Light Dependence of Chloroplast Replication and Starch Metabolism in the Moss Polytrichum commune

Spores of Polytrichum conwtuine were grown on a mineral salt solution with or without sucrose and exposed to continuous white light, continuous darkness, red light and/or far-red light. With sucrose, germination and filament growth occurred in all conditions, Without sucrose, germination and filament growth occurred only in light. Two phytochrome mediated responses of the chloroplasts were demonstrated. Chloroplast replication occurred in continuous white light and red light of 15 min/6 hours. In continuous darkness and in far red light of 15 min/6 hours, the size of the chloroplasts increased; but no replication occurred. Both the chloroplast replication and chloroplast size were red, far-red light reversible. When changed from one continuous light environment to another, a lag period occurred before the chloroplasts responded to the new environment. Electron micrographs of sections and in vivo staining of the chloroplasts with iodine solution demonstrated that the change in size of the chloroplasts was at least partially due to the synthesis and degradation of starch.     

Light- and sucrose-dependent gene expression in photomixotrophic cell suspension cultures and protoplasts of rape (Brassica napus L.)

Light-dependent gene expression was analysed in photomixotrophic cell suspension cultures of rape (Brassica napus L.) growing in media containing either 2.0% or 0.6% sucrose. During growth in darkness phytochrome type I and NADPH-protochlorophyllide oxidoreductase (Pchlide reductase) accumulated in both cell culture lines to a similar extent. Illumination with continuous white, blue or red light, but not with far-red light, resulted in disappearance of both chromoproteins within 24 h in both cell cultures. Further analysis showed that the phytochrome system of rape cell cultures reacts in a similar way to that of re-etiolated dicotyledonous plants, showing rapid P_fr destruction and rapid P_fr dark reversion. In contrast, the light-dependent expression of genes encoding the major chlorophyll a- and b-binding protein (CAB) and the re-accumulation of chlorophyll were found to be strongly dependent on sucrose concentration in culture media. Whereas cells grown in darkness in medium containing 2.0% sucrose showed, after exposure to continuous white light, a very weak re-induction of CAB mRNA, CAB protein and chlorophyll accumulation, the cells in medium containing 0.6% sucrose reacted very strongly. It was also possible to demonstrate that phytochrome (by high irradiance response, HIR, and by low fluence response, LF) and the blue/UV-A receptor are involved in the light-dependent gene expression of CAB. Similar to complete cells, protoplasts derived from the two different cell cultures showed an almost identical sucrose concentration-dependent and light-quality-dependent regulation of CAB mRNA accumulation. As the dark-grown photomixotrophic cells and protoplasts reflect some typical photoregulatory characteristics known from dark-grown plants it is supposed that this system will be an excellent tool for studying biochemical and molecular biological aspects of light-dependent signal transduction in cells of higher plants.     

Effects of Colchicine and Light on Cell Form in Fern Gametophytes. Implications for a Mechanism of Light-induced Cell Elongation

Spores of the fern, Onoclea sensihilis L., suffer a disruption of normal development when they are cultured on media containing colchicine. Cell division is inhibited, and the spores develop into giant spherical cells under continuous white fluorescent light. In darkness only slight cell expansion occurs. Spherical cell expansion in the light requires continuous irradiation. Photosynthesis does not seem to be involved, since variations in light intensity do not affect the final cell diameter; the addition of sucrose to the medium does not permit cell expansion in darkness; and the inhibitor DCMU does not block the light-induced cell expansion. Continuous irradiation of colchicine-treated spores with blue, red or far-red light produces different patterns of cell expansion. Blue light permits spherical growth, similar to that found under white light, whereas red and far-red light promote the reestablishment of polarized filamentous growth. Although ethylene is unable to induce polarized cell expansion in colchicine-treated spores in darkness or white and blue light, it enhances filamentous growth which already is established by red or far-red irradiation. Both red and far-red light increase the elongation of normal filaments (untreated with colchicine) above that of dark-grown plants, but under all 3 conditions the rates of volume growth are identical. Light, however, does cause a decrease in the cell diameters of irradiated filaments. These data are used to construct an hypothesis to explain the promotion of cell elongation in fern protonemata by red and far-red light. The model proposes light-mediated changes in microtubular orientation and cell wall structure which lead to restriction of lateral cell expansion and enhanced elongation growth.     

