New race of <Emphasis Type="Italic">Phytophthora vignae</Emphasis> f. sp. <Emphasis Type="Italic">adzukicola</Emphasis>, the causal agent of Phytophthora stem rot of the adzuki bean

 A new race of Phytophthora vignae f. sp. adzukicola, designated race 4, is reported from central and western Hokkaido, Japan. The isolates obtained from diseased plants of a new cultivar, cv. Syumari, which is resistant to races 1, 2, and 3, were determined to be a new race by the pathogenic reaction on a set of differential adzuki bean cultivars (cv. Erimo-shozu, cv. Kotobuki-shozu, cv. Noto-shozu, cv. Urasa-shimane, and cv. Syumari). Received: March 7, 2002 / Accepted: August 13, 2002     

Interspecies nuclear transfer embryos reconstructed from cat somatic cells and bovine ooplasm

This study was conducted to investigate the developmental capacity of domestic cat-bovine reconstructed embryos via interspecies somatic cell nuclear transfer (iSCNT) and to observe the mitochondrial DNA (mtDNA) content of the iSCNT embryos. The iSCNT embryos were generated using mixed-breed domestic cat fibroblasts as donor cells and enucleated bovine oocytes as the recipient cytoplasm. When the developmental capacities of iSCNT embryos and parthenogenic bovine embryos were compared, there was no difference (P>0.05) in the rates of cleavage and development to the 8-cell stage (86.6 vs. 84.0% and 32.2 vs. 36.2%, respectively). However, in contrast to development of parthenogenic embryos to the morula and blastocyst stages, no iSCNT embryos (0/202) developed beyond the 8-cell stage. For mtDNA analysis, iSCNT embryos at the 1-cell, 2-cell, 4-cell and 8-cell stages were randomly selected. Both cat and bovine mtDNA quantification analysis were performed using quantitative PCR. The levels of both cat and bovine mtDNA in cat-bovine iSCNT embryos varied at each stage of development. The cat mtDNA concentration in the iSCNT embryos was stable from the 1-cell to 8-cell stages. The bovine mtDNA in the iSCNT embryos at the 8-cell stage was significantly lower than that at the 4-cell stage (P<0.05). No difference in the proportions of cat mtDNA in the iSCNT embryos was found in any of the observed developmental stages (1- through 8-cell stages). In conclusion, bovine cytoplasm supports domestic cat nucleus development through the 8-cell stage. The mtDNA genotype of domestic cat-bovine iSCNT embryos illustrates persistence of heteroplasmy, and the reduction in mtDNA content might reflect a developmental block at the 8-cell stage.     

Seasonal variation of paulownia witches’-broom phytoplasma in paulownia trees and distribution of the disease in the Tohoku district of Japan

Seasonal variation of paulownia witches’-broom (PWB) phytoplasma within different organs (leaves, branch and trunk bark and roots) in paulownia trees was investigated by the amplification of a PWB-specific DNA fragment by the polymerase chain reaction (PCR). In leaf samples, PWB phytoplasma was first detected in June and the incidence gradually increased. On the other hand, the PWB was detected at relatively low incidence in branch bark, trunk bark and roots and the incidence did not change among seasons. A survey of PWB in 27 fields in the Tohoku district of Japan showed that malformed flower buds were observed in paulownia trees in almost all of the fields. PWB-phytoplasma was also detected by PCR from paulownia trees in almost all of the fields in Iwate and Fukushima Prefectures. The frequencies of trees in which phytoplasma was detected by PCR were higher than those in which symptoms were observed. These results indicated that PCR amplification of a PWB-specific DNA fragment is an effective tool for practical diagnose and that PWB is widely distributed in the Tohoku district of Japan.     

Rotylenchus uniformis (Thorne) on carrots

Rotylenchus uniformis (Thorne) causes small orange lesions in the cortex of carrot roots and also orange discoloration of the stele in the vicinity of these lesions. The tolerance limit of carrots toR. uniformis was about 30 nematodes per 5 g soil in pot trials at about 17°C and 2 nematodes per 5g soil at 5° to 10°C. However, this temperature is too low for normal development of the plant. Field observations in the literature suggest that carrot yields are not reduced at densities ofR. uniformis up to 20 nematodes per 5 g soil. No field data are available for higher densities. From the frequency distribution of different densities ofR. uniformis in soil samples from carrot fields, and assuming a tolerance limit of 16 nematodes per 5 g soil, the area where losses occur is estimated to be between 10 and 15% of the total area under carrots. The total reduction of yield by the nematode would then be less than 1%. If the tolerance limit is 32 nematodes per 5 g soil, the damage would be negligible.Carrot, cauliflower andPhaseolus beans are among the best hosts ofR. uniformis.     

