Phototropism in Hypocotyls of Radish : II. Role of cis- and trans-Raphanusanins, and Raphanusamide in Phototropism of Radish Hypocotyls

When etiolated radish (Raphanus sativus var. hortensis f. gigantissimus Makino) hypocotyls were subjected to a continuous unilateral illumination with white fluorescent light (0.1 watt per square meter), the growth rate at the lighted side was strongly inhibited for the first 2 hours, while that at the shaded side showed no change. After 2.5 hours growth on the lighted side recovered gradually, while that on the shaded side was slightly inhibited. The neutral growth inhibitors, cis- and trans-raphanusanins and raphanusamide, were determined in the lighted and shaded sides from 1 hour before until 2 hours after the start of unilateral illumination. In the lighted side, cis- and trans-raphanusanins increased by 0.5 hour after the start of illumination, reached 3 to 3.5-fold greater concentrations than in the shaded side after 1 hour, and then decreased gradually. Raphanusamide increased in the lighted side to a 3-fold greater concentration than that in the shaded one 2 hours after the start of the illumination. Unilateral applications of cis- and trans-raphanusanins and raphanusamide suppressed the growth of the hypocotyl on the applied side more than that on the opposite one, causing the hypocotyls to bend towards the site of application. The data suggest that phototropic curvature in radish is caused by the light-induced synthesis of growth-inhibiting cis- and trans-raphanusanins, and raphanusamide at the site of illumination.     

Phototropism in Hypocotyls of Radish : I. Isolation and Identification of Growth Inhibitors, cis- and trans-Raphanusanins and Raphanusamide, Involved in Phototropism of Radish Hypocotyls

Three growth inhibitors which might be involved in phototropism of Sakurajima radish (Raphanus sativus var. hortensis f. gigantissimus Makino) hypocotyls, were isolated as crystalline forms from light-exposed radish seedlings and identified as cis- and trans-raphanusanins and 6-methoxy-2,3,4,5-tetrahydro-1,3-oxazepin-2-one (designated raphanusamide). The cis- and trans-raphanusanins inhibited growth of etiolated radish hypocotyls at concentrations higher than 1.5 micromolar, raphanusamide at concentrations higher than 20 micromolar.     

Phototropism in Hypocotyls of Radish: IV. Flank Growth and Lateral Distribution of cis- and trans-Raphanusanins in the First Positive Phototropic Curvature

The first positive phototropic curvature induced by a pulse of unilateral white irradiation (0.1 watt per square meter, 30 seconds) of etiolated and de-etiolated Sakurajima radish (Raphanus sativus var hortensis f. gigantissimus Makino) hypocotyls was analyzed in terms of differential growth and growth inhibitor contents of the hypocotyls. In both etiolated and de-etiolated hypocotyls, the growth rates at the lighted sides were suppressed whereas those at the shaded ones showed no change. De-etiolation treatment induced a larger difference between the growth rates at the lighted and shaded sides of the hypocotyls, resulting in a larger curvature of de-etiolated seedlings than of etiolated ones. The contents of growth inhibitors, cis- and trans-raphanusanins, increased in the lighted but not in the shaded halves of the hypocotyls of etiolated seedlings. In de-etiolated seedlings, the two inhibitors increased due to the de-etiolation treatment. When de-etiolated seedlings were exposed to a pulse of unilateral irradiation the level of the two inhibitors remained high along the lighted side for 1 h following the light pulse, whereas at the shaded side the contents of the inhibitors abruptly decreased upon transfer to the dark, the difference between their amounts in the lighted and shaded sides being larger than in etiolated seedlings. Another growth inhibitor, raphanusamide, did not respond to the phototropic stimulus, although its amounts increased by the de-etiolation treatment. These data suggest that cis- and trans-raphanusanins are involved in the first positive phototropic response of radish hypocotyls, and that de-etiolation magnifies the phototropic response through induction of a larger lateral gradient of the raphanusanins in the hypocotyls by the phototropic stimulus.     

