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1.
A purified preparation of arginine decarboxylase fromCucumis sativus seedlings displayed ornithine decarboxylase activity as well. The two decarboxylase activities associated with the single
protein responded differentially to agmatine, putrescine andPi. While agmatine was inhibitory (50 %) to arginine decarboxylase activity, ornithine decarboxylase activity was stimulated
by about 3-fold by the guanido arnine. Agmatine-stimulation of ornithine decarboxylase activity was only observed at higher
concentrations of the amine. Inorganic phosphate enhanced arginine decarboxylase activity (2-fold) but ornithine decarboxylase
activity was largely uninfluenced. Although both arginine and ornithine decarboxylase activities were inhibited by putrescine,
ornithine decarboxylase activity was profoundly curtailed even at 1 mM concentration of the diamine. The enzyme-activated
irreversible inhibitor for mammalian ornithine decarboxylase,viz. α-difluoromethyl ornithine, dramatically enhanced arginine decarboxylase activity (3–4 fold), whereas ornithine decarboxylase
activity was partially (50%) inhibited by this inhibitor. At substrate level concentrations, the decarboxylation of arginine
was not influenced by ornithine andvice-versa. Preliminary evidence for the existence of a specific inhibitor of ornithine decarboxylase activity in the crude extracts
of the plant is presented. The above results suggest that these two amino acids could be decarboxylated at two different catalytic
sites on a single protein. 相似文献
2.
A simple, reproducible and rapid protocol for the purification of arginine decarboxylase fromCucumis sativus seedlings has been standardised. The purification steps involved ion-exchange chromatography on diethylaminoethyl-cellulose
followed by gel filtration on Sephadex G-l 50. The purified enzyme preparation migrated as a single stainable band on Polyacrylamide
gels at both basic and acidic pH, but under denaturing and reducing conditions on sodium dodecyl sulphate-polyacrylamide gels
resolved into polypeptides of molecular weight 48,000,44,000 and 15,000. However, in the absence of 2-mercaptoethanol on electrophoresis
on sodium dodecyl sulphate-polyacrylamide gels, the enzyme moved as single band with a molecular weight of 150,000. Evidence
was obtained to indicate that these three polypeptides were probably derived from a single larger molecular weight enzyme.
On storage of the purified protein, the 48,000 species was preferentially degraded to smaller polypeptides. The preliminary
data suggested that the 48,000 and 44,000 species shared many common tryptic peptides as revealed by finger printing of the
[125I ]-labelled protein. The purified enzyme was a glycoprotein and had aK
m
of 0.5 mM for arginine. Its activity was stimulated by dithiothrietol and pyridoxal phosphate. EDTA did not inhibit the enzyme
activity. Mn2+ at 1 mM stimulated arginine decarboxylase activity but was inhibitory at higher concentration 相似文献
3.
Arginine decarboxylase (arginine carboxy-lyase EC 4.1.1.19) of Cucumis sativus cotyledons, has a pH optimum of 8.3 and a temperature optimum of 40°. Among the various plant hormones administered to excised cotyledons in culture, benzyladenine and its riboside were most effective in increasing the arginine decarboxylase activity and putrescine content. The enzyme activity and putrescine content were significantly increased on acid feeding of the cotyledons and decreased by KCl treatment. The KCl effect could be only partially reversed by benzyladenine. Abscisic acid inhibited cotyledon growth and also reduced arginine decarboxylase and putrescine levels. This effect was overcome by cytokinins. The half life of the enzyme using cycloheximide was 3.7 hr. Dibutyryl cyclic AMP and 5′-AMP also marginally stimulated the enzyme and putrescine levels. Mixing experiments indicate that there is neither a non-dialysable activator nor inhibitor of the enzyme. 相似文献
4.
