Light Induced Concentration Changes of Adenosine-Triphosphate in Phycomyces Sporangiophores

The concentrations of extractable adenosine triphosphate (ATP) following the induction of positive light-growth responses in Phycomyces sporangiophores by blue light stimuli have been measured by means of the luciferin-luciferase assay. The ATP concentration in the light-sensitive growing zone increases 30 to 50% within 30 seconds after the start of a light stimulus and returns to the normal adapted level within 1 minute after stimulation. The ATP concentration is constant for any level of light adaptation and is uniform along the length of sporangiophores even though the light sensitivity is confined to a growing zone less than 5 mm long. These results suggest that one of the initial biochemical steps after a light stimulus is the production of extractable ATP.     

Tropic Responses of Phycomyces Sporangiophores to Gravitational and Centrifugal Stimuli

A low-speed centrifuge was used to study the tropic responses of Phycomyces sporangiophores in darkness to the stimulus of combined gravitational and centrifugal forces. If this stimulus is constant the response is a relatively slow tropic reaction, which persists for up to 12 hours. The response is accelerated by increasing the magnitude of the gravitational-centrifugal force. A wholly different tropic response, the transient response, is elicited by an abrupt change in the gravitational-centrifugal stimulus. The transient response has a duration of only about 6 min. but is characterized by a high bending speed (about 5°/min.). An analysis of the distribution of the transient response along the growing zone shows that the active phase of the response has a distribution similar to that of the light sensitivity for the light-growth and phototropic responses. Experiments in which sporangiophores are centrifuged in an inert dense fluid indicate that the sensory mechanism of the transient response is closely related to the physical deformation of the growing zone caused by the action of the gravitational-centrifugal force on the sporangiophore as a whole. However, the response to a steady gravitational-centrifugal force is most likely not connected with this deformation, but is probably triggered by the shifting of regions or particles of differing density relative to one another inside the cell.     

A Kinetic Model for Adaptation and the Light Responses of Phycomyces

A kinetic model is described consisting of two sequential first order processes connected by two parallel reaction pathways, one of which is light-catalyzed. A change in light flux changes the rate constant of the light-dependent process, whereupon the levels of two chemical intermediaries readjust. The model's output duplicates all the main features of the cell's light-growth and dark-growth responses except their latent periods. An asymmetric modification of the model reproduces the two types of phototropic inversion discovered by Reichardt and Varjú and by Dennison. Simple exponential equations describe these responses of the model, as well as the theoretical course of its light and dark adaptation. It is concluded that adaptation in Phycomyces consist in the photocatalytic adjustment of the level of a metabolic reservoir.     

The Light Growth Response of Phycomyces

With the help of an automated tracking system we have studied the characteristics of the transient light growth response of Phycomyces. The response shows a sharply defined latency. The Q₁₀ of the reciprocal latency is 2.4. Response patterns at different peaks of the action spectrum are the same. The gradual variation of response magnitude over a wide range of adapted intensifies parallels that of phototropism. The responses to saturating stimuli exhibit a strong oscillation with a constant period of 1.6 min and variable damping. The growth responses to sinusoidally varying light intensities show a system bandwidth of 2.5 x 10^-3 Hz. The linear dependence of phase shift on frequency is largely attributable to the latency observed with pulse stimuli. In the high intensity range a previously suspected increase of the steady-state growth rate with intensity has been confirmed. The light growth responses of mutants selected for diminished phototropism have been investigated. Many of these mutants have sizable but grossly distorted growth responses.     

Absence of Significant Light-Induced Changes in cAMP Levels in Sporangiophores of Phycomyces blakesleeanus

We were unable to find transient changes in the amount of cAMP or cGMP that had been proposed to mediate the light-growth response in sporangiophores of Phycomyces blakesleeanus.     

Role of Nitric Oxide Synthase in the Light-Induced Development of Sporangiophores in Phycomyces blakesleeanus

Blue light controls the development of sporangiophores in the zygomycete Phycomyces blakesleeanus Burgeff. Light represses the production of microsporangiophores and enhances the development of macrosporangiophores. Inhibition of the biosynthesis of tetrahydrobiopterin, a cofactor of NO synthase, inhibits this photomorphogenesis. Light induces production of citrulline from arginine in the mycelium and in sporangiophores. The citrulline-forming activity is dependent on NADPH, independent of calcium, and inhibited by NO synthase inhibitors. It is reduced in tetrahydrobiopterin-depleted mycelium. Light induces emission of NO from the developing fungus in the same order of magnitude as citrulline formation from arginine. The NO donor sodium nitroprusside can replace the light effect on sporangiophore development, and inhibitors of NO synthase repress it. We suggest that a fungal NO synthase is involved in sporangiophore development and propose its participation in light signaling.     

The Effects of Paraffin Oil on Phototropic and Geotropic Responses in Avena Coleoptiles

Experiments are described which were designed to test the significanceof the coleoptile tip as the site of reception of light stimulusleading to negative photo-tropic response under paraffin oil.The results show clearly that the tip is of paramount importancein this respect. Further experiments in which the coleoptiletips were bisected in a plane at right angles to the light rayslend support to the hypothesis that negative phototropism underoil is related tolateral transport of material in the tip. Lastly,experiments which show that the geotropic response of coleoptilesis not reversed by immersion in oil are described. These findingsare discussed in relation to certain hypotheses concerning themechanism of negative phototropism under oil.     

