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1.
Cell suspension cultures of parsley and soybean were incubated for 44 to 48 h with14C-labeled DDT or Kelthane; autoclaved cultures were used as controls. Most of the radioactivity became associated with the cells, and metabolites were isolated by a sequential extraction procedure. The metabolites amounted to 0.6 to 2.2% of the applied pesticide. Relatively non-polar metabolites were identified as DDE in the case of DDT, and remained unidentified in the case of Kelthane. Polar metabolites were also isolated and are as yet unidentified. They were chromatographically different from the known and less polar metabolites of DDT and Kelthane reported from animal and insect studies. [DDT-1,1,1-Trichloro-2,2-bis-(4-chlorophenyl)-ethane; Kelthane=(1,1-bis-(4-chlorophenyl)-2,2,2-trichloro-ethanol; DDE=1,1-Dichloro-2,2-bis-(4-chlorophenyl)-ethylene.]Abbreviations DDT
1,1,1-Trichloro-2,2-bis-(4-chlorophenyl)-ethane
- Kelthane
(1,1-bis-(4-chlorophenyl)-2,2,2-trichloro-ethanol
- DDE
1,1-Dichloro-2,2-bis-(4-chlorophenyl)-ethylene
- DDA
2,2-bis-(4-chlorophenyl)-acetic acid
- DDOH
2,2-bis-(4-chlorophenyl)-ethanol
- DDD
1,1-Dichloro-2,2-bis-(4-chlorophenyl)-ethane
- DBP
4,4-Dichloro-benzophenone
- DDMU
1-Chloro-2,2-bis-(4-chlorophenyl)-ethylene
- DDM
Bis-(4-chlorophenyl)-methane
- FW-152
1,1-Bis-(4-chlorophenyl)-2,2-dichloro-ethanol
- SDS
sodium dodecylsulphate 相似文献
2.
Kimitoshi Fukunaga Yurie Mine Ken-ichi Samejima Makoto Yoshimoto Katsumi Nakao Yoshiaki Sugimura 《Biotechnology letters》2002,24(14):1157-1160
Modified Candida rugosa lipase was co-lyophilized with two gemini-type amphiphiles, l- and d-2-(3-bis-[3-(2,3,4,5,6-pentahydroxy-hexanoylamino)-propyl]-carbamoyl -propionylamino)-pentanedioic acid didodecyl ester or dodecanoic acid 2-[(3-bis-[3-(2,3,4,5,6-pentahydroxy-hexanoylamino)-propyl]-carbamoyl -propionyl)-(2-dodecanoyloxy-ethyl)-amino]-ethyl ester. Enzymatic activities of the modified lipases in the transesterification between racemic 2,2-dimethyl-1,3-dioxolane-4-methanol and vinyl butyrate in cyclohexane were enhanced as much as by 37-78, 1.5–5- and 41–83-fold of magnitude relative to that of native enzyme, respectively. The lack of significant enhancement of the enzymatic activity, only in the case of the d-isomeric amphiphile-modified lipase, was considered from the topological view of the amphiphile. 相似文献
3.
Trigonelline (TRG), which act as a cell cycle regulator and a compatible solute in response to salinity and water-stress, is the N-methyl conjugate of nicotinic acid the formation of which is catalyzed by S-adenosyl-L-methionine nicotinic acid-N-methyltransferase. The enzyme was purified 2650-fold from soybean (Glycine max L.) leaves with a recovery of 4 %. The purification procedure included ammonium sulfate (45 – 60 %) precipitation, linear gradient DEAE-Sepharose chromatography, adenosine-agarose affinity chromatography, hydroxyapatite chromatography and gel filtration (Sephacryl-S-200). The purified enzyme preparation showed a major band with a molecular mass of 41.5 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis that is related to the enzyme activity. The native enzyme had a molecular mass of about 85 kDa as estimated by gel filtration. The Km values for S-adenosyl-L-methionine and nicotinic acid were 31 and 12.5 M, respectively. The purified enzyme showed optimum activity at pH 6.5 and temperature of 40 – 45 °C. High concentration of dithiothreitol (10 mM) and glycerol (20 %) stabilize the enzyme during purification and storage. Hg2+ strongly inhibits enzyme activity. 相似文献
4.