Arginine decarboxylase (polyamine synthesis) mutants of Arabidopsis thaliana exhibit altered root growth 总被引:1,自引:0,他引:1
Mark B. Watson Kimberly K. Emory Ruth Marie Piatak & Russell L. Malmberg 《The Plant journal : for cell and molecular biology》1998,13(2):231-239
Putrescine and polyamines are produced by two alternative pathways in plants. One pathway starts with the enzyme arginine decarboxylase; the other with ornithine decarboxylase. The authors developed an in vivo screening strategy to identify mutants with low levels of arginine decarboxylase activity. The screen requires both a primary screen of the M2 generation and a secondary screen of the M3 generation. The method used was to screen 15 000 EMS-mutagenized M2 seedlings for low levels of arginine decarboxylase (ADC) activity and identified seven mutants that fall into two complementation groups. These mutants have from 20% to 50% of wild-type enzyme activity. Morphological alterations common among the mutants include increased levels of lateral root branching. The authors obtained a double mutant combining the alleles with the lowest activities from the two complementation groups; this has lower ADC enzyme activity and putrescine levels than either of the single mutants. The double mutant has highly kinked roots that form a tight cluster; it also has narrower leaves, sepals, and petals than either single mutant or wild-type, and delayed flowering. These results suggest there may be more than one ADC gene in Arabidopsis, and that ADC and polyamine levels play roles in root meristem function and in lateral growth of leaf-homolog organs. 相似文献
5.
M. M. Anisimov V. V. Logachev G. N. Likhatskaya V. V. Makhan'kov N. I. Uvarova 《Biology Bulletin》2003,30(3):287-290
We studied the effect of extractive substances (ES) and total glycoside fraction (TGF) from Panax ginseng on plant cell growth. The seeds of cucumber Cucumis sativus L., KIT variety, were used for the biological assay. The tested substances inhibited the primary root growth in Cucumis sativus seedlings. Actively growing seedlings were most sensitive to this effect. 相似文献
6.
Among the various amines administered to excisedCucumis sativus cotyledons in short-term organ culture, agmatine (AGM) inhibited arginine decarboxylase (ADC) activity to around 50%, and putrescine was the most potent entity in this regard. Homoarginine (HARG) dramatically stimulated (3- to 4-fold) the enzyme activity. Both AGM inhibition and HARG stimulation of ADC were transient, the maximum response being elicited at 12 h of culture. Mixing experiments ruled out involvement of a macromolecular effector in the observed modulation of ADC. HARG-stimulated ADC activity was completely abolished by cycloheximide, whereas AGM-mediated inhibition was unaffected. Half-life of the enzyme did not alter on treatment with either HARG or AGM. The observed alterations in ADC activity are accompanied by change in Km of the enzyme. HARG-stimulated ADC activity is additive to that induced by benzyladenine (BA) whereas in presence of KCl, HARG failed to enhance ADC activity, thus demonstrating the overriding influence of K+ on amine metabolism. 相似文献
7.
Cucurbitaceae are characterized by a high copy number for nuclear ribosomal RNA genes. We have investigated the genomic ribosomal DNA (rDNA) of four closely related species of this family with respect to structure, length heterogeneity, and evolution. InCucumis melo (melon) there are two main length variants of rDNA repeats with 10.7 and 10.55kb.Cucumis sativus (cucumber) shows at least three repeat types with 11.5, 10.5, and 10.2kb.Cucurbita pepo (zucchini) has two different repeat types with 10.0 and 9.3kb. There are also two different repeat types inCucurbita maxima (pumpkin) of about 11.2 and 10.5kb. Restriction enzyme mapping of the genomic rDNA of these four plants and of cloned repeats ofC. sativus shows further heterogeneities which are due to methylation or point mutations. By comparison of the restriction enzyme maps it was possible to trace some evolutionary events in the family ofCucurbitaceae. Some aspects of regulation and function of the middle repetitive rRNA genes (here between 2000 and 10000 copies) are discussed. 相似文献
8.
Homoarginine decarboxylase has been purified ca 110-fold from Lathyrus sativus seedlings and resolved from arginine decarboxylase by DEAE-Sephadex column chromatography. The enzyme was less active than arginine decarboxylase and was highly labile. This preparation decarboxylated l-lysine in addition to L-homoarginine. The purified enzyme preparation had an absolute requirement for exogenous Mn2+ or Fe2+ for both the enzyme activities. The pH and temperature optima for decarboxylation of both homoarginine and lysine were the same viz. 8·4 and 41° respectively. The Km value l-homoarginine was 3·33 mM and for l-lysine was 0·88 mM. Arginine and homoarginine decarboxylases appear to be different and separable entities having different physico-chemical characteristics, despite the fact that their respective guanido amino acid substrates undergo similar metabolic conversion to guanido- and diamines in this plant system. 相似文献
9.
