Factors affecting the pattern of the dose response curve of clubroot disease caused by Plasmodiophora brassicae

The severity of clubroot disease caused by Plasmodiophora brassicae Woron. depended on the density of resting spores in soil, soil type, soil pH, and host susceptibility. The dose response curve (DRC) was determined to indicate the relationship between the disease index and these parameters for predicting the disease severity for each field. DRC patterns varied with the soil types and even among soils of the same type depending on the sampling areas. Disease incidence was lower in the soils adjusted to a higher pH than in those at the initial pH. DRC patterns were also influenced by plant species or cultivars. The DRC is useful for estimating the potential disease severity in agricultural fields with different soils and different plants and essential for the development of an integrated pest management (IPM) strategy for the control of clubroot disease.     

Operational efficiencies and costs of an arm roll forwarder: A case study at Nasu in Tochigi Prefecture, Japan

We investigated a developed arm roll forwarder at Nasu in Tochigi Prefecture, Japan. An arm roll forwarder can only load a steel container that has been fully loaded with logs beforehand, and can later unload such a container. Such a forwarder can shorten the loading and unloading times and improve operational efficiency. We examined two operation systems with an arm roll forwarder and a forwarder. In the first system the loading was done with a grapple-loader. In the second system the loading was done with a processor. The loading times of an arm roll forwarder were significantly less than those of a forwarder. Because the optimal cycle times (excluding the waiting times for an arm roll forwarder) were significantly reduced, the costs of using an arm roll forwarder are lower, although the loading capacity was small and the hourly operation cost was high. The maximum operational efficiencies varied depending on forwarding distances. The second operation system with an arm roll forwarder exhibited the best operational efficiency within a 1,580-m forwarding distance, and beyond that distance it exhibited the highest operational efficiency when a forwarder was used. Similarly, the cost of operation of the system with an arm roll forwarder was the lowest within a 1,130-m forwarding distance, and beyond that distance the cost was the lowest when using the forwarder. Therefore, the arm roll forwarder is effective within a certain forwarding distance.     

Increased Cellular Distribution of Vimentin and Ret in the Cingulum of Rat Offspring After Developmental Exposure to Decabromodiphenyl Ether or 1,2,5,6,9,10-Hexabromocyclododecane

Abstract: To determine effects of developmental exposure to brominated flame retardants (BFRs), weak thyroid hormone disruptors, on white matter development, white matter-specific global gene expression analysis was performed using microdissection techniques and microarrays in male rats exposed maternally to decabromodiphenyl ether (DBDE), one of the representative BFRs, at 10, 100 or 1000 ppm. Based on previous gene expression profiles of developmental hypothyroidism and DBDE-exposed cases, vimentin⁺ immature astrocytes and ret proto-oncogene (Ret)⁺ oligodendrocytes were immunohistochemically examined after developmental exposure to representative BFRs, i.e., DBDE, 1,2,5,6,9,10-hexabromocyclododecane (HBCD; 100, 1000 or 10,000 ppm) and tetrabromobisphenol A (TBBPA; 100, 1000 or 10,000 ppm). Vimentin⁺ and Ret⁺ cell populations increased at ≥ 100 ppm and ≥ 10 ppm DBDE, respectively. Vimentin⁺ and Ret⁺ cells increased at ≥ 1000 ppm HBCD, with no effect of TBBPA. The highest dose of DBDE and HBCD revealed subtle fluctuations in serum thyroid-related hormone concentrations. Thus, DBDE and HBCD may exert direct effects on glial cell development at ≥ middle doses. At high doses, hypothyroidism may additionally be an inducing mechanism, although its contribution is rather minor.     

Gentian Kobu-sho-associated virus: a tentative, novel double-stranded RNA virus that is relevant to gentian Kobu-sho syndrome

A virus-like dsRNA of about 23 kbp was detected in gentian plants showing Kobu-sho syndrome including stunting, shortened internodes, and tumors on stems, nodes and roots. Nucleotide sequence analysis has suggested that this dsRNA is related to Pestivirus species but not to any other plant viruses. It was protected from externally added RNase, suggesting that the dsRNA is encapsidated. The dsRNA was co-extracted in a crude homogenate of glutaraldehyde-fixed tissue with the virus-like particles that have been associated previously with Kobu-sho syndrome in gentian (Usugi et al. Jpn J Phytopathol 76:21–24, 2010). The RNA sequence was detected in more than 99 % of Kobu-sho gentian individuals but in less than 20 % of apparently healthy gentian individuals from fields affected with Kobu-sho. Thus, we propose naming the virus Gentian Kobu-sho-associated virus.     

Comparison of PCR assays for detection and quantification of Phomopsis sclerotioides in plant and soil

We developed a real-time PCR assay using a TaqMan probe (TM-qPCR) for specific detection and quantification of Phomopsis sclerotioides, causal agent of black root rot of cucurbit crops. The design of the primer sets and hybridization probe was based on the internal transcribed spacer region of the ribosomal DNA. The TM-qPCR assay was compared with a conventional, standard PCR (sPCR) assay and on a quantitative real-time PCR (SG-qPCR) assay based on SYBR Green I. The TM-qPCR assay had a detection limit of ca. 0.4 fg of P. sclerotioides DNA, which was approximately 100 times more sensitive than the sPCR assay and almost equivalent to the SG-qPCR assay. The TM-qPCR and SG-qPCR assays both were able to detect various quantities of P. sclerotioides DNA from diseased plants and infested soils, including DNA levels that were not detectable by the sPCR assay. However, the TM-qPCR was advantageous for samples containing PCR-inhibiting substances because its multiplex real-time PCR function allows the adjustment of cycle threshold values with an internal control. Based on the high specificity and sensitivity required for analyzing DNA in natural samples, the newly developed TM-qPCR assay was the most reliable tool for rapidly detecting and quantifying P. sclerotioides in plant and soil samples.     

