A New and Simple Technique for the Isolation of Plasma Membrane Lipids from Root-Tips

It has been suggested that plasma membrane (PM) lipids play a major role in aluminum (Al) tolerance; however, no direct investigations have been carried out using PM lipids from root-tips. Here we report a new technique for PM isolation as an alternative to the laborious two-polymer phase partitioning method that is commonly applied, as follows: 1) separation of protoplasts from 1-cm root-tip portions by enzymatic digestion, 2) attachment of the purified protoplasts to glass plates coated with polylysine, 3) preparation of PM ghosts by successive burst of the attached protoplasts using three separate buffer solutions (25 mM PIPES, 5 mM EDTA, and 2 mM MgCl₂, at pH 7.0) with slow stirring for 60 s. The PMs were confirmed to be devoid of organelle membranes by fluorescence microscopy, thin layer chromatography (TLC) and western blot analysis. The PM lipids obtained were found to be useful for studies on their differential permeability and lipid composition between lines of triticale or cultivars of maize under Al stress.     

Elevated CO2 concentrations increase leaf nitrate reduction by strengthening sink activity in soybean plants

An experiment was conducted to examine the effect of CO₂ enrichment on the nitrate uptake, nitrate reduction activity, and translocation of assimilated-N from leaves at varying levels of nitrogen nutrition in soybean using ¹⁵N tracer technique. CO₂ enrichment significantly increased the plant biomass, apparent leaf photosynthesis, sugar and starch contents of leaves, and reduced-N contents of the plant organs only when the plants were grown at high levels of nitrogen. A high supply of nitrogen enhanced plant growth and increased the reduced-N content of the plant organs, but its effect on the carbohydrate contents and photosynthetic rate were not significant. However, the combination of high CO₂ and high nitrogen levels led to an additive effect on all these parameters. The nitrate reductase activity increased temporarily for a short period of time by CO₂ enrichment and high nitrogen levels. ¹⁵N tracer studies indicated that the increase in the amount of reduced-N by CO₂ enrichment was derived from nitrate-N and not from fixed-N of the plant. To examine the translocation of reduced-N from the leaf in more detail, another experiment was conducted by feeding the plants with ¹⁵NO₃-N through a terminal leaflet of an upper trifoliated leaf under depodding and/or CO₂ enrichment conditions. The export rate of ¹⁵N from the terminal leaflet to other plant parts decreased by depodding, but it increased by CO₂ enrichment. CO₂ enrichment increased the percentage of plant ¹⁵N in the stem and / or pods. Depodding increased the percentage of plant ¹⁵N in the leaf and stem. The results suggested that the increase in the leaf nitrate reduction activity by CO₂ enrichment was due to the increase of the translocation of reduced-N from leaves through the strengthening of the sink activity of pods and / or stem for reduced-N.     

Effects of Ozone and/or Excess Soil Nitrogen on Growth, Needle Gas Exchange Rates and Rubisco Contents of Pinus Densiflora Seedlings

The effects of ozone (O₃) and excess soil nitrogen (N), singly and in combination, on growth, needle gas exchange rates and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) contents of Pinus densiflora seedlings were investigated. One-year-old seedlings were grown in 1.5-L pots filled with brown forest soil with 3 levels of N supply (0, 100 or 300 mg N·L^?1 fresh soil volume). The seedlings were exposed to charcoal-filtered air or 60±5 nL·L^?1 O₃ (8 hours a day) in naturally-lit phytotrons for 173 days from 22 May to 11 November. The exposure to O₃ or high N supply to the soil caused a significant reduction in the dry weights of the seedlings. Although no significant interactive effects of O₃ and excess soil N were detected on the dry weight growth of the seedlings, the whole-plant dry weight of the O₃-exposed seedlings grown in the soil treated with 300 mg N·L^?1 was greatly reduced compared with the control value. Ozone reduced net photosynthetic rate at 350 µmol·mol^?1 CO₂ (A ₃₅₀ ), carboxylation efficiency (CE) of photosynthesis and Rubisco content without a significant change in the gaseous phase diffusive conductance to CO₂ (gs) of the needles. The excess soil N reduced the A ₃₅₀ , CE, gs and Rubisco content of the needles. These results suggest that the reduction in the dry weight growth of Pinus densiflora seedlings induced by the exposure to O₃ and/or excess soil N was caused by reduction in the net photosynthetic rate mainly due to the decrease of Rubisco quantity in the chloroplasts.     

The Process of Rice Domestication: A New Model Based on Recent Data

Rice domestication involved a complex process of selection by ancient humans, leading to the development of a new species, Oryza sativa, during the last 10,000 years. Recent rice genome research and the cloning of domestication-related genes have provided novel knowledge about the domestication process, although recent reviews have revealed more complexity than previously suspected. Some of this additional complexity may arise from a poor fit between the new data and previous models. DNA divergence observed in existing landraces, cultivars, and wild relatives can now be determined accurately, so the order of major DNA changes such as single nucleotide polymorphisms during the domestication process can be abducted owing to relatively low mutation rates. Therefore, a new model to explain the selection process during rice domestication, such as the one proposed for further discussion in this paper, is needed to accommodate the new DNA evidence.     