The multifunctional enzyme, putrescine synthase has been purified fromCucumis sativus and characterized. This enzyme harbours agmatine iminohydrolase, ornithine transcarbamylase, putrescine transcarbamylase
and carbamate kinase activities, whose concerted action results in agmatine → putrescine conversion. The enzyme resolved into
two aggregation forms, enzyme aggregated and enzyme monomer upon electrophoresis at pH 8.3. Evidence has been provided by
two-dimensional gel electrophoresis that both enzyme aggregated and enzyme monomer comprise of identical polypeptide chains.
Under non-reducing conditions on sodium dodecyl sulphate-polyacrylamide gel electrophoresis, the protein moves as a single
150 KDa polypeptide; however, in the presence of 2-mercaptoethanol on sodium dodecyl sulphate-polyacrylamide gel elec trophoresis,
it migrates as 3 polypeptides of molecular weight 48,000, 44,000 and 15,000. The enzyme undergoes age-dependentin vivo proteolytic degradation from a 66 KDa polypeptide (primary translational product), through 48 KDa polypeptide to 44 KDa species
and finally to small molecular weight peptides.
Preliminary results of this work were presented at Golden Jubilee and Annual General Body Meetings of Society of Biological
Chemists (India) and the Second Congress of Asian and Ocean Biochemists (1980) held at Bangalore, 1981,Indian J. Biochem. Biophys.,18, 113. 相似文献
10.
Exposure of cucumber seedlings (Cucumis sativus L.) to chilling temperature resulted in injuries such as increased leakage of cellular materials, loss of water and wilting. In addition, the development of the seedlings after the exposure to chilling was impaired. Abscisic acid applied to the seedlings prior to chilling significantly ameliorated these injuries. 相似文献
11.
Physiological and biochemical changes related to methyl jasmonate (MeJA)-induced chilling tolerance of rice (Oryza sativa L. cv. Taichung Native 1) seedlings were investigated. Treatment of whole plants with 10 mmol m?3 MeJA for 48 h before chilling (5 °C) was optimal for the induction of chilling tolerance. MeJA greatly improved the survival ratio of chilled seedlings and ameliorated chilling injury such as demolition of membrane structure (estimated by electrolyte leakage). MeJA also prevented water loss in chilled seedlings by reducing the opening of stomata and decreasing the root bleeding rate. Putrescine and spermine levels in shoots increased but spermidine levels decreased on exposure to MeJA. In roots, putrescine levels also increased and spermidine levels increased transiently on exposure to MeJA. Activities of arginine decarboxylase (ADC; EC 4.1.1.19) and S-adenosylmethionine decarboxylase (SAMDC; EC 4.1.1.50) in both shoots and roots increased on exposure to MeJA, while the activity of ornithine decarboxylase (ODC; EC 4.1.1.17) remained unchanged. The MeJA-induced putrescine increase was inhibited by 50 mmol m?3α-difluoromethylarginine (DFMA), an irreversible inhibitor of ADC, but not by 50 mmol m?3α-difluoromethylornithine (DFMO), an irreversible inhibitor of ODC. The effect of MeJA on the induction of chilling tolerance was also reduced by 50 mmol m?3 DFMA. The effects of DFMA were partly prevented by 1 mol m?3 putrescine. This indicates that putrescine accumulation is required for the induction of chilling tolerance of rice seedlings by MeJA. 相似文献
12.
