Development of a new selection method and quality improvement of sugary-1 rice mutants

Brown rice of sugary-1 mutants has a wrinkled character because of the presence of phytoglycogen instead of starch in the inner part of the endosperm. Because the wrinkled phenotype was used as a sole selection marker for progeny of the sugary-1 strain, identification of mutant seeds with improved appearance is very difficult. We found that sugary-1 varieties contained not only phytoglycogen but also free glucose in the endosperm, and these were positively correlated. In the segregated F₂ seeds that resulted from crossing Hokurikutou237 (sugary-1) and Koshihikari strains, glucose and phytoglycogen were also significantly correlated. Thus, we identified new sugary types with improved appearance from these progeny using glucose measurements. The F₄ seeds of the improved strain had moderate phytoglycogen contents and seed germination characteristics. Native-PAGE showed that pullulanase activity in the improved strain increased in developing seeds compared with Hokurikutou237, although isoamylase activity was extremely low and similar to that in sugary-1 types. The new selection method in this study efficiently aids the development of improved sugary rice types that lack the wrinkled phenotype.     

Development of cleaved amplified polymorphic sequence (CAPS)-based markers for identification of sweetpotato cultivars

To develop a simple and reliable method to identify sweetpotato cultivars, we designed cleaved amplified polymorphic sequence (CAPS)-based markers and used them to perform genotyping of Japanese sweetpotato cultivars. In order to screen the CAPS-based markers, 13 primer pairs were designed from the exon sequences of 11 sweetpotato genes to amplify fragments containing an intron. By digesting the amplified products with 8 restriction enzymes having different recognition sites, a total of 27 polymorphic marker fragments were obtained. Genotyping of 60 Japanese sweetpotato cultivars using these markers suggested that the markers can effectively distinguish sweetpotato cultivars. Among the genes used for primer design, the gene encoding the dihydroflavonol 4-reductase (DFR) showed the largest degree of polymorphism. To our knowledge, this is the first report on the development of CAPS-based markers in sweetpotato.     

Changes in size of soil seed bank in Robinia pseudoacacia L. (Leguminosae), an exotic tall tree species in Japan: Impacts of stand growth and apicultural utilization

This study examined the factors that influence the size of a soil seed bank in Robinia pseudoacacia stands. We proposed two hypotheses: the amount of buried seeds of R. pseudoacacia is proportional to stand biomass rather than stand age, and apicultural utilization influences the size of the soil seed bank. R. pseudoacacia generally produces seeds with various degrees of physical dormancy. In addition, this tree is short-lived, and fecundity declines after 30–40 years in relation to tree vigor. However, R. pseudoacacia is intensively used in apiculture as an important honey source, and supply of honeybees by beekeepers could influence seed crops. We investigated 25 plantations in Hokkaido (Central, Hidaka, and Oshima) and in Nagano, Japan, where many naturally regenerated R. pseudoacacia stands occur. We found great variation in the size of the soil seed bank among stands, ranging from 13,757 to 6.4 seeds m⁻² per plot. A generalized linear mixed model revealed that both basal area (BA_Rp) and apicultural utilization best explained the size of the soil seed bank. Both the positive effect of BA_Rp and small contribution of stand age to the model implied that the soil seed bank is transitory, and will not persist for very long time. The large contribution of apicultural utilization to the size of the soil seed bank implied pollinator limitation under natural conditions. R. pseudoacacia often has large flower crops, and native pollinators can not keep up with the demand for pollination. Thus, the supply of honeybees by beekeepers should improve the pollination success of R. pseudoacacia. From these results, we conclude that we must consider stand history with respect to apicultural utilization if the land is harvested or a stand declines.     

In vivo immunomodulatory effects of dietary purple sweet potato after immunization in chicken