Oscillatoria amphigranulata is a fast-growing (3 doublings/day) cyanobacterium isolated from sulfide hot springs in New Zealand. Photosynthesis, as measured by incorporation of [14C]-HCO
3
-
, was initially inhibited by 0.3–1.5 mM sulfide at pH 7.9–8.1. However, conversion to sulfide-dependent anoxygenic photosynthesis occurred in about 2 h or less under light intensities of 3–14 klx. Under the stimulation of higher light intensity (8–14 klx) a partial recovery of oxygenic photosynthesis also occurred. It was concluded that oxygenic photosynthesis was responsible for 21–42% of the total incorporation at sulfide concentrations of 1.0–0.3 mM, respectively. This contribution was suppressed at 1.5 mM sulfide and not elicited under lower light intensities (3–7 klx). As judged by the inhibitory effect of 10 g/ml chloramphenicol protein synthesis was required for attainment of both anoxygenic photosynthesis and photosystem II recovery. Sulfide could not be replaced by thiosulfate, elemental sulfur or dithionite as electron donors in photosynthesis, but elemental sulfur could serve as the sole assimilatory source of sulfur. Oxygenic photosynthesis was inhibited by DCMU [3-(3,4-dichlorophenyl)-1,1-dimethylurea] or DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone), but sulfide relieved the effect of either inhibitor in adapted cells, indicating that electrons derived from sulfide enter the photosynthetic electron transport chain at a point beyond plastoquinone.Uncommon abbreviations DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- DBMIB
2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone
- DSPD
disalicyclidene propanediamine
- DNP-INT
2-4-dinitrophenyl ether of 2-iodo-4-nitrothymol
- TMPD
N,N,N,N-tetramethyl-p-phenylenediamine
- PPO
2,5-diphenyloxazole
- POPOP
1,4-bis-2-(5-phenyl oxzolyl) benzene 相似文献
5.
1. Cooperation between phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) and cinnamic acid hydroxylases was investigated using microsomal fractions from cotyledons of cucumber (Cucumis sativus L.). The interpretations were based on experiments which demonstrate a limited exchange between the pool of cinnamic acid formed by the membrane-bound phenylalanine ammonia-lyase and the cinnamic acid pool external to the enzyme-membrane system. 2. The extent of cooperation between the microsomal enzymes was proved to be influenced by treatment of the cotyledons with light. On exposure to UV-light, which is known to enhance greatly the soluble phenylalanine ammonia-lyase activity in cell cultures, differential effects on the levels of microsomal and soluble phenylalanine ammonia-lyase, and of cinnamic acid hydroxylases, were observed. The time course of the enzyme activities and their cooperation in vitro after treatment of the cotyledons with light were studied. 3. The extent of cooperation in vitro was found to vary depending on the concentration of L-phenylalanine. 4. Homogenates obtained from etiolated cotyledons of Cucumis sativus in the absence of Mg2+ were fractionated by sucrose density gradient centrifugation and examined for phenylalanine ammonia-lyase, cinnamic acid o-hydroxylase, cinnamic acid o-hydroxylase, and several marker enzymes. Ammonia-lyase activity was highest in fractions with 25% sucrose, in which primarily smooth endoplasmic reticulum is localized. Hydroxylase activities co-occur with phenylalanine ammonia-lyase in these fractions (density=1.100 g/cm3), and also in fractions at higher densities (d=1.12–1.13 and 1.15 g/cm3).Abbreviations PAL
L-phenylalanine ammonia-lyase
- Tris
tris-(hydroxymethyl)aminomethane
- EDTA
ethylenediamine tetraacetic acid
- ATPase
ATP phosphohydrolase 相似文献
6.
