration of glucose by the alga was stimulated by low intensities of white light. This response was not immediate, but was clearly present after the third day of incubation.

Malonate and cyanide were inhibitory to growth of C. vulgaris on inorganic medium or glucose medium under 300 ft-c of white light. These data suggested that succinic dehydrogenase and cytochrome oxidase systems were present.

Substances inhibitory to growth were excreted into the medium under dark-growth conditions, and 2 of these substances were indentified as formic and acetic acids.

The evidence suggested that respiration of glucose cannot proceed for an extended period of time in darkness. The reason for this is postulated to be the lack of a cytochrome or a cytochrome precursor.

     

Photocontrol of the germination of onoclea spores: I. Action spectrum

Light stimulates the germination of spores of the fern Onoclea sensibilis L. At high dosages, broad band red, far red, and blue light promote maximal germination. Maximal sensitivity to these spectral regions is attained from 6 to 48 hours of dark presoaking, and all induced rapid germination after a lag of 30 to 36 hours. Maximal germination is attained approximately 70 hours after irradiation. Dose response curves suggest log linearity. The action spectrum to cause 50% germination shows that spores are most sensitive to irradiation in the red region (620-680 nm) with an incident energy less than 1000 ergs cm⁻²; sensitivity decreases towards both shorter and longer wavelengths. Although the action spectrum is suggestive of phytochrome involvement, photoreversibility of germination between red and far red light has not been demonstrated with Onoclea spores. An absorption spectrum of the intact spores reveals the presence of chlorophylls and carotenoids. Since the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea does not inhibit germination, it is concluded that photosynthesis does not play a role in the germination process.     

Influence of Light Intensity on Sorbitol Metabolism, Growth, and Chlorophyll Content of Malus Tissue Cultures

Apple stem callus cultures of Malus pumila Mill., cultivars,McIntosh and Cortland, and of Malus robusta Rehd. No. 5 weregrown under light intensities of o, 850, 3350, and 7800 lx ona medium with sorbitol as the carbon source. Cortland callusgrew better in dark whereas growth of Robusta was not influencedby light. Light did not influence growth of McIntosh callusbetween o and 3350 lx, although slight stimulation in growthoccurred at 7800 lx. The chlorophyll content of the culturesincreased with light intensity, although that of Cortland wasvery low at all intensities. Tissue analysis of cultures grownin light at 7800 lx and in dark indicated that the growth ofthese callus tissues as influenced by light was directly relatedto their sorbitol and carbohydrate content. McIntosh and Cortlandshowed opposite trends in the levels of starch, sorbitol, sucrose,glucose, and total carbohydrate in light and dark. Similar tothe growth response of Robusta, the levels of these carbohydratesin this callus were apparently not influenced by light. Sucrose,accumulating in concentrations of between 45 and 62 per centof total carbohydrate, tended to be the predominant carbohydrateconstituent in both light- and dark-grown cultures.     

Increased photoreversal of ultraviolet injury by flashing light

1. Photoreversal of ultraviolet (UV) injury was studied in the ciliate protozoan Tetrahymena pyriformis (geleii) strain W, cultured in the absence of other living organisms. The division pattern of progeny of single animals was followed in hanging drop preparations. 2. A sublethal dose of 450 ergs/mm.² of monochromatic UV of wave length 2654 A produces a lag before the first division followed by a period of cessation of fission after the second division. This cessation sometimes lasts as long as 6 weeks, during which time the animals become smaller and rounder and more opaque. Organisms about to resume division increase in size and transparency; after a few divisions the animals regain their normal division rate. 3. The effect of UV ranging in intensity from 5 to 15 ergs/mm.²/sec. was found to obey the reciprocity law quite well for the UV effect on the division pattern of T. pyriformis. However, the same dose at lower and at higher intensities was less effective. 4. The effect of a dose of UV delivered at high intensity (19 ergs/mm.²/sec.) could be increased by flashing the light, indicating that the system became saturated in the continuous light. 5. A photoreversing dose of monochromatic blue light of wave length 4350 A was found to be more effective when delivered as continuous light at a low intensity, or as intermittent light at a high intensity, rather than as continuous light at the high intensity—indicating that a dark mechanism participates in photoreversal. 6. The time for the dark reaction was determined to be of the order of a few hundredths of a second in experiments in which different lengths of dark period were used while maintaining a constant light period of 0.0025 second. 7. For Colpidium colpoda the efficiency of a given dose of photoreversing light was increased by flashing the light. 8. The present experiments are interpreted in terms of data available in the literature.     

