• BMB Reports
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• 제31권1호
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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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• 제37권4호
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• pp.445-452
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• 1994

Methyl jasmonate(MeJA)가 녹두(Phaseolus radiatus L.) 하배축과 잎에서 에틸렌 생성에 미치는 영향을 조사하였다. 녹두 하배축 조직에서의 에틸렌 생성은 MeJA의 농도에 비례해서 감소했으며 그 억제 효과는 3시간 이후부터 현저하게 나타났다. 1-Aminocyclopropane-1-carboxylic acid (ACC) oxidase의 활성에 대한 MeJA의 억제 효과 역시 3시간 이후부터 나타났으며 ACC synthase의 활성에는 별 효과가 없었다. $450\;\mu\textrm{M}$ MeJA가 포함된 배지에서 배양시킨 녹두 하배축 조직에서 옥신 유발 에틸렌 생성은 대조구에 비하여 현저히 억제되었는데 이러한 옥신 유발 에틸렌 생성에 대한 MeJA의 억제 기작을 알아보기 위하여 ACC synthase의 활성과 ACC oxidase의 활성을 조사하였다. MeJA는 옥신이 처리된 조직내 ACC양과 ACC synthase 활성 및 ACC oxidase의 활성을 감소시켰다. 따라서 옥신 유발 에틸렌 생성에 대한 MeJA의 억제 효과는 MeJA가 ACC synthase의 활성 뿐만 아니라 ACC oxidase의 활성을 감소시킴으로써 나타난다고 볼 수 있다. 하배축과는 반대로 $4.5\;\mu\textrm{M}$로 처리한 잎 조직에서의 에틸렌 생성은 대조구에 비해 약 50% 촉진되었으며 옥신에 의해 유발된 에틸렌 생성량은 MeJA에 의해 증가되었다. 잎에서의 MeJA에 의한 에틸렌의 합성 증가는 ACC oxidase의 활성 증가에 기인하였으며 ACC synthase의 효소활성은 변화가 없어TEk. 따라서 MeJA는 녹두의 서로 다른 조직에서 에틸렌 합성에 대해 서로 상반된 효과를 나타내었다.

• Journal of Plant Biology
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• 제38권3호
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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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• 제39권1호
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• pp.31-40
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• 1996

Arabidopsis thaliana의 에틸렌 관련 돌연변이체인 에틸렌 과다 생성 돌연변이체(eto1-1)와 에틸렌에 대한 반응이 둔감한 돌연변이체(etr1-3과 ein2-1)의 생리·생화학적 특성을 분석하였다. 2∼3주된 Arabidopsis 식물로부터 얻은 성숙한 잎을 재료로 이용하였다. 에틸렌 생성량은 eto1-1은 야생형의 2배, etr1-3은 야생형의 4배, 그리고 ein2-1은 야생형의 4.5배 더 많았다. eto1-1에서의 ACC synthase와 ACC oxidase 활성은 야생형과 비슷하였으나 ACC 함량은 야생형보다 4.5배 더 많았다. ACC-N-malo-nyltransferase의 활성은 eto1-1이 야생형보다 3배 더 높았으며 SAM synthetase의 활성은 야생형보다 1.5배 더 높았다. 이들 결과로부터 eto1-1로의 변형은 SAM에서 ACC로 전환하는 과정 이전에 있음을 추측할 수 있다. etr1-3과 ein2-1에서 ACC synthase의 활성은 야생형보다 높았으나 ACC oxidase의 활성은 야생형보다 낮았다. 그러나 SAM synthetase 활성은 etr1-3에서는 야생형과 비슷하였고 ein2-1에서는 야생형보다 1.7배 높았다. 이것은 etr1-3과 ein2-1이 에틸렌에 대한 반응에 결함이 있기 때문에 그것으로 인하여 자가 조절이 되지 않았기 때문으로 추정된다. etr1-3의 ACC 함량은 야생형보다 2.3배 더 많았으며 ACC N-malonyltransferase의 활성은 야생형보다 3.9배 더 높았다. 그리고 ein2-1의 ACC 함량은 야생형보다 1.7배 더 많았으며 ACC N-malonyltransferase의 활성이 촉진된 것으로 추정된다. In vitro kinase assay를 한 결과 eto1-1과 ein2-1에서 36 kDa 단백질의 인산화를 관찰할 수 있었다.

• 원예과학기술지
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• 제29권4호
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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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• 제36권3호
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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.

• 원예과학기술지
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• 제19권4호
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• pp.545-549
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• 2001

에틸렌의 수용체와의 결합을 방해함으로써 에틸렌의 작용을 방해하는 물질은 1-MCP의 처리를 통하여 단감 과실의 과육 연화에 대한 에틸렌의 영향을 밝히기 위한 실험을 수행하였다. 1-MCP와 에틸렌의 처리에 따른 경도 변화의 양상은 cellulase, PG, PME, ${\beta}$-galactosidase 등의 세포벽 분해 효소의 활성 변화 양상과 유사하였으며, 이들 효소의 활성은 1-MCP의 처리에 의해 저하되었다. 이러한 결과는 에틸렌의 영향에 의한 각종 세포벽 분해 효소의 활성화가 저장 후 단감 과실의 과육 연화 현상과 밀접한 관련이 있음을 보인다. 그러나 연화 지연 효과와는 달리 저장 후의 단감 과실에서 1-MCP는 에틸렌의 생성, 즉 ACC의 축적 및 ACC oxidase 활성을 억제하지 못하였으며, 이러한 사실은 저장 후 단감 과실에 있어서 과육의 연화 현상이 과실의 에틸렌 생성량 그 자체보다는 과실의 에틸렌에 대한 반응성에 보다 밀접하게 관련되어 있음을 보인다.

• Journal of Plant Biology
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• 제35권4호
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• pp.409-414
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• 1992

Quercetin(3,5,7,3'-4'-OH-flavone)이 보리(Hordeum vulgare var. hexastichon ASCHERS.) 자엽초 조직에서 옥신유발 에틸렌 생성을 현저히 촉진시켰다. $3{\times}10^{-5}\;M$의 quercetin에서 배양한 보리 자엽초 조직에서 옥신 유발 에틸렌 생성은 4시간 이후부터 증가되기 시작하여 8시간 이후에는 200%까지 증가되었다. Quercetin은 내재옥신인 indol-3-acetic acid(IAA)에 의한 에틸렌 생성은 촉진하나 합성 옥신인 2,4-dichlorophenoxyacetic acid(2,4-D)나 naphthaleneacetic acid(NAA)에 의한 에틸렌 생성에는 아무런 영향을 주지 않았다. 또한, 에틸렌 전구체인 1-aminocyclopropen-1-carboxylic acid(ACC) 유발 에틸렌 생성에도 촉진효과가 없었다. 이러한 결과는 에틸렌 생성에 대한 quercetin의 효과는 ACC 전단계에 작용한다는 것을 의미한다. Quercetin이 IAA 대사에 어떠한 영향을 미치는지 확인한 결과 조직을 quercetin에 1시간 배양시켰을 때 조직내 IAA oxidase 효소 활성이 90% 감소되었으며, IAA conjugation에는 별 영향을 미치지 않았다. 따라서, 옥신 유발 에틸렌 생성에 대한 quercetin의 촉진작용은 quercetin이 IAA oxidase의 활성을 억제시켜 높아진 IAA 수준이 옥신 유발 에틸렌 생성을 증가시키는 것으로 생각된다.

• Journal of Plant Biology
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• 제37권3호
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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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• 제39권4호
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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.