Determination of molecular weight of agars and effect of the molecular weight on the glass transition

A novel procedure to determine the molecular weight (MW) and MW distributions for various agars is described. The MW values of commercial agars, an agarose, an agaropectin, and hydrolyzed agaroses were determined by size exclusion chromatography-low angle laser light scattering, using 4.0 M guanidine hydrochloride as eluent to avoid gelation. The MW for the commercial agars was between 106 400 and 243 500 with polydispersity between 1.283 and 6. 600. The MW of the agarose separated from a commercial agar was lower than that of the agaropectin. To prepare agaroses with different MW values, the obtained agarose was hydrolyzed. The MW of the agarose decreased with hydrolysis time, and the polydispersity, on the contrary, increased. The glass transition temperature (T(g)) of agarose with different MW values and that of agaropectin were measured by differential scanning calorimetry. The T(g) of the agarose was higher than that of the agaropectin with higher MW. The T(g) of agarose increased with MW.     

Bacterial Communities Responsible for the Decomposition of Rice Straw Compost in a Japanese Rice Paddy Field Estimated by DGGE Analysis of Amplified 16S rDNA and 16S rRNA Fragments

To estimate the succession and phylogenetic composition of the bacterial communities responsible for the decomposition of rice straw compost under flooded conditions during the cultivation period of paddy rice, denaturing gradient gel electrophoresis (DGGE) analyses targeting 16S rDNA and 16S rRNA, followed by sequencing were conducted in a Japanese paddy field. The DGGE bands of the bacterial communities in the rice straw compost were significantly more numerous in the DNA samples than in the RNA samples. Although the band number of the DNA samples was almost constant throughout the period, RNA samples showed fewer DGGE bands after mid-season drainage than before it. Thus, about 81% of the bacteria present in rice straw compost were considered to be metabolically "active" before mid-season drainage and about 62% after it. The changes in the DGGE patterns of bacterial DNA and RNA before and after mid-season drainage, respectively, were also revealed by cluster analysis and principal component analysis of the DGGE patterns. These results indicated that the bacterial communities of rice straw compost incorporated into flooded paddy fields changed gradually along with the decomposition, except for the period of mid-season drainage, but that they were influenced by mid-season drainage. Members of β-, γ- and δ-Proteobacteria, Cytophaga-Flavobacterium-Bacteroides (CFB) group, Chlorobia, Verrucomicrobia, Chloroflexi, Spirochaetes, Firmicutes (clostridia) and Actinobacteria were present during the decomposition of rice straw compost. Characteristic "active" bacteria among them were as follows: Clostridium, Acinetobacter (γ-Proteobacteria) and β-Proteobacteria before mid-season drainage, Flavobacterium, Chondromyces , Chlorflexi and δ-Proteobacteria after mid-season drainage, and Spirochaeta and myxobacteria throughout the period.     

Polymorphism of rRNA gene loci in the dog

We examined the number of ribosomal gene (rDNA) loci in the metaphase spreads of 54 dogs by FISH method. We found that in 16 dogs (30%) one or two loci were missing. The total number of rDNA loci was varied from 5 to 7 in males and from 4 to 6 in females. As the male dog consistently bears the rDNA on the Y chromosome, the polymorphism of the rDNA locus was ascribed to the absence of autosomal rDNA loci. Indeed, the frequency of polymorphism is almost equivalent in both sexes in the beagle. In one female beagle dog, remarkable intense fluorescence signals were observed at the four autosomal loci, indicating the in situ amplification of rDNA.     