This study was intended to determine the modulatory effects of dietary supplementation of purple sweet potato ( Ipomoea batats Poir., PSP) on the immune response of chickens. PSP was included in a basal starter diet by 1% (PSP^L) or 3% (PSP^H) and continually fed. Newcastle disease (NDV) vaccine, Brucella abortus (BA) and sheep red blood cells (SRBC) were used for chicken immunization. Antibody titers against these antigens were used to estimate humoral immunity. Concanavalin A (Con A)-induced proliferations of splenocytes, thymocytes and peripheral blood lymphocytes (PBL), ratios of CD4- and CD8-single positive and CD4-CD8-double negative (DN) cells in splenocytes, were both used to indicate cellular immunity. Relative weights of spleen, thymus and bursa and white blood cell (WBC) counts were studied. PSP^H increased anti-NDV ( P  < 0.05), anti-BA ( P  < 0.01) and anti-SRBC titers ( P  < 0.05) in response to secondary immunization, whereas PSP^L increased titers of anti-BA ( P  < 0.05) and anti-SRBC ( P  < 0.01). Proliferations of splenocytes and thymocytes were augmented with PSP^L ( P  < 0.05). PSP^H-treated chickens had lower ( P  < 0.05) ratios of CD4-sigle positive lymphocytes. Proliferation of PBL, weights of lymphoid organs and WBC counts were not affected. These results suggest that dietary PSP supplementation could enhance the immune response after immunization in chickens.     

Chlamydial species among wild birds and livestock in the foothills of Mt. Afadjato,Ghana

The members of family Chlamydiaceae have a broad host range and cause many kinds of diseases in humans and animals. Several cases of Chlamydiaceae being detected in atypical hosts have been reported recently. Consequently, cross-species monitoring of Chlamydia in wildlife and livestock is pertinent for public health, animal hygiene and wildlife conservation. In this study, we conducted molecular surveillance of Chlamydia in wild birds and livestock around a small village in the foothills of Mt. Afadjato, Ghana where direct contact between wildlife and livestock occurs. Among 29 captured wild birds and 63 livestock, 5 sheep, 30 goats and 28 chickens, the positive ratios of Chlamydia were 24.1%, 40.0%, 43.3% and 26.9%, respectively. Chlamydia pecorum was detected in wild birds, goats, sheep and chickens. On the basis of the variable domain 2 region of ompA, several samples from different hosts showed identical sequences and were phylogenetically located to the same clusters. In addition, using ompA, C. psittaci, C. abortus and C. gallinacea were also detected in this small habitat. Further genetic and pathogenic analyses of the chlamydial distribution in this area, which represents the interface of wild and domestic animal interactions, may improve our knowledge of their transmission among different hosts.     

Effect of exudate released from seeds and seedling roots of common bean (Phaseolus vulgaris L.) on proliferation of Rhizobium sp. (Phaseolus)

Effects of seed and root exudates obtained from common bean on the proliferation of Rhizobium sp. (Phaseolus) were examined in a combination of three plant cultivars with three Rhizobium strains. In the first experiment, seed or root exudate was mixed with an Andosol soil extract, and bacterial proliferation in the mixture was traced. Seed exudate was prepared from hydroponic solution used in seed imbibition for 24 h, and a series of root exudates was prepared from a hydroponic solution collected every 24 h from the initiation of rooting up to 96 h after rooting. Regardless of the common bean cultivars and Rhizobium strains used, Rhizobium population markedly increased of the 24 h of culture in the mixture containing seed exudates, whereas a negligible increase was detected in the mixture with root exudates. The mixture containing root exudates collected within a period of 72–96 h after initial rooting (96–120 h after seed imbibition) exerted an inhibitory effect on Rhizobium proliferation. The seed exudates contained large amounts of sugars, amino acids, nitrogen, phosphorus, potassium, and magnesium compared to any root exudates. In the second experiment, Rhizobium was inoculated directly to common bean seeds sowed in a vermiculite bed which was sterilized and moistened with a plant nutrient solution. Compared with the control (without seed), a remarkable increase in the number of bacterial cells was observed in all the combinations of plant cultivars and Rhizobium strains 24 h after sowing. These results reveal that seed exudates of common bean have a substantial potential to promote Rhizobium proliferation, and that root exudates in a particular period of culture contain some inhibitory factors.     

