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Phytochrome A Mediates the Promotion of Seed Germination by Very Low Fluences of Light and Canopy Shade Light in Arabidopsis.

Seeds of the wild type (WT) and of the phyA and phyB mutants of Arabidopsis thaliana were exposed to single red light (R)/far-red light (FR) pulses predicted to establish a series of calculated phytochrome photoequilibria (Pfr/P). WT and phyB seeds showed biphasic responses to Pfr/P. The first phase, i.e. the very-low-fluence response (VLFR), occurred below Pfr/P = 10-1%. The second phase, i.e. the low-fluence response, occurred above Pfr/P = 3%. The VLFR was similarly induced by either a FR pulse saturating photoconversion or a subsaturating R pulse predicted to establish the same Pfr/P. The VLFR was absent in phyA seeds, which showed a strong low-fluence response. In the field, even brief exposures to the very low fluences of canopy shade light (R/FR ratio < 0.05) promoted germination above dark controls in WT and phyB seeds but not in the phyA mutant. Seeds of the phyA mutant germinated normally under canopies providing higher R/FR ratios or under deep canopy shade light supplemented with R from light-emitting diodes. We propose that phytochrome A mediates VLFR of A. thaliana seeds.

Journal Article↗

Photocontrol of Peanut (Arachis hypogaea L.) Embryo and Ovule Development in Vitro.

The purpose of the present study was to determine the nature of the photoreceptor that controls the light-dependent development of the peanut ovule and embryo, and to determine whether this photoreceptor activity is located in the ovular or the embryonic tissue. Peanut ovules were aseptically excised from surface sterilized gynophores (pegs) and cultured in darkness, continuous far red (FR), or given 15 minutes daily irradiations with white (W), red (R), blue (B); FR, R/FR, FR/R light for 21 days. Increases in the volume of ovules cultured in dark, continuous FR, FR, or R/FR were significantly greater than for ovules cultured in W, R, FR/R, or B radiation. Halving the ovule or removing the micropyle did not affect the relative change in ovule volume in response to different light treatments. When the light sources were altered after the first 10 days of culture, the ovules exposed first to darkness or FR radiation began to increase in ovule volume which then ceased after subsequent exposure to W, R, or B radiation. Likewise, if W, R, or B radiation was given during the first 10 days, ovule development was inhibited, and could be stimulated by exposure to darkness or FR light. Growth of embryos, when removed from the influence exerted by ovular tissue, was unaffected by the light. However, when embryos remained attached to the ovular tissue, their growth was stimulated in darkness or by FR light and inhibited by W, R, or B light. The R/FR reversibility indicates that the photoreceptor that regulates ovular growth is phytochrome and that the maternal ovular tissue appears to be the site of photoreception, which may then transmit some developmental signal to the embryo.

Journal Article↗

A Semidian Rhythm in the Flowering Response of Pharbitis nil to Far-Red Light: II. The Involvement of Phytochrome.

The semidian ( approximately 12 h) periodicity in the effect of far-red (FR) interruptions of the light period preceding inductive darkness on flowering in Pharbitis nil appears to be mediated by phytochrome: (a) promotion by interruptions 2 hours before inductive darkness (-2 hours) and inhibition at -8 hours are greater the higher the proportion of FR/R+FR during the interruption; (b) brief FR exposures followed by darkness are even more effective than FR throughout; (c) the effect of brief FR is reversed by subsequent R; (d) R interruptions of an FR background are most promotive at -8 hours, when FR is most inhibitory. Promotive FR interruptions at -2 or -14 hours shorten the critical dark period whereas inhibitory FR interruptions at -8 hours lengthen it. We conclude that the semidian rhythm is controlled by a ;timing pool' of phytochrome FR absorbing form (Pfr) which disappears rapidly in darkness: four different estimates from our experiments indicate that Pfr was reduced to the level set by FR within 20 to 45 minutes in darkness. However, flowering may also be influenced by a ;metabolic pool' of Pfr with a delayed loss in darkness, the time of which can be advanced or retarded by shifting the semidian rhythm.

Journal Article↗

The effect of temperature, photoperiod, and light quality on gluconasturtiin concentration in watercress (Nasturtium officinale R. Br.).