Carbon skeletons for amide synthesis during ammonium nutrition in tomato and wheat roots

Heavy application of ammonium nitrogen to plant roots results in the conversion of ammonium nitrogen to the nitrogen of amides, glutamine, and asparagine, which are stored in roots or translocated to shoot. Since the net synthesis of such amides requires the supply of corresponding carbon skeletons, the carbon metabolism in amide synthesis in response to ammonium supply was investigated in tomato and wheat roots. The content of major primary amino acids was determined in tomato and wheat roots during a 4-d period of ammonium nutrition after 1-d culture in nitrogen-free nutrient solution. Ammonium supply led to a continuous increase in the asparagine content in wheat roots, whereas in tomato roots, the glutamine content increased 1 d after ammonium supply and thereafter the glutamine content was higher than the asparagine content. The amounts of amino acids synthesized from glucose-¹⁴C increased while the amounts of organic acids decreased in tomato roots by the supply of ammonium nitrogen for 1 d, compared to the roots that did not receive nitrogen. In tomato roots, the proportion of labeled glutamine was higher than that of labeled asparagine and the C5 amino acids were more strongly labeled than the C4 amino acids. These findings were different from the previous ones in wheat roots where the proportion of asparagine was found to be extremely high (Koga and Ikeda 2000: J. Fac. Agr. Kyushu Univ45, 7–13). To examine the in vivo asparagine synthesis, aspartate-'4C was fed to the roots. The labeling of asparagine, which was the most strongly labeled amino acid among the free amino acids, was remarkably strong in wheat roots whereas the labeling of glutamine was also pronounced in tomato roots. These results indicate that the ability to replenish carbon skeletons for amide synthesis in ammonium nutrition is different between tomato and wheat roots.     

Photosynthetic Carbon Metabolism in Corn Leaves Treated with Methionine Sulfoximine and Ammonium

The physiological role and significance of organic acids in the metabolic parts of plant which are involved in the respiratory or photosynthetic chain has not yet been elucidated apart from that of particular accumulation of specific organic acids in some storage tissues. Large fluctuations of the organic acid concentration in plants, often reflected by some minor changes in the growing conditions and even by the sampling time, may complicate the interpretation of the results obtained.

It has been considered that one of the most probable roles of organic acid accu-mulation in plant is to adjust the cation-anion balance in plant sap, and to take part in the transport of metallic cations in plant (1–3). It may also be suggested that the excess supply of sodium chloride affects appreciably the organic acid metabolism in plant through the compensation of the disequilibrium between the amount of inorganic cations and anions in plant saps caused by the uptake of a larger quantity of sodium ion (4).

The authors are currently carrying out a series of investigations about the physiological responses of crop plants to the excess supply of sodium chloride from the view point of species differences, and some of the results have been reported previously (5, 6). Recent development in high pressure liquid ion chromatography has enabled a more reliable and sensitive determination of small quantities of organic acids. In the present report the authors investigated th.e changes in the concentration of organic acids in bleeding and leaf saps of cucumber and tomato seedlings grown at different levels of sodium chloride, in relation to the concentration of inorganic cations and anions.     

Responses to nitrogen sources and regulatory properties of root phosphoenolpyruvate carboxylase

Time course of changes in extractable root phosphoenolpyruvate carboxylase (PEP C) activity was investigated in wheat, barley, and tomato plants fed with different nitrogen sources. Ammonium-fed plants exhibited a 2–2.5-fold higher PEPC activity than nitrate-fed plants at 7 d after the onset of nitrogen supply. Western blot analysis revealed that the amounts of PEPC subunit proteins increased gradually as reflected in the extractable PEPC activity. These results suggest that the increase in PEPC activity may be due to de novo protein synthesis. PEPC was SO-fold purified from tomato roots after several chromatographic steps. Metabolite effects on the partially purified enzyme were also investigated under optimal or suboptimal conditions in terms of pH and concentrations of phosphoenolpyruvate. Organic acids and acidic amino acids inhibited the enzyme activity, while hexose phosphates stimulated it. Glutamine and asparagine produced in the course of ammonium assimilation hardly affected the activity.     