Function of Light in the Light-induced Geotropic Response in Zea Roots

The light doses just above the threshold energy value for inducing the geotropic responsiveness in the roots of Zea mays L., cv. Golden Cross Bantam 70, caused a drastic rise in the NADPH level and a drop in the NADP level in 2-millimeter root tips. Some reducing agents lowered the threshold energy value up to about one-third of the control. From these results, we deduce that light may exert two functions in the geotropic response of Zea primary roots, one being the photochemical transformation of a photoreceptor and the other being the induction of a reduction state in the tissue.     

The Lens Effect and Phototropism of Phycomyces

Normally, the dioptrics in air of the cylindrical sporangiophore of Phycomyces blakesleeanus confer on the distal side a focusing advantage of about 30 per cent for unilateral stimuli of parallel light. This advantage can be nullified or reversed to produce negative curvatures by means of diverging light stimuli. A thin cylindrical glass lens was positioned 0.15 mm from the light-adapted growing zone with its long axis parallel to the long axis of the sporangiophore. A 3 minute blue stimulus was given and the lens removed. Reproducible negative curvatures were observed with a maximum of 13 degrees occurring within 8 minutes after the beginning of the stimulus. Experiments in air were done in a water-saturated atmosphere to minimize avoidance responses due to the proximity of the lens. The data support Buder's conclusion that the focusing advantage is the principal mechanism which produces the response differential necessary for phototropism. When the lens advantage is small, the attenuation becomes important in determining the direction of the response. Data obtained from sporangiophores immersed in inert liquids indicate that the attenuation is about 14 per cent. Therefore, whenever the focusing advantage is less than 14 per cent, negative curvatures are produced by unilateral stimuli.     

The Effects of Red, Far-red, and Blue Light on the Geotropic Response of Coleoptiles of Zea mays

The effects of red, far-red, and blue light on the geotropicresponse of excised coleoptiles of Zea mays have been investigated.Seedlings were grown in darkness for 5 or 6 days, exposed tovarious light treatments, and then returned to darkness fordetermination of the geotropic response. The rate of response of the coleoptiles is decreased after theyhave been exposed to red light (620–700 mµ, 560ergs cm^–2sec^–1 for the 24 hrs, but not for the 4hrs, preceding stimulation by gravity. Furthermore, their rateof response is greatly reduced if they are exposed to red lightfor 10 min and then returned to darkness for 20 hrs before geotropicstimulation. At 25° C an interval of 6 to 8 hrs elapses between a 10-minexposure to red light and the first detectable decrease in thegeotropic response of the coleoptile. This interval can be lengthenedby exposing the seedlings to low temperatures (0° to 2°C) after the light treatment but cannot be greatly shortenedby increasing the duration of exposure to red light. Using a standard procedure of exposing 5-day-old etiolated seedlingsto light for various times, replacing them in darkness for 20hrs and then determining the response of the coleoptiles to4 hrs geotropic stimulation, it has been found that: (a) Exposureto red light for 15 sec significantly decreases the geotropiccurvature of the coleoptiles and that further reduction occurson increasing the length of the light treatment to 2 and 5 min.(b) Far-red light has no effect on the geotropic response ofthe coleoptiles but it can completely reverse the effect ofred light. After repeated alternate exposure to red and far-redlight the geotropic response of the coleoptile is determinedby the nature of the last exposure, (c) Complete reversal ofthe effect of red light by far-red radiation only occurs whenexposure to far-red follows immediately after exposure to red.The reversing effect of far-red radiation is reduced if a periodof darkness intervenes between the red and far-red light treatments,and is lost after a dark interval of approximately 2 hrs. The effect of red light on the rate of geotropic response ofthe coleoptiles is independent of their age and length at thetime of excision. Blue light acts in a similar way to red light, but the seedlingsare less sensitive to blue than to red light. Coleoptiles grown throughout in a mixture of continuous, weak,red, and far-red light have a lower rate of geotropic responsethan etiolated coleoptiles.     

Growth and Geotropic Responses of Avena Coleoptiles to 4-Amino-3,5,6-Trichloropicolinic Acid

The elongation and geotropic responses of coleoptile sections of Avena sativa L. to various concentrations of 4-amino-3,5,6-trichloropicolinic acid (Tordon) proved to be qualitatively similar to those previously reported for 2,3,6-trichlorobenzoic acid (TCBA). Tordon stimulated growth in a range of concentrations from 1 × 10^?6 to 1 × 10^?4M but higher concentrations were inhibitory. Geotropic curvature was extensively depressed by 1 × 10^?5 and 1 × 10^?4M Tordon, concentrations which accelerated elongation. A similar differential effect has been reported for TCBA and other auxins. Several other picolinic acids and related compounds were tested, but only very slight responses were noted.     

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.