Effects of Light Illumination on the Population of Translatable mRNA in Radish Cotyledons during Dark-Induced Senescence

The population of translatable mRNA in radish cotyledons hasbeen shown to change strikingly during dark-induced senescenceby analyzing cell-free translation products from polyadenylatedRNA using two-dimensional electrophoresis (Kawakami and Watanabe1988, Plant Cell Physiol. 29: 33–42). In the present study,the translatable mRNAs in the cotyledons were studied afterre-illumination of seedlings which had been induced to senesceby dark treatment. All of the mRNA species which showed relativeincrease or decrease during a 24-h dark treatment returned totheir initial levels after a subsequent 24-h illumination withwhite light. When seedlings were kept in the dark for 48 h,21 mRNA species increased, but the increase in some of themwas no longer reversible; 9 out of the 21 mRNA species remainedat the same level even after 48 h of re-illumination. All ofthe 18 mRNA species which decreased during 48 h in the darkreturned to their initial levels upon re-illumination. The mRNAspecies that showed irreversible increase were also found toaccumulate in cotyledons senesced after a lengthy growth periodunder light. There were also some mRNA species whose contentwas not affected by dark treatment, but increased transientlyafter re-illumination of the seedlings. These observations arediscussed in relation to the progress of senescence in the cotyledons. (Received September 24, 1987; Accepted December 19, 1987)     

Negative Phototropism in Vaucheria terrestris Regulated by Calcium III. The Role of Calcium Characterized by Use of a High-Power Argon-Ion Laser as the Source of Unilateral Blue Light

A tip-growing Xanthophycean algal coenocyte, Vaucheria terrestrissensu Gtz, is able to change the sign of its phototropic responsefrom positive to negative as a result of its ability to sensethe fluence rate (=intensity) of unilateral blue light (BL).The mechanism that determines the sign of phototropism was investigatedusing a high-power argon-ion laser (457.9 nm) as a source ofvery strong unilateral BL. The fluence-response relationshipwas determined by changing both the fluence rate and the durationof irradiation. Positive phototropic bending was induced whenthe fluence rate of BL from the laser was below 60 W m^–2.The positive bending obeyed the reciprocity law and was notaffected by the concentrations of external Ca²⁺ ions between0.4 mM and 4.4 mM. The positive curvature decreased when thealga was exposed to a unilateral pulse of BL with a durationof 10–300 s at fluence rates higher than 60 W m^–2.The alga finally showed a deep negative curvature when eitherthe fluence rate or the duration of irradiation was furtherincreased. The inversion of the phototropic response and developmentof the negative phototropic response was greatly enhanced inthe presence of 4.4 mM Ca²⁺ ions. However, the mechanism thatdetermine the sign of phototropism seemed to require a BL pulseof longer than several seconds, even when the fluence rate wassufficiently high. The role of cytoplasmic Ca²⁺ ions in positiveand negative phototropic responses is discussed. ¹This study was carried out as part of NIBB Cooperative ResearchProgram for the Okazaki Large Spectrograph (89-513 and 90-518). ²Part of this study was reported at the XXXII Yamada Conferenceon Plant Cell Walls as Biopolymers with Physiological Functions,May 5–8, 1992, Osaka (Kataoka and Watanabe 1992).     

Phototropism and Protein Phosphorylation in Higher Plants: Unilateral Blue Light Irradiation Generates a Directional Gradient of Protein Phosphorylation Across the Oat Coleoptile

Blue light induces the phosphorylation of a 116 kDa oat protein found in plasma membrane preparations from coleoptile tips. We developed a very sensitive in vitro method that allowed us to determine the tissue distribution of protein phosphorylation after applying unilateral and bilateral blue light pulses in vivo. We found that following unilateral in vivo irradiation the degree in phosphorylation of the 116 kDa protein is significantly higher at the irradiated than at the shaded side of the coleoptile tip. This asymmetry can be considered as previously missing criterion that protein phosphorylation represents an early event within the transduction chain for phototropism.     

