• Title/Summary/Keyword: Dark incubation

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Effects of Paraquat on Ascorbic Acid and Malondialdehyde Contents, and Superoxide Dismutase Activity in Spinach Chloroplasts under Light and Dark

  • Won-Hyuck Park;Yang-Hee Chang
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.81.1-86
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    • 1992
  • The amounts of ascorbic acid in chloroplasts treated with light and light+paraquat (PQ) were reduced by 81 and 82% of initial level, respectively at 24 hr at incubation. And those treated with dark and dark+PQ were decreased by W and 55% of the original level, respectively. Malondialdehyde (MDA) contents at 24 hr of dark and dark+PQ treatment were increased by 6 and 31% of the initial level, respectively. When chloroplasts were treated with light and light+PQ, MDA contents after 24 hr were increased by 88 and 146% of the initial level, respectively. SOD activities treated with light and light+PQ were increased by 10 and 20% of the initial level, respectively for 3 hr and thereafter reduced by 46 and 49% of the original level, respectively at 24 hr. However, the SOD activities treated with dark and dark+PQ were decreased by 37 and 30% of the initial level, respectively. It is considered that PQ triggers the oxidation of ascorbic acid, the induction of lipid peroxidation and the inactivation of SOD under light so that PQ has inhibitors effect on the pathway of plant metabolism. Key word: ascorbic acid, malondialdehyde, superoxide dismutase, paraquat, lipid peroxidation.

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Effects of Paraquat on Ascorbic Acid and Malondialdehyde Contents, and Superoxide Dismutase Activity in Spinach Chloroplasts under Light and Dark

  • Park Won-Hyuck;Chang Yang-Hee
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.1 no.2
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    • pp.81-86
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    • 1997
  • The amounts of ascorbic acid in chloroplasts treated with light and light+paraquat (PQ) were reduced by 81 and $82\%$ of initial level, respectively at 24 hr at incubation. And those treated with dark and dark+PQ were decreased by 46 and $55\%$ of the original level, respectively. Malondialdehyde (MDA) contents at 24 hr of dark and dark+PQ treatment were increased by 6 and $31\%$ of the initial level, respectively. When chloroplasts were treated with light and light+PQ, MDA contents after 24 hr were increased by 88 and $146\%$ of the initial level, respectively. SOD activities treated with light and light+PQ were increased by 10 and $20\%$ of the initial level, respectively for 3 hr and thereafter reduced by 46 and $49\%$ of the original level, respectively at 24 hr. However, the SOD activities treated with dark and dark+PQ were decreased by 37 and $30\%$ of the initial level, respectively, It is considered that PQ triggers the oxidation of ascorbic acid, the induction of lipid peroxidation and the inactivation of SOD under light so that PQ has inhibitory effect on the pathway of plant metabolism.

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The Effect of Light on the Production of Reserpine in Cultured Rauwolfia serpentina Cells

  • Yamamoto, Osamu
    • Natural Product Sciences
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    • v.2 no.2
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    • pp.90-95
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    • 1996
  • When reserpine-producing cell strains of Rauwolfia serpentina were transferred from the dark to the light irradiation, the production of reserpine was extremely enhanced whereas the cell growth was suppressed. In an incubation period of 20 days, the most effective culture condition for reserpine production was the combination of 8 days of dark culture and following 12 days of light culture. The time courses of both cell growth and reserpine production were measured in vitro in order to clarify the effect of wave length range of light on the biosynthesis of reserpine. Although the growth of cultured cells which had been incubated under continuous red, yellow, and green lights, respectively, was similar to that of the cultured cells subcultured in the dark. The cells cultured under red light irradiation produced less reserpine than dark-grown cultures. Both blue and near-ultraviolet light inhibited the growth of cultured cells. The production of reserpine was strikingly enhanced by blue light, but was strongly inhibited by near-ultraviolet light.

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Decrease of Photochemical Efficiency Induced by Methyl Viologen in Rice(Oryza sativa L.) Leaves is Partly due to the Down-Regulation of PSII

  • Kim, Jin-Hong;Lee, Choon-Hwan
    • Journal of Photoscience
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    • v.9 no.3
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    • pp.65-70
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    • 2002
  • In the rice leaves treated with methyl viologen (MV), the photochemical efficiency of PSII (or $F_{v/}$F $m_{m}$) was significantly decreased, and significant portion of the photoinactivation process was reversible during the dark-recovery. The dark-reactivation process was relatively slow, reaching its plateau after 2-2.5 h of dark incubation. The damaged portion of functional PSII was 13%, based on the value of I/ $F_{o}$- I/ $F_{m}$ after this dark-recovery period. The reversible photoinactivation process of PSII function in the MV-treated leaves consisted of a xanthophyll cycle-dependent development of NPQ and a xanthophyll cycle-independent process. The latter process was reversible in the presence of nigericin. As well as the increase in the values of Chl fluorescence parameters, the epoxidation process during the dark-recovery after the MV-induced photooxidation was very slow. These results suggest that the photooxidative effect of MV is partly protected by the down-regulation of PSII before inducing physical damages in core proteins of PSII.I.I.I.I.

