Terrein is a bioactive fungal metabolite whose effects are almost unknown. In this study, we found for the first time that terrein has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 microM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrein treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrein reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.
Oxytricha nova telomeric DNA contains guanine-rich short-tandem repeat sequences (GGGGTTTT) n and terminates as a single strand at the 3'-end. This single-stranded overhang forms a novel DNA structure, namely, G-quadruplex, comprising four quartets. In this study, we investigated the structures and dynamics of unimolecular Oxytricha nova ( O. nova) telomeric G-quadruplexes by performing single molecule fluorescence resonance energy transfer (FRET) spectroscopy and bulk circular dichroism (CD) measurements. We observed that unimolecular O. nova G-quadruplexes exhibit structural polymorphism according to monovalent cations. In the presence of Na (+), only antiparallel conformation is detected, which was demonstrated in previous studies; however, in the presence of K (+), they fold into two different conformations, a parallel conformation and an antiparallel one different from that induced by Na (+). Furthermore, these G-quadruplexes show extremely high stability in their dynamics when compared with human G-quadruplexes. While human telomeric G-quadruplexes that possess three quartets display fast dynamic behavior (<100 s) at low K (+) concentrations or high temperatures, O. nova G-quadruplexes maintain their conformational state for a long time (>1000 s), even at the lowest K (+) concentration and the highest temperature investigated. This high stability is primarily due to an extra quartet that results in additional cation coordination. In addition to cation coordination, we propose that other factors such as base stacking and the size of the thymine loop may contribute to the stability of O. nova G-quadruplexes; this is based on the fact that the O. nova G-quadruplexes were observed to be more stable than the human ones in the presence of Li (+), which is known to greatly destabilize G-quadruplexes because of imprecise coordination. This extreme stability of four-quartet G-quadruplexes enables telomere protection even in the absence of protective proteins or in the case of abrupt environmental changes, although only a single G-quadruplex structure can be derived from the short single-stranded overhang.
Timely detection of herbicide resistance at an early stage of crop cultivation is essential to help farmers find alternative solutions to manage herbicide resistance in their fields. In this study, maximum quantum yield of PS II [F v /F m = (F m -F o )/F m ] was measured at the 4-5 leaf stage to discriminate between herbicideresistant and susceptible biotypes of Echinochloa species. The differences in F v /F m between herbicide-resistant and susceptible Echinochloa spp. were consistent with the whole-plant assay based on I 50 (herbicide doses causing a 50% inhibition of F v /F m ) and GR 50 (herbicide doses causing a 50% reduction in plant fresh weight) values and R/S ratios (herbicide resistance index), regardless of the mode of action of the tested herbicides. A PS II inhibitor caused the fastest inhibition of F v /F m , compared with ACCase and ALS inhibitors, after herbicide treatment. The required time for discrimination between herbicide-resistant and susceptible Echinochloa spp. was 64 h after PS II inhibitor treatment, much shorter than those of ACCase and ALS inhibitor-treated plants, which required 168 and 192 h respectively. The leaf chlorophyll fluorescence assay provided reliable diagnostics of herbicide resistance in Echinochloa spp. with significant time savings and convenient measurement in field conditions compared with the conventional whole-plant assay.
Summary The effects of herbicide dose on rice‐weed competition were investigated to develop a combined model, which can be utilised to estimate an optimum herbicide dose for a given weed density in paddy rice cultivation. Field studies were conducted in Suwon for rice‐Echinochloa crus‐galli competition and Iksan for rice‐Eleocharis kuroguwai during 2007. The competitive effect of the weeds E. crus‐galli and E. kuroguwai decreased with increasing doses of flucetosulfuron and azimsulfuron, respectively, in the same manner as the standard dose–response curve. The combination of the rectangular hyperbolic model and the standard dose–response curve adequately described the complex effects of herbicide dose and weed competition on rice yield. Parameter estimates were used with the model to predict rice yield and estimate the doses of flucetosulfuron and azimsulfuron required to restrict rice yield loss caused by E. crus‐galli and E. kuroguwai, respectively, to an acceptable level. For a rice yield of 5.0 t ha−1, the model recommended flucetosulfuron doses of 8.7, 13.4 and 20.1 g a.i. ha−1 when infested with E. crus‐galli at 12, 24 and 48 plants m−2 respectively. For a rice yield of 5.2 t ha−1, the model recommended azimsulfuron doses of 3.9, 7.5 and 12.6 g a.i. ha−1 when infested with E. kuroguwai at 24, 48 and 96 plants m−2 respectively. The theoretical outputs of the combined model appear robust and indicate there are opportunities for reduced herbicide use in the field. These now require evaluation under field conditions.
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