Autotoxic potential of cucurbit crops 总被引:20,自引:1,他引:20
Soil sickness is often observed in cucurbit crops such as Citrullus lanatus, Cucumis melo and Cucumis sativus, but not in cucurbit crops such as Cucurbita moschata, Lagenaria leucantha and Luffa cylindrica. Results showed that root aqueous extracts of Citrullus lanatus, Cucumis melo and Cucumis sativus were autotoxic, but those of Cucurbita moschata, Momordica charantia and Luffa cylindrica were less autotoxic to the radicle elongation of respective species. Plant growth of Citrullus lanatus, Cucumis melo and Cucumis sativus were greatly inhibited by autotoxic substances released from powered root tissue at a rate of 1 g per seedling. Root exudates
of Citrullus lanatus, Cucumis melo and Cucumis sativus were autotoxic to radicle elongation and seedling growth of respective species. However, root exudates of Citrullus lanatus did not inhibit radicle elongation of Cucurbita ficifolia, which is commonly used as rootstock for the grafting of Citrullus lanatus, Cucumis melo and Cucumis sativus to decrease soil-borne diseases in commercial production. It seems possible to overcome autotoxicity in cucurbit crops by
grafting on Cucurbita ficifolia.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
13.
Among the various amines administered to excisedCucumis sativus cotyledons in short-term organ culture, agmatine (AGM) inhibited arginine decarboxylase (ADC) activity to around 50%, and putrescine was the most potent entity in this regard. Homoarginine (HARG) dramatically stimulated (3- to 4-fold) the enzyme activity. Both AGM inhibition and HARG stimulation of ADC were transient, the maximum response being elicited at 12 h of culture. Mixing experiments ruled out involvement of a macromolecular effector in the observed modulation of ADC. HARG-stimulated ADC activity was completely abolished by cycloheximide, whereas AGM-mediated inhibition was unaffected. Half-life of the enzyme did not alter on treatment with either HARG or AGM. The observed alterations in ADC activity are accompanied by change in Km of the enzyme. HARG-stimulated ADC activity is additive to that induced by benzyladenine (BA) whereas in presence of KCl, HARG failed to enhance ADC activity, thus demonstrating the overriding influence of K+ on amine metabolism. 相似文献
14.
Control by ethylene of arginine decarboxylase activity in pea seedlings and its implication for hormonal regulation of plant growth 总被引:4,自引:3,他引:1
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Activity of arginine decarboxylase in etiolated pea seedlings appears 24 hours after seed imbibition, reaches its highest level on the 4th day, and levels off until the 7th day. This activity was found in the apical and subapical tissue of the roots and shoots where intensive DNA synthesis occurs. Exposure of the seedlings to ethylene greatly reduced the specific activity of this enzyme. The inhibition was observed within 30 min of the hormone application, and maximal effect—90% inhibition—after 18 hours. Ethylene at physiological concentrations affected the enzyme activity; 50% inhibitory rate was recorded at 0.12 microliters per liter ethylene and maximal response at 1.2 microliters per liter. Ethylene provoked a 5-fold increase in the Kmapp of arginine decarboxylase for its substrate and reduced the Vmaxapp by 10-fold. However, the enzyme recovered from the inhibition and regained control activity 7 hours after transferral of the seedlings to ethylene-free atmosphere. Reducing the endogenous level of ethylene in the tissue by hypobaric pressure, or by exposure to light, as well as interfering with ethylene action by treatment with silver thiosulfate or 2,5-norbornadiene, caused a gradual increase in the specific activity of arginine decarboxylase in the apical tissue of the etiolated seedlings. On the basis of these findings, the possible control of arginine decarboxylase activity by endogenous ethylene, and its implication for the hormone effect on plant growth, are discussed. 相似文献
15.
Dr. C. Ramachandran W. A. Brandenburg A. P. M. den Nijs 《Plant Systematics and Evolution》1985,151(1-2):31-41
Infraspecific cytogenetical variation was studied in a diverse collection of five non-cultivated and cultivatedCucumis sativus accessions. The individual chromosomes of different accessions could be identified by the C-banding pattern and chromosome
measurements. About 40–50% of the genomic area are made up of heterochromatin inC. sativus. The non-cultivated accessions exhibit more heterochromatin and lower chiasma frequencies per pollen mother cell than cultivated
accessions. There is infraspecific variation in C-banding pattern, karyomorphology and multinucleolate cells. The use of C-banding
in infraspecific classification is discussed. 相似文献
16.
