6─氨基己酸二聚体水解酶基因在异源宿主中的表达

将起源于黄色菌（Ｆｌａｖｏｂａｃｔｅｒｉｕｍｓｐ．ＫＩ７２）的６─氨基己酸二聚体水解酶（ＥⅡ）基因以质粒ｐＴＳ１２０９为载体，转入假单孢菌ＰｓｅｕｄｏｍｏｎａｓｐｕｔｉｄａＮＴ１１２６中，得到具有ＥⅡ高表达活性转基因株，表明以假单胞菌为宿主进行ＥⅡ酶分子育种的可行性，并为ＥⅡ酶尚处于进化中的推论提供了旁证。     

<Emphasis Type="Italic">Spongospora subterranea</Emphasis> soil contamination and its relationship to severity of powdery scab on potatoes

We investigated soil contamination by Spongospora subterranea f. sp. subterranea (Sss) and disease severity of powdery scab in 29 potato fields in Hokkaido, Japan, using a hydroponic culture method with tomato seedlings as bait plants. The quantity of Sss infection on the roots of bait plants was evaluated using the polymerase chain reaction (PCR) and expressed in terms of the infection potential in the soil. The infection potential was positively correlated with the disease severity of harvested tubers, whereas the spore ball density determined using PCR had an indistinct relationship with disease severity. The infection potential can be useful in evaluating soil contamination and in applying countermeasures against powdery scab.     

Time-dependent changes in protein phosphorylation patterns in rat brain synaptosomes caused by deltamethrin

Effects of deltamethrin, a powerful pyrethroid insecticide, on the protein phosphorylation and dephosphorylation processes during depolarization in rat brain synaptosomes were studied by using [³²P]phosphoric acid as a starting radiotracer and high external concentration of potassium ions or veratridine (10^?-5 M) as depolarizing agents. At the onset of depolarization there was a quick rise in phosphorylation in various synaptic proteins for about 15–30 s followed by a gradual decline in levels of phosphorylation. The effect of deltamethrin (10^?-7 M) on this system was found to be dependent on the length of preincubation of the synaptosome with the pesticide prior to depolarization. At an early stage (0–3 min preincubation period) it caused a modest suppression of protein phosphorylation activities. When the period of deltamethrin preincubation was extended to 5–20 min, however, it caused a significant increase in protein phosphorylation throughout the depolarization period. At the later stage of the action of deltamethrin (e.g. preincubation period of 30–40 min), deltamethrin-treated synaptosomes no longer responded to the depolarization signal to raise the level of phosphorylation on many proteins. These results indicate that deltamethrin's actions on the synaptic process are complex. Depending on the length of exposure, its effects on protein phosphorylation responses in intact synaptosomes could be either stimulatory or inhibitory. To study the cause of deltamethrin-induced synaptic block at the later stage, effects of deltamethrin on protein kinases were studied by using lysed synaptic membranes with [gamma-³²P]ATP. Deltamethrin was shown to inhibit calcium–calmodulin-dependent protein phosphorylation activities at 10^?-7 M when given directly to the enzyme source 10 min prior to the addition of [³²P]ATP. Such an observation helps to explain the inhibitory action of deltamethrin on protein phosphorylation which occurs at the late stage of its action (i.e. preincubation time > 20 min).     

Current classification of Ralstonia solanacearum and genetic diversity of the strains in Japan

Ralstonia solanacearum is the causal organism of bacterial wilt of more than 200 species representing 50 families of plants in tropical, subtropical, and warm temperate regions in the world. Traditionally classified into five races based on differences in host range, R. solanacearum has also been grouped into six biovars on the basis of biochemical properties. With recent developments in molecular biology, various DNA-based analyses have been introduced and used to confirm that this binary system does not completely represent the diversity within R. solanacearum strains. Therefore, a new hierarchical classification scheme has been suggested, which defines R. solanacearum as a species complex and reorganized the concept of the species as a monophyletic cluster according to a phylogenetic analysis based on genomic sequence data. Here we discuss the current bacterial wilt situation and genetic relationships based on the recent classification system of Japanese R. solanacearum strains as well as worldwide strains. We also review the genetic, biochemical, and pathological characteristics of R. solanacearum strains, in particular, those affecting potato and Zingiberaceae plants as distinctly important pathogens in relation to continuously problematic and recent emergent diseases in Japan.     