The effects of different growth regimes on gluconasturtiin concentration in watercress (Nasturtium officinale R. Br.) were investigated. Watercress plantlets at the 5th mature leaf stage (ca. 2 weeks old) were exposed to different day and night temperatures, to long (16 h) or short (8 h) days, to red (R) or far-red (FR) light given during the main long day photoperiod, and finally to R or FR light given at the end of the main photoperiod. Watercress plants grown under long days contained a 30-40% higher gluconasturtiin concentration and had a higher fresh weight than watercress plants grown under short days. Watercress plants grown under long days and temperatures of 15 or 10 degrees C had at least a 50% higher gluconasturtiin concentration, but a lower fresh weight, than that of plants grown at 20 or 25 degrees C. Watercress plants grown under metal halide light enriched with R light had approximately a 25-40% higher concentration of gluconasturtiin as compared to the FR-enriched plants. Likewise, a brief R light exposure at the end of the main photoperiod resulted in approximately a 25% or higher concentration of gluconasturtiin as compared to a FR end-of-day exposure. These data indicate that the concentration of gluconasturtiin in watercress can be significantly increased by growing plants at lower temperatures, under long days, and by exposure to R light.

Chromatography, High Pressure Liquid↗

DNA binding by the coliphage 186 repressor protein CI.

The cI gene of coliphage 186 maintains lysogeny and confers immunity to 186 infection by repressing the major early promoter, p(R), and the promoter for the late transcription activator gene, p(B). Gel mobility shirt and DNase I footprinting show that CI protein binds to the DNA at p(R) and p(B) and also to sites approximately 300 base pairs upstream and downstream of p(R), called FL and FR. Mutations which cause virulence reduce CI binding to p(R). The biochemical and genetic data identify three CI operators at p(R), two at p(B), and single operators at FL and FR. The operators at the p(B), FL, FR, and central p(R) sites are inverted repeat sequences, separated by 5 base pairs (Type A) or, in the case of p(R), by 4 base pairs (Type A'). A different inverted repeat operator sequence (Type B) is proposed for the binding sites on each side of the central site at p(R). Thus, CI appears to recognize two distinct DNA sequences. CI binds cooperatively to adjacent operators, and binding at p(R) is strongly dependent on these cooperative interactions. A high order CI multimer appears to be the active DNA binding species, even at single operators.

Amino Acid Sequence↗

Long-lived Intermediates in Phytochrome Transformation I: In Vitro Studies.

Irradiation of phytochrome solutions with a high-intensity mixed red and far red light source causes measurable absorbancy increases at 543 nm. Evidence is presented that these absorbancy increases are caused by accumulation of intermediates on the P(R) to P(FR) pathway with relatively slow thermal decay constants. Kinetic analysis of the decay signals is consistent with the interpretation that the signals represent simultaneous independent and parallel decay of 2 species by first order kinetics to P(FR). If actinic light intensity is kept constant and exposure time changed, the relative amounts of the 2 components change, with proportionately more of the rapidly decaying species present following short exposure times. If the amount of the intermediates is decreased by decreasing actinic light intensity at constant exposure time, however, the relative amounts of the 2 remain constant. The Q(10) for intermediate decay following illumination is approximately 2.0, while that for complete phototransformation of the pigment in either direction is very close to 1.0. Incomplete transformation of P(R) to P(FR), caused by overlapping absorption of the 2 forms, is shown by the presence of intermediates (indicating cycling of the pigment) in continuous red light. Such intermediates do not appear in continuous far red, indicating a rate of pigment cycling below detection by the available instrumentation.

Journal Article↗

The Arabidopsis COG1 gene encodes a Dof domain transcription factor and negatively regulates phytochrome signaling.

Light is a critical environmental factor that influences almost all developmental aspects of plants, including seed germination, seedling morphogenesis, and transition to reproductive growth. Plants have therefore developed an intricate network of mechanisms to perceive and process environmental light information. To further characterize the molecular basis of light-signaling processes in plants, we screened an activation tagging pool of Arabidopsis for altered photoresponses. A dominant mutation, cog1-D, attenuated various red (R) and far-red (FR) light-dependent photoresponses. The mutation was caused by overexpression of a gene encoding a member of the Dof family of transcription factors. The photoresponses in Arabidopsis were inversely correlated with the expression levels of COG1 mRNA. When the COG1 gene was overexpressed in transgenic plants, the plants exhibited hyposensitive responses to R and FR light in a manner inversely dependent on COG1 mRNA levels. On the other hand, transgenic lines expressing antisense COG1 were hypersensitive to R and FR light. Expression of the COG1 gene is light inducible and requires phytochrome A (phyA) for FR light-induced expression and phytochrome B (phyB) for R light-induced expression. Thus, the COG1 gene functions as a negative regulator in both the phyA- and phyB-signaling pathways. We suggest that these phytochromes positively regulate the expression of COG1, a negative regulator, as a mechanism for fine tuning the light-signaling pathway.