The Influence of Low Intensities of Light Pollution on Bat Communities in a Semi-Natural Context

Anthropogenic light pollution is an increasingly significant issue worldwide. Over the past century, the use of artificial lighting has increased in association with human activity. Artificial lights are suspected to have substantial effects on the ecology of many species, e.g., by producing discontinuities in the territories of nocturnal animals. We analyzed the potential influence of the intensity and type of artificial light on bat activity in a semi-natural landscape in France. We used a species approach, followed by a trait-based approach, to light sensitivity. We also investigated whether the effect of light could be related to foraging traits. We performed acoustic surveys at sites located along a gradient of light intensities to assess the activity of 15 species of bats. We identified 2 functional response groups of species: one group that was light-tolerant and one group that was light-intolerant. Among the species in the latter group that appear to be disadvantaged by lighting conditions, many are rare and threatened in Europe, whereas the species from the former group are better able to thrive in disturbed habitats such as lighted areas and may actually benefit from artificial lighting. Finally, several methods of controlling light pollution are suggested for the conservation of bat communities. Recommendations for light management and the creation of dim-light corridors are proposed; these strategies may play an important role in protecting against the impact of light pollution on nocturnal animals.     

Somatic Mutational Transients. III. Response by Two Genes in a Clone of Nicotiana to 24 Roentgens of Gamma Radiation Applied at Various Intensities

Somatic effects of radiation intensity on the mutable V and stable R genes were detected in plants of a heterozygous clone (v(S3)/v(s), R/r) subjected to the same dose at four rates. The effects were compared by counting speckled and purple sectors in flowers from irradiated and control plants. Response curves were estimated from the mutant sector averages, observed over a period of transient response for successive mature flowers. A structure for integrating the mutational contributions from different flowers was provided by models. The average control mutation rates are 8,110 per 10(7) cells for the V gene, and 49.45 for R. At a constant intensity of 4320 roentgens/hour, average induced mutation rates per 10(7) cells per roentgen for V increase from 194 (at 24 r total dose) to 1,116 (at 3 r dose); the corresponding rates for R increase from 7.24 to 27.65. With these responses as standards, both genes at corresponding total doses yield lower rates at lower intensities. For the series of intensities 1.2, 0.6, 0.3 and 0.15 roentgens/hour, the decreases in mutation rate for the V gene are, respectively, 66, 148, 315, and 617 per 10(7) cells per roentgen. The corresponding decrements for R are 4.86, 8.70, 14.61, and 23.51. These effects are non-linear functions of intensity for both genes, but V is at least 13 times as sensitive as R. Radiation operating to extinguish a buffering system against final mutation can account for the dose and dose-rate effects observed.     

Negative Phototropism in Vaucheria terrestris Regulated by Calcium I. Dependence on Background Blue Light and External Calcium Concentration

In the presence of 1–8 mM Ca²⁺ unilateral 10–15min irradiation with blue light can elicit negative phototropicbending in the tip-growing coenocytic fresh water alga, Vaucheriaterrestris (Xanthophyceae), when it is simultaneously irradiatedwith strong blue or green background light. By changing wavelength,fluence rate, and duration of background light and holding thoseof unilateral light (456 nm, 1.7 Wm^–2) negative phototropicresponse was analyzed: the wavelength of the background lighthad to be shorter than 540 nm; red light (660 nm) was ineffectiveeven at very high fluence rates (100W.m^–2). The negativebending was strongly and specifically dependent on the externalCa²⁺ concentration. Other divalent cations, Mg²⁺ and Ba²⁺, wereeither toxic or quite ineffective; Sr²⁺ could partly supportthe growth, but mediated neither positive nor negative phototropicbending. The rate of tip-growth was not significantly alteredbetween 10⁶M and 10^-6 mM Ca²⁺. Pre-irrradiation with the backgroundlight slightly increased the negative curvature; whereas thatwithout subsequent simultaneous irradiation does not cause negativebending, but rather increases the positive curvature. Three-mindelayed start of background light did not cause negative bendingany more. The present results strongly suggest that blue light elicitsan influx of Ca²⁺ at the apex of Vaucheria and that the increasedcytoplasmic Ca²⁺ regulates the sensitivity and direction ofphototropic response. (Received June 23, 1988; Accepted September 7, 1988)     

光照强度对金果榄光合特性和生长品质的影响

目的:探讨不同光照强度对金果榄植株光合特性和生长的影响.方法:设置光照强度为自然光强的100％、52.7％、33.2％和15.8％四种处理,测定其光合速率等生理指标、叶绿素、生物产量及活性成分含量的变化.结果:1.低光照强度处理的光合速率均高于100％自然光照处理,33.2％处理最高,显著地高于52.7％处理和15.8...     