The increase in extractable phenylalanine ammonia-lyase (PAL;EC 4.3.1.5.) activity induced in French bean cell suspension cultures in response to treatment with autoclaved ribonuclease A was inhibited by addition of the phenylpropanoid pathway intermediates cinnamic acid, 4-coumaric acid or ferulic acid. The effectiveness of inhibition was in the order cinnamic acid>4-coumaric acid>ferulic acid. Cinnamic acid also inhibited the PAL activity increase induced by dilution of the suspensions into an excess of fresh culture medium. Addition of low concentrations (<10-5M) of the pathway intermediates to cultures at the time of application of ribonuclease gave variable responses ranging from inhibition to 30–40% stimulation of the PAL activity measured at 8 h. Following addition of pathway intermediates to cultures 4–5 h after ribonuclease treatment, rapid increases followed by equally rapid declines in PAL activity were observed. The cinnamic acid-stimulated increase in enzyme activity was unaffected by treatment with cycloheximide at a concentration which gave complete inhibition of the ribonuclease-induced response. However, cycloheximide completely abolished the subsequent decline in enzyme activity. Treatment of induced cultures with -aminooxy--phenylpropionic acid (AOPPA) resulted in increased but delayed rates of enzyme appearance when compared to controls not treated with the phenylalanine analogue. The results are discussed in relation to current views on the regulation of enzyme levels in higher plants.Abbreviations AOPPA
-aminooxy--phenylpropionic acid
- PAL
L-phenylalanine ammonia-lyase (EC 4.3.1.5)
- AOA
-aminooxyacetic acid 相似文献
7.
The regulation of purified glutathione S-transferase from rat liver microsomes was studied by examining the effects of various sulfhydryl reagents on enzyme activity with 1-chloro-2,4-dinitrobenzene as the substrate. Diamide (4 mM), cystamine (5 mM), and N-ethylmaleimide (1 mM) increased the microsomal glutathione S-transferase activity by 3-, 2-, and 10-fold, respectively, in absence of glutathione; glutathione disulfide had no effect. In presence of glutathione, microsomal glutathione S-transferase activity was increased 10-fold by diamide (0.5 mM), but the activation of the transferase by N-ethylmaleimide or cystamine was only slightly affected by presence of glutathione. The activation of microsomal glutathione S-transferase by diamide or cystamine was reversed by the addition of dithiothreitol. Glutathione disulfide increased microsomal glutathione S-transferase activity only when membrane-bound enzyme was used. These results indicate that microsomal glutathione S-transferase activity may be regulated by reversible thiol/disulfide exchange and that mixed disulfide formation of the microsomal glutathione S-transferase with glutathione disulfide may be catalyzed enzymatically in vivo. 相似文献
8.
The characteristics of the conversion of 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene by pea (Pisum sativum L.) epicotyls and by pea epicotyl enzyme are compared. Of the four stereoisomers of 1-amino-2-ethylcyclopropane-1-carboxylic acid (AEC), only (1R,2S)-AEC is preferentially converted to 1-butene in pea epicotyls. This conversion is inhibited by ACC, indicating that butene production from (1R,2S)-AEC and ethylene production from ACC are catalyzed by the same enzyme. Furthermore, pea epicotyls efficiently convert ACC to ethylene with a low K
m (66 M) for ACC and do not convert 4-methylthio-2-oxo-butanoic acid (KMB) to ethylene, thus demonstrating high specificity for its substrate. In contrast, the reported pea epicotyl enzyme which catalyzes the conversion of ACC to ethylene had a high K
m (389 mM) for ACC and readily converted KMB to ethylene. We show, moreover, that the pea enzyme catalyzes the conversion of AEC isomers to butene without stereodiscrimination. Because of its lack of stereospecificity, its low affinity for ACC and its utilization of KMB as a substrate, we conclude that the reported pea enzyme system is not related to the in-vivo ethylene-forming enzyme.Abbreviations ACC
1-Amino cyclopropane-1-carboxylic acid
- AEC
1-amino-2-ethylcyclopropane-1-carboxylic acid
- EFE
ethylene-forming enzyme
- KMB
4-methylthio-2-oxobutanoic acid 相似文献
9.
