Abstract
1-Aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase are key enzymes in the ethylene biosynthetic pathway in plant tissues, and in vitro assay of their activities is indispensable for analysis, especially, for studying the action mechanism of inhibitors of ethylene biosynthesis. The enzymes can be obtained from plant tissues that are producing ethylene abundantly, such as ripening fruit- and senescing flower tissues, but it is necessary to separate the enzymes from co-extracted ACC by partial purification, making the procedure laborious and time-consuming. Here, we describe the production of the enzymes in Escherichia coli cells from corresponding cDNAs, and the procedures for assay of activities of the enzymes.
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References
Abeles FB, Morgan PW, Saltveit ME Jr (1992) Ethylene in plant biology, 2nd edn. Academic Press, New York
Iturriagagoittia-Bueno T, Gibson EJ, Schofield CJ, John P (1996) Inhibition of 1-aminocyclo-propane-1-carboxylate oxidase by 2-oxoacids. Phytochemistry 43:343–349
Smith JJ, Ververidis P, John P (1992) Characterization of the ethylene-forming enzyme partially purified from melon. Phytochemistry 31:1485–1494
Yang SF, Hoffman NE (1984) Ethylene biosynthesis and its regulation in higher plants. Annu Rev Plant Physiol 35:155–189
ten Have A, Woltering EJ (1997) Ethylene biosynthetic genes are differentially expressed during carnation (Dianthus caryophyllus L.) flower senescence. Plant Mol Biol 34:89–97
Woodson WR, Park KY, Drory A, Larsen PB, Wang H (1992) Expression of ethylene biosynthesis pathway transcripts in senescing carnation flowers. Plant Physiol 99:526–532
Fujino DW, Reid MS, Yang SF (1980) Effects of aminooxyacetic acid on post-harvest characteristics of carnation. Acta Hort 113:59–64
Baker JE, Wang CY, Lieberman M, Hardenburg RE (1977) Delay of senescence in carnations by rhizobitoxine analogue and sodium benzoate. Hort Sci 12:38–39
Satoh S, Esashi Y (1980) α-Aminoisobutyric acid: A probable competitive inhibitor of conversion of 1-aminocyclopropane-1-carboxylic acid to ethylene. Plant Cell Physiol 21:939–949
Satoh S, Esashi Y (1983) α-Aminoisobutyric acid, propyl gallate and cobalt ion and the mode of inhibition of ethylene production by cotyledonary segments of cocklebur seeds. Physiol Plant 57:521–526
Serrano M, Romojaro F, Casas JL, Del Rio JA, Acosta M (1990) Action and mechanism of α-aminoisobutyric acid as a retardant of cut carnation senescence. Sci Hort 44:127–134
Midoh N, Saijou Y, Matsumoto K, Iwata M (1996) Effects of 1,1-dimethyl-4-(phenylsulfonyl)semicarbazide (DPSS) on carnation flower longevity. Plant Growth Regul 20:195–199
Kosugi Y, Matsuoka A, Higashi A, Toyohara N, Satoh S (2014) 2-Aminooxyisobutyric acid inhibits the in vitro activities of both 1-aminocyclopropane-1-carboxylate (ACC) synthase and ACC oxidase in ethylene biosynthetic pathway and prolongs vase life of cut carnation flowers. J Plant Biol 57:218–224
Satoh S, Kosugi Y, Sugiyama S, Ohira I (2014) 2,4-Pyridinedicarboxylic acid prolongs the vase life of cut flowers of spray carnations. J Japan Soc Hort Sci 83:72–80
Lizada MCC, Yang SF (1979) A simple and sensitive assay for l-aminocyclopropane-1-carboxylic acid. Anal Biochem 100:142–147
Kosugi Y, Shibuya K, Tsuruno N, Iwazaki Y, Mochizuki A, Yoshioka T, Hashiba T, Satoh S (2000) Expression of genes responsible for ethylene production and wilting are differently regulated in carnation (Dianthus caryophyllus L.) petals. Plant Sci 158:139–145
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Ana Biochem 72:248–254
Harada H, Murakoshi Y, Torii Y, Tanase K, Onozaki T, Morita S, Masumura T, Satoh S (2011) Analysis of genomic DNA of DcACS1, a 1-aminocyclopropane-1-carboxylate synthase gene, expressed in senescing petals of carnation (Dianthus caryophyllus) and its orthologous gene in D. superbus var. longicalycinus. Plant Cell Rep 30:519–527
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Satoh, S., Kosugi, Y. (2017). Escherichia coli-Based Expression and In Vitro Activity Assay of 1-Aminocyclopropane-1-Carboxylate (ACC) Synthase and ACC Oxidase. In: Binder, B., Eric Schaller, G. (eds) Ethylene Signaling. Methods in Molecular Biology, vol 1573. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6854-1_5
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DOI: https://doi.org/10.1007/978-1-4939-6854-1_5
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