Changes in Levels of Intermediates of the C(4) Cycle and Reductive Pentose Phosphate Pathway under Various Light Intensities in Maize Leaves

The rate of CO₂ assimilation and levels of metabolites of the C₄ cycle and reductive pentose phosphate pathway in attached leaves of maize (Zea mays L.) were measured over a range of light intensity from 0 to 1,900 microEinsteins per square meter per second under a saturated CO₂ concentration of 350 microliters per liter and a limiting CO₂ concentration of 133 microliters per liter. The level of ribulose 1,5-bisphosphate (RuBP) stayed almost constant (around 60 nanomoles per milligram chlorophyll [Chl]) from low to high light intensities under 350 microliters per liter. Levels of 3-phosphoglycerate (PGA) increased from 100 to 650 nanomoles per milligram Chl under 350 microliters per liter CO₂ with increasing light intensity. The calculated RuBP concentration of 6 millimolar (corresponded to 60 nanomoles per milligram Chl) was about two times above the estimated RuBP binding-site concentration on ribulose bisphosphate carboxylase-oxygenase (Rubisco) of ~2.6 millimolar in maize bundle sheath chloroplasts in the light. The ratio of RuBP/PGA increased with decreasing light intensity under 350 microliters per liter CO₂. These results suggest that RuBP carboxylation is under control of light intensity possibly due to a limited supply of CO₂ to Rubisco through the C₄ cycle whose activity is highly dependent on light intensity. Pyruvate level increased with increasing light intensity as long as photosynthesis rate increased. A positive relationship between levels of PGA and those of pyruvate during steady-state photosynthesis under various conditions suggests that an elevated concentration of PGA increases the carbon input into the C₄ cycle through the conversion of PGA to PEP and consequently the level of total intermediates of the C₄ cycle can be raised to mediate higher photosynthesis rate.     

Comparative Analysis Between Pseudomonas aeruginosa Genotypes and Severity of Symptoms in Patients with Unilateral or Bilateral Otitis Externa

Random amplified polymorphic DNA (RAPD) analysis was done on 32 isolates of Pseudomonas aeruginosa. These isolates were obtained from 22 patients who presented to the emergency room in a major medical center in Beirut, Lebanon, during a 5-month period with the diagnosis of either unilateral or bilateral otitis externa. Patients had yellowish to greenish discharge, moderate to severe external auditory canal swelling, moderate to severe pain, and periauricular cellulitis. None of these patients had intrinsic predisposing factors. An ear swab was obtained from both ears of patients, cultured on trypticase soy agar. P. aeruginosa was identified on the basis of pyocyanine production and API identification kits. RAPD analysis was done by using two primers (10 mer and 21 mer primers) and appropriate PCR conditions on extracted DNA. Our data have shown 23 RAPD patterns (A–W) distributed among the 32 P. aeruginosa isolates. RAPD patterns were reproducible. Twenty of 32 isolates were recovered from 10 patients with bilateral otitis externa. The remaining 12 of 32 isolates were recovered from 12 different patients with unilateral otitis externa. Eleven RAPD patterns (A,B,C,D,E,F,H,I,R,U,V) were associated with severe clinical symptoms, including severe pain, severe external auditory canal swelling, periauricular cellulitis, and a yellowish discharge. The remaining RAPD patterns were not associated with severe infections. This denotes a possible association between certain genotypes and severity of symptoms. Received: 6 July 2000 / Accepted: 5 September 2000     

Unilateral and Bilateral Cortical Resection: Effects on Spike-Wave Discharges in a Genetic Absence Epilepsy Model

Research Question

Recent discoveries have challenged the traditional view that the thalamus is the primary source driving spike-and-wave discharges (SWDs). At odds, SWDs in genetic absence models have a cortical focal origin in the deep layers of the perioral region of the somatosensory cortex. The present study examines the effect of unilateral and bilateral surgical resection of the assumed focal cortical region on the occurrence of SWDs in anesthetized WAG/Rij rats, a well described and validated genetic absence model.