Dieter Strack 《Planta》1982,155(1):31-36
Protein preparations from cotyledons of red radish (Raphanus sativus L. var. sativus) catalyzed the the formation of depsides between cinnamic acids and L-malate, using 1-O-acyl glucose conjugates as the donors. This activity showed an absolute acceptor specificity towards L-malate and a pronounced donor specificity with 1-sinapoylglucose (1-O-sinapoyl--D-glucose). Maximal rate of sinapoyl-L-malate formation was found to be at pH 6.3, and there was no requirement for metal ions or sulfhydryl group reagents. The K
m values were found to be 0.46 mM for 1-sinapoylglucose and 54 mM for L-malate. Protein extracts obtained from seedlings at different stages of seedling development did not significantly differ with respect to the properties of the enzymatic activity. Appearance and development of extractable activities correlated well with the in vivo transacylation kinetics of 1-sinapoylglucose to sinapoyl-L-malate during seedling growth. Maximal activity was extracted from 10–14-d-old seedlings and found to be at 67 pkat pair-1 of cotyledons. This new enzymatic activity in phenylpropanoid metabolism refers to an enzyme which can be classified as 1-sinapoylglucose: L-malate sinapoyltransferase (SMT) (EC 2.3.1.-).Abbreviations DTE
dithioerythriol
- HPLC
high performance liquid chromatography
- IAA
indoleacetic acid
- ME
2-mercaptoethanol
- Mes
2-(N-morpholino)ethanesulfonic acid
- Mops
3-(N-morpholino)propanesulfonic acid
- SMT
1-O-Sinapoyl--D-glucose: L-malate sinapoyltransferase 相似文献
10.
Tarun Bhowmik Mark Lueck J. L. Steele 《Journal of industrial microbiology & biotechnology》1993,12(1):35-41
Summary
d-(–)-Lactate dehydrogenase (LDH) was purified to homogeneity from a cell-free extract ofLactobacillus helveticus CNRZ 32. The native enzyme was determined to have a molecular weight of 152 000 and consisted of four identical subunits of 38 000. This enzyme was NAD dependent fructose 1,6-diphosphate (FDP) and ATP independent. It was most active on pyruvate followed by -hydroxypyruvate as substrates. TheK
m values for pyruvate andd-(–)-lactate were 0.64 and 68.42 mM respectively, indicating that the enzyme has a higher affinity for pyruvate. The enzyme activity was completely inhibited byp-chloromercuribenzoate (1 mM) and partially by iodoacetate, suggesting the involvement of the sulfhydryl group (-SH) in catalysis. Optima for activity by the purified enzyme were pH 4.0 and 50–60°C. Limited inhibition ofd-(–)-LDH was observed with several divalent cations. Additionally, HgCl2 was observed to strongly inhibit enzyme activity. The purified enzyme was not affected by dithiothreitol or any of the metal chelating agents examined. 相似文献
11.
Glutathione reductase activity was detected and characterized in heterocysts and vegetative cells of the cyanobacterium Nostoc muscorum. The activity of the enzyme varied between 50 and 150 nmol reduced glutathione· min-1·mg protein-1, and the apparent Km for NADPH was 0.125 and 0.200 mM for heterocysts and vegetative cells, respectively. The enzyme was found to be sensitive to Zn+2 ions, however, preincubation with oxidized glutathione rendered its resistance to Zn+2 inhibition. Nostoc muscorum filaments were found to contain 0.6–0.7mM glutathione, and it is suggested that glutathione reductase can regenerate reduced glutathione in both cell types. The combined activity of glutathione reductase and isocitrate dehydrogenase in heterocysts was as high as 18 nmol reduced glutathione·min-1·mg protein-1. A relatively high superoxide dismutase activity was found in the two cell types; 34.2 and 64.3 enzyme units·min-1·mg protein-1 in heterocysts and vegetative cells, respectively.We suggest that glutathione reductase plays a role in the protection mechanism which removes oxygen radicals in the N2-fixing cyanobacterium Nostoc muscorum.Abbreviations DTNB
5-5-dithiobis-(2-nitrobenzoic acid)
- EDTA
ethylenediaminetetra-acetic acid
- GR
glutathione reductase (EC1.6.4.2)
- GSH
reduced glutathione
- GSSG
oxidized glutathione
- OPT
O-phtaldialdehyde
- SOD
superoxide dismutase (EC 1.15.1.1) 相似文献
12.
