Toxicity of the antifouling biocide Sea-Nine 211 to marine algae, crustacea, and a polychaete

We evaluated the acute toxicity of the antifouling biocide Sea-Nine 211 to the algae Chaetoceros calcitrans, Dunaliella tertiolecta, Tetraselmis tetrathele, and Skeletonema costatum, the crustacea Tigriopus japonicus and Portunus trituberculatus, and the polychaete Perinereis nuntia. The algae, and especially the diatoms C. calcitrans and S. costatum, were sensitive to Sea-Nine 211 toxicity, with the average acute toxicity values being 0.32, 3.9, 1.6, 0.22, 1.6, 12, and 27 μg/l for C. calcitrans, D. tertiolecta, T. tetrathele, S. costatum, T. japonicus, P. trituberculatus, and P. nuntia, respectively. A sediment toxicity test for Sea-Nine 211 using the polychaete P. nuntia revealed demonstrated that the 14-day median lethal concentration was 110 μg/kg dry-wt sediment and that growth was the most sensitive indicator. The chronic toxicity values of Sea-Nine 211 for the diatoms C. calcitrans and S. costatum were within the range of reported Sea-Nine 211 concentrations in seawater in coastal Japan, and the toxicity values for P. nuntia were within the reported concentrations in sediment. Based on these results, Sea-Nine 211 may have toxic effects on some sensitive species residing in the coastal areas of Japan, but the ecological risk posed by Sea-Nine 211 would appear to be confined to a limited area of Japanese coastal waters.     

Simple prediction of crude protein content in vegetation growing on abandoned cultivated land

The accuracy of two simple methods was compared for the prediction of crude protein (CP) content of above‐ground plant material of mixed‐species composition on abandoned cultivated land in Japan. The first method is based on standard CP values (in g kg^?1 dry matter) for individual species (STV method) as listed in the literature. The second procedure (GLM method) was an application of the generalized linear model using the relative above‐ground biomass of monocots and legumes, total herbage mass, and day of year. Predictions were made at the quadrat scale, and for surveyed sites based on average of values for five or six quadrats in a single survey. A ‘leave‐site‐out’ method was adopted for model validation of the generalized linear model. The observed values of CP content ranged between 21·5 and 161·9 g kg^?1 dry matter (DM). With the STV method, the values of root mean square error (RMSE indicates average estimation error) were 50·9 at the quadrat level and 53·8 at the surveyed‐site level (both g kg^?1 DM). When a ‘leave‐site‐out’ validation was carried out, the RMSE‐values for the GLM method were 23·2 at the quadrat level and 13·2 at the surveyed‐site level (both g kg^?1 DM). We therefore propose adoption of the GLM method for the purpose of estimating the CP content in herbage on abandoned sites.     

Relation between pathogenicity and genetic variation within <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

The relation between diversity of pathogenicity on clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa subsp. pekinensis) bred in Japan and DNA polymorphisms in 17 populations of Plasmodiophora brassicae from cruciferous plants was examined by inoculation tests and random amplified polymorphic DNA (RAPD) analysis using 18 arbitrary primers. Four pathotypes (A–D) were identified after inoculation of six CR cultivars of Chinese cabbage in the 17 populations from cruciferous crops. A relatively high level of genetic diversity was also detected among these populations in the RAPD analysis. Although the four pathotypes could not be clearly differentiated using the RAPD data, most populations of three pathotypes had a consistent location on the dendrogram. All pathotype B (virulent on five cultivars except Utage 70) and D (avirulent on all cultivars) populations, which were common in incompatible interactions with cv. Utage 70, were located in a single subcluster. All five pathotype C populations (virulent only on cv. Utage 70) except for one population grouped in another single subcluster. Because four pathotype A populations (virulent on all six cultivars, races 4 and 9) fell in different subclusters, the populations may be genetically polyphyletic. Populations from cruciferous weed Cardamine flexuosa differed remarkably from those from cruciferous crops in pathogenicity on common cultivars of Chinese cabbage and turnip and C. flexuosa, but they grouped in a single cluster with all race 9 populations from crops. Race 9 populations from crops may thus be closely related to populations from the weed rather than to races 1 and 4 from crops.     

Involvement of circadian clock in crowing of red jungle fowls (Gallus gallus)

The rhythmic locomotor behavior of flies and mice provides a phenotype for the identification of clock genes, and the underlying molecular mechanism is well studied. However, interestingly, when examining locomotor rhythm in the wild, several key laboratory‐based assumptions on circadian behavior are not supported in natural conditions. The rooster crowing ‘cock‐a‐doodle‐doo’ is a symbol of the break of dawn in many countries. Previously, we used domestic inbred roosters and showed that the timing of roosters' crowing is regulated by the circadian clock under laboratory conditions. However, it is still unknown whether the regulation of crowing by circadian clock is observed under natural conditions. Therefore, here we used red jungle fowls and first confirmed that similar crowing rhythms with domesticated chickens are observed in red jungle fowls under the laboratory conditions. Red jungle fowls show predawn crowing before light onset under 12:12 light : dim light conditions and the free‐running rhythm of crowing under total dim light conditions. We next examined the crowing rhythms under semi‐wild conditions. Although the crowing of red jungle fowls changed seasonally under semi‐wild conditions, predawn crowing was observed before sunrise in all seasons. This evidence suggests that seasonally changed crowing of red jungle fowls is under the control of a circadian clock.     