Marine Plankton Algae Grown with Light-Dark Cycles.

The diatoms Ditylum brightwellii and Nitzschia turgidula were grown in semi-continuous culture under various combinations of light intensity, temperature and daylength (photoperiod). Growth was strongly limited by light intensities below 0.03 cal/em². min in both species. Above this intensity, light saturation of growth was rapidly approached in Nitzschia but only gradually so in Ditylum. The growth rate in continuous light was never significantly higher than with 16 hours of light plus 8 hours of dark. In Ditylum, continuous light above 0.03 cal/cm². min caused a strong inhibition of growth at all temperatures. The chlorophyll concentration in the cells was greater the shorter the photopceriod. In cultures synchronised by different combinations of light intensity and photoperiod, cell division generally took place in the light. Synchrony was best under short photoperiods of bright light. Time courses are shown for chlorophyll synthesis and photosynthesis in synchronised cultures.     

Efficiency and pattern of UV pulse laser-induced RNA-RNA cross-linking in the ribosome

Escherichia coli ribosomes were irradiated with a KrF excimer laser (248 nm, 22 ns pulse) with incident pulse energies in the range of 10–40 mJ for a 1 cm² area, corresponding to fluences of 4.5 to 18 × 10⁹ W m^–2, to determine strand breakage yields and the frequency and pattern of RNA–RNA cross- linking in the 16S rRNA. Samples were irradiated in a cuvette with one laser pulse or in a flow cell with an average of 4.6 pulses per sample. The yield of strand breaks per photon was intensity dependent, with values of 0.7 to 1.3 × 10^–3 over the incident intensity range studied. The yield for RNA–RNA cross-linking was 3 × 10^–4 cross-links/photon at the intensity of 4.5 × 10⁹ W m^–2, an ~4-fold higher yield per photon than obtained with a transilluminator. The cross-link yield/photon decreased at higher light intensities, probably due to intensity-dependent photoreversal. The pattern of cross-linking was similar to that observed with low intensity irradiation but with four additional long-range cross-links not previously seen in E.coli ribosomes. Cross- linking frequencies obtained with one laser pulse are more correlated to internucleotide distances than are frequencies obtained with transilluminator irradiation.     

Production of <Emphasis Type="Italic">Cocconeis neothumensis</Emphasis> (Bacillariophyceae) biomass in batch cultures and bioreactors for biotechnological applications: light and nutrient requirements

Cocconeis neothumensis, a benthic diatom living as epiphyte on Posidonia oceanica leaves, was hypothesised to synthesize secondary metabolites inducing apoptosis in the androgenic gland of the protandric shrimp Hippolyte inermis. The optimization of C. neothumensis cultures is a primary aim in order to identify and characterise the active compounds produced by this diatom. Light intensity and concentration of nutrients, such as silicates and selenium, are of major importance in determining the growth rate of diatoms and the maximum produced biomass. Thus, we evaluated the growth performances of C. neothumensis cultures at different light intensities, corresponding to those recorded in Posidonia oceanica meadows in April, when the induction of sexual reversal in Hippolyte inermis occurs; the growth performances were also determined in relation to different selenium and silicate availability in the culture medium and the possible interactions between these two micronutrients were evaluated. Two methods of cultivations were compared: batch cultures in Petri dishes and cultures in a bioreactor with a continuous medium flow in the system. C. neothumensis showed a faster growth at low light intensities, although with a good acclimation capacity in the range from 60 to 140 μmol photons m⁻² s⁻¹. The presence of selenium in the medium improved both the exponential growth rate and the maximum cell density. The same results were evidenced for a silicate concentration double that in Guillard f/2 medium. On the other hand, a simultaneous increase of silicate concentration and the presence of selenium led to a loss of the positive effects detected with single nutrients. This result suggests either a possible antagonism in selenium and silicate uptake or a negative interaction between these two micronutrients in Cocconeis. The yields of the two tested cultivation methods calculated in terms of diethyl ether extract dry weight per unit of substrate area were highest for the bioreactor.     