An H+ ATPase at the plasma-membrane of guard cells is thought to establish an electrochemical gradient that drives K+ and Cl– uptake, resulting in osmotic swelling of the guard cells and stomatal opening. There are, however, conflicting results regarding the effectiveness of the plasma-membrane H+-ATPase inhibitor, vanadate, in inhibiting both H+ extrusion from guard cells and stomatal opening. We found that 1 mM vanadate inhibited light-stimulated stomatal opening in epidermal peels of Commelina communis L. only at KCl concentrations lower than 50 mM. When impermeant n-methylglucamine and HCl (pH 7.2) were substituted for KCl, vanadate inhibition was still not observed at total salt concentrations50 mM. In contrast, in the absence of Cl–, when V2O5 was used to buffer KOH, vanadate inhibition of stomatal opening occurred at K+ concentrations as high as 70 mM. Partial vanadate inhibition was observed in the presence of the impermeant anion, iminodiacetic acid (100 mM KHN(CH2CO2H)2). These results indicate that high concentrations of permeant anions prevent vanadate uptake and consequently prevent its inhibitory effect. In support of this hypothesis, an inhibitor of anion uptake, anthracene-9-carboxylic acid, partially prevented vanadate inhibition of stomatal opening. Other anion-uptake inhibitors (1 mM 4,4-diisothiocyanatostilbene-2,2-disulfonic acid, 1 mM 4-acetamido-4-isothiocyanostilbene-2,2-disulfonic acid, 200 M Zn2+) were not effective. Decreased vanadate inhibition at high Cl–/vanadate ratios may result from competition between vanadate and Cl– for uptake. Unlike metabolic inhibitors, vanadate did not affect the extent of stomatal closure stimulated by darkness, further indicating that the observed action of vanadate represents a specific inhibition of the guard-cell H+ ATPase.Abbreviations DIDS
4,4-diisothiocyanatostilbene-2,2-disulfonic acid
- FC
fusicoccin
- SITS
4-acetamido-4-isothiocyanostilbene-2,2-disulfonic acid
We thank Drs. R.T. Leonard (University of California, Riverside, USA) and K.A, Rubinson (Yellow Springs, Oh., USA) for helpful comments on the research, Janet Sherwood (Harvard University) for excellent plant care, and Angela Ciamarra, Anne Gershenson, Gustavo Lara (Harvard University) and Orit Tal (Hebrew University) for valuable technical assistance. This research was supported by a grant from the National Science Foundation (DCB-8904041) to S.M.A. 相似文献
13.
Incidence of Fusarium wilt in Cucumis sativus L. is promoted by cinnamic acid, an autotoxin in root exudates 总被引:4,自引:1,他引:4
The effects of Fusarium oxysporum f. sp. cucumerinum, the pathogen causing Fusarium wilt in cucumber and cinnamic acid, a principal autotoxic component in the root exudates of cucumber (Cucumis sativus L.), on plant growth, Photosynthesis and incidence of Fusarium wilt in cucumber were studied in order to elucidate the interaction of autotoxins and soil-borne pathogens in the soil sickness. F. oxysporum. f. sp. cucumerinum (FO) and cinnamic acid (CA) at 0.1 or 0.25 mM significantly decreased net photosynthetic rate, stomatal conductance and the quantum yield of Photosystem II photochemistry (PSII), followed by a reduction of plant biomass production, but did not induce photoinhibition. Pretreatment with CA before inoculation with FO increased the effectiveness of FO, together with a slight photoinhibition. CA pretreatment significantly increased percentage of plants affected by Fusarium wilt, browning index of vascular bundle and Fusarium population in the nutrient solution. All these results indicate that CA enhanced Fusarium wilt by predisposing cucumber roots to infection by FO through a direct biochemical and physiological effect. It is likely that soil sickness results from an interaction of many factors.