INTERACTION OF LIGHT QUALITY AND NUCLEASES IN THE GROWTH OF THE GAMETOPHYTES OF ASPLENIUM NIDUS

Appropriate concentrations of ribonuclease A and B selectively inhibited initiation of two-dimensional morphology in the gametophytes of Asplenium nidus, grown under a photoperiod of 5½ hr white light. Filamentous growth was promoted in such sporelings, the individual cells of which were significantly longer than corresponding cells of the control. Higher concentrations of enzymes were required to inhibit two-dimensional growth in the gametophytes grown in blue light. Concentrations of ribonuclease A or B which inhibited two-dimensional growth in white light promoted growth in length of the protonema in red light. Growth modifications in the sporelings induced by deoxyribonuclease in different light conditions were similar to those induced by the ribonucleases. The results lend further support to the postulated role of RNA in the regulation of two-dimensional growth in fern gametophytes.     

Light-susceptibility of camptothecin production from<Emphasis Type="Italic">in vitro</Emphasis> cultures of<Emphasis Type="Italic">Camptotheca acuminata</Emphasis> Decne

Production of camptothecin (CPT) from callus cultures ofCamptotheca acuminata Decne was affected by light and culture conditions. Among the culture media tested, modified B5 medium containing 3% (w/v) sucrose, 2 mg/L 2,4-D, 2 times of MS medium vitamins, 500 mg/L casein hydrolysate, 250 mg/L myo-inositol, 0.05% (w/v) activated charcoal, and 0.15% (w/v) gelite was used for callus induction. The highest cell growth and CPT production were obtained in dark and green light condition, respectively. Photoperiod has no effect on cell growth and CPT production. Both cell growth and CPT production were also influenced by combination ratio of red and blue light. Cell growth and CPT production were the highest in the ratio of red and blue light 90∶10.     

Der Einfluss hellroter und dunkelroter Strahlung auf die geotropische Reaktion der Keimlinge vonSinapis alba L

Summary Our experiments indicate that the negative geotropic reactivity of dark-grown mustard seedlings (Sinapis alba L.) is greatly increased when the seedlings are irradiated with visible light. Obviously in irradiated seedlings the geotropic reaction is more rapid and intense. The results of this paper point out that light controls the geotropic behaviour in exactly the same way as morphogenesis. Apparently the increased geotropic reactivity of the irradiated seedlings is only one manifestation of the basic metabolic changes which occur in the dark-grown seedling after the absorption of light and which finally lead to photomorphogenesis.In earlier papers we have been able to show that photomorphogenesis in these mustard seedlings is controlled by two basic photoreactive systems: the reversible red far — red system and the so called blue far — red system. The same, probably, is true for the control of the geotropic reactivity under our standard experimental conditions.

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THE RELATIONSHIP BETWEEN THE PROMOTION OF ELONGATION OF FERN PROTONEMATA BY LIGHT AND GROWTH SUBSTANCES

Germinating spores of the fern Onoclea sensibilis L. were grown in darkness, so that they developed as filaments (protonemata). Brief daily exposure of the filaments to red, far-red or blue light increased the rate of filament elongation. Filament elongation was also promoted by indoleacetic acid. When filament elongation was promoted with both indoleacetic acid and exposure to light, the growth promotions caused by red and far-red light were additive to auxin-induced growth. Blue light promoted elongation only at sub-optimal concentrations of auxin. Elongation induced by guanine was additive to red- and far-red-induced elongation. Gibberellic acid had no effect on elongation under any condition. Blue-light-induced elongation resembled auxin-induced elongation in its requirement for exogenous sucrose and sensitivity to inhibition by parachlorophenoxyisobutyric acid. Red and far-red light were active regardless of the presence or absence of sucrose and promoted elongation at a concentration of parachlorophenoxyisobutyric acid which completely inhibited blue-light-induced elongation.     

