Homeostasis of brassinosteroids (BRs) is essential for normal growth and development in higher plants. We examined responsiveness of 11 BR metabolic gene expressions to the decrease or increase of endogenous BR contents in Arabidopsis (Arabidopsis thaliana) to expand our knowledge of molecular mechanisms underlying BR homeostasis. Five BR-specific biosynthesis genes (DET2, DWF4, CPD, BR6ox1, and ROT3) and two sterol biosynthesis genes (FK and DWF5) were upregulated in BR-depleted wild-type plants grown under brassinazole, a BR biosynthesis inhibitor. On the other hand, in BRexcessive wild-type plants that were fed with brassinolide, four BR-specific synthesis genes (DWF4, CPD, BR6ox1, and ROT3) and a sterol synthesis gene (DWF7) were down-regulated and a BR inactivation gene (BAS1) was up-regulated. However, their response to fluctuation of BR levels was highly reduced (DWF4) or nullified (the other eight genes) in a bri1 mutant. Taken together, our results imply that BR homeostasis is maintained through feedback expressions of multiple genes, each of which is involved not only in BR-specific biosynthesis and inactivation, but also in sterol biosynthesis. Our results also indicate that their feedback expressions are under the control of a BRI1-mediated signaling pathway. Moreover, a weak response in the mutant suggests that DWF4 alone is likely to be regulated in other way(s) in addition to BRI1 mediation.

The antimutagenic activity of n-hexane extracts from eight strains of daikon (Raphanus sativus; Japanese white radish) have been examined using the UV-induced mutation assay of Escherichia coli B/r WP2. A correlation was found between the potency of antimutagenicity and the amount of 4-(methylthio)-3-butenyl isothiocyanate (MTBITC) in their n-hexane extracts. Because the pure MTBITC also showed antimutagenicity, MTBITC is presumably the active antimutagen principle in n-hexane extracts of daikon. Among the eight strains of daikon studied, Aokubi, the improved common strain in Japan, contained 71.0 micromol of MTBITC in 100 g of fresh daikon. In contrast, Karami and Momoyama, which are original wild strains, contained much more MTBITC (363.5 and 168.0 micromol/100 g, respectively). In addition, phenethyl isothiocyanate was found in a lesser amount (5-33 nmol/100 g) in eight strains of daikon, and allyl isothiocyanate and benzyl isothiocyanate were not detectable in any strains (<3 nmol/100 g). The amount of total isothiocyanate in grated daikon was 7.0 times higher than that in cut daikon measured after 30 min of cooking. Through eating habits, humans might be able to consume substantial amounts of the antimutagen MTBITC from dishes using the grated form of wild strains of daikon. Therefore, it is possible to substantially increase the intake of the antimutagenic ingredient of daikon (i.e., MTBITC) by changing food preferences and preparation procedures (i.e., using the grated form of the wild strains).

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