Mos and the mitogen-activated protein kinase do not show cytostatic factor activity in early mouse embryos

Mos and the mitogen-activated protein kinase (MAPK) cascade have been established as crucial regulators of second meiotic metaphase arrest, the so-called CSF arrest, in mammalian oocytes. They are also thought to play a role in regulating mitotic metaphase arrest of early mammalian embryos. In the present study, we examined whether mitotic arrest is induced in early mouse embryos by activation of extracellular signal-regulated kinases (ERKs), which are major MAPKs in mouse eggs, and their substrate, p90Ribosomal S6 kinase (RSK), as reported in Xenopus embryos. Wild-type Mos (wt-Mos), degradation-resistant Mos mutant (P2G-Mos) or constitutive active mutant of MAPK/ERK kinase, MEK (SDSE-MEK), was expressed in early mouse embryos by injecting the respective expression vectors into the pronucleus of fertilized eggs, and the developmental rates were then examined up to 72 h after insemination. Expression of P2G-Mos and SDSE-MEK succeeded in activating ERKs and RSK in developing mouse embryos, while wt-Mos failed to activate them in spite of expression of mos mRNA, indicating that the wt-Mos protein is unstable in early mouse embryos. Although the activated levels of ERKs and RSK in the vector-injected embryos were comparable to those of meiotically arrested mouse oocytes, their developmental rates were identical to those of the control embryos. These results suggest that activation of MAPK and RSK does not induce mitotic arrest in early mouse embryos. The present study indicates that there are large physiological differences between early mouse embryos and mouse oocytes and that CSF arrest of mouse eggs in mitosis should be discussed separately from that in meiosis.     

Finding of a highly efficient ZFN pair for Aqpep gene functioning in murine zygotes

The generation efficiencies of mutation-induced mice when using engineered zinc-finger nucleases (ZFNs) have been generally 10 to 20% of obtained pups in previous studies. The discovery of high-affinity DNA-binding modules can contribute to the generation of various kinds of novel artificial chromatin-targeting tools, such as zinc-finger acetyltransferases, zinc-finger histone kinases and so on, as well as improvement of reported zinc-finger recombinases and zinc-finger methyltransferases. Here, we report a novel ZFN pair that has a highly efficient mutation-induction ability in murine zygotes. The ZFN pair induced mutations in all obtained mice in the target locus, exon 17 of aminopeptidase Q gene, and almost all of the pups had biallelic mutations. This high efficiency was also shown in the plasmid DNA transfected in a cultured human cell line. The induced mutations were inherited normally in the next generation. The zinc-finger modules of this ZFN pair are expected to contribute to the development of novel ZF-attached chromatin-targeting tools.     

Use of soil nonexchangeable potassium by paddy rice with clay structural changes under long-term fertilizer management

To prove the hypothesis that paddy rice utilizes soil nonexchangeable potassium (neK) and causes associated structural changes in clay minerals, K status and clay mineralogy of 22 surface soils from three paddy fields under long-term fertilizer management for 51–93 years were investigated. Soil neK content was determined as the difference between 1 mol L⁻¹ hot HNO₃ extractable K and 1 mol L⁻¹ ammonium acetate exchangeable K. Clay mineralogy was identified by X-ray diffraction (XRD). The radiocesium interception potential (RIP), an index of frayed edge sites in the interlayer sites of 2:1 type clay minerals, was also determined. The neK contents under the -K and NPK treatments were considerably lower than those under the unfertilized treatment in all the fields, indicating the exploitation of soil neK by rice. XRD analysis of the clay samples revealed 7% shift from the 1.0 peak to 1.4 nm one under the -K treatment compared with the unfertilized one, and the amounts of neK were negatively correlated with those of RIP (p < .01), suggesting the expansion of interlayer spaces of the 2:1 type phyllosilicates such as mica due to the release of neK. In addition, the neK content positively correlated with K balance of the long-term experiments (p < .05). The differences of neK between unfertilized K and -K treatments corresponded to 22–157 kg K ha⁻¹, or 0.42–1.68 kg K ha⁻¹ year⁻¹. In conclusion, utilization of considerable amount of soil neK under K depleted conditions should be considered to establish sustainable K management for paddy rice.     

Effects of application of lime nitrogen and dicyandiamide on nitrous oxide emissions from green tea fields

Abstract

The aim of this study was to assess the mitigating effects of lime nitrogen (calcium cyanamide) and dicyandiamide (DCD) application on nitrous oxide (N₂O) emissions from fields of green tea [Camellia sinensis (L.) Kuntze]. The study was conducted in experimental tea fields in which the fertilizer application rate was 544 kg nitrogen (N) ha^?1 yr^?1 for 2 years. The mean cumulative N₂O flux from the soil between the canopies of tea plants for 2 years was 7.1 ± 0.9 kg N ha^?1 yr^?1 in control plots. The cumulative N₂O flux in the plots supplemented with lime nitrogen was 3.5 ± 0.1 kgN ha^?1, approximately 51% lower than that in control plots. This reduction was due to the inhibition of nitrification by DCD, which was produced from the lime nitrogen. In addition, the increase in soil pH by lime in the lime nitrogen may also be another reason for the decreased N₂O emissions from soil in LN plots. Meanwhile, the cumulative N₂O flux in DCD plots was not significantly different from that in control plots. The seasonal variability in N₂O emissions in DCD plots differed from that in control plots and application of DCD sometimes increased N₂O emissions from tea field soil. The nitrification inhibition effect of lime nitrogen and DCD helped to delay nitrification of ammonium-nitrogen (NH₄⁺-N), leading to high NH₄⁺-N concentrations and a high ratio of NH₄⁺-N /nitrate-nitrogen (NO₃^–-N) in the soil. The inhibitors delayed the formation of NO₃^–-N in soil. N uptake by tea plants was almost the same among all three treatments.     