Structure and functions of the placenta in common minke (Balaenoptera acutorostrata), Bryde’s (B. brydei) and sei (B. borealis) whales

The structure and functions of placentas were examined in 3 species of rorqual whales, common minke (Balaenoptera acutorostrata), Bryde’s (B. brydei) and sei (B. borealis) whales, with the aim of confirming the structural characteristics of the chorion, including the presence of the areolar part, and clarifying steroidogenic activities and fetomaternal interactions in the placentas of these whales. Placentas were collected from the second phase of the Japanese Whale Research Program under Special Permit in the North Pacific (JARPN II). Histological and ultrastructural examinations revealed that these whale placentas were epitheliochorial placentas with the interdigitation of chorionic villi lined by monolayer uninucleate cells (trophoblast cells) and endometrial crypts as well as folded placentation by fold-like chorionic villi. Moreover, well-developed pouch-like areolae were observed in the placentas, and active absorption was suggested in the chorionic epithelial cells of the areolar part (areolar trophoblast cells). Berlin blue staining showed the presence of ferric ions (Fe³⁺) in the uterine glandular epithelial cells and within the stroma of chorionic villi in the areolar part. An immunohistochemical examination revealed tartrate-resistant acid phosphatase (TRAP; known as uteroferrin in uteri) in the cytoplasm of glandular cells and areolar trophoblast cells. This result suggested that, in cetaceans, uteroferrin is used to supply iron to the fetus. Furthermore, immunoreactivity for P450scc and P450arom was detected in trophoblast cells, but not in areolar trophoblast cells, suggesting that trophoblast cells synthesize estrogen in whale placentas. Therefore, we herein immunohistochemically revealed the localization of aromatase and uteroferrin in cetacean placentas during pregnancy for the first time.     

Vertical Changes in Bacterial Communities in Percolating Water of a Japanese Paddy Field as Revealed by PCR-DGGE

Percolating water was sampled from the plow layer and subsoil layer in a Japanese paddy field, and the bacterial communities were compared together with floodwater by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) targeting a partial 16S rRNA gene and subsequent sequencing. The number of DGGE bands ranged from 16 to 28 with no significant differences among the sampling sites and times. Only 2 bands were common for the three sources of water samples. DGGE bands specific for the floodwater samples and percolating water samples from the plow layer were identified, while percolating water samples from the subsoil layer did not show specific bands but displayed common bands to those of the floodwater samples (7 bands) and percolating water samples from the plow layer (1 band). Cluster analysis of the DGGE banding patterns showed a distinct clustering in the samples of percolating water from the plow layer and a closer relationship between the others. These results suggest that the bacterial communities in percolating water changed during downward movement through the plow layer and subsoil layer. Sequences of the DGGE bands specific for the samples of percolating water from the plow layer showed a close relationship with anaerobic bacteria such as iron-reducers or uncultured bacterial DNA isolated from environments that are considered to be less oxic. On the other hand, the sequences of the bands specific for the samples of floodwater and percolating water from the subsoil layer showed a close relationship with uncultured bacterial DNA isolated from freshwater environments.     

Nodulation during vegetative growth of soybean stage does not affect the susceptibility to red crown rot caused by Calonectria ilicicola

Soybean cultivar Enrei and its derivative lines, non-nodulating En1282 and supernodulating Sakukei 4, were grown in an experiment field naturally infested with the red crown rot fungus, Calonectria ilicicola, to determine whether nodulation elicits the disease. Infection frequency by the fungus invariably increased with time and reached about 100% 6 weeks after sowing. Disease severity was invariably slight until 8 weeks after sowing. At harvest, however, disease severity was lowest in En1282 and highest in Sakukei 4. Enrei was intermediately susceptible. Sakukei 4 produced many more nodules than Enrei, while En1282 completely failed to nodulate. For three other pairs of nodulating and non-nodulating isogenic lines, all non-nodulating lines had lower severity than their nodulating counterparts at harvest. Pot experiments with Enrei and its derivatives revealed that inoculation with Bradyrhizobium sp. did not affect disease severity during the vegetative stage of plant growth, i.e., 6 weeks after sowing. These results suggested that the production of nodules in the vegetative stage is not, at least directly, related to susceptibility to the disease, but physiologic changes after anthesis vary depending on the nodulation traits in the respective soybean lines, resulting in different levels of susceptibility to red crown rot.