Modulation by sphingosine of phosphorylation of substrate proteins by protein kinase C in nuclei from cow mammary gland

Protein kinase C (PKC) is an enzyme activated by diacylglycerols such as 1-oleoyl-2-acetyl-sn-glycerol (OAG), phospholipids (in particular phosphatidylserine; PS) and Ca2+, which regulate a wide variety of intracellular functions by phosphorylating multiple substrate proteins and enzymes. The effect of sphingosine, the backbone moiety of sphingolipids, on PKC activity and phosphorylation of endogenous proteins catalyzed by PKC was investigated in nuclei of cow mammary gland. Sphingosine inhibited nuclear PKC activity when lysine-rich histone was used as the substrate. The sphingosine inhibition of the PKC activity was reversed by the excess addition of PS, but not by OAG or Ca2+. Several nuclear proteins, including 56-kDa, 43-kDa, 38-kDa and 36-kDa proteins, were shown to be substrates for PKC. Of the substrate proteins, the 38-kDa and 36-kDa proteins were identified as annexin I, the Ca2+/phospholipid-binding protein; the 56-kDa and 43-kDa proteins have not yet been identified. Sphingosine inhibited phosphorylation of the 56-kDa protein and the 36-kDa annexin I, whereas it enhanced that of the 43-kDa protein. The 38-kDa annexin I species was unaffected by sphingosine. As with the PKC activity, inhibition by sphingosine of phosphorylation of the 56-kDa protein and 36-kDa annexin I was reversed by the excess addition of PS, but not by OAG or Ca2+. In addition, by the excess addition of PS and not by OAG or Ca2+, the sphingosine-enhanced phosphorylation of the 43-kDa protein was reversed and returned to near the level in the absence of sphingosine. It is suggested that sphingosine is involved in the regulation of PKC-dependent phosphorylation in the nucleus by modulating the association of PKC or its substrates, particularly annexin I, with membrane phospholipids in cow mammary gland.     

Changes in vitamin A added to a fermented total mixed ration prepared with reed canarygrass (Phalaris arundinacea L.)

A fermented total mixed ration (TMR) was prepared by adding a vitamin premix containing vitamin A and enzyme to reed canarygrass roughage. The vitamin A levels were then determined after 30 days of fermentation at 20 or 30°C. The vitamin A contents had decreased in roughage fermented at both storage temperatures, and decreased further when an enzyme supplement was included. Because the majority of the added vitamin A was destroyed during fermentation and storage, the addition of vitamin A at the beginning of preparation of fermented TMR is not recommended.     

Nutritional parameters in the blood of dams during late gestation and immediately after calving,in the umbilical vein at calving,and in the blood of calves immediately following birth in Holstein heifers pregnant with either Holstein or beef breed fetuses

Dairy cattle management lacks consideration of fetal breed, the effect of which on fetal growth and nutrition are unclear. We investigated blood parameters in 12 late-pregnant Holstein heifers with similar (Holstein, n = 5) or different (Japanese Black [n = 4] or F1 cross [n = 3]; Holstein × Japanese Black) fetus breeds and in their umbilical cords and calves. Samples were obtained from dams 1 week before calving (−1 week) and immediately after calving, from the umbilical vein at calving, and from calves immediately after birth. Dams with beef fetuses had higher serum glucose levels (−1 week; p < .05) than those with Holstein fetuses. Plasma total amino acid, total essential amino acid, total nonessential amino acid, and other amino acid concentrations were lower in the umbilical veins of dams with calves of the beef breeds than in those of the Holstein breeds (p < .05). Furthermore, serum glucose and plasma amino acid levels were lower in the beef calves than in the Holstein calves (p < .05). Overall, nutrient supply from dams to beef fetuses was lower than that to Holstein fetuses. Our findings may facilitate feeding management of dairy cattle pregnant with beef breeds for appropriate fetal growth and nutrition.     

Nitrate Loss for Denitrification during High Frequency Research in Floodplain Groundwater of the Tama River

This study aims to identify the possible roles of a floodplain inbiogeochemical nitrogen cycles, based on analysis of groundwaterdynamics of inorganic nitrogen species. Groundwater samples collected from boreholes made of poly vinyl chloride located onthe floodplain in the middle reach of the Tama River, Nagata district were analyzed for dissolved organic carbon, dissolved oxygen, inorganic nitrogen compounds, major anions and the stableisotope ratio of nitrate-nitrogen.Dissolved oxygen in groundwater samples collected from marked borehole (G-5) was low (50 ± 16 μM) and nitrate decreasedfrom 63 μM to < 10 μM during our research period. Concentrations of nitrate, nitrite and nitrous oxide decreased one after the other. These decreases in concentration, combinedwith ¹⁵N-enrichment of groundwater nitrate suggest denitrification as a significant nitrate sink, yielding an isotopic fractionation of nitrate-nitrogen with an enrichment factor (ε= –17.9‰) that is comparable tothose in various soil and groundwater systems. This study provesthat floodplains can perform as nitrogen sinks for groundwaterin river catchments.