Microwave pyrolysis of cellulosic materials for the production of anhydrosugars

Large-scale microwave rapid pyrolysis of cellulosic materials has been investigated. Levoglucosan (1,6-anhydro--d-glucopyranose) was obtained from a larch log as the main anhydrosugar in 2.6% yield on the basis of dry wood weight. This yield would be much higher than that obtainable by conventional pyrolysis in the largescale reaction. Levoglucosenone (1,6-anhydro-3,4-dideoxy--D-glycero-hex-3-enopyranos-2-ulose) was found to be produced in one-quarter the amount of levoglucosan. Other anhydrosugars, such as mannosan (1,6-anhydro--D-mannopyranose), galactosan (1,6-anhydro--d-galactopyranose), and xylosan (1,4-anhydro--d-xylopyranose), were also confirmed to be produced as minor components depending on the proportion of the monosaccharide content in the larch. When microwave pyrolysis of used papers and filter papers was performed, the yields of levoglucosan were about 6% and 12%, respectively, suggesting that a higher content of cellulose gives a larger amount of levoglucosan.     

Genetic Diversity in AA and CC Genome Oryza Species in Southern South Asia

The CC genome of Oryza is found in nine species of Oryza that are distributed on all continents having a tropical climate. Three diploid Oryza species with CC genome are found in Africa and Asia to Papua New Guinea. In southern South Asia these three CC genome diploid species can be found, O. eichingeri and O. rhizomatis in Sri Lanka and O. officinalis in India. AA genome wild relatives of rice are also found in the same geographic region. Germplasm of both diploid CC and AA genome Oryza germplasm has recently been collected from Sri Lanka. AFLP analysis was used to compare the genetic diversity of the two Oryza genomes from a similar geographic region in southern South Asia. In addition, the diploid CC Oryza germplasm was also analyzed by RAPD and SSR methodologies and the combined results were analyzed. The results show that in southern South Asia the diploid CC genome species have a high level of genetic diversity compared to the diploid AA genome species. Molecular marker analysis revealed that populations of O. rhizomatis from northern and southeastern Sri Lanka are genetically differentiated. One accession of O. rhizomatis was aligned with O. eichingeri. This accession was collected from the site of O. rhizomatis that is the closest to a population of O. eichingeri. O. eichingeri showed lower genetic diversity than the other two diploid Oryza CC genome species. O. officinalis accessions from Assam, India, and China were genetically less diverged from O. eichingeri and O. rhizomatis than two accessions of O. officinalis from Kerala state, India. The first two authors contributed equally to this research     

Mapping a new source of resistance to powdery mildew in mungbean

Both restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP) analyses were employed to map a new source of resistance to powdery mildew in mungbean. Disease scores of an F₂ population derived from the cross between a moderately resistant breeding line VC1210A and a susceptible wild relative (Vigna radiata var. sublobata, accession TC1966) showed a continuous distribution and was treated as a quantitative trait. Although no significant quantitative trait loci (QTL) that can explain the variation was detected by QTL analysis based on the reconstructed RFLP linkage map, new marker loci associated with resistance were discovered by AFLP analysis. The RFLP loci detected by two of the cloned AFLP bands are associated with resistance and constitute a new linkage group. A major resistance quantitative trait locus was found on this linkage group that accounted for 64.9% of the variation in resistance to powdery mildew. One of the probes developed in this study has the potential to assist in breeding for powdery mildew resistance in mungbean.     

The formation process of button ulcers in pigs experimentally infected with a subgenotype 2.1 isolate of classical swine fever virus

We evaluated the role of classical swine fever virus (CSFV) in the formation of button ulcers in the mucosa of the gastrointestinal tract. Histopathological and immunohistochemical analyses of pigs experimentally infected with a subgenotype 2.1 isolate of CSFV, which was isolated in Japan in 2019, revealed follicular necrosis in the submucosal mucosa-associated lymphoid tissue and herniation of crypts as factors that contribute to the development of button ulcers during CSFV infection. These findings indicate that CSFV induces follicular necrosis and is one of the causative agents of button ulcers in pigs.     

Systems for making NIAS Core Collections, single-seed-derived germplasm, and plant photo images available to the research community

The National Institute of Agrobiological Sciences (NIAS) Genebank coordinates the conservation of plant, microorganism, and animal genetic resources related to food and agriculture in Japan. It also coordinates the distribution of genetic resources in the public domain for research, breeding, and educational purposes. To operate the NIAS Genebank efficiently, we have developed a genetic resources database, data management software, and web-based data retrieval systems to make the data available worldwide. This article describes the NIAS Genebank’s Core Collections of global and Japanese soybean (Glycine max (L.) Merr.), Japanese azuki bean (Vigna angularis (Willd.) Ohwi et Ohashi), and Japanese wheat (Triticum aestivum L. s. l.), all of which are available through the Genebank website. This article also describes new features of the NIAS Genebank database, such as the ability to select single-seed-derived germplasm of soybean in the plant search system and to download photographic data on accessions. By using the downloaded plant image PDF files, users can obtain detailed passport and agronomic information by clicking on the image of an accession of interest.     