Aluminum-tolerance of carrot (daucus carota L.) plants regenerated from selected cell cultures

Abstract

Aluminum-tolerant variants from cell cultures of carrot (Daucus carota L. cv. MS Yonsun) were selected by exposing them to excess ionic aluminum. The medium contained A1CI₃ and 0.1 mM phosphate at pH 4.0. Under ionic aluminum stress (0.5-1.0 mM), 282 calli were selected. Twenty-eight fertile plants were regenerated from 7 selected calli. Based on the root elongation tests of the seedlings, the seeds displayed aluminum-tolerance. In the case of the cell line H7-08, the selection occurred at a frequency of ca. 6 × 10^-7 when the materials were plated on agar medium containing 0.5 mM aluminum.     

QTLs for phosphorus deficiency tolerance detected in upland NERICA varieties

Phosphorous (P) deficiency is a major yield limiting factor in rice (Oryza sativa L.) production. The interspecific New Rice for Africa (NERICA) varieties combine general stress tolerance from African cultivated rice (Oryza glaberrima Steud) with characteristics associated with high yield from O. sativa. However, little is known about their ability to tolerate P deficiency. Here, we examined the variation for tolerance to P deficiency among the 18 upland NERICAs and their parents in multi‐year field experiments. The good performance under P deficiency of the O. glaberrima parent CG 14 and some NERICAs suggested that these tolerant NERICAs contain loci associated with P deficiency tolerance inherited from CG 14. Additionally, four QTL clusters for P deficiency tolerance were detected on chromosomes 4, 6 and 11 using F₃ lines derived from the cross between the P deficiency tolerant variety NERICA10 and a Japonica‐type sensitive variety ‘Hitomebore’. These QTLs represent the first step in identifying stress tolerance genes from O. glaberrima that could subsequently be used to enhance P deficiency tolerance in O. sativa.     

ATP depletion inhibits retraction of chromatophores induced by GABA in cephalopod Todarodes pacificus

Color changes in cephalopods are generated by the expansion or retraction of chromatophores located under the dermis. The behavior of the chromatophores is regulated by neurotransmitters; l-glutamate (l-Glu) is an excitatory transmitter that causes the chromatophores to expand. To date, serotonin [5-hydroxytryptamine (5-HT)] is the only neurotransmitter known to stimulate retraction of chromatophores. We found that the chromatophores in the Japanese squid Todarodes pacificus were regulated by γ-aminobutyric acid (GABA), an inhibitory neurotransmitter, and that GABA caused expanded chromatophores to retract. We also found that adenosine 5′-triphosphate (ATP) concentrations in skin samples remained stable at their initial values for more than 24 h after the death of each squid; therefore, the chromatophores could respond to both l-Glu and GABA during that period. Furthermore, we attempted to reduce the levels of ATP by storing skin sample in sodium azide solution. The chromatophores in sodium azide-treated skin samples were induced to expand by l-Glu, but these expanded chromatophores could not be induced to retract by GABA. Based on these observations, we conclude that ATP is essential for retraction, but not expansion, of chromatophores.     

Toxicity and absorption of dietary leucomalachite green in Nile tilapia Oreochromis niloticus

Farmed fish are exposed to risks from feed-borne chemical contamination, such as leucomalachite green (LMG) in fish meal. Consequently, the use of malachite green is prohibited in aquacultural practice. An improved understanding of the toxicity of dietary LMG provided to farmed fish is needed in order to manage risk. Oreochromis niloticus specimens were fed experimental diets containing 0, 100, 500, and 2500 μg/kg LMG for 28 days. On sampling days 7, 14, and 28, fish in the exposed groups had detectable levels of LMG. Accumulation levels were approximately 12.2 % (in the liver) and 6.2 % (in the muscle) of the LMG concentration in the feed (104, 510, and 2200 μg/kg). Hematocrit and mean corpuscular hemoglobin concentrations in the 500 and 2500 μg/kg groups were significantly elevated at 7 and 14 days. Hemoglobin in the group that received the highest dose was significantly higher than that in the control group. Significant increases in the activities of alanine aminotransferase and alkaline phosphatase were also detected in the group receiving the highest dose. Total cholesterol concentrations in all of the exposed groups were significantly higher than those in the control group. These observations of toxicity were dose dependent. Histological changes in gills and livers were observed in LMG-exposed fish.     

Modeling the effects of N application on growth,yield and plant properties associated with the occurrence of chalky grains of rice

The objective of this study was to propose a model for explaining rice responses to a wide range of N application rates in various growth attributes associated with the occurrence of chalky grains. We improved the sub-model for N uptake process of a previous rice model which was originally developed for explaining genotypic and environmental variations in the whole growth processes, considering the difference in the rate of N loss from the plant-soil system between indigenously supplied soil mineral N and fertilizer N. A total of 80 growth datasets of cultivar ‘Koshihikari’ grown at Shiga prefecture, Japan, in 2010 was utilized for the calibration and validation of the model. The rice growth model well explained the above-ground biomass growth (RMSD = 78.7 g m^?2) and rough dry grain yield (RMSD = 83.2 g m^?2) for the validation data-set, simultaneously. The simulated carbohydrate content available per single spikelet was negatively correlated with the observed percentage of the milky-white grain which includes white-cored grain (r = ?.77, p < .001) for all the data-sets of calibration and validation. On the other hand, the observed percentage of the sum of white-back and white-base grains was closely correlated with the simulated plant N content available per single spikelet (r = ?.59, p < .001). It was suggested that the present rice growth model would rationally explain the effects of N application on the occurrence of the chalky grains through the dynamic change of the carbohydrate content and plant N content available per single spikelet.