Soil chemical properties of an Indonesian red acid soil as affected by land use and crop management

In the middle terrace area of south Sumatra, Indonesia, where red acid soils poor in crop productivity are widely distributed, the effects of cropping pattern and cultivation techniques on physico-chemical properties of soil were investigated. Five patterns for cassava cropping, including monoculture, a rotation with annual food crops, and three intercroppings with differences in the combination with annual crops and in the planting density, were evaluated in Experiment I. In Experiment II, eight plots composed of the combinations of two tillage methods (no-tillage or conventional tillage), the presence or absence of surface mulch from crop residues, and two rates of chemical fertilizers were established for a maize–soybean–cowpea sequential cropping pattern. At the end of 3 years, there was no difference in total C and total N concentrations among the plots in Experiment I irrespective of the mulch treatment using crop residues. Soil organic matter (SOM) concentration was not affected even in the no-tillage plot where the maximum crop residues (20 t ha⁻¹) was given as surface mulch with the increased root residues due to higher rates of fertilizers (Experiment II). In Experiment I, available P concentration was highest in an intercropping with higher fertilizer rates and lowest cassava planting density. In Experiment II, an increase in available P was attained by mulching and the higher rate of fertilizers, and a minor positive effect of fertilizer was also observed in exchangeable Mg and K concentrations. Surface mulch resulted in less clay fraction compared with the non-mulch plots in both the experiments, suggesting its effect on the maintenance of soil particle distribution. An additional finding suggested no prominent influence of cassava monoculture on the level of SOM in this area based on the comparison with other major land uses, including secondary forest, rubber plantation, and mixed cultivation of fruits with crops. Nevertheless, the introduction of crop residue mulch and higher rates of fertilizers are recommended for sustaining soil quality and achieving higher crop yields.     

Characterization of Rehmannia mosaic virus isolated from chili pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis>) in Japan

In 2010, severe necrotic mosaic disease and fruit distortion were observed on greenhouse-grown chili pepper (Capsicum annuum cv. Fushimi-amanaga) plants in Kyoto Prefecture, Japan. Electron microscopic imaging and genomic RNA sequencing indicated that the virus responsible was a new isolate of Rehmannia mosaic virus (ReMV), which had not been previously reported in Japan. Although ReMV systemically infected many Solanaceae species, including chili pepper and tomato (Solanum lycopersicum), tobamovirus-resistance genes from species of Capsicum (L ^1a , L ² , L ³ , and L ⁴ ) and tomato (Tm-1, Tm-2, and Tm-2 ^a ) conferred resistance against ReMV.     

Porencephaly in a cynomolgus monkey ( macaca fascicularis )

Porencephaly was observed in a female cynomolgus monkey (Macaca fascicularis) aged 5 years and 7 months. The cerebral hemisphere exhibited diffuse brownish excavation with partial defects of the full thickness of the hemispheric wall, and it constituted open channels between the lateral ventricular system and arachnoid space. In addition, the bilateral occipital lobe was slightly atrophied. Histopathologically, fibrous gliosis was spread out around the excavation area and its periphery. In the roof tissue over the cavity, small round cells were arranged in the laminae. They seemed to be neural or glial precursor cells because they were positive for Musashi 1 and negative for NeuN and GFAP. In the area of fibrous gliosis, hemosiderin or lipofuscin were deposited in the macrophages, and activated astroglias were observed extensively around the excavation area.     

Immunomodulatory effects of Alliums and Ipomoea batata extracts on lymphocytes and macrophages functions in White Leghorn chickens: In vitro study

We previously described that supplementary garlic, onion and purple sweet potato (PSP) enhance humoral immune response in White Leghorn chickens. In the present in vitro study, we investigated the effects of garlic (GE), onion (OE) and PSP (PSPE) extracts on proliferation, interleukin (IL)‐2 and interferon (INF)‐γ gene expression of stimulated lymphocytes. The effects on microbicidal activity, reactive oxygen species (ROS) and nitric oxide (NO) productions of stimulated peritoneal macrophages were studied as well. The results showed that GE augmented Concanavalin A (ConA)‐induced splenocytes (4, 8 and 16 µg/mL) and thymocytes (2, 4 and 8 µg/mL) proliferations, and gene expression of IL‐2 (8 and 16 µg/mL) and INF‐γ (16 µg/mL). None of the examined extracts had mitogenic effect nor stimulated bursacytes response to phorbol 12‐myristate 13‐acetate (PMA). Macrophages exhibited superior microbicidal activity and ROS production with GE at 4 and 8 µg/mL and with OE at 25.6 µg/mL. None of the extracts showed stimulatory effects on NO production. The extracts showed concentration‐dependent inhibitory effects on all measured parameters at higher concentrations. Taken together, it is likely that garlic has direct stimulatory effects on immune cell functions, whereas the in vitro inhibitory effects of onion and PSP were likely attributed to high flavonoid contents.