Physicochemical properties of akabane virus: A member of the simbu arbovirus group of the family bunyaviridae

The multiplication of Akabane virus was not inhibited in the presence of 5-iodo-2′-deoxyuridine, indicating the presence of RNA. The virus was considered to have an envelope, as it was sensitive to ether and chloroform. It was readily inactivated by deoxycholate and trypsin, but was not precipitated by protamine sulphate. The virus was very labile at pH 3 and also rather heat-labile. Akabane virus was readily filtered through membrane filters of 200 or 100-nm pore size, but not through 50-nm filters. Equilibrium centrifugation in a CsCl density gradient gave a peak of infectivity and hemagglutinin at a density of 1.22 g/ml. The peak fractions thus obtained contained numerous virus particles, roughly spherical, variable in size, 70 to 130 nm in diameter, and mostly having a ragged, closely adherent envelope with projections, when examined, following phosphotungstic acid negative staining, in an electron microscope.     

Responses to the Standard for Exchange of Nonclinical Data (SEND) in non-US countries

The Standard for the Exchange of Nonclinical Data (SEND), adopted by the US FDA, is part of a set of regulations and guidances requiring the submission of standardized electronic study data for nonclinical and clinical data submissions. SEND is the nonclinical implementation of SDTM (Study Data Tabulation Model), the standard electronic format for clinical regulatory submissions to FDA. SEND, SDTM, and the associated Controlled Terminology have been developed by CDISC (Clinical Data Interchange Standards Consortium). In order to successfully implement SEND, interdisciplinary contributions between sponsors and CROs, need a model for task allocation. This is being undertaken by the Pharmaceutical Users Software Exchange (PhUSE). Because SEND is currently the preferred submission format of the US FDA only and will become required by it starting in December 2016, only American academic societies and companies are actively involved. An exception to this is the INHAND initiative, which leads the way in standardizing terminology for toxicological pathology. On the other hand, international globalization of other clinical and nonclinical practices is not feasible because there are substantial differences between the US and non-US countries in CRO involvement in drug development. Thus, non-US countries must consider and develop approaches to SEND that meet their needs. This paper summarizes the activities of the major organizations involved in SEND development and implementation, discusses the effective use of SEND, and details a compliance scheme (research material of the Showa University School of Medicine) illustrating how pharmaceutical companies can complete a large amount of work up to an FDA application with the effective utilization of CROs and solution providers.     

Adventitious bud formation and plantlet regeneration from leaves of fig (Ficus carica L.)

Summary

Leaf fragments of fig (Ficus carica L. ‘Masui Dauphine’) regenerated from in vitro shoot culture were excised and inoculated on MS medium supplemented with different combinations of 2, 4-D, TDZ, and 0.5 mM phloroglucinol. Addition of 2, 4-D induced root formation directly on the explant, and the presence of phloroglucinol significantly increased root formation. When a combination of 2, 4-D and TDZ was added to MS medium containing phloroglucinol, the explants started to produce adventitious buds at the edges. The addition of phloroglucinol was effective in inducing adventitious bud formation. Excised shoots were rooted successfully in MS medium that was either hormone free or supplemented with 1.0 mg l^–1 indolebutyric acid. Regenerated plantlets were successfully established in soil after a short period of acclimatization. This is the first protocol of organogenesis and plant regeneration from vegetative organs of Ficus carica L.     

Establishment of reliable methods of in vitro pollen germination and pollen preservation of Brassica rapa (syn. B. campestris)

A simple and reliable method for evaluating the viability of Brassica pollen was established in which the in vitro germination rate of pollen was adopted as the index of the viability of pollen grains. Pollen grains were preincubated in an atmosphere in which the relative humidity (RH) was fixed to 52% or 66% at 20 °C for 5 hours. They were cultured for 16 hours at 25 °C in a liquid Kwack's medium (1964) supplemented with 20% sucrose, and the pH was adjusted to 8.0. They were then observed under a microscope and the number of germinating and unchanged pollen grains were counted. The germination rate of pollen was improved and stabilized by preincubation and the use of a high pH medium. More than 90% of the freshly harvested pollen grains of Brassica rapa (syn. B. campestris) germinated constantly in these conditions Undehisced anthers were collected from flowers at anthesis and dehydrated by incubation at 20 °C for 16–24 hours in an atmosphere where the RH was fixed to 15% or 32%. They were put into a plastic vial and preserved in a freezer at -20 °C. The germination percentage of the preserved pollen was scored at intervals during preservation. The germination rate of the pollen grains preserved at -20°C for 1 year was higher than 50% and the pollen proved to be efficient for seed set. Most of the seeds germinated normally. This revised version was published online in August 2006 with corrections to the Cover Date.     

