Identification of a 4-Deoxy-l-erythro-5-hexoseulose Uronic Acid Reductase,FlRed, in an Alginolytic Bacterium Flavobacterium sp. Strain UMI-01

In alginate-assimilating bacteria, alginate is depolymerized to unsaturated monosaccharide by the actions of endolytic and exolytic alginate lyases (EC 4.2.2.3 and EC 4.2.2.11). The monosaccharide is non-enzymatically converted to 4-deoxy-l-erythro-5-hexoseulose uronic acid (DEH), then reduced to 2-keto-3-deoxy-d-gluconate (KDG) by a specific reductase, and metabolized through the Entner–Doudoroff pathway. Recently, the NADPH-dependent reductase A1-R that belongs to short-chain dehydrogenases/reductases (SDR) superfamily was identified as the DEH-reductase in Sphingomonas sp. A1. We have subsequently noticed that an SDR-like enzyme gene, flred, occurred in the genome of an alginolytic bacterium Flavobacterium sp. strain UMI-01. In the present study, we report on the deduced amino-acid sequence of flred and DEH-reducing activity of recombinant FlRed. The deduced amino-acid sequence of flred comprised 254 residues and showed 34% amino-acid identities to that of A1-R from Sphingomonas sp. A1 and 80%–88% to those of SDR-like enzymes from several alginolytic bacteria. Common sequence motifs of SDR-superfamily enzymes, e.g., the catalytic tetrad Asn-Lys-Tyr-Ser and the cofactor-binding sequence Thr-Gly-x-x-x-Gly-x-Gly in Rossmann fold, were completely conserved in FlRed. On the other hand, an Arg residue that determined the NADPH-specificity of Sphingomonas A1-R was replaced by Glu in FlRed. Thus, we investigated cofactor-preference of FlRed using a recombinant enzyme. As a result, the recombinant FlRed (recFlRed) was found to show high specificity to NADH. recFlRed exhibited practically no activity toward variety of aldehyde, ketone, keto ester, keto acid and aldose substrates except for DEH. On the basis of these results, we conclude that FlRed is the NADH-dependent DEH-specific SDR of Flavobacterium sp. strain UMI-01.     

Mechanism of the rice hull‐induced germination of Monochoria vaginalis seeds in darkness

The seeds of Monochoria vaginalis require light to germinate. However, they are able to germinate in darkness in the presence of rice hulls. The mechanism of this phenomenon was investigated in this study, in which only the unsterilized seeds of Monochoria were induced to germinate by the presence of a rice hull extract. The rice hull extract was able to be replaced by either a mixture of amino acids and phosphate or a yeast extract. The culture media in which germination was observed always showed turbidity, indicating the propagation of bacteria, which were identified as Bacillus sp., Pedobacter sp., Pantoea sp. and Sphingomonas sp. Treatment with the individual bacterial species or combinations of these bacteria induced Monochoria seed germination when added to the media containing the rice hull extract. It was concluded that the rice hull extract accelerates the propagation of bacteria, which in turn digest the seed coat of Monochoria, facilitating its germination.     

Recent advances in the high-density rotifer culture in Japan

The success of rapidly growing aquaculture industry depends on the steady supply of fish seeds. Appropriate food for initial larval stages is critical for mass scale fish seed production. Live food found better over artificial food for fish larvae culture. Rotifers have been found to be the best live food for feeding fish larvae in early life stage. Attempts have been made to develop viable techniques for the production of rotifer through batch, continuous, and semi-continuous culture methods. In order to feed increased number of fish larvae, rotifer need to be cultured under high-density method. Various efforts have been made for increasing culture density of rotifer. In Japan, stable high-density culture of rotifer has been developed in commercial scale at a rate of 20,000–30,000 ind. ml^?1. Later on, ultra-high-density rotifer culture (160,000 ind. ml^?1) was found successful, which can fulfill the increased demand for rotifer as fish larval food. Furthermore, a scope of alternative use of rotifer can be explored. The development of the high-density rotifer culture method in Japan has been reviewed. The considerations of the associated nutritional requirements, microbial aspects, and prospects of high-density culture have been discussed.     

Exposure to blue LED light before the onset of darkness under a long‐day photoperiod alters melatonin secretion,feeding behaviour and growth in female dairy calves

The effect of blue LED on melatonin secretion, feeding behaviour and growth was addressed in Holstein female dairy calves. In Exp.1, six animals (8 weeks old, 97 ± 4.1 kg BW) were exposed to yellow or blue LED for 2 hr before darkness over 7 days under a long‐day photoperiod (LDPP). In Exp. 2, six animals (8 weeks old, 88.5 ± 4.8 kg BW) were exposed to blue light from a white LED all daytime or a yellow LED for 2 hr before the darkness of LDPP (blue light cut) over 3 weeks. In Exp. 1, blue light mildly suppressed melatonin secretion during the 2‐hr treatment but did not affect the timing of the nightly melatonin rise. However, the rise in nighty melatonin levels was higher with yellow than blue LED. In Exp. 2, white LED completely suppressed melatonin secretion during the 2‐hr treatment, but plasma melatonin concentrations were similar during the darkness. Grass hay intake, rumination time, frequency of water intake and body weight gain were higher in animals exposed to the yellow rather than the white LED. Overall results indicate that exposure to blue light from white LEDs under an LDPP suppresses melatonin secretion and might negatively impact the development of female dairy calves.     