Growth, greening, and phytochrome in etiolated spirodela (lemnaceae)

Two species of Spirodela were grown aseptically in a simple mineral medium containing sucrose. Weak red light (15 erg cm⁻² sec⁻¹) enhanced dark growth of S. oligorrhiza, whereas weak far red light (15 erg cm⁻² sec⁻¹) when given after the red light reduced this effect.     

The effect of light quality on the growth and development of in vitro cultured Doritaenopsis plants

The influence of light quality on growth and development of in vitro grown Doritaenopsis hort. (Orchidaceae) plants was investigated. Growth parameters like leaf and root fresh/dry mass and leaf area were highest with plants grown under red plus blue light emitting diodes (LEDs). Leaf length was greater with the plants grown under red LED. Carbohydrate (starch, sucrose, glucose and fructose) and leaf pigment (chlorophylls and carotenoids) biosynthesis of the plants was significantly increased in plants grown under red plus blue LEDs compared to red or blue LED and fluorescent light treatments. This study suggests that the production of quality Doritaenopsis plants is possible by culturing the plants in vitro under a mixture of blue plus red light sources.     

Effects of Light on the Growth and Development of the Liverwort, Sphaerocarpos donnellii Aust

Fragments of thalli of the liverwort, Sphaerocarpos donnellii Aust., inoculated into liquid medium containing sucrose and mineral salts, attain a much greater dry weight after 9 days growth in continuous white light than in darkness. Light causes this difference by increasing the rate of growth of the plants. This growth response is mediated by the pigment systems of photosynthesis and phytochrome. An inhibitor of photosynthesis, DCMU, at concentrations which inhibit light-mediated CO₂ fixation, decreases the growth rate of light-grown but not dark-grown plants. Light still slightly increases the growth rate of plants in the presence of DCMU. This latter response is mediated by phytochrome, since it can be effected by a 2 minute exposure to low intensity red light every 12 hours, and far-red light reverses the effect of red. The increased growth rate effected by red light is related to a change in the morphology of the plants. Dark-grown plants form compact balls of tissue consisting of lobes. These lobes are rounded and thick and exhibit an abnormal callus-type growth, with few well-defined meristematic regions. Plants grown in red light form fluffy balls of tissue. The lobes of these plants have a morphology more typical of Sphaerocarpos in nature. They are 2 cell layers thick, flattened, and have numerous well-defined meristematic areas. The greater number of meristems allows for the increased growth rate of the plants grown in red light.     

Response of excised Avena coleoptile segments to red and far-red light

Summary In seeking a simple, red light-promoted straight growth test in which phytochrome assays may be conducted without interference by protochlorophyll, the response of excised Avena coleoptile segments to red and far-red light was re-examined. The elongation of apical (non-decapitated) segments is promoted by a brief exposure to red light, and this effect is almost completely nullified by an immediately subsequent exposure to far-red light. Although growth promotion by red light occurs in distilled water alone, the effect is greater on a medium consisting of 0.02 M phosphate buffer, pH 6.2 to 6.4, with 1 to 2% sucrose. Over the pH range 4.5 to 7.4, dark-growth decreases with increasing pH, but the absolute increment brought about by red light is nearly constant. Elongation appears to be entirely the result of increased cell size.Contrary to previous reports, similar results can be obtained with subapical (decapitated) coleoptile segments, although the absolute magnitude of the response is reduced.Research carried out at Brookhaven National Laboratory under the auspices of the U.S. Atomic Energy Commision.     

