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In this study, we identified and evaluated the genetic relationships among Cinnamomum plants, which are used in traditional medicine. We also attempted to monitor the distribution of traditional medicines derived from Cinnamomum cassia by using DNA barcoding and a species-specific DNA marker. Plants of the genus Cinnamomum, and in particular C. cassia, are commonly used as medicinal herbs in the form of Cinnamomi Ramulus, Cinnamomi Cortex, and Cassiae Cortex Interior. However, it is difficult to distinguish among different Cinnamomum species based on morphological features, and so to overcome this limitation, nucleotide sequences of the internal transcribed spacer (ITS) region of Cinnamomum DNA were determined and compared. On the basis of the discrepancy in determined ITS sequences, a 408-bp product, amplified by the primer pair CC F1/CC R3, was developed as a C. cassia-specific DNA marker. Using the developed DNA marker in combination with the ITS 2 nucleotide sequence, we monitored imported and commercially supplied medicinal products derived from Cinnamomum plants in markets in Korean, China, and Japan. The results revealed that most of the specimens monitored were derived from C. cassia.Electronic supplementary materialThe online version of this article (doi:10.1007/s13258-016-0476-5) contains supplementary material, which is available to authorized users.

Exogenous ACC-induced ethylene production in mung bean hypncotyl segments was severely inhibited by the treatment of spermine in a 6 hr incubation. In protoplasts acquired from the same tissues and pretreated with ACC, the concentration of cytosolic Ca 2~ was increased after the addition of exogenous Ca :+. However, this increase of Ca z+ concentration was strongly suppressed by spermine. In previous studies, an artifical increase of Ca 2+ influx by the treatment of a Ca2 § stimulated ACC-induced ethylene production. The inhibitory effect of spermine on ACC-induced ethylene production was more prominent than that of putrescine which has fewer NH ~ § groups than spermine. In addition, spermine more prominently suppressed ACC-induced ethylene production in protoplasts in which tewer Ca z~ ions were released from CaZ § organelles. Also, the amount of transcript of ACC-oxidase which converts ACC to ethylene was decreased by the treatment of spermine. However, this reduction resulted only through the suppression of ethylene production in a 2 hr incubation of mung bean segments. On the basis of these results, we suggest that there is a coupling of ACC-uptake to the increase of cylosolic Ca 2~ concentration. In addition, the reduction of exogenous ACC-induced ethylene production by spermine could have resulted, at least partially, from reducing the Ca 2~ influx which stimulates A('C-oxidase activity.

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Purified malformin A1 (cyclo-D-Cys-D-Cys-L-VaI-D-Leu-L-Ile), a cyclicpentapeptide toxin from Aspergillus niger, was applied to the hypocotyl segments of mung bean (Vigna radiata L.) seedlings to investigate its role in regulating ethylene biosynthesis. Production of ethylene was induced by treating the plants with 0.1 mM indole-3-acetic acid (IAA). When 0.1 pM malformin A1 was then applied, ethylene production increased and the activities of two key enzymes for its biosynthesis, 1-aminocyclopropane-l-carboxylic acid (ACC)-synthase (ACS) and ACC-oxidase (ACO), were also stimulated. However, at levels of I or 10 laM malformin A1, both ethylene production and enzymatic activities were significantly reduced. In the case of ACO, in vitro activity was regulated by malformin A1, independent of ACS activity or the influence of IAA. Furthermore, the conjugate form of ACC, N-malonyl ACC, was significantly promoted by treatment with 0.1 I~M malformin A1. These data suggest that malformin A1 can modulate ethylene production through diverse paths and that its effect depends on the concentration of the treatment administered.