Effect of nitrate fertilization of gametophytes of the kelp Undaria pinnatifida on growth and maturation of the sporophytes cultivated in Matsushima Bay, northern Honshu, Japan

In northern Japan, the early production of high-quality specimens of kelps Undaria pinnatifida prior to the outbreak of pinhole disease is needed. To address this need, we tested the effect of nitrate fertilization of gametophytes on the growth and maturation of sporophytes using experimental and control gametophytes derived from sporophytes cultivated in Matsushima Bay, northern Honshu, Japan. From mid-August to October 2008, experimental gametophytes were cultured on a rope indoors at a nitrate concentration of 1 mg/L at the optimal seawater temperature of 20 °C, whereas control gametophytes were cultured without the addition of nitrate in normal seawater at an average temperature of 23.5 °C during this period in the past 30 years. Next, the seedlings were cultivated in Matsushima Bay until March 2009. For the experimental kelps, the total length, stipe length, stipe width, sporophyll length, sporophyll width, and dry weight of the blade, midrib, and sporophyll increased significantly from December to January compared to those of control kelps. Matured sporophylls of the experimental kelps were more than those of the control kelps from early December to late January. Significant higher photosynthesis occurred at seawater temperatures of 10 and 15 °C, as well as higher uptake rates of NO₃–N of the experimental kelps in December and January. These results show that nitrate fertilization of the gametophytes makes it possible to harvest the high-priced blade, midrib, and sporophyll in January, which is earlier than the traditional harvest date.     

Spatial variations in nitrous oxide and nitric oxide emission potential on a slope of Japanese cedar (Cryptomeria japonica) forest

To quantify the spatial variation and spatial structure of nitrous oxide (N₂O) and nitric oxide (NO) emission from forest soils, we measured N₂O and NO emission rates from surface soil cores taken at 1 m intervals on a cross-line transect (65 m × 20 m) on a slope of Japanese cedar ( Cryptomeria japonica ) forest in a temperate region of central Japan and analyzed the spatial dependency of N oxide gas emissions using geostatistics. We divided N₂O emission into N₂O from denitrification and N₂O from nitrification using the acetylene inhibition method. According to the geostatistical analysis, N₂O emission rates on the slope had large spatial variation and weak spatial dependency. This weak spatial dependency was caused by the inordinately high N₂O emissions on the slope, which were derived mainly from denitrification. In contrast, NO emission rate on the slope had large spatial variation, but strong spatial dependency and a distinct spatial distribution related to slope position, that is, high in the middle of the slope and low in the shoulder and the foot of the slope. The CN ratio and water-filled pore space were the dominant factors controlling NO emission rate on a slope. Our results suggest that spatial information about topographic factors helps to improve the estimation of both N₂O emission and NO emission from forest soils.     

Biological roles of monoterpene volatiles derived from rough lemon (<Emphasis Type="Italic">Citrus jambhiri</Emphasis> Lush) in citrus defense

Volatile compounds of plants, including monoterpenes, are a possible source of signal molecules that induce defense systems to protect plants from tissue damage. Volatile compounds from rough lemon leaves were trapped by solid-phase microextraction fibers in sealed vials, and subsequent gas chromatography–mass spectrometry and gas chromatography analyses identified the profile of the major components, mainly various monoterpenes. Among several monoterpenes examined, citral, citronellal, and linalool significantly inhibited the spore germination and hyphal growth of Alternaria alternata. The effect of linalool was fungistatic, while the effects of citral and citronellal were partially fungicidal. Wounding of rough lemon leaves induced a significant increase in release of monoterpenes. The release of linalool was the most abundant and was 14.5 times that of unwounded rough lemon leaves. Unlike the wounding treatment, microbe attack did not significantly change monoterpene releases, and there was statistically no difference found in the peak areas from microbe-treated and untreated leaves. Linalool, limonene, and β-pinene also had insect-repellant effects on wild-type Drosophila melanogaster. Expression patterns of defense-related genes in rough lemon and rice significantly changed after treatment with vapors of monoterpene volatiles. Taking these results together, monoterpene volatiles are likely to play roles in the defense of rough lemon against microbe and insect pathogens.     