Methods

Male WAG/Rij rats were used: 9 in the resected and 6 in the control group. EEG recordings were made before and after craniectomy, after unilateral and after bilateral removal of the focal region.

Results

SWDs decreased after unilateral cortical resection, while SWDs were no longer noticed after bilateral resection. This was also the case when the resected areas were restricted to layers I-IV with layers V and VI intact.

Conclusions

These results suggest that SWDs are completely abolished after bilateral removal of the focal region, most likely by interference with an intracortical columnar circuit. The evidence suggests that absence epilepsy is a network type of epilepsy since interference with only the local cortical network abolishes all seizures.     

The Influence of Chlorophyll on the Spectral Control of Elongation Growth in Chenopodium rubrum L. Hypocotyls

The inhibitory effectiveness of various monochromatic wavelengthsbetween 399 and 802 nm on hypocotyl elongation growth in light-grownChenopodium rubrum L. seedlings has been studied. The responsesof normal light-grown seedlings and chlorophyll-free light-grownseedlings were compared. Both types of seedling responded moststrongly to the blue and red waveband although a distinct peakof red light effectiveness was not observed in normal greenseedlings. The presence of chlorophyll also correlates witha lower inhibitory effectiveness of most wavelengths in the400–700 nm waveband. Photon fluence-rate response curves were not parallel; whereasthe plants were very sensitive to changes in fluence-rate inthe blue waveband, a much less marked fluence-rate dependencywas observed in the red and far-red wavebands. (Received September 10, 1981; Accepted April 26, 1982)     

温度与光强对高山杜鹃催花期间花芽营养物质积累的影响

以高山杜鹃为材料,研究了在3个昼/夜温度(6℃/0℃、22℃/16℃、30℃/24℃)和3个光照强度(2 000/10lx、7 500/5 500lx、10 000/8 000lx)组合成的9个光温催花条件下,高山杜鹃花芽内可溶性糖、淀粉、可溶性蛋白和游离氨基酸等营养物质的变化以及对始花期的影响。结果表明:随着昼夜温度的升高和光照强度的加强,高山杜鹃花芽内的可溶性糖和淀粉含量增加,且在整个催花过程中持续上升;可溶性蛋白含量变化整体呈先升后降的趋势,前期随温度光强增加而增加,后期则相反;游离氨基酸含量的整体变化趋势与可溶性蛋白含量相反,表现为先降后升。在30℃/24℃、10 000/8 000lx的处理下,高山杜鹃最早进入始花期,比目标日期(春节)提前18d开花。可见,适当高温和高光强(22℃/16℃~30℃/24℃,7 500/5 500~10 000/8 000lx)处理能显著提高高山杜鹃花芽内可溶性糖、淀粉、可溶性蛋白和游离氨基酸等营养物质的积累和代谢,促进了其花芽发育,有效缩短了催花时间,使植株提前进入始花期,从而达到提前上市的目的。     

Distribution of type III collagen in the pulp parenchyma of the human developing tooth. Light and electron microscope immunotyping

The distribution of collagen type III throughout the pulp tissue from human developing tooth was studied using specific antibodies, immuno-fluorescence as well as immuno-peroxidase labelling for electron microscopy. Our results indicate that type III and type I collagen are present in the pulp. The staining intensity seems to correlate with the relatively high proportions of type III collagen biochemically found in pulp. In addition, type III collagen and reticulin fibres are similarly distributed, except that the Von Korff fibres were never detected with anti-type III collagen antibodies. Correspondingly, at the ultrastructural level, type III collagen appears as fine, branched filaments or electron dense material distributed throughout the tissue and particularly in close association with the plasma membrane of pulp fibroblasts. In contrast, type I collagen appears as typical coarse cross banded fibres.