New severe strains of Melon necrotic spot virus: symptomatology and sequencing

New strains of Melon necrotic spot virus (MNSV), designated MNSV-YS and MNSV-KS, caused much more severe growth retardation on melon plants than MNSV-NH, which was previously reported as the most severe strain of MNSV in Japan. MNSV-YS spread much more quickly than MNSV-NH in infected plants, and induced more severe growth retardation, even though the appearance of necrotic lesions on inoculated cotyledons was much slower. MNSV-KS had properties intermediate between those of the other two strains. The results suggest that faster-spreading strains can multiply more rapidly as a result of lower levels of activity in inducing necrotic lesions in melon plants. The complete sequences of MNSV-YS and MNSV-KS were determined, and an RT–PCR–RFLP method based on these sequences was successfully developed to detect and discriminate between the three strains.     

Partitioning of photosynthates originating from bent shoots in the arching and high-rack culture systems of cut rose production

The arching and high-rack culture systems were developed and patented by Japanese rose growers. Both culture systems have bent canopies (lower bent shoots). In the arching culture system, shoots sprouting from the crown are harvested as cut flowers. However, the high-rack culture system also has a bent canopy originating from the mother stem (upper bent shoots) and flower stems sprout and is harvested at the top of each mother stem. Partitioning of photosynthates originating from bent shoots in arching and high-rack culture systems of rose production was investigated to elucidate how carbohydrates are re-allocated from the bent shoots in different culture systems of roses. At the flowering stage in both culture systems, 50–70% of ¹³C-photosynthates originated from bent shoots were exported to other parts within 72 h after ¹³CO₂ feeding to the bent shoot. In the arching culture system, photosynthates from lower bent shoots were partitioned mainly to the roots and crown. Similarly, in the high-rack culture system, between 71 and 86% of the exported carbon from the bent shoots were allocated to below the point of bending (roots + crown + mother stems) and only 9–28% was allocated to flowering shoots above the point of bending. In both culture systems, photosynthate translocation from the lower bent shoot directly to flowers was low. Accordingly, bent shoots in rose plants acted as a source of photosynthates, independent of culture system. The height of the bent shoots determined for a great deal in the re-allocation of the photosynthates, and provides a partial explanation for difference in production of cut roses.     

An H5N1 highly pathogenic avian influenza virus that invaded Japan through waterfowl migration

In 2010, an H5N1 highly pathogenic avian influenza virus (HPAIV) was isolated from feces of apparently healthy ducks migrating southward in Hokkaido, the northernmost prefecture of Japan. The H5N1 HPAIVs were subsequently detected in domestic and wild birds at multiple sites corresponding to the flyway of the waterfowl having stopovers in the Japanese archipelago. The Hokkaido isolate was genetically nearly identical to H5N1 HPAIVs isolated from swans in the spring of 2009 and 2010 in Mongolia, but less pathogenic in experimentally infected ducks than the 2009 Mongolian isolate. These findings suggest that H5N1 HPAIVs with relatively mild pathogenicity might be selected and harbored in the waterfowl population during the 2009-2010 migration seasons. Our data provide "early warning" signals for preparedness against the unprecedented situation in which the waterfowl reservoirs serve as perpetual sources and disseminators of HPAIVs.     

MgO-induced defence against bacterial wilt disease in Arabidopsis thaliana

Bacterial wilt, caused by Ralstonia solanacearum, is a devastating soilborne disease in plants that limits the production of many crops worldwide. Although management of bacterial wilt has so far been unsuccessful, enhancing host resistance to the pathogen may be an effective control strategy. Recently, magnesium oxide (MgO) was found to induce defence responses against R. solanacearum in tomato plants. Here, the mechanisms underlying MgO-induced defence responses against R. solanacearum (MgO-i DARS) were investigated using Arabidopsis thaliana as a host plant. MgO-i DARS was confirmed in A. thaliana mutants deficient in jasmonic acid or ethylene signalling pathways as well as in the wildtype (Col-0) plants. In contrast, no MgO-i DARS was found in A. thaliana mutants deficient in the salicylic acid (SA) production (sid2-2) and signalling pathways (tga1-1 and npr-1). MgO treatment led to significant accumulation of SA in both roots and shoots of Col-0. The SA biosynthesis gene isochorismate synthase 1 (ICS1) was induced in roots and shoots of A. thaliana treated with MgO. An NADPH oxidase gene respiratory burst oxidase homolog D (AtRbohD) was up-regulated in both roots and shoots of Col-0 treated with MgO. No MgO-i DARS was observed in A. thaliana mutants deficient in AtRbohD. These results suggest that SA and RBOHD-mediated ROS are pivotal for MgO-i DARS in A. thaliana.     

In vitro fertilization and development of in vitro matured bovine follicular oocytes

Bovine follicular oocytes matured in vitro were fertilized in vitro using epididymal spermatozoa from five different bulls and then cultured to the blastocyst stage in vitro. The fertilization rate, based on one pair of pronuclei and presence of one sperm tail, ranged from 55.2 to 64.3%. Embryo development (cleavage to blastocyst stage) ranged from 21.4 to 31.0% of the cultured ova reaching 8 cells at 3 to 4 d after insemination to 1.3 to 3.7% reaching hatched blastocysts at 9 to 10 d. It is concluded that individual variation among bulls is not a significant factor in fertilization and development rates of bovine follicular oocytes when epididymal spermatozoa are used.     

