• BMB Reports
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• v.31 no.1
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• pp.70-76
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• 1998

The final step in ethylene biosynthesis is catalyzed by the enzyme 1-aminocyclopropane-1-carboxylate (ACC) oxidase. ACC oxidase was extracted from mung bean hypocotyls and its biochemical characteristics were determined. In vitro ACC oxidase activity required ascorbate and $Fe^{2+}$, and was enhanced by sodium bicarbonate. Maximum specific activity (approximately 20 nl ethylene $h^{-1}$ mg $protein^{-1}$) was obtained in an assay medium containing 100 mM MOPS (pH 7.5), $25\;{\mu}M$ $FeSO_4$, 6 mM sodium ascorbate, 1 mM ACC, 5 mM sodium bicarbonate and 10% glycerol. The apparent $K_m$ for ACC was $80{\pm}3\;{\mu}M$. Pretreating mung bean hypocotyls with ethylene increased in vitro ACC oxidase activity twofold. ACC oxidase activity was strongly inhibited by metal ions such as $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, and $Mn^{2+}$, and by salicylic acid. Inactivation of ACC oxidase by salicylic acid could be overcome by increasing the $Fe^{2+}$ concentration of the assay medium. The possible mode of inhibition of ACC oxidase activity by salicylic acid is discussed.

• Journal of Plant Biology
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• v.37 no.4
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• pp.445-452
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• 1994

Effects of methyl jasmonate (MeJA) on ethylene production in mungbean (Phaseolus radiatus L.) hypocotyl and leaf segments were studied. Ethylene production in mungbean hypocotyl segments was decreased in proportion to MeJA concentrations and $450\;\mu\textrm{M}$ of MeJA showed 50% inhibitory effect. This inhibitory effect appeared after 3 h of incubation period and continued for 24 h. Inhibition of ethylene production by MeJA was due to the decrease in 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase activity. However, MeJA treatment had no effect on ACC content and ACC synthase activity. MeJA also inhibited auxin-induced ethylene production in hypocotyls. To investigate the mechanisms of the inhibitory effect of MeJA on the auxin-induced ethylene production, ACC synthase and ACC oxidase activity were examined after MeJA treatment. MeJA decreased the ACC content and ACC synthase activity as weD as ACC oxidase activity in the auxin-treated tissue. These results suggest that the inhibition of MeJA on auxin-induced ethylene production is not due to the direct inhibitory effect of MeJA on the ACC synthase, but to the inhibition of the ability of IAA to promote the synthesis of ACC synthase. In contrast, ethylene production from the detached mungbean leaves was stimulated by MeJA. The rate of ethylene production increased approximately 65% over the control after 12 h of incubation period by $4.5\;\mu\textrm{M}$ MeJA. When MeJA was applied to detached leaves along with IAA, the effect of MeJA appeared to be additive. In an effort to elucidate mechanisms of MeJA action on auxin-induced ethylene production in the leaf tissue, enzyme activities of ACC synthase and ACC oxidase were examined. MeJA stimulated ACC oxidase activity but did not affect ACC synthase activity in leaf tissue. Together, these results suggest that MeJA plays different roles in the ethylene production in the different mungbean tissues.issues.

• Journal of Plant Biology
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• v.38 no.3
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• pp.235-242
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• 1995

Effects of methyl jasmonate (MeJA) on ethylene production in tomato(Lycopersicon esculentum Mill.) hypocotyl segments and fruits were studied. Ethylene production in tomato hypocotyl segments was inhibited by the increasing concentratons of MeJA, and 450 $\mu$M of MeJA showed 50% inhibitory effect. Time course data indicate that this inhibitory effect of MeJA appeared after 3 h of incubation period and continued until 24 h. Inhibition of ethylene producton by MeJA was due to the decrease in 1-aminocyclopropane-1-carboxylic acid(ACC) synthase activity. However, MeJA treatment had no effect on ACC oxidase activity and the accumulaton of ACC oxidase mRNAs. MeJA also inhibited auxin-induced ethylene production by decreasing in ACC synthase activity. In contrast, MeJA stimulated ethylene production in tomato fruits. When 30 $\mu$L/mL MeJA was treated in a gaseous state, ethylene production doubled and this stimulating effect continued until 4 days. To investigate the mechanisms of MeJA on ethylene production, ACC synthase and ACC oxidase activities were examined after MeJA treatment. MeJA increased the activities of both ACC synthase and ACC oxidase, and induced ACC oxidase mRNA accumulation. These data suggest that MeJA plays distinct roles in the ethylene production in different tomato tissues. It is possible that MeJA affects differently the mechanisms of signal transuction leading to the ethylene biosynthesis.