Purification of curcumin, demethoxycurcumin, and bisdemethoxycurcumin by high-speed countercurrent chromatography

Curcuminoids are substances of great interest because of their important pharmacological activities, particularly anti-inflammatory, anticarcinogenic, and anti-Alzheimer's activities. In this study, we report the first procedure and effect of processing for the high, efficient, and useful purification of curcumin, demethoxycurcumin, and bisdemethoxycurcumin from turmeric powder. Purification involves high-speed countercurrent chromatographic (HSCCC) separation of these curcuminoids using a simple two-phase solvent system composed of n-hexane/chloroform/methanol/water (5/10/7.5/2.5, v/v). The HSCCC-fractionated effluent peaks indicated that the peak resolutions were 1.7 between curcumin and demethoxycurcumin and 2.1 between demethoxycurcumin and bisdemethoxycurcumin for 25 mg of loaded turmeric powder. These purified substances were analyzed by liquid chromatography-tandem mass spectrometry with scan and daughter scan negative modes, and the wide absorbance from 200 to 500 nm was monitored by photodiode array detection. The separation yielded 1.1 mg of curcumin, 0.6 mg of demethoxycurcumin, and 0.9 mg of bisdemethoxycurcumin (>98% purity). Moreover, the antioxidant effect of curcuminoids was measured by a 1,1-diphenyl-2-picrylhydrazil assay. The order of antioxidant activity was purified curcumin > purified demethoxycurcumin > purified bisdemethoxycurcumin > turmeric powder. Curcumin, demethoxycurcumin, and bisdemethoxycurcumin can be used for various evaluations of their pharmacological activities.     

Quantitative PCR assay for the detection of the parasitic ciliate <Emphasis Type="Italic">Cryptocaryon irritans</Emphasis>

We developed a quantitative PCR assay for detecting the parasitic ciliate Cryptocaryon irritans, which causes “white spot disease” in marine fishes, from the natural environment. A specific primer set for C. irritans was designed and its high specificity was confirmed in silico: almost all of the sequences deposited in the GenBank nucleotide database were covered, 22/23 for the forward primer and 7/7 for the reverse primer. We estimated that there were 3,415.9 rRNA gene copies per genome of C. irritans. In artificial mixture experiments to validate whether the qPCR assay is applicable to natural samples, the estimated copy numbers showed significantly positive correlations with the number of theronts added (p < 0.001). When we applied this qPCR assay to natural samples collected bimonthly from surface and bottom seawaters at an aquaculture site (water depth, 10 m) from May 2009 to March 2010, we only detected C. irritans (112.0 ± 6.3 cells/l) in the surface seawater sample in November. This qPCR assay is a useful tool for detecting C. irritans rapidly and quantitatively in natural environments; it could also help advance our understanding of the ecology of C. irritans, as well as facilitate the diagnosis of the disease.     

DaizuBase,an integrated soybean genome database including BAC-based physical maps

Soybean [Glycine max (L) Merrill] is one of the most important leguminous crops and ranks fourth after to rice, wheat and maize in terms of world crop production. Soybean contains abundant protein and oil, which makes it a major source of nutritious food, livestock feed and industrial products. In Japan, soybean is also an important source of traditional staples such as tofu, natto, miso and soy sauce. The soybean genome was determined in 2010. With its enormous size, physical mapping and genome sequencing are the most effective approaches towards understanding the structure and function of the soybean genome. We constructed bacterial artificial chromosome (BAC) libraries from the Japanese soybean cultivar, Enrei. The end-sequences of approximately 100,000 BAC clones were analyzed and used for construction of a BAC-based physical map of the genome. BLAST analysis between Enrei BAC-end sequences and the Williams82 genome was carried out to increase the saturation of the map. This physical map will be used to characterize the genome structure of Japanese soybean cultivars, to develop methods for the isolation of agronomically important genes and to facilitate comparative soybean genome research. The current status of physical mapping of the soybean genome and construction of database are presented.     

AFLP analysis of diploid species in the genus Vigna subgenus Ceratotropis

A representative set of germplasm of genusVigna subgenus Ceratotropis specieswas used to analyze genetic diversity using amplified fragment lengthpolymorphism (AFLP) methodology. The germplasm was selected based onmorphological diversity and geographic origin and includes 18 out of the 21species in the subgenus Ceratotropis. Genetic diversitywithin and between taxa was measured using information generated by AFLP bandpolymorphism from which Jaccard's similarity coefficient and nucleotidediversity were estimated. The data were also used to generate phenograms tovisualize relationships among analyzed accessions. All species in sectionsCeratotropis and Aconitifoliae arewell separated. However, most of the species in sectionAngulares show a high level of similarity suggesting a lowlevel of genetic differentiation. In section Angulares onespecies, V. trinervia, is welldifferentiated from the other species and represents a separate evolutionarybranch between section Ceratotropis and other species insection Angulares. The relationships between the newlydescribed species V. aridicola,V. exilis, V.nepalensis and V.tenuicaulis and other species in the subgenusCeratotropis are described for the first time based on AFLPdata.