Microbial reduction mechanism of ferric iron in paddy soils (Part I)

In the previous paper¹⁾ the authors proved that the ferric iron reduction in submerged paddy soils is entirely or mostly effected by the activities of microorganisms in the soil. Prior to the study on the ferric iron reducing microorganisms isolated from the soil, the present paper reports the research for the microbial mechanism of iron reduction. working with the soil itself.     

The reduction mechanism of manganese in paddy soils

Hydroquinone method manganese (soluble in pH 7.0, 1 N-ammonium acetate solution containing 0.2 percent hydroquinone) and microbially active manganese (soluble in pH 7.0, 1 M-magnesium sulfate solution after flooding soils with or without Chinese milk vetch for 12 or 20 days respectively at 30°C) of 22 paddy soils were determined. The amounts of manganese reduced with sodium oxalate under acid conditions (oxalate method manganese (a) and (b), the former was determined under more rigorous conditions than the latter) were also determined and compared with hydroquinone method manganese and microbially active manganese.

Their levels of many soil samples representing soil groups were also determined to examine the dlfferences In amounts of active manganese among soil groups. The results obtained are as follows.

The relationship between microbially and chemically active manganese. 1) The amounts of microbially active manganese in soils were 48 to 68 mg Mn per 100 g oven-dried soil and these were increased by the addition of Chinese milk vetch. 2) The amounts of hydroquinone method manganese were less than microbially active manganese, and the amounts of oxalate method manganese (b) were larger than microbially active manganese. The amounts of oxalate method manganese (a) were the largest of all the types of manganese. 3) There were high correlations between the amounts of various types of active manganese described in 2). The levels of microbially and chemically active manganese. 1) The amounts of microbially active manganese lay between the amounts of chemically active manganese determined by the hydroqulnone method and by the oxalate method (b) In all soil samples representing soil groups. High correlations were found between these types of active manganese. 2) The hydroquinone method was considered to be unsuitable for quantitatively determining the amounts of chemicallY active manganese in soils of high organic matter content. 3) In both cases of microbially active manganese and chemically active manganese, tha widest range and the largest amount determined were both observed in strongly gley soila. The averages of theae types of active manganese were high in strongly gley soils, pea, and muck soils, and black soils. The differences among soil groups were smalle1 than the differences among soil samples, and little tendency was observed in the differences among soil groups.

From these findings described above it is suggested that the oxalate methoo (b) is more appropriate than the hydroqulnone method for determining chemically active manganese as an index of microbially active manganese.     

Effect of flutianil on the morphology and gene expression of powdery mildew

Flutianil, a fungicide effective only on powdery mildew, was previously reported to affect the host cell''s haustorial formation and nutrient absorption. Studies were conducted to investigate flutianil''s primary site of action on Blumeria graminis morphology using transmission electron microscope (TEM) observation and RNA sequencing (RAN-seq) techniques. TEM observation revealed that flutianil caused the extra-haustorial matrix and fungal cell wall to be obscured, without remarkable changes of other fungal organelles. RNA-seq analysis indicated that, unlike other powdery-mildew fungicides, flutianil did not significantly affect the constantly expressed genes for the survival of B. graminis. Genes whose expression is up- or downregulated by flutianil were found; these are the three sugar transporter genes and various effector genes, mainly expressed in haustoria. These findings indicate that the primary site of action of flutianil might be in the haustoria.     

An Improved Synthesis of 7, 8-Epoxy-1,3,11-cembratriene- 15R(α), 16-diol,a Cembranoid of Marine Origin with a Potent Cancer Chemopreventive Activity

An effective method for the synthesis of 7,8-epoxy-1,3,11-cembratriene- 15R(α),16-diol and its in vitro Epstein-Barr Virus Early Antigen (EBV-EA) Activation Chemopreventive Assay are reported. This semisynthetic product is a new cembranoid with a potent tumor inhibitory activity that is expected to be a lead compound for a new class of chemopreventive agents of marine origin.     

An index for evaluating the flood-prevention function of paddies

Urbanization is expanding in and around small to medium-sized cities as well as in large cities, and the expansion includes low-lying areas where the flood risk is high. Flood risk in small watersheds has been evaluated using precise mathematical models. However, these models cannot be applied to larger basins due to the complexity of data collection and analysis. As a result, quantitative evaluations of flood risk at macro scales and the flood-prevention function of paddies around urban areas have not been made. This paper proposes a method and an index that can be used to evaluate the flood-prevention function of paddies on a regional scale. In addition, the use of this system for basin-wide management is illustrated by an example for the Kinu and Kokai rivers, which are tributaries of the Tone River of Japan. The applicability of the approach is evaluated using data from a major flood that occurred in the study area.