Growth of Tsuru-rindo (Tripterospermum japonicum) culturedin vitro under various sources of light-emitting diode (LED) irradiation

We studied the effects of light generated by LEDs on the growth of Tsururindo (Tripterospermum japonicum) shoots. Apical shoots (2–3 cm long) were cultured on MS basal media supplemented with 3% sucrose, and were maintained for four weeks under five different light qualities: F (fluorescent lamp), red LED (R), 70% red + 30% blue LED (R7B3), 50% red + 50% blue (R5B5), or blue LED (B). Rooting was promoted by red light (100%) but was inhibited by blue light. Plant growth, as defined by root number, fresh weight, and chlorophyll content, was generally healthier for cultures irradiated with mixed LEDs, and was the best under R7B3. Ventilation resulted in more rapid apical shoot growth and rooting compared with control plants, when both were treated with the R7B3 system. We demonstrated here that plant growth can be controlled by using LEDs to adjust for the most effective irradiation conditions, compared with the performance observed when conventional fluorescent lamps are utilized.     

Effect of Far Red and Red Light on a Pelletable Peroxidase Activity in Extracts from Spinach Leaves

The effect of in vitro irradiations on ihe pelletability of peroxitlase activity was tested in extracts prepared from leaves of light-grown spinach (Spinacia oleracea). It appeared that far red light increased the pelletable activity and that red light reversed this effect. This phytochrome-mediated effect occurred in a particulate fraction which banded at 39% (w/w) sucrose on an isopycnic density gradient.     

RNA and protein metabolism in the particulate fractions of the gametophytes of bracken fern during growth in red and blue light

Summary The metabolism of RNA and protein in the gametophytes of bracken fern (Pteridium aquilinum) is affected by the quality of light in which they are grown. When sporelings were grown as two-dimensional gametophytes in blue light, particulate fractions separated from the sporelings exhibited greater incorporation of uridine-³H and leucine-³H into RNA and protein, respectively, than those from sporelings grown as one-dimensional protonema in red light. After various periods of exposure of gametophytes to red or blue light in the presence of uridine-³H, the nuclei-rich fraction showed the highest specific activity in RNA, and irrespective of incubation time, blue light was more effective than red light. The possibility that enhanced synthesis of RNA in the nucleus in response to blue light is significantly related to the morphological growth pattern of the gametophytes, is discussed.     

Photoaffinity labeling of proteins in bovine testis nuclear extract

A binary system of photoaffinity reagents for selective affinity labeling of DNA polymerases has been developed. The photoreactive probe was formed in nuclear extract, using an end-labeled oligonucleotide containing a synthetic abasic site. This site was incised by apurinic/apyrimidinic endonuclease and then dNMPs carrying a photoreactive adduct were added to the 3(') hydroxyl using base-substituted arylazido derivatives of dUTP or dCTP. This results in the synthesis of photoreactive base excision repair (BER) intermediates. The photoreactive group was then activated, either directly (UV light exposure 320nm) or in the presence of the sensitizer of dTTP analog containing a pyrene group (Pyr-dUTP) under UV light 365nm. DNA polymerase beta was the main target crosslinked by photoreactive BER intermediates in this nuclear extract. In contrast, several proteins were labeled under the conditions of direct activation of arylazido group.     

Flowering of Cultivated Green and SAN 9789-Treated Chenopodium rubrum Plants Exposed to White,Blue, and Red Light

Green plants and plants devoid of photosynthetic pigments were compared with regard to their ability to flower under various growth conditions. Green plants of Chenopodium rubrum L. and plants treated with norflurazon SANDOZ-9789 (SAN) were grown on sucrose-containing media with or without hormones (GA₃, BA, IAA, ABA) under short-day photoperiodic or continuous illumination with white, blue, or red light. Green and SAN-treated albino plants produced flowers only under short-day conditions. The flowering of green plants was independent of the presence of sucrose and hormones in the medium as well as of the light quality. The albino plants produced flowers under white and blue light but did not flower in red light. The addition of GA₃ or BA to the medium induced flowering of albino plants exposed to red light. The functional interaction of photoreceptors in the flowering control is discussed.     