Comparisons of hyphal growth of Raffaelea quercivora among four Japanese Fagaceae species

Japanese oak wilt causes widespread oak mortality in Japan. Possible differences in susceptibility to the causal fungus, Raffaelea quercivora, may be due to vessel arrangements in host trees. To clarify whether constitutive defence mechanisms including vessel arrangements or induced defence mechanisms are the main determinants of host susceptibility, we inoculated the fungus into living seedlings or sterilized stem segments of four Japanese fagaceous species. In seedlings, water conductance was assessed with dye. In both seedlings and stem segments, hyphal growth was examined by fluorescence microscopy. In seedlings, the area of non‐conductive sapwood in stem cross sections and hyphal growth differed significantly among species. In the susceptible species Quercus crispula and Quercus serrata, hyphal growth was significantly and positively correlated with the proportion of non‐conductive sapwood. In stem segments, hyphal growth was not significantly different among species or between vessel arrangements and was similar to or greater than that in seedlings. These results suggest that the extent of sapwood colonization by R. quercivora could be used as a marker for susceptibility and that susceptibility is determined mainly by induced defence responses.     

Antifungal activities of hyoscyamine and scopolamine against two major rice pathogens: <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis> and <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

The antifungal activities of hyoscyamine and scopolamine, major alkaloids extracted from the desert plant Hyoscyamus muticus, against two rice pathogens, Magnaporthe oryzae and Rhizoctonia solani, were studied. The minimum inhibitory concentration of hyoscyamine that resulted in distinctive inhibition (MIC₅₀) was 1 μg/ml for both fungi. Exposure to hyoscyamine caused the leakage of electrolytes from the mycelia of both fungi. Hyoscyamine (>1 μg/ml) irreversibly delayed or inhibited conidial germination and appressorium formation in M. oryzae grown on polystyrene plates. Hyoscyamine effectively inhibited the attachment of conidia to the surface of rice (Oryza sativa) leaves and inhibited appressorium formation on the leaves. A high concentration of scopolamine (1000 μg/ml) also delayed or inhibited conidial germination in M. oryzae, but conidial germination was restored after washing the conidia with water. Antifungal activity of hyoscyamine was reduced by scopolamine. Magnaporthe oryzae infection was significantly suppressed (by >95%) in leaves of intact rice plants treated with hyoscyamine (10 μg/ml). Moreover, 10 μg hyoscyamine/ml significantly reduced the disease severity index for sheath blight to ≤0.2, when compared with the disease index of control plants (>7.0). Hyoscyamine (>20 μg/ml) completely inhibited sclerotial germination and development of R. solani by delaying the initiation, maturation, and melanization of the sclerotia. These results suggest that tropane alkaloids may be useful for controlling blast and sheath blight diseases of rice and for studying the mechanisms that regulate conidial germination in M. oryzae and sclerotial germination and development in R. solani.     

Choanephora rot of floral tops of <Emphasis Type="Italic">Hyoscyamus muticus</Emphasis> caused by <Emphasis Type="Italic">Choanephora cucurbitarum</Emphasis>

Floral rot of Egyptian henbane (Hyoscyamus muticus L.) was found on potted plants in a greenhouse in Yamaguchi city, Japan, in the late summer of 2008 and 2009. The symptoms were identical to those of rots caused by Choanephora species. The pathogen was isolated and identified as C. cucurbitarum (Berkeley and Ravenel) Thaxter. This new disease was named Choanephora rot (Kougai-kabi-byo) of Egyptian henbane.     

Effect of height-to-diameter ratio on stem stiffness of sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) cultivars

The lumber from sugi plantations in Japan displays large intra- and intertree variation in mechanical properties, even within a stand. These variations seem to be induced by the effects of the characteristics of cultivars as well as the effects of growth traits on mechanical properties. Therefore, the effects of growth traits on mechanical properties per cultivar need to be precisely examined. In this study, we focused on the effects of growth traits, especially height-to-diameter ratio (H/D ratio), on stem stiffness per cultivar. Sixteen cultivars were classified into three groups according to the relationships between stem stiffness and growth traits. In cultivars that showed a close correlation between growth traits and stem stiffness, it was assumed that stem stiffness could be controlled to a certain extent by silvicultural practices using the H/D ratio as an indicator. In cultivars that showed a weak correlation between growth traits and stem stiffness, selecting cultivars for the production of logs with higher mechanical properties seemed to be effective; in this study, Kumotoshi, Tanoaka, and Edanaga were found to be suitable. Tree age and site index may be important factors for producing wood with higher mechanical properties.