Antiangiogenic activity of brown algae fucoxanthin and its deacetylated product, fucoxanthinol

The antiangiogenic effects of fucoxanthin and a deacetylated product, fucoxanthinol, were examined. Fucoxanthin significantly suppressed HUVEC proliferation and tube formation at more than 10 microM, but it had no significant effect on HUVEC chemotaxis. The formation of blood vessel-like structures from CD31-positive cells was evaluated using embryonic stem cell-derived embryoid bodies. Fucoxanthin effectively suppressed the development of these structures at 10-20 microM, suggesting that it could suppress differentiation of endothelial progenitor cells into endothelial cells involving new blood vessel formation. Fucoxanthin and fucoxanthinol suppressed microvessel outgrowth in an ex vivo angiogenesis assay using a rat aortic ring, in a dose-dependent manner. These results imply that fucoxanthin having antiangiogenic activity might be useful in preventing angiogenesis-related diseases.     

Practical method combining loop-mediated isothermal amplification and bait trap to detect <Emphasis Type="Italic">Pythium helicoides</Emphasis> from hydroponic culture solutions

Two detection methods combining loop-mediated isothermal amplification (LAMP) and a bait trap were developed to detect Pythium helicoides in greenhouses containing roses, miniature roses, and poinsettias in hydroponic culture systems. In “Bait-LAMP”, a crude extract derived from perilla seeds as the bait was used in the LAMP reaction, whereas in the “Bait culture-LAMP”, a crude extract of mycelia grown out from perilla seeds onto Pythium-selective medium served as the bait. The two methods are simple and rapid for practical monitoring of P. helicoides in hydroponic culture systems.     

Thermogravimetric evaluation of chitin degradation in soil: implication for the enhancement of ammonification of native organic nitrogen by chitin addition

Degradation of chitin, which is an aminopolysaccharide used as a soil amendment, has been often monitored in soil via its degradation products such as carbon dioxide and ammonium. We report here the applicability of thermogravimetry to measure the amount of chitin added to soil. The maximum pyrolysis rate of the upland surface soil of Brown Forest soil supplemented with chitin was strongly correlated with added chitin content (r = 0.999) when the content exceeded 6.0 g kg^?1. The maximum pyrolysis rates of chitin-added soil (around 385°C) was distinctive from those of soil supplemented with cellulose, chitosan, N-acetylglucosamine, and N,N’-diacetylchitobiose (around 340°C, 300°C, 200°C, and 240°C, respectively), indicating the specific detection of chitin. Soil incubation study demonstrated that 60 g kg^?1 chitin added to the soil declined exponentially (r = 0.993) within days and could not be detected at 90 days after the addition of chitin. Total carbon (C) content also decreased within days whereas total nitrogen (N) remained almost constant over the 90 days. The amount of ammonium-N increased in the initial 30 days after the addition of chitin and reached about 3.6 g kg^?1, which corresponded to the amount of N in the added chitin (4.1 g kg^?1) while the amount of nitrite-N and nitrate-N were below 2.0 and 15 mg kg^?1, respectively. Comparison of the measured ammonium-N and total-C contents with those calculated from the measured chitin-content implied that addition of chitin enhanced degradation of native organic compounds in soil.     

Cryopreservation of male and female gonial cells by vitrification in the critically endangered cyprinid honmoroko <Emphasis Type="Italic">Gnathopogon caerulescens</Emphasis>

We investigated the feasibility of cryopreservation of spermatogonia and oogonia in the critically endangered cyprinid honmoroko Gnathopogon caerulescens using slow-cooling (freezing) and rapid-cooling (vitrification) methods. Initially, we examined the testicular cell toxicities and glass-forming properties of the five cryoprotectants: ethylene glycol (EG), glycerol (GC), dimethyl sulfoxide (DMSO), propylene glycol (PG), and 1,3-butylene glycol (BG), and we determined cryoprotectant concentrations that are suitable for freezing and vitrification solutions, respectively. Subsequently, we prepared the freezing solutions of EG, GC, DMSO, PG, and BG at 3, 2, 3, 2, and 2 M and vitrification solutions at 7, 6, 5, 5, and 4 M, respectively. Following the cryopreservation of the testicular cells mainly containing early-stage spermatogenic cells (e.g., spermatogonia and primary spermatocytes), cells were cultured for 7 days and immunochemically stained against germ cell marker protein Vasa. Areas occupied by Vasa-positive cells indicated that vitrification led to better survival of germ cells than the freezing method, and the best result was obtained with 5 M PG, about 50% recovery of germ cells following vitrification. In the case of ovarian cells containing oogonia and stage I, II, and IIIa oocytes, vitrification with 5 M DMSO resulted the best survival of oogonia, with equivalent cell numbers to those cultured without vitrification. The present data suggest that male and female gonial cells of the endangered species G. caerulescens can be efficiently cryopreserved using suitable cryoprotectants for spermatogonia and oogonia, respectively.