Some Factors Affecting the Anthocyanin Formation by Populus Cells in Suspension Culture

In order to elucidate the mechanism of anthocyanin formation by Populus cells in suspension culture, the favourable conditions for anthocyanin formation were investigated. The influence of some factors affecting the anthocyanin formation, i.e. light, sucrose and riboflavin etc. were also examined. Light irradiation and high sucrose concentrations brought about a marked increase of PAL activity, which increased rapidly at the lag phase preceding the anthocyanin formation. The effect of blue light on anthocyanin formation was markedly superior to other kinds of monochromatic light (green and red) or white light. Riboflavin was effective only under light exposure. It was inferred that light, sucrose, riboflavin and PAL activity etc. were closely related with the anthocyanin formation. Especially, light and sucrose cooperated in the increase of PAL activity which was a key enzyme in the biosynthesis of anthocyanin.     

Separation of cell enlargement and division in bean leaves

A method is presented for inducing cell enlargement in intact leaves and leaf strips of Phaseolus vulgaris L. without the complication of cell division. Primary bean leaves complete cell division and stop growing after 10 d in dim red light. Transfer to white light induces expansion (50% in 24 h) which is entirely the consequence of cell enlargement. Leaf strips from red-light-grown seedlings placed in white light and provided external solutes (10 mM KCl+10 mM sucrose) expand at the same rate as intact leaves in the light. This system makes possible future investigation of the mechanism of leaf cell enlargement.     

Stomatal Opening and Cell Enlargement in Response to Light and Phytohormone Treatments in Primary Leaves of Red-Light-Grown Seedlings of Phaseolus vulgaris L.

Cell enlargement in primary leaves of bean seedlings grown for10 days in dim red light in response to different light andphytohormone treatments was studied. On day 10, bean leaf discswere floated on 1% sucrose with, or without, phytohormones fordifferent periods (up to 24 h) under dim red light, or discswere floated in sucrose solution and irradiated with white orblue light. Cell enlargement was enhanced by continuous whiteand blue light and by benzyladenine, kinetin and gibberellicacid. When seedlings were grown for 8 days under dim red light afterwhich a 2-day dark period was interposed (for the accumulationof inactive phytochrome), cell enlargement was enhanced by a5-min irradiation with red light. This growth induction wasfar-red reversible. The conditions under which cell enlargement was promoted, alsoinduced the opening of the stomata. Red light induced a far-redreversible transient stomatal opening. Based on the kineticsof stomatal opening and cell enlargement we formulated the hypothesisthat cell enlargement in leaves in response to light and phytohormonesis mediated by the stomatal response to these factors. (Received September 30, 1983; Accepted February 27, 1984)     

Regulation of expression of the cucumber isocitrate lyase gene in cotyledons upon seed germination and by sucrose

A 6.5 kb cucumber genomic DNA fragment containing the icl gene was introduced into Nicotiana plumbaginifolia and shown to direct isocitrate lyase (ICL) mRNA synthesis in transgenic seedlings upon germination, in a temporally regulated manner. Two putative icl promoter fragments, of 2900 and 572 bp, were subsequently linked to the GUS reporter gene and introduced into N. plumbaginifolia. Both constructs directed GUS expression after transgenic seed germination, and although the 572 bp fragment gave only 1% of the activity of the 2900 bp fragment, it directed expression in the same cotyledon-specific and temporally regulated pattern. Seedlings were transferred to darkness after 18 days growth in the light, to induce a starvation response. The 2900 bp construct was activated by starvation and repressed by exogenous sucrose, whereas the 572 bp construct was not starvation-responsive. To localize the region of the 2900 bp promoter fragment which is responsible for regulation by sucrose, further deletions were make, linked to GUS, and assayed in a cucumber protoplast transient assay system. Constructs with promoters of 2900, 2142 and 1663 bp were activated by starvation and repressed by sucrose, but promoters of 1142 and 572 bp showed no such response. We conclude that the icl gene promoter contains at least two distinct cis-acting elements, one required for the response to sucrose and the other which participates in expression upon seed germination.