GROWTH RATES OF PSEUDOPEDINELLA PYRIFORME (CHRYSOPHYCEAE) IN RESPONSE TO 75 COMBINATIONS OF LIGHT,TEMPERATURE AND SALINITY1

Clonal cultures were established of single cells of Pseudopedinella pyriforme Carter isolated from the Chesapeake Bay. Axenic cultures were incubated at 5, 10 and 15 C; in 2 1/2, 5, 7 1/2, 10 and 15‰; and at 143, 285, 428, 571 and 714 μW/cm² white light. Maintainable growth rates were calculated from absorbance readings taken for 3 to 5 days from acclimated cultures. The maximum number of doublings per day was 0.9 at 2 1/2‰ salinity, 571 μW/ cm² and 15 C. A salinity of 5 ‰ produced the fastest growth at most light intensities up to 428μW/cm² and temperatures of 5, 10 and 15 C.     

Toxin Production by Microcystis aeruginosa as a Function of Light in Continuous Cultures and Its Ecological Significance

The effects of light intensity and light quality on toxin production by Microcystis aeruginosa were examined in continuous cultures. Light intensity had a pronounced effect on toxicity and the toxin production rate. Toxicity and the toxin production rate increased with light intensity up to an intensity of about 40 microeinsteins m^-2 s^-1 and decreased at higher light intensities, while the ratio of toxin to protein was constant at intensities of more than 40 microeinsteins m^-2 s^-1. Light quality had only slight effects on toxicity. The results of our laboratory experiments were supported by the results of field work in which we examined toxin production at different depths in a lake. Our observations explain the mixed pattern of high and low toxicity found in a surface bloom of M. aeruginosa. Our findings also indicate that production of the peptide toxin can be uncoupled from general protein synthesis.     

Interplay between the Reactions to Light and to Gravity in Phycomyces

Sporangiophores of Phycomyces do not grow directly towards a horizontal beam of light, but equilibrate at an angle of about 30° above the horizontal. After describing several related observations, this paper suggests that the dioptric properties of an obliquely illuminated cylindrical lens, illustrated by a dummy cell, as well as a negative geotropic response, play major roles in determining the direction of growth. The shift of the equilibrium direction of growth towards the vertical, or a purely geotropic response, over a tenfold range of very low intensities (around 10⁶ quanta/cm² sec., or 10^-13watt/cm²) has been studied, and an action spectrum made, measuring the quantum fluxes producing a standard intermediate equilibrium direction of growth at different wavelengths. This may differ from the action spectra at higher intensities in lacking conspicuous maxima from 370 to 490 mµ. However, in the ultraviolet it parallels the other spectra, although without showing the much higher quantum efficiency of ultraviolet relative to visible light previously noted. Possible interpretations are discussed.     

Effect of irradiation intensity on cell growth and kalata B1 accumulation in <Emphasis Type="Italic">Oldenlandia affinis</Emphasis> cultures

Light irradiation had remarkable effects on callus growth of Oldenlandia affinis with an optimum intensity of 35 μmol m⁻² s⁻¹. Biosynthesis of kalata B1, the main cyclic peptide in O. affinis, was induced and triggered with rising irradiation intensities. The highest concentration of kalata B1, 0.49 mg g⁻¹ DW characterised by the maximum productivity of 3.88 μg per litre and day was analysed at 120 μmol m⁻² s⁻¹, although callus growth was repressed. The light saturation point was established to be 35 μmol m⁻² s⁻¹, where kalata B1 productivity was in a similar order (3.41 μg per day) due to the higher growth index. O. affinis suspension cultures were shown to accumulate comparable specific kalata B1 concentrations in a delayed growth associated production pattern. These were dependent on irradiation intensity (0.16 mg g⁻¹ at 2 μmol m⁻² s⁻¹; 0.28 mg g⁻¹ at 35 μmol m⁻² s⁻¹). The batch cultivation process resulted in a maximum productivity of 27.30 μg per litre and day with culture doubling times of 1.16 d⁻¹. Submers operation represented a 8-fold product enhancement compared to callus cultivation.