Abbreviations: CA – cinnamic acid; CFU – colony forming units; Ci – intercellular CO2 concentration; FO –Fusarium oxysporum (Schlechtend.:Fr) f. sp. cucumerinum (Owen) Snyder & Hansen; Fv/Fm– maximal quantum efficiency of PS II; Fv/Fm– the efficiency of excitation capture by open PS II centers; Gs – stomatal conductance; NPQ – non-photochemical quenching coefficient; Pn – net photosynthetic rate; PS II – Photosystem II; PS II– the quantum yield of PS II photochemistry; qP – photochemical quenching coefficient. 相似文献
14.
d-Ribulose-1,5-bisphosphate carboxylase/oxygenase has been purified 80-fold from malate-grown Thiocapsa roseopersicina by salting out the enzyme from the high-speed supernatant between 68–95% saturation with respect to (NH4)2SO4, gelfiltration through Sephadex G-100, and DEAE-cellulose chromatography followed by sedimentation into a 14–34% glycerol gradient. The specific activity of enzyme for the carboxylase reaction was 2.45 mol RuBP-dependent CO2 fixed/min · mg protein (at pH 8.0 and 30° C) and for the oxygenase reaction was 0.23 mol RuBP-dependent O2 consumed/min · mg protein (at pH 8.6, and 25° C). The enzyme, which was ultracentrifugally homogeneous in the presence of 4 and 10% v/v glycerol, was stable for at least one year at-80° C in the presence of 10% glycerol. S20, w values obtained in the presence of 4 and 10% glycerol were 19.3 and 16.2, respectively. The enzyme contained both large (53,000-daltons) and mixed small subunits (15,000- and 13,500-daltons).Borate-dependent inactivation of the enzyme by 2,3-butadione, which was greatly reduced in the presence of the product 3-phosphoglycerate, suggested that one or more arginines are at the active site.Abbreviations DTT
dithiotreitol
- RuBP
d-ribulose-1,5-bisphosphate
- SDS
sodium dodecylsulfate
- TCA
trichloroacetic acid
- TEMBDG
buffer (pH 8.0 at 25°C) containing 20 mM Tris, 1 mM disodium EDTA · 2 H2O, 10 mM MgCl2·6 H2O, 50 mM NaHCO3, 0.1 mM DTT and 10% glycerol (v/v) 相似文献
15.
Prolyl hydroxylase, which is responsible for the hydroxylation of peptidyl proline residues, has been isolated and purified from the green alga Chlamydomonas reinhardii. The enzyme, which appears to be loosely associated with microsomal membranes, was released into solution by sonication in the presence of detergent. Purification was achieved by ion-exchange chromatography followed by affinity chromatography using the immobilized substrate poly-L-proline. Apart from its differing substrate specificity the enzyme appears to possess similar molecular characteristics to prolyl hydroxylase isolated from animal tissues: the active enzyme is a tetramer of about 240–250 kDa and nonidentical monomers of 65 and 60 kDa. The monomers are capsule shaped having a dimension of 12×7 nm.Abbreviations Da
dalton
- DEAE
diethylaminoethyl
- DTT
dithiothreitol
- Hepes
4-(2-hydroxymethyl)-1-piperazine ethanesulfonic acid
- -KGA
-ketoglutarate
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecyl sulfate 相似文献
16.