Amino Acid Sequence↗

Photochemical and Nonphotochemical Reactions of Phytochrome in vivo.

The nonphotochemical reactions of phytochrome in the coleoptiles of dark-grown corn seedlings were studied at 3 temperatures: 14 degrees , 24 degrees , and 34 degrees . The data obtained show that the destruction of P(fr) is the only measurable reaction occurring; reversion of P(fr) to P(r) was not found. The Q(10)'s (2.7 and 3.5) and zero order kinetics found for the destruction reaction are consistent with the hypothesis that the reaction is enzyme-mediated.In vivo action spectra for phytochrome transformation in the coleoptiles of darkgrown corn seedlings were obtained which agree qualitatively with those obtained by other workers for phytochrome-mediated physiological responses and in vitro action spectra. In vivo conversion of phytochrome by blue light, as determined from spectrophotometric measurements of phytochrome itself, is reported. Action peaks for P(r) were found at 667 mmu and in the blue in the region of 400 mmu, with a broad shoulder from 590 mmu to 640 mmu. Action peaks for P(fr) were found at 725 mmu and in the blue in the region of 400 mmu with a minor peak at 670 mmu, and a broad shoulder from 590 mmu to 640 mmu. The ratio of the quantum efficiencies of P(r) at 667 mmu and P(fr) at 725 mmu (Phi(r667)/Phi(fr725)) was estimated to be 1.0.

Journal Article↗

Impedance cardiography: a potential monitor for hemodialysis.

BACKGROUND: Impedance cardiography (ICG) technology has improved dramatically, and at least one device now can give a measurement of fluid status by using thoracic fluid content (TFC), along with cardiac output (CO) and cardiac index (CI). With a built-in sphygmomanometer cuff, it can also provide blood pressure (BP) and systemic vascular resistance index (SVRI). A currently available small portable ICG that provides reliable measures of fluid status could be an ideal noninvasive monitor for hemodialysis (HD), with the potential of helping avoid significant hemodynamic instability during HD. METHODS: A case series of patients with chronic renal failure was studied while undergoing HD using ICG (BioZ, CardioDynamics, Int. Corp., San Diego, CA). Parameters recorded at 15-min intervals included TFC, CI, BP (systolic, diastolic, and mean arterial), SVRI, and heart rate. Using the Pearson method, the percentage changes in each of the parameters during the HD session were correlated to the amount of fluid removed (FR), normalized to body weight. RESULTS: Forty-one patients were enrolled, but six patients were excluded due to incomplete data; therefore, 35 patients (13 men and 22 women) formed the basis of the analysis. The age range was 28 to 87 (mean 55.1 +/- 16.1) years. The amount of FR was 2.88 +/- 1.13 L (37.3 +/- 14.6 ml/kg). TFC decreased in all patients during the HD session (average reduction 12.7 +/- 8 kohms(-1)); whereas all other hemodynamic parameters showed both increases and decreases. The correlation of change in TFC with FR was moderate (r = 0.579, P = 0.0003); other hemodynamic parameters showed a poor correlation with FR. Neither the standard hemodynamic parameters nor the ICG device's special parameters were able to identify the five patients in this series who experienced significant hemodynamic instability or intradialytic hypotension. CONCLUSION: TFC, measured easily and noninvasively using ICG, correlates with the amount of fluid removed during HD. In comparison with the other hemodynamic parameters measured, TFC changed most consistently with fluid removal. Whether or not serial TFC measurements in a given patient at different HD sessions can guide the extent of FR will require additional study. This compact, easily operated, and nonobtrusive ICG device with the capability for continuously providing the standard hemodynamic parameters plus CO, TFC, and standard limb lead electrocardiography could replace current monitoring systems.

Adult↗

Maize leaves turn away from neighbors.