• Journal of Plant Biology
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• v.39 no.1
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• pp.31-40
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• 1996

The physiological and biochemical characterizations of the ethylene-related mutants in Arabidopsis thaliana - ethylene overproducing mutant (eto1-l) and ethylene insensitive mutants (etrl-3, ein2-l) - were detailed in this studies. Two or three week.old mature rosette leaves (before bolting) were used as the plant materials. Ethylene productions of eto1-l, etrl-3, and ein2-l mutants were about 200%, 400%, and 450% compared to that of wild type, respectively. ACC synthase and ACC oxidase activities of eto1-l mutant were similar to those of wild type. ACC content and ACC N-malonyltransferase activity, however, were 4.5 times and 3 times higher than those of wild type, respectively. SAM synthetase activity increased by 50% in eto1-l mutant plant. These results indicated that the alteration in the eto1-l mutant occured before the step of the conversion of SAM to ACe. In etrl-3 and ein2-l mutants, ACC synthase activities increased, but ACC oxidase activities decreased. ACC content and ACC N-malonyltransfcrase activity were 2 times higher than those of wild type. SAM synthetase activity in etrl-3 is similar to those of wild type, while it increased by 73% in ein2-l. These results showed that the block in ethylene action affected the autoregulation of ethylene biosynthesis, so that ACC synthase activity was not autoinhibited and ACC oxidase activity was not auto stimulated by ethylene. When the leaf tissues were used for in vitro kinase assay, a cytosolic protein (approximately 36 kDa) was phosphorylated only in eto1-l and ein2-l mutants.utants.

• Horticultural Science & Technology
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• v.29 no.4
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• pp.326-335
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• 2011

When bougainvilleas are subjected to indoor low-light conditions, flower bracts regularly abscise. This study elucidates the effects of plant growth regulators on bract longevity of potted bougainvillea. Potted 'Taipei Red' bougainvillea in four different bract development stages were treated with 1-MCP (1-methylcyclopropene), NAA (1-naphthaleneacetic acid), SNA (sodium salt of naphthaleneacetic acid), IBA (indolebutyric acid), BA (6-benzylaminopurine), $KH_2PO_4$ (potassium dihydrogen phosphate), Put (diamine putrescine), SA (salicylic acid), or STS (silver thiosulfate) and were moved to indoor low-light conditions after treatments. Experimental results indicate that 1-MCP, NAA, SNA, BA, Put, and SA prolonged bract longevity, and this effect increased as bract stage increased. The effect of STS was significant in early bract stages and decreased as bract stages increased. Additionally, 1-MCP, NAA, SNA, BA, Put, SA, and STS treatment significantly reduced endogenous ACC (1-aminocyclopropene-1-carboxylate) content and ACC oxidase activity, suggesting that the inhibition of ethylene production was achieved via physiological metabolism. However, treatment with IBA or $KH_2PO_4$ had no effect on the bract longevity at any stage. In the combined chemical treatments, NAA + STS or NAA + SA were effectively for prolonging bract longevity and contained less protein or chlorophyll degradation, decrease ACC oxidase or ethylene production than the control. In conclusion, we propose that combined chemical treatment significantly prolonged the bract longevity and more effectively than single chemical treatment at any stage.

• Journal of Plant Biology
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• v.36 no.3
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• pp.233-239
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• 1993

Effect of Ca2+ on the conversion of 1-aminocycloprophane-1-carboxylic acid (ACC) to ethylene was studied with 2.5 day-old mung bean hypocotyl segments. The conversion of ACC in these tissues was inhibited by plasmolysis and sulfosuccinimidyl (hydroxyphenyl) propionate (sulfo-SHPP). The ACC induced ethylene production in HC (high calcium)-tissue grown on the Ca2+ added medium was greater than that in N (normal)-tissue. HC-tissue had a lower inhibition rate of ACC conversion by EGTA and Ca2+ -channel blockers than N-tissue. The rates of the ACC conversion by both kinds of tissues were stimulated by the Ca2+ ionophore A23187. From these results, we suggests a mechanism for the stimulative effect of Ca2+ on the conversion of ACC to ethylene as follows; in some tissues where ACC conversion is linked with plasma membrane, Ca2+ may be transported from apoplast through Ca2+ -channel into the cytoplasm ad stimulate ACC-oxidase activity.