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Sperm nuclear DNA fragmentation and chromatin structure in one-day-old ejaculated sperm

  • Jee, Byung-Chul;Suh, Chang-Suk;Shin, Mi-Sun;Lee, Hee-Jun;Lee, Jae-Ho;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.2
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    • pp.82-86
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    • 2011
  • Objective: To evaluate sperm nuclear DNA fragmentation and chromatin structure after 18 hours' incubation at room temperature. Methods: Twenty-eight male partners who participating IVF treatment were prospectively included in this study. Ejaculated sperm count and motility were assessed. The sperm was then immediately processed by the conventional swim-up method. After utilization of some of the sample for routine clinical use, the remainder of each of the samples was divided into two aliquots. One aliquot was immediately assessed for sperm nuclear DNA fragmentation (TUNEL assay) and chromatin structure (toluidine blue [TB] staining). The other aliquot was incubated at room temperature for 18 hours and then assessed by two methods. Only dark-TB sperms were considered as having abnormal chromatin structure. Data before and after extended incubation were compared using a paired Student's $t$-test. Results: Before and after extended culture, nuclear DNA fragmentation assessed by TUNEL was $4.9{\pm}4.7%$ and $7.0{\pm}6.4%$, respectively ($p$=0.008). The proportion of abnormal chromatin structure (dark-TB sperm) was $8.2{\pm}5.6%$ and $10.3{\pm}6.5%$ ($p$ <0.001), before and after incubation, respectively. Conclusion: After 18 hours' incubation at room temperature, sperm nuclear DNA and chromatin structure were significantly affected. The IVF practitioner should bear this information in mind when performing delayed insemination, especially for $in$ $vitro$ maturation cycles.

Leaf Senescence in a Stay-Green Mutant of Arabidopsis thaliana: Disassembly Process of Photosystem I and II during Dark-Incubation

  • Oh, Min-Hyuk;Kim, Yung-Jin;Lee, Choon-Hwan
    • BMB Reports
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    • v.33 no.3
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    • pp.256-262
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    • 2000
  • In this study the disassembly process of chlorophyII (ChI)protein complexes of a stay-green mutant (ore10 of Arabidopsis thaliana) was investigated during the dark incubation of detached leaves. During this dark-induced senescence (DIS), the Chi loss was delayed in the mutant, while the photochemical efficiency of photosystem II (PSII) or Fv/Fm was accelerated when compared with the wild type (WT) leaves. This indicates that the decrease in Fv/Fm is a separate process and not causally-linked to the degradation of Chi during DIS of Arabidopsis leaves. In the native green gel electrophoresis of the Chi-protein complexes, which was combined with an additional twodimensional SDS-PAGE analysis, the delayed senescence of this mutant was characterized by the appearance of an aggregate at 1 d or 2 d, as well as very stable light harvesting complex II (LHCII) trimers until 5 d after the start of DIS. The polypeptide composition of the aggregates varied during the whole DIS at 5 d. Dl protein appeared to be missing in the aggregates. This result supports the idea of a faster depletion of functional PSH in the mutants compared with WT, as suggested by the earlier reduction of Fv/Fm and the stable Chl a/b ratio in the mutants. At 5 d, the WT leaves also often showed aggregates, but the polypeptide composition was different from those of ore10. The results presented suggest that the formation of aggregates, or stable LHCII trimers in the stay-green mutants, is a way to structurally protect Chi-protein complexes from serious proteolytic degradation. Detailed disassembly processes of Chi-protein complexes in WT and ore10 mutants are discussed.

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The Role of $Ca^{2+}$ in Retardation Effects of Benzyladenine on the Senescence of Wheat (Triticum aestivum L.) Leaves

  • Hong, Kee-Jong;Jin, Chang-Duck;Hong, Young-Nam
    • Journal of Plant Biology
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    • v.39 no.2
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    • pp.113-121
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    • 1996
  • The role of Ca2+ on benzyladenine (BA)-induced senescence retardation in mature wheat (Triticum aestivum L.) primary leaves was investigated. When an extracellular calcium chelator, ethylene glycol-bis-($\beta$-aminoethylether)-N, N'-tetraacetic acid (EGTA) together with BA, was applied to senescing leaves for 4 days of dark incubation, the content of chlorophyll and soluble protein decreased rapidly. And, the content of malondialdehyde (MDA), known to be a degradation product of membrane lipids, increased compared with the BA alone control. The BA-EGTA combination also caused the stimulation of protease and RNase activity and a rapid loss of catalase activity owing to the decling of BA effects. In the case of treatment with only intracellular calcium antagonist 3, 4, 5-trimethoxybenzoic acid 8-(diethylamino) octyl ester (TMB-8) without the BA addition, the chlorophyll content at day 4 after dark incubation decreased in paralled with the increasing concentration of the antagonist. In addition, the chlorophyll content at 10-5 M calcium ionophore A23187 treatment in the absence of BA was similar to that of the BA alone treatment. These results suggest that calcium may mediate the retardation effect of BA on leaf senescence by acting as a second messenger and that the calcium input from cell organelles, as well as the calcium inflow from intercellular spaces and cell walls, may be involved in modulating cytosolic calcium levels related to BA action.