Activation with ethanol improves embryo development of ICSI-derived oocytes by regulation of kinetics of MPF activity

Developmental potential of bovine embryos that are not artificially activated after intracytoplasmic sperm injection (ICSI) is generally very low. In this study, we investigated effects of artificial activation with ethanol on kinetics of maturation promoting factor (MPF) activity (p34(cdc2) kinase activity) and development of bovine oocytes following ICSI. Treatment of oocytes with ethanol at 4 h after ICSI improved their first cleavage and further preimplantation development (51% vs. 13%, 14% vs. 4%: treatment with vs. without ethanol, respectively). MPF activity of oocytes was lowered until at least 2 h after ICSI. In oocytes without activation after ICSI, MPF activity temporarily elevated at 6 h after ICSI, whereas this phenomena was not observed in the oocytes treated with ethanol. Furthermore, MPF activity was elevated 20 h after ICSI in oocytes activated with ethanol, whereas this elevation of MPF activity was not shown in oocytes without activation. These results indicate that the stimulus of sperm was sufficient to lower MPF activity of oocytes following ICSI, and moreover the activation treatment of bovine oocytes with ethanol after ICSI served to maintain the low levels of MPF activity until the next cell cycle started.     

Effect of alkali extraction on the lignin monomeric composition inEucalyptus

The effect of alkali extraction on the lignin monomeric composition was examined inEucalyptus camaldulensis andE. globulus by thioacidolysis using extractive-free samples as a control. Results showed that the effect onEucalyptus is different among species and among sample positions in the trunk, although a small amount of lignin is solubilized during the extraction in all samples. In addition, it was proved that lignin extracted by the alkali extraction is not always guaiacyl-rich, probably relating to the original lignin monomeric composition, which depends on the sample species or the sample position in the trunk.     

Inhibition activity of essential oils obtained from Japanese trees against <Emphasis Type="Italic">Skeletonema costatum</Emphasis>

The growth inhibition activities of essential oils obtained from Cryptomeria japonica, Chamaecyparis obtusa, and Pinus thunbergii were examined against the bacillariophyceae Skeletonema costatum, also known as red tide plankton. The essential oils were extracted from the heartwood, leaves, and bark of these typical indigenous Japanese conifers. The essential oils from C. japonica bark and P. thunbergii heartwood possessed strong growth inhibition activity. The chemical compositions of these essential oils were analyzed by gas chromatography/flame ionization detection (GC-FID) and gas chromatography/mass spectrometry (GC-MS). α-Terpineol and longifolene were the main components of the essential oil from P. thunbergii heartwood. The C. japonica bark essential oil was mainly composed of α-terpineol, δ-cadinene, isophyllocladene, and ferruginol. Ferruginol and longifolene showed more potent growth inhibition against S. costatum than hinokitiol (β-thujaplicine), which is known to be a strong antifungal compound among wood components. Ferruginol and longifolene were important factors for the growth inhibition activity of the essential oils from C. japonica bark and P. thunbergii heartwood, respectively. These results suggest the possibility of using C. japonica bark and P. thunbergii heartwood for the control of red tide plankton.     

Quinone profiling of bacterial populations developed in the surface layer of volcanic mudflow deposits from Mt. Pinatubo (the Philippines)

The diversity of bacterial populations developed in the surface layer (0-0.25 m depth) of volcanic mudflow deposits from the Mt. Pinatubo volcano (the Philippines) was investigated using quinone profiling. Samples were collected from sites (named N and S1) that had been hit repeatedly by mudflows during successive rainy seasons after the violent eruption of 1991 and also from sites (F1 and F3) covered by mudflow in 1991 but with no deposition in following years. The total microscopic count ranged from 10⁸ g⁻¹ (N and S1 sites) to 3.9×10⁹ g⁻¹(site F3). In the N sample only three quinone species were detected, while the quinone profiles of samples from sites S3 and F3 showed higher diversity. Tetrahydrogenated menaquinone with eight isoprenoid units [MK-8 (H₄)] was the predominant quinone species in the sample from site N, while MK-8, MK-8 (H₂), MK-8 (H₄), MK-9 (H₄) and MK-9 (H₈) were found as major quinones in the sample from site F3. Because these MK species are known to be the major respiratory quinones of the Actinobacteria, this bacterial group is expected to predominate in the land with primary vegetation recovery following the impact of the volcanic mudflow.