• Horticultural Science & Technology
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• v.19 no.4
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• pp.545-549
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• 2001

The relationship between ethylene action and flesh softening in post-storage persimmon fruits was investigated by treating the fruits with 1-MCP. The patterns of firmness change caused by various treatments with 1-MCP and ethylene were similar to those of changes in cell wall-degrading enzyme activities, including cellulase, PG, PME, and ${\beta}$-galactosidase. Moreover, the activities of these enzymes were inhibited by 1-MCP treatment. These results show that the cell wall-degrading enzymes influenced by ethylene account for the flesh softening process of post-storage persimmon fruit. The ethylene production of fruits, as measured by ACC content and ACC oxidase activity, nevertheless, was not influenced by 1-MCP treatments. It is suggested that the flesh softening phenomena in post-storage persimmon fruits is correlated to the ethylene responsiveness of tissue rather than the ethylene production rate per se.

• Journal of Plant Biology
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• v.35 no.4
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• pp.409-414
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• 1992

Effect of quercetin, a kind of natural plant flavonoids, on auxin-induced ethylene production in barley coleoptiles was studied. Auxin-induced ethylene production was apparently stimulated by quercetin. This stimulatory effect of quercetin appeared after 4 h of incubation period. Ethylene production was stimulated 200% over the control after 8 h of incubation by $3{\times}10^{-5}\;M$ quercetin. The quercetin effect was most prominent at $10^{-4}\;M$ of IAA. Ethylene production induced by the synthetic auxin, 2,4-D and NAA, was not significantly affected by quercetin. Also ACC-based ethylene production was unaffected by the flavonoid. In an effort to elucidate mechanisms of quercetin action on auxin-induced ethylene production, the effect of quercetin on 1M metabolism was studied. Data obtained from these experiments indicate that quercetin treatment resulted in about 90% inhibition of IAA oxidase activity. IAA ($3{\times}10^{-5}\;M$) conjugation was found to be not affected by quercetin. This results suggest that the stimulatory effect of quercetin on auxin-induced ethylene production may be due to the fact that quercetin inhibits 1M oxidase activity, thus increasing the free IAA level.

• Journal of Plant Biology
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• v.37 no.3
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• pp.271-276
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• 1994

The possibility that 1-aminocyclopropane-1-carboxylic acid (ACC)-uptake may be dependent on the H+-gradient established across the plsma membrane was tested in protoplasts isolated from 2.5 day old mungbean hypocotyls. The ACC-induced ethylene production was inhibited when the H+-gradient was collapsed by the treatment with carbonycyamide-p-trifluro-methoxy-phenylhydrazone (FCCP). Moreover, the treatment with o-vanadate, a specific inhibitor of plasma membrane H+-ATPase, caused the inhibition of ethylene production. The ACC-induced ethylene production was inhibited by the treatemnt with verapamil (Ca2+-channel blocker), or ethylene glycol-bis($\beta$-aminoethyl ether) N, N, N', N'-tetraacetic acid (EGTA) (Ca2+-chelator). In contrast, the ehtylene production was stimulated by the application of A23187 (Ca2+ ionophore). The inhibitory effect of EGTA in the ethylene producton was magnified in the presence of A23187. From these results, we suggest that the external Ca2+ influx to the cytosol resulted in the stimulatin of ACC oxidase activity after ACC-uptake resulting from a H+-gradient across the plasma membrane.

• Journal of Plant Biology
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• v.39 no.4
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• pp.295-300
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• 1996

The physiological effects of oligogalacturonic acid (OGA:D. P. 6-7), a product of acid hydrolysis of polygalacturonic acid (PGA), on ethylene biosynthesis in mung bean (Vigna radiata W.) hypocotyl segments was studied. Among PGA, OGA and monogalacturomic acid (MGA), only OGA stimulated ethylene production in mung bean hypocotyl segments, and the most effective concentraton of OGA was 50$\mu\textrm{g}$/mL. Time course data indicated that this stimulatiion effect of OGA appeared after 90 min incubation period and continued until 24 h. When indol-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) were treated with OGA to investigate the mechanism of OGA on ethylene production, they did not show synergistic effects on ethylene production. The stimulation of ethylene production by OGA was due to the increase of in vivo ACC synthase activity, but OGA treatment had no effect of in vivo ACC oxidase activity. The effect of aminoethoxy vinyl glycine (AVG) and Co2+, the inhibitor of ethylene synthesis, was siminished a little by the OGA, but the treatment of Ca2+, known to increase ACC, with OGA did not increase the ethylene production, this effect seems to be specific for Ca2+ because other divalent cation, Mg2+, did not show the inhibition of OGA-indyuced ethylene production. It is possible that the OGA adopts a different signal transduction pathway to the ethylene bioxynthesis.