First report of Moroccan pepper virus on lisianthus (Eustoma grandiflorum) in Japan

In sap from plants of lisianthus (Eustoma grandiflorum) with necrotic stunt symptoms in Yamaguchi Prefecture in Japan, spherical particles with a diameter of about 30 nm were observed with transmission electron microscopy. We sequenced the complete genome of an isolate using the tombusvirus-specific primers and found that it shared about 96 % nucleotide identities with Moroccan pepper virus (MPV) in the genus Tombusvirus. The isolate reproduced necrotic stunt symptoms on lisianthus plants after mechanical inoculation. This is the first report of MPV on lisianthus in Japan.     

Changes in the soil C and N contents,C decomposition and N mineralization potentials in a rice paddy after long-term application of inorganic fertilizers and organic matter

A long-term experiment on combined inorganic fertilizers and organic matter in paddy rice (Oryza sativa L.) cultivation began in May 1982 in Yamagata, northeastern Japan. In 2012, after the 31^st harvest, soil samples were collected from five fertilizer treatments [(1) PK, (2) NPK, (3) NPK + 6 Mg ha^?1 rice straw (RS), (4) NPK + 10 Mg ha^?1 rice straw compost (CM1), and (5) NPK + 30 Mg ha^?1 rice straw compost (CM3)], at five soil depths (0–5, 5–10, 10–15, 15–20 and 20–25 cm), to assess the changes in soil organic carbon (SOC) content and carbon (C) decomposition potential, total nitrogen (TN) content and nitrogen (N) mineralization potential resulting from long-term organic matter addition. The C decomposition potential was assessed based on the methane (CH₄) and carbon dioxide (CO₂) produced, while the N mineralization potential was determined from the potassium chloride (KCl)-extractable ammonium-nitrogen (NH₄⁺-N), after 2, 4, 6 and 8 weeks of anaerobic incubation at 30°C in the laboratory. Compared to NPK treatment, SOC in the total 0–25 cm layer increased by 67.3, 21.0 and10.8%, and TN increased by 64.2, 19.7 and 10.6%, in CM3, RS and CM1, respectively, and SOC and TN showed a slight reduction in the PK treatment by 5.2 and 5.7%, respectively. Applying rice straw compost (10 Mg ha^?1) instead of rice straw (6 Mg ha^?1) to rice paddies reduced methane production by about 19% after the soils were measured under 8 weeks of anaerobic incubation at 30°C. Soil carbon decomposition potential (Co) and nitrogen mineralization potential (No) were highly correlated with the SOC and TN contents. The mean ratio of Co/No was 4.49, lower than the mean ratio of SOC/TN (13.49) for all treatments, which indicated that the easily decomposed organic matter was from soil microbial biomass and soil proteins.     

Suppressive effect of acetylated glyceride (Bemidetach™EC) on <Emphasis Type="Italic">Tomato yellow leaf curl virus</Emphasis> transmitted by the whitefly <Emphasis Type="Italic">Bemisia tabaci</Emphasis> on greenhouse tomato plants

Acetylated glyceride (Bemidetach?EC)—a food additive—repels adult sweet potato whiteflies (Bemisia tabaci) and inhibits their mating behavior. We evaluated the effects of acetylated glyceride spraying of greenhouse-grown tomato plants on infestation with B. tabaci and the occurrence of Tomato yellow leaf curl virus (TYLCV) disease under commercial-like conditions. The abundance of adult B. tabaci was significantly reduced by three sprayings of acetylated glyceride, and the TYLCV incidence was significantly suppressed to less than 30% of that in the untreated control. These results suggest that acetylated glyceride sprays suppress the secondary spread of TYLCV in greenhouse-grown tomatoes by lessening B. tabaci adult density.     

Development of vaccine strains of H5 and H7 influenza viruses

To establish vaccine strains of H5 and H7 influenza viruses, A/duck/Hokkaido/Vac-1/04 (H5N1) [Vac-1/04 (H5N1)], A/duck/Hokkaido/Vac-3/07 (H5N1) [Vac-3/07 (H5N1)], and A/duck/Hokkaido/ Vac-2/04 (H7N7) [Vac-2/04 (H7N7)] were generated from non-pathogenic avian influenza viruses isolated from migratory ducks. Vac-1/04 (H5N1) and Vac-3/07 (H5N1) were generated by genetic reassortment between H5N2 or H5N3 virus as an HA gene provider and H7N1 or H6N1 viruses as an NA gene provider. Vac-2/04 (H7N7) was a genetic reassortant obtained using H7N7 and H9 N2 viruses to give high growth character of the H9N2 virus in chicken embryonated eggs. The results of sequence analyses and experimental infections revealed that these H5N1 and H7N7 reassortant viruses were non-pathogenic in chickens and embryos, and had good growth potential in embryonated eggs. These viruses should be useful to develop vaccines against H5 and H7 highly pathogenic avian influenza viruses.