Pathogenic characters of Japanese potato strains of Ralstonia solanacearum

Ralstonia solanacearum (Rs) strains in phylotypes I and IV isolated from potato in Japan were investigated for pathogenicity on potato, tomato, eggplant, Solanum integrifolium, tobacco, groundnut, and pumpkin. The strains were divided into 17 types based on differences in their pathogenicity on the tested plants. Particularly, the pathogenicity of most phylotype I strains on eggplant was distinctly different from that of the phylotype IV strains. When nine potato varieties (included two breeding lines) were inoculated with several Rs strains, phylotype IV strains were highly virulent on the breeding lines that are regarded as resistant to phylotype I strains.     

Watercore in Japanese pear. III. Changes in the activities of some enzymes relating to the degradation of cell walls and the accumulation of sugar

The relationship between cellular breakdown and the activities of some cell-wall degrading enzymes (endocellulase, exocellulase, polygalacturonase, pectin methylesterase and β-galactosidase) was investigated in the watercored tissue of Japanese pear fruit, Pyrus serotina Rehder var. ‘Culta’ Rehder, cultivar ‘93-3’. In watercored tissue at an early stage, endocellulase activity, especially neutral form, was higher than that in healthy tissue. As the symptom of the watercore developed, polygalacturonase and β-galactosidase activities in the watercored tissue increased compared with those in healthy tissue. This indicated that in tissues at an early stage of watercore, these cell-wall degrading enzymes were activated, and operative in the breakdown of the cell wall. However, exocellulase and pectin methylesterase were not activated in the watercored tissue. The breakdown of cellular tissue was also supported by the fact that the DNA and protein contents decreased in the watercored tissue. Invertase activity decreased in the watercored tissue, compared with healthy tissue, and it was suggested that its decrease could be one of the causes of sucrose accumulation in the watercored tissue.     

Antiobese effects of novel saponins from edible seeds of Japanese horse chestnut (Aesculus turbinata BLUME) after treatment with wood ashes

Recently, we have identified novel saponins from edible seeds of Japanese horse chestnut ( Aesculus turbinata BLUME) after processing the natural seeds with wood ashes to remove bitterness. We attempted to determine anti-obesity effects of those saponins from edible seeds as well as natural seeds. The purified individual components of saponins from natural and edible seeds inhibited pancreatic lipase in vitro. The potency was in the order of escins > desacylescins > deacetylescins. Escins Ib and IIb as well as deacetylescins Ib and IIb with the angeloyl moiety were more potent than the corresponding Ia and IIa series with the tigloyl moiety. Moreover, in vivo anti-obesity effects of the saponin fractions were monitored for 8 weeks in mice fed high-fat diets. Saponin fractions from both seeds significantly attenuated the elevation in body weight, the mass of peritoneal adipose tissues, and plasma triacylglycerol, which was accompanied by higher contents of undigested fats in feces without changes in food intake, indicating the effective inhibition of fat digestion in vivo. Taken together, saponin fractions including desacylescins and deacetylescins from edible seeds are potentially useful for the development of nutraceutical foods with anti-obesity effects and more attenuated bitter taste.     

Electron microprobe analysis of zinc and other elements within and around rice root grown in flooded soil

Electron microprobe analysis was applied to the localization of zinc, phosphorus, sulfur, and calcium within and around the rice root grown in the flooded soil of high zinc content (300 ppm).

A method for the preparation of the section of the undisturbed soil with the rice root in place is described. Having been dehydrated with ethanol, the sample became free of water soluble ions and, therefore, the insoluble fractions of the elements were mainly localized. The results obtained were as follows:

1) A longitudinal view of the root in the soil was successfully shown as a secondary electron image.

2) The compaction effect of the expanding root on the orientation of the soil particles was not observed.

3) The distribution patterns of the elements in the apical region of the root differed distinctly from those in the basal region. In the latter region, the elements were accumulated in the cells of the epidermis, exodermis, endodermis, and marginal layers of stele. Zinc and phosphorus appeared to be in particles at the same sites in this region. In the apical region, all the elements but calcium were highly and evenly distributed in the central part of the root and scarcely found in marginal layers. Calcium was not significantly detected anywhere in this region.

4) In the basal region, all the elements, whether they were actively absorbed or not, were found to be accumulated at the same sites.

5) Flooding of the soil seemed not to affect the distribution of the elements within the root.