Augmentation of H2 photoproduction in Rhodopseudomonas palustris by N-heterocyclic aromatic compounds 总被引:1,自引:0,他引:1
Increases of 23- (5.6 mmol acetylene reduced mg dry wt–1) and 16- (4 mmol acetylene reduced mg dry wt–1) fold in nitrogenase activity and 12- (671 l H2 mg dry wt–1 h–1) and 6- (349 l mg dry wt–1 h–1) fold in H2 photoproduction in Rhodopseudomonas palustris JA1 over 24 h were achieved with pyrazine 2-carboxylate (3 mM) and 3-picoline (3 mM), respectively, and were higher than earlier reports of enhancement (1.5 to 5- fold) in biological H2 production using various alternative methods. 相似文献
17.
Spinach (Spinacia oleracea L.), broad bean (Vicia faba L.) and beech (Fagus sylvatica L.) plants were exposed to ozone at concentrations often measured in air during the summer months (120–300 g·m–3) and antioxidants were determined in the leaf tissue and in the aqueous phase of the cell wall, the apoplasm. Concentrations of both reduced ascorbate (AA) and its oxidized form, dehydroascorbate (DHA), showed the tendency to increase transiently in the apoplasm of spinach leaves 6–24 h after starting fumigation with ozone. In beech leaves, apoplasmic AA and DHA increased 3–7 d after beginning of treatment. At the very high concentration of 1600 g O3·m–3, an increase of apoplasmic AA was already measured after 1 d in beech leaves. Apparently, spinach and beech leaves respond to oxidative stress by increasing AA transport into the apoplasm and by accelerating DHA export. In contrast to these observations, DHA accumulated during 3 d of fumigation with only 120 g O3·m–3 in the apoplasm of broad bean leaves, while AA contents did not increase. After termination of fumigation, the extracellular redox state of ascorbate normalized within 1 d. Glutathione could not be detected in the apoplasm of any of the three leaf species. Intracellular AA changed its redox state in response to exposure to elevated concentrations of ozone. After 4–6 weeks of fumigation with 200–300 g O3·m–3 an increase of intracellular DHA was measured in beech leaves. At the same time, chlorophyll contents decreased and characteristic symptoms of ozone damage could be observed. However, no significant change in the redox state of apoplasmic ascorbate could be detected in beech leaves. Evidently, detoxification of ozone by apoplasmic AA was insufficient to protect the leaf tissue. Fumigation with a high ozone concentration (1600 g·m–3) caused an appreciable increase in the cellular contents of the oxidized forms of ascorbate and glutathione in beech leaves. Whereas in spinach leaves intracellular antioxidant contents and redox states were not altered during fumigation with 120–240 g O3·m–3, in broad bean leaves the intracellular DHA concentration increased and intracellular ascorbate became more oxidized after fumigation of the plants with 120 g O3·m–3. Apparently, broad bean leaves are more sensitive to ozone than beech and spinach leaves.Abbreviations AA
ascorbate, reduced form
- DHA
ascorbate, oxidized form (dehydroascorbate)
- FW
fresh weight
- GSH
glutathione, reduced form
- GSSG
glutathione, oxidized form
- IWF
intercellular washing fluid
- Vair
intercellular air space volume of leaves
- Vapo
apoplasmic water volume of leaves
This work was supported within the Sonderforschungsbereich 251 of the University of Würzburg. 相似文献
18.