Developmental changes of phyllochron in near-isogenic lines of rice (Oryza sativa L.) with different growth durations

The repeated elements called phytomers, which consist of leaf, node, internode and axillary bud, play an important role in the development of modular organization in plants. Rice has the striking feature that the rate of the phytomer production is closely synchronized with the rate of leaf emergence (phyllochron). We examined developmental changes of phyllochron by using 10 near-isogenic lines (NILs) showing diversified growth durations in rice. The NILs were established by backcrosses with a strain practically insensitive to photoperiod, and they consisted of early- or late-flowering NILs whose differences of growth duration were caused by the combinations of alleles at 6 loci. The developmental patterns of phyllochron were evaluated by means of a quartic polynomial, which fitted well in most cases. The results indicated that phyllochron greatly changed during development, especially in late-flowering NILs as well as the recurrent parents, although the fluctuation of phyllochron was not so marked in the early-flowering NILs. Thus, the developmental change of phyllochron was highly dependent upon the genotypes and/or growth duration; however, it was associated with neither floral initiation nor temperature, indicating that the change of phyllochron might reflect internal or physiological changes which occur during the life cycle of rice. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of the effects of five QTL regions on Fusarium head blight resistance and agronomic traits in spring wheat (Triticum aestivum L.)

Fusarium head blight (FHB) is an important disease of wheat (Triticum aestivum L.). The aim of this study was to determine the effects of quantitative trait locus (QTL) regions for resistance to FHB and estimate their effects on reducing FHB damage to wheat in Hokkaido, northern Japan. We examined 233 F₁-derived doubled-haploid (DH) lines from a cross between ‘Kukeiharu 14’ and ‘Sumai 3’ to determine their reaction to FHB during two seasons under field conditions. The DH lines were genotyped at five known FHB-resistance QTL regions (on chromosomes 3BS, 5AS, 6BS, 2DL and 4BS) by using SSR markers. ‘Sumai 3’ alleles at the QTLs at 3BS and 5AS effectively reduced FHB damage in the environment of Hokkaido, indicating that these QTLs will be useful for breeding spring wheat cultivars suitable for Hokkaido. Some of the QTL regions influenced agronomic traits: ‘Sumai 3’ alleles at the 4BS and 5AS QTLs significantly increased stem length and spike length, that at the 2DL QTL significantly decreased grain weight, and that at the 6BS QTL significantly delayed heading, indicating pleiotropic or linkage effects between these agronomic traits and FHB resistance.     

A split-root apparatus for examining the effects of hydraulic lift by trees on the water status of neighbouring crops

We describe a split-root system for examining the effects of hydraulic lift by trees on crop growth. In this system, upper lateral tree roots were grown in a container set on the ground through which the taproot of the tree could penetrate into the moist soil below. The container, with a radius of 0.5 m and a height of 0.20 m, consisted of two compartments divided by a waterproof barrier. A markhamia tree (Markhamia lutea (Benth.) Schumann) and upland rice (Oryza sativa (L.)) plants were planted in one compartment, with only rice plants planted in the other compartment. Irrigation of the container was ceased at the start of the experiment. The stomatal conductance of the rice plants in the associated side, in which both trees and rice plants were grown, declined more rapidly during the first drying period than in the rice-only compartment, suggesting that there was competition for water between the tree and the crop plants. However, during the later drying period, the rice plants in the associated side were green and viable, while those in the rice-only side became desiccated. Rice roots were seen intermingling with tree roots, and the soil water content in the associated site tended to be higher than in the rice-only side. It is likely that hydraulic lift occurred in the associated side and that water that had been transferred to the surface roots was released into the soil, enabling the rice plants in this compartment to remain viable. This novel system is useful for examination of the effects of hydraulic lift by trees on the growth of neighbouring crops. This revised version was published online in June 2006 with corrections to the Cover Date.     

Absolute configuration of new bisaboranoids from Yaku-sugi (Cryptomeria japonica) wood

The absolute configuration of bisabolanoids isolated from Yaku-sugi wood were compared with those of dihydrocarvone and dihydrocarveol isomers based on their¹³C nuclear magnetic resonance data and optical rotations.     

Construction and characterization of a BAC library of soybean

We have constructed a soybean (Glycine max (L.) Merrill) bacterial artificial chromosome (BAC) library from green leaf protoplasts of the cultivar, Misuzudaizu. The library contains 53,760 clones with an average insert size of 116 kb. About 2.9% chloroplast DNA origin was revealed by PCR and colony hybridization. Apart from 2.8% clones having no insert, this library represents 5.2 genome equivalents. With this genome coverage, the probability of having any DNA sequence represented in the library is higher than 99.5%. Three-dimensional pools of the BAC library in combination with the use of a high efficiency genome scanning (HEGS) electrophoresis facilitate rapid and efficient PCR-based screening. An average of five positive clones were identified after screening the BAC library with SSR and STS markers. BAC-end walking was performed for three SSR associated BACs. This library will provide a good resource for positional cloning of agronomically and biologically important QTL genes that Misuzudaizu possesses.