Terence A. Smith 《Phytochemistry》1979,18(9):1447-1452
Arginine decarboxylase activity in the shoots of seedlings was high in oats, intermediate in barley and low in rice, maize, wheat and rye. After partial purification, the arginine decarboxylase from the shoots of potassium deficient oat seedlings was separated into two fractions, A (MW 195 000) and B (MW 118 000), by gel chromatography. On gel electrophoresis, the mobilities of these fractions were respectively 0.12 and 0.55 relative to bromophenol blue at pH 9.5. Fraction A was twice as active as fraction B in extracts of seedlings grown with both normal and potassium deficient nutrition, despite the greater activity ( × 5) of the potassium deficient plants. The properties of the two fractions were similar with respect to pH optimum (7–7.5), Km (3 × 10 ?5M) and the effect of inhibitors. Fraction A was purified to apparent homogeneity by DEAE-cellulose chromatography. The enzyme was specific for l-arginine and it was strongly inhibited by NSD 1055, d-arginine and canavanine. Mercaptoethanol and dithiothreitol stimulated the enzyme by ca 50% and p-chloromercuribenzoate was an inhibitor. Pyridoxal phosphate stimulated activity by ca 30% and EDTA stimulated activity by 30%. Ca2+ and Mg2+ inhibited the enzyme by 50% at ca 20 mM. Putrescine and the polyamines showed only moderate inhibition at 10 mM, but agmatine reduced activity to 30% at this concentration. 相似文献
17.
HALVOR AARNES 《Physiologia plantarum》1974,32(4):400-402
Regulation of aspartate kinase activity was examined in some higher plant seedlings (Helianthus annuus, Raphanus sativus, Cucutnis sativus, Sinapis alba), a green (Chlorellapyrenoidosa), and a blue-green alga (Anacystis nidulans). In Cucumis sativus a concerted feedback regulation by L-lysine and L-threonine was indicated. In all the other species examined, aspartate kinase was inhibited by both L-lysine and L-threonine and it depended upon the species which of them was the strongest inhibitor. Only slight effects of L-leucine, L-isoIeucine and L-valine were observed, except in the Sinapis alba enzyme. 相似文献
18.
Gioconda San-Blas Felipe San-Blas Françoise Sorais Belisario Moreno José Ruiz-Herrera 《Archives of microbiology》1996,166(6):411-413
Putrescine and spermidine were the only polyamines found inParacoccidioides brasiliensis, a dimorphic fungus pathogenic for humans. Free polyamines (putrescine>spermidine) increased during the first 24 h of yeast growth, with a second peak at 42 h, and also during the first 12 h of mycelium-to-yeast transition (spermidine>putrescine). Conjugated and bound polyamines were also quantified. 1,4-Diamino-2-butanone decreased free putrescine and spermidine accumulation by inhibiting the activity of ornithine decarboxylase. The increase in free polyamines corresponds to bud emergence in yeast growth and to the mycelium-to-yeast transition ofP. brasiliensis.Abbreviations
DAB
1,4-Diamino-2-butanone
-
Y
Yeasts
-
M
Mycelia
-
ODC
Ornithine decarboxylase 相似文献
19.
20.
Cytochemical demonstration of malate synthase and glycolate oxidase in microbodies of cucumber cotyledons 总被引:6,自引:5,他引:1
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The cytochemical localizations of malate synthase (glyoxysomal marker) and glycolate oxidase (peroxisomal marker) have been examined in cotyledon segments and sucrose-gradient fractions from germinated cucumber (Cucumis sativus L.) seedlings. The seedlings were grown in the dark for 4 days, transferred to 4 hours of continuous light, then returned to the dark for 24 hours. Under these conditions, high specific activities for both glyoxysomal and peroxisomal enzymes are maintained in cotyledon homogenates and microbody-enriched fractions. Electron cytochemistry of the marker enzymes reveals that all or virtually all the microbodies observed in cotyledonary cells and sucrose-gradient fractions contain both enzymes. The staining in gradient fractions was determined from scoring a minimum of 600 photographed microbodies for each enzyme. After correcting for the number of particles stained for catalase reactivity (representing true microbodies), 94 and 97% of the microbodies were found stained for malate synthase and glycolate oxidase activity, respectively. 相似文献