A model for explaining genotypic and environmental variation in vegetative biomass growth in rice based on observed LAI and leaf nitrogen content

The objectives of this study are to propose a model for explaining the genotypic and environmental variation in above-ground biomass growth via photosynthesis and respiration processes from transplanting to heading for different rice genotypes grown under a wide range of environments, and to identify the physiological traits associated with genotypic difference in the biomass growth based on model analysis. Cross-locational experiments were conducted with nine different rice genotypes at eight locations in Asia covering a wide climate range under irrigated conditions with sufficient nitrogen application. The crop growth rate observed during the period from transplanting to heading ranged from 3.4 to 19.4 g m⁻² d⁻¹ among the genotypes grown at the eight locations. About one-third of the data sets were utilized for model calibration and the remaining sets were used for model validation. An above-ground biomass growth model was developed by integrating processes of single leaf photosynthesis as a function of stomatal conductance and leaf nitrogen content, growth and maintenance respiration and crop development. To rigorously examine the validity of this process model, measured data were input as external variables for leaf area index (LAI) development and leaf nitrogen content per unit leaf area. The model well explained the observed dynamics in above-ground biomass growth (R² = 0.95*** for validation dataset) of nine rice genotypes grown under a variety of environments in Asia. The model simulation suggested that genotypic difference in the biomass growth was closely related to the difference in the stomatal conductance and leaf nitrogen content, as well as to LAI. This paper proposes the model structure, algorithms and all parameter values contained in the model, and discuss its effectiveness as a component of a more comprehensive model for simulating dynamics of biomass growth, LAI development and nitrogen uptake as a function of genotypic coefficients and environments.     

Structure and properties of syndiotactic polystyrene fibers prepared in high-speed melt spinning process

High-speed melt spinning of syndiotactic polystyrene was carried out using high and low molecular weight polymers, HMs-PS and LMs-PS, at the throughput rates of 3 and 6 g/min. The effect of take-up velocity on the structure and properties of as-spun fibers was investigated. Wide angle X-ray diffraction (WAXD) patterns of the as-spun fibers revealed that the orientation-induced crystallization started to occur at the take-up velocities of 2–3 km/min. The crystal modification wasα-form. Birefringence of as-spun fibers showed negative value, and the absolute value of birefringence increased with an increase in the take-up velocity. The cold crystallization temperature analyzed through the differential scanning calorimetry (DSC) decreased with an increase in the take-up velocity in the low speed region, whereas as the melting temperature increased after the on-set of orientation-induced crystallization. It was found that the fiber structure development proceeded from lower take-up velocities when the spinning conditions of higher molecular weight and lower throughput rate were adopted. The highest tensile modulus of 6.5 GPa was obtained for the fibers prepared at the spinning conditions of LMs-PS, 6 g/min and 5 km/min, whereas the highest tensile strength of 160 MPa was obtained for the HMs-PS fibers at the take-up velocity of 2 km/min. Elongation at break of as-spun fibers showed an abrupt increase, which was regarded as the brittle-ductile transition, in the low speed region, and subsequently decreased with an increase in the take-up velocity. There was a universal relation between the thermal and mechanical properties of as-spun fibers and the birefringence of as-spun fibers when the fibers were still amorphous. The orientation-induced crystallization was found to start when the birefringence reached — 0.02. After the starting of the orientation-induced crystallization, thermal and mechanical properties of as-spun fibers with similar level of birefringence varied significantly depending on the processing conditions.     

Determination of the Chemical Structures of Tandyukisins B–D,Isolated from a Marine Sponge-Derived Fungus

Tandyukisins B–D (1–3), novel decalin derivatives, have been isolated from a strain of Trichoderma harzianum OUPS-111D-4 originally derived from the marine sponge Halichondria okadai, and their structures have been elucidated on the basis of spectroscopic analyses using 1D and 2D NMR techniques. In addition, their chemical structures were established by chemical transformation. They exhibited weak cytotoxicity, but selective growth inhibition on panel screening using 39 human cancer cell lines.     

Effect of an Introduced Phytoene Synthase Gene Expression on Carotenoid Biosynthesis in the Marine Diatom Phaeodactylum tricornutum

Carotenoids exert beneficial effects on human health through their excellent antioxidant activity. To increase carotenoid productivity in the marine Pennales Phaeodactylum tricornutum, we genetically engineered the phytoene synthase gene (psy) to improve expression because RNA-sequencing analysis has suggested that the expression level of psy is lower than other enzyme-encoding genes that are involved in the carotenoid biosynthetic pathway. We isolated psy from P. tricornutum, and this gene was fused with the enhanced green fluorescent protein gene to detect psy expression. After transformation using the microparticle bombardment technique, we obtained several P. tricornutum transformants and confirmed psy expression in their plastids. We investigated the amounts of PSY mRNA and carotenoids, such as fucoxanthin and β-carotene, at different growth phases. The introduction of psy increased the fucoxanthin content of a transformants by approximately 1.45-fold relative to the levels in the wild-type diatom. However, some transformants failed to show a significant increase in the carotenoid content relative to that of the wild-type diatom. We also found that the amount of PSY mRNA at log phase might contribute to the increase in carotenoids in the transformants at stationary phase.