In commercial crops, maize (Zea mays) plants are typically grown at a larger distance between rows (70 cm) than within the same row (16-23 cm). This rectangular arrangement creates a heterogeneous environment in which the plants receive higher red light (R) to far-red light (FR) ratios from the interrow spaces. In field crops, the hybrid Dekalb 696 (DK696) showed an increased proportion of leaves toward interrow spaces, whereas the experimental hybrid 980 (Exp980) retained random leaf orientation. Mirrors reflecting FR were placed close to isolated plants to simulate the presence of neighbors in the field. In addition, localized FR was applied to target leaves in a growth chamber. During their expansion, the leaves of DK696 turned away from the low R to FR ratio signals, whereas Exp980 leaves remained unaffected. On the contrary, tillering was reduced and plant height was increased by low R to FR ratios in Exp980 but not in DK696. Isolated plants preconditioned with low R/FR-simulating neighbors in a North-South row showed reduced mutual shading among leaves when the plants were actually grouped in North-South rows. These observations contradict the current view that phytochrome-mediated responses to low R/FR are a relic from wild conditions, detrimental for crop yield.

Adaptation, Physiological↗

Characteristics of three strains of feline fibrosarcoma virus grown in cat and marmoset monkey cells.

Two strains of feline fibrosarcoma virus (ST-FeSV and GA-FeSV) were found to induce tumors in cats and marmosets, and to transform feline and marmoset cells in vitro after primary inoculation. A third strain (SM-FeSV) failed to induce tumors or transform marmoset cells after primary inoculation; however, when SM-FeSV-injected marmoset cultures were passed 26 times in vitro, the cell cultures released infectious virus which transformed marmoset fibroblasts but still failed to induce tumors in marmosets. ST-FeSV induced mainly round-cell type transformation (r foci), GA-FeSV induced predominantly mixed round-fusiform cell type transformation (fr foci), and SM-FeSV induced r and fr type foci with a higher proportion of fusiform cells in the fr foci than seen with GA-FeSV. Transforming virus was obtained from r or mixed r/fr foci of ST-FeSV but not from fr foci; heat treatment changed the virus from producing almost exclusively r type foci to inducing an increased number of fr foci. Passage of FeSV in cat cells yielded viruses with a higher ratio of infectivity for feline vs marmoset cells, while passage of FeSV in marmoset cells yielded virus with a relatively higher infectivity ratio for marmoset cells; the three strains differed in the degree of change in the infectivity ratio. Despite the alteration of host range of SM-FeSV propagated in marmoset fibroblasts, the virus retained feline P-30 antigen by CF and FA assays. Neutralization tests did not indicate but also did not exclude an alteration of the surface antigens of ST-FeSV or SM-FeSV propagated in marmoset fibroblasts. The alteration of the relative infectivity of FeSV during passage in marmoset cells may be due to: (1) the selection of a variant present in the original heterogenous uncloned population; (2) mutation; or (3) recombination with some marmoset genetic material, possibly an as yet unidentified endogenous marmoset virus.

Animals↗

Spectroscopic detection of a phytochrome-like photoreceptor in the myxomycete Physarum polycephalum and the kinetic mechanism for the photocontrol of sporulation by Pfr.

Sporulation of the true slime mold Physarum polycephalum (Myxomycetales) can be triggered by the far-red/red reversible Physarum phytochrome. Physarum plasmodia were analyzed with a purpose-built dual-wavelength photometer that is designed for phytochrome measurements. A photoreversible absorbance change at 670 nm was monitored after actinic red (R) and far-red (FR) irradiation of starved plasmodia, confirming the occurrence of a phytochrome-like photoreceptor in Physarum spectroscopically. These signals were not found in growing plasmodia, suggesting the Physarum phytochrome to be synthesized during starvation, which makes the cells competent for the photoinduction of sporulation. The photoconversion rates by R and FR light were similar in the phytochromes of Physarum and etiolated oat shoots. In dark-grown Physarum plasmodia that had not been preexposed to any light only R induced a detectable absorbance change while FR did not. This indicates that most (at least 90%) of the photoreversible pigment occurs in the red-absorbing form. Since the effectiveness of FR in triggering sporulation was enhanced by preirradiation with R, it is concluded that at least part of the Pr can be photoconverted to the active Pfr photoreceptor species. We propose a kinetic mechanism for the photocontrol of sporulation by photoconversion of Pfr, which may also hold for the high-irradiance response to FR in Arabidopsis and Cuscuta.