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Optimal culture methods for plant regeneration via shoot organogenesis in the 'Fuji' apple (사과 '후지'의 기관형성을 통한 식물체 재생에 효율적인 배양방법)

  • Yoon Kyung Lee;Youngju Kwon;Yong Joon Yang
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.176-182
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    • 2023
  • Plant regeneration protocols for adventitious shoot organogenesis from apple (Malus domestica 'Fuji') leaf explants were developed in the present study. The effects of dark incubation periods in the early stages of culture, pre-treatment methods, the number of explants per culture container, the type of culture containers, and the orientation of the explants on culture media were evaluated to determine the optimal shoot regeneration conditions for 'Fuji' apple leaf explants. Light incubation of explants produced minimal response. However, dark incubation of explants for 4 weeks during the initial culture period enhanced shoot regeneration frequency. Comparing the number of explants per container, a higher percentage of shoot regeneration was obtained with nine explants per container compared with four explants per container. Pre-treatment, before culture, by dipping explants in a liquid regeneration medium containing 40 g/L of sorbitol for 2 hours produced the highest shoot formation rate, and the time of shoot formation was accelerated. The percentage of shoot regeneration and number of shoots per regenerating explant reached a maximum of 87.5% and 4.7, respectively. The regenerated shoots were elongated and rooted on a rooting medium of 1/4 MS with 0.2 mg/L IBA. The plantlets were successfully acclimatized, and the regenerated plants produced normal phenotypes.

The Parallel Measurements of Stomatal Apertures and Apoplastic pH on Guard Cells from Epidermal Strips and Intact Leaves of Commelina communis

  • Lee, Joon-Sang
    • Korean Journal of Environmental Biology
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    • v.20 no.1
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    • pp.78-84
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    • 2002
  • The effects of light and darkness on stomatal aperture and guard cell apoplastic pH in the intact leaf and in the isolated epidermal strips of Commelina communis have been investigated. Stomata in the intact leaf opened wide in the light. In contrast, stomata in the isolated epidermal strips did not respond clearly to light. To eucidate the relationship between the stomatal aperture and the guard cell apoplastic pH, apoplastic pH was measured. In the light the guard cell wall of intact leaf was acidified by pH 1.9 units, falling from pH 7.3 to pH 5.4 in the first 10 minutes. On the contrary, apoplastic pH of isolated epidermal strips changed slowly from pH 7.3 to pH 6.9 at 20 min. Stomata in the intact leaf closed rapidly in the dark. On the other hand, stomata in the isolated epidermal strips failed to close in dark. There was a slow increase in apoplastic pH on transfer to the dark after incubation for 1.5 h in the light and the level observed before the experiment was regained after around 40 min. When the isolated epidermal strips were transferred to the dark, apoplastic pH maintained a uniform level of around pH 7.2-7.4. These results indicate that the mechanism of stomatal opening and closing from isolated epidermal strips and intact leaves could be different.

Production and Isolation of Chlamydospores in Cylindrocaupon destructans Causing Root Rot of Panax quinquefolium (미국삼(Panan quinquefolium)에서 분리한 뿌리썩음병균 Cylindrocarpon destructans의 후막포자 생성 및 분리)

  • 조대휘;유연현
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.304-309
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    • 1998
  • Incubation condition affecting the chlamydospore formation and isolation from mycelia and conidia of Cylindrocarpon destructanse (isolate ACY-9701), isolated from the root rot lesion of the American ginseng (Panax quinquefolium) was investigated. Chlamydospores were formed from mycilia but not from conidia on the Czapek-Dox agar without carbon or nitrogen source after 20 days incubation at 2$0^{\circ}C$. In the medium added with nitrogen and carbon sources, immatured chlamy-dospore-like cells were formed from microconidia and mycelia as well. Immatured chlamydospore-like cells were formed from mycelia as well as microconidia In corn, kidney bean, and pea root extracts after 20 days incubation at 20"C, while typical chlamydospores were formed from both of them in the root extract of Panax quinquefolium. The 3.6 log chlamydospore/mm" was converted from microconidia in the medium, which was equal to 2.5% conidia formed. Under the light condition (251.1 pmol/m" sec, 12 hrs dark and light cycle), 4.2 log/mm" of chlamydospores were converted from interracially or terminal cells of macroconidia, which was 4.0% of macroconidia produced on Potato dextrose agar (PDA). When mycelia and microconidia were stored at -7$0^{\circ}C$ for 32 days and incubated on PDA after thawing at room temperature to isolate chlamydospores from them, microconidia and mycelia were still alive. Meanwhile, microconidial lysis was found after heating them at 32$^{\circ}C$ for 7 days, but the chlamydospores converted from macroconidia were not lysed up to 13 days at 32"C. to 13 days at 32"C.ot;C.

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