The levels of both tightly and loosely bound Mn in chloroplasts from fresh, cold and dark stored as well as illuminated leaves of Lycopersicon esculentum Mill. were studied in relation to Hill reaction activity. The tightly bound Mn pool represents one third of the total Mn content in chloroplasts isolated from the fresh leaves, and its level does not change following cold storage and illumination of leaves. Upon cold and dark storage of leaves the loss from the chloroplasts of about 40%–50% of the total amount of Mn is accompanied by an almost complete inactivation of the Hill reactions, as studied with water as an electron donor, as well as by the appearance of an EPR signal characteristic of free Mn2+ ions. Following illumination of such leaves, the restroration of Hill reaction activity is accompanied by an increase in the total Mn content in chloroplasts of up to 70%–80% of the Mn level measured in the fresh leaves and by disappearance of the EPR signal. In contrast, aging in the cold of isolated chloroplasts does not affect their Mn content. The addition of manganese does not result in the restoration of Hill reaction activity in chloroplasts from cold stored leaves but causes a restoration of this activity inhibited by linolenic acid. The data suggest that the loosely bound Mn pool (extractable with Tris) can be differentiated into two fractions: (1) one functionally inactive in electron transport and (2) one responsible for restoration of Hill reaction activity. Mn of the latter fraction (about 45% of the total Mn content) probably originates from the free Mn ions present in the interior of the chloroplasts following the cold and dark storage of leaves and from Mn reincorporated into chloroplasts from the cytoplasm. Incorporation of Mn from both these sources into thylakoid membrane to form a functionally active, loosely bound Mn pool proceeds during the illumination of leaves and results in the restoration of Hill reaction activity inhibited following the storage of leaves in dark and cold.Abbreviations Chl
chlorophyll
- DCIP
2,6-dichlorophenolindophenol
- Diquat
1,1-ethylene 2,2-dipiridylium dibromide
- EPR
electron paramagnetic resonance
- FFA
free fatty acid
- MV
methyl viologen, N,N-dimethyl-4,4 dipyridyldihydrochloride
- Tris
tris-(hydroxymethyl) aminomethane 相似文献
19.
Calcium-mediated changes in peroxidase and O-diphenol oxidase activities of cotton fibres (Gossypium spp.) and its possible relationship to ABA 总被引:3,自引:0,他引:3
A. S. Basra R. S. Sarlach Rewa Dhillon-Grewal C. P. Malik 《Plant Growth Regulation》1992,11(2):159-164
This work is an investigation of the influence of ABA and calcium on soluble and wall-bound activities of peroxidase and O-diphenol oxidase of cotton fibres at the stages of primary and secondary wall development, during incubation of intact fibres for 3h at 28°C. ABA (10 M) caused marked inhibition of enzyme activities in both the fractions, whereas calcium (1 mM) was promotory. The incorporation of 1 mM EGTA (a calcium chelator) and chlorpromazine (10 g cm–3) (a calmodulin antagonist) resulted in decreased enzyme activities suggesting regulation of enzyme synthesis and/or secretion to the cell wall by calcium-calmodulin. As a general trend, the relative effect of Ca2+ on the activity of peroxidase in the wall was much greater than on the soluble activity, but this was not true of O-diphenol oxidase. It is inferred that ABA inhibits enzymic activities by inhibiting calmodulin synthesis or its mobilization to sites of action.Abbreviations ABA
abscisic acid
- EGTA
ethylene glycol-bis( amino-ethyl ether)N,N tetraacetic acid
- DAA
days after anthesis 相似文献
20.
M. Barghchi 《Plant Cell, Tissue and Organ Culture》1988,15(3):233-244
Procedures were developed for micropropagation of Alnus cordata through in vitro axillary shoot multiplication of axillary bud explants cultured in Murashige & Skoog (MS) medium. Establishment of cultures from plants grown in the field was very difficult due to bacterial contamination and phenolic oxidation in explants causing severe browning. Explants were first cultured on an MS medium containing 4.4 M 6-benzyladenine and 87.6 mM sucrose (initiation medium) for 7 days and then transferred to an MS medium containing 1.1 M 6-benzyladenine and 333 mM glucose (multiplication medium) for a further 20–25 days. It was necessary to transfer cultures from initiation medium to multiplication medium after 7 days to minimize excessive callus growth, abnormally thick and brittle leaves, inhibition of shoot elongation, and senescence. Shoot multiplication comparable to the above method was achieved by culture of axillary bud explants in MS medium supplemented with 1.1–4.4 M 6-benzyladenine and 333 mM glucose 4–5 weeks after culture. Shoots rooted in MS medium (1/2 x macro-nutrients) supplemented with 1.2–4.9 M indolebutyric acid. Also, 98% rooting was achieved when cultures were treated with 625 mgl-1 indolebutyric acid for 24 h at the end of the shoot production stage and rooted in vivo as mini-cuttings. Plantlets established well in soil. 相似文献