Animals↗

Aroma of fresh strawberries is enhanced by ripening over red versus black mulch.

Strawberry (Fragaria ananassa Duch) is a high-value food crop, the aroma of which is important in consumer satisfaction. We hypothesized that the concentration of fresh strawberry aroma compounds could be enhanced by growing the berries over a red plastic mulch that was formulated to reflect more far-red (FR) and red light (R) and a higher FR/R photon ratio than is reflected by standard black plastic mulch. Strawberries of the cultivars "Chandler" and "Sweet Charlie" were grown in trickle-irrigated field plots over the two colors of mulch. The berries were harvested when ripe, and aroma compounds were isolated by dynamic headspace sampling. Entrained compounds were quantified by gas chromatography and identified by gas chromatography/mass spectroscopy. Twenty-three compounds were identified, and most of them were aliphatic esters. Strawberries that ripened over the red mulch during periods of sunny weather had significantly higher concentrations of aroma compounds than berries grown over the black plastic mulch. Total levels of aroma compounds from berries grown over red plastic mulch were higher for both Chandler and Sweet Charlie. We postulate that FR and R in light reflected from the red mulch acted through the natural phytochrome system to modify gene expression enough to result in greater concentration of aroma compounds in fresh strawberries.

Agriculture↗

Transduction of folate receptor cDNA into cervical carcinoma cells using recombinant adeno-associated virions delays cell proliferation in vitro and in vivo.

Although folate receptors (FRs) mediate folate uptake into cells, the independent role of FRs in cell proliferation remains unclear. We tested the hypothesis that transduction of FR cDNA in sense or antisense orientation using recombinant adeno-associated virus modulated FR expression and altered proliferation of cervical carcinoma cells (which constitutively overexpress FR genes). We determined that the integration of recombinant adeno-associated virions was not site specific. When compared with untransduced cells, sense and antisense FR cDNA-transduced cells exhibited an increase and decrease in FR mRNA and FR expression on the cell surface, respectively. However, when compared with antisense FR cDNA-transduced and untransduced cells, sense FR cDNA-transduced cells exhibited statistically significant (a) increased in total FRs, (b) smaller colonies, (c) lowered cell proliferation in vitro, and (d) less tumor volume with dramatic prolongation of tumor doubling times (225.6 h vs. 96 h) after transplantation into nude mice. Finally, (f) using single cell-derived transduced clones, an inverse relationship between cell proliferation and FR expression was established (r = 0.90, P < 0.001). Thus, transduction of sense/antisense FR cDNA into cervical carcinoma cells modulated expression of FRs and had an impact on cell proliferation in vitro and in vivo.

Animals↗

The first density functional study on the

The hetero-Diels-Alder reactions of 1,2-diaza-1,3-butadiene with ethylene, methyl vinyl ether, and methyl acrylate have been investigated theoretically with the aid of density functional theory using the Becke3LYP/6-31G(d) computational level. In the reactions with substituted alkenes, the transition states are concerted but asynchronous; the shortest bond-forming distance involves the nonsubstituted carbon of the alkene. In agreement with the experimental results, the reaction with methyl vinyl ether proceeds with high endo stereoselectivity and with complete regioselectivity to form the 6-methoxy cycloadduct. The conformational study of the transition states shows a sharp s-trans preference. In contrast, the [4 + 2]-cycloaddition of 1,2-diaza-1,3-butadiene with methyl acrylate have been found to occur with low regio- and stereoselectivity but with a s-cis preference in the transition structures.

Journal Article↗

Diastereoselective cycloadditions of 1,3-thiazolium-4-olates with chiral 1,2-diaza-1,3-butadienes

A series of 2-(N-methyl)benzylamino-1,3-thiazolium-4-olates (2-aminothioisomunchnones) react with chiral 1,2-diaza-1,3-butadienes derived from carbohydrates to afford a diastereomeric mixture of (4R,5S)- and (4R,5R)-4,5-dihydrothiophenes. These substrate-controlled cycloadditions are chemoselective, regiospecific, and proceed with a high facial diastereoselection. A theoretical rationale at semiempirical level does justify the stereochemical outcome observed in the experiments.

Journal Article↗