Characterization of an Alginate Lyase,FlAlyA, from Flavobacterium sp. Strain UMI-01 and Its Expression in Escherichia coli

A major alginate lyase, FlAlyA, was purified from the periplasmic fraction of an alginate-assimilating bacterium, Flavobacterium sp. strain UMI-01. FlAlyA showed a single band of ~30 kDa on SDS-PAGE and exhibited the optimal temperature and pH at 55 °C and pH 7.7, respectively. Analyses for substrate preference and reaction products indicated that FlAlyA was an endolytic poly(mannuronate) lyase (EC 4.2.2.3). A gene fragment encoding the amino-acid sequence of 288 residues for FlAlyA was amplified by inverse PCR. The N-terminal region of 21 residues except for the initiation Met in the deduced sequence was predicted as the signal peptide and the following region of six residues was regarded as propeptide, while the C-terminal region of 260 residues was regarded as the polysaccharide-lyase-family-7-type catalytic domain. The entire coding region for FlAlyA was subjected to the pCold I—Escherichia coli BL21(DE3) expression system and ~eight times higher yield of recombinant FlAlyA (recFlAlyA) than that of native FlAlyA was achieved. The recFlAlyA recovered in the periplasmic fraction of E. coli had lost the signal peptide region along with the N-terminal 3 residues of propeptide region. This suggested that the signal peptide of FlAlyA could function in part in E. coli.     

Assessment of criteria for improvement in Theileria orientalis sergenti infection tolerance

We examined the relationships between calf productivity, anemia and Theileria orientalis sergenti infection from records of 109 Japanese shorthorn calves during a grazing period and we quantified the relative importance of the T. orientalis sergenti infection‐resistance criteria used. A decrease in average parasitemia (AvePara) and an increase in average packed cell volume (AvePCV) would cause a decrease in the number of drug treatments. An increase in AvePCV would decrease the number of drug treatments more efficiently than a decrease in AvePara. An increase in AvePCV would cause an increase in dairy gain, while a decrease in AvePara would cause a decrease in dairy gain. Therefore, an improvement in the capacity to control anemia development would improve T. orientalis sergenti infection resistance more efficiently than an improvement in the capacity to control parasitemia development.     

An outbreak of bovine ephemeral fever in Nagasaki Prefecture in 1988

In October, 1988, bovine ephemeral fever (BEF) occurred in Nagasaki Prefecture and throughout Kyushu island, with the exception Miyazaki Prefecture. The first outbreak occurred in Hirado-shi on October 17. The total number of diseased cattle was 24 in 24 farms in Nagasaki Prefecture. The clinical findings were mainly sudden fever, anorexia, and instability in standing. The serum neutralizing antibodies against BEF virus (BEFV) rose in all infected cattle. Twelve strains of the virus were isolated in HmLu-1 cell cultures made directly from the heparinized blood of 17 infected cattle. The buffy coat was mainly collected from the samples and washed three times with phosphate buffered saline. These isolates were all neutralized by an antiserum against BEFV (Yamaguchi strain). With the aid of an electron microscope, a representative of isolates named Hirado-9 with a length of 150 nm was seen in the sample of infected HmLu-1 cell cultures. Both Hirado-9 and Yamaguchi strains reacted with antisera. The outbreak of BEF in 1988 was the first since 1971 in Nagasaki Prefecture. The result proved that BEFV can be easily isolated in HmLu-1 cell culture from the washed blood cells of infected cattle.     

Solute effects on the interaction between water and ethanol in aged whiskey

The hydrogen-bonding structure of water-ethanol in whiskey was examined on the basis of (1)H NMR chemical shifts of the OH of water and ethanol. Phenolic acids and aldehydes (gallic, vanillic, and syringic acids; vanillin and syringaldehyde) exhibited their structure-making effects regardless of the presence or absence of 0.1 or 0.2 mol dm(-3) acetic acid. The OH-proton chemical shifts were measured for 32 malt whiskey samples of a distillery, aged for 0-23 years in five different types of casks. The OH-proton chemical shift values of the whiskies shifted toward the lower field in proportion to their contents of total phenols. It can be concluded that the strength of the hydrogen bonding in aged whiskies is directly predominated by acidic and phenolic components gained in oak wood casks and not dependent on just the aging time.     

Importance of rubberwood in wood export of Malaysia and Thailand

We examined the contribution of rubberwood to the timber export markets of Malaysia and Thailand. In Malaysia, rubberwood has grown from 26% of total exported wood products in 1998 to 35% in 2007. A high proportion of furniture products (80%) is rubberwood, whereas the contribution of rubberwood to other wooden products is less than 20%. Only 10% of sawn timber and logs is rubberwood. In Thailand, rubberwood contributes to around 60% of total exported wood products, arising from a high share of not only furniture products (70%) but also other wood products (around 50%) and sawn timber and logs, which have increased in share from 40% in 1998 to 79% in 2007. We conclude that the high proportion of rubberwood products in the wood export markets of these two countries is a result of: (1) scarcity of raw wood materials because of strict controls on the logging of natural forests; and (2) governmental support to rubberwood production, including financial support to rubber planters and technical assistance to downstream timber processors.     

Estimation of rubberwood production in Cambodia

We estimated the production of rubberwood from 1996 to 2011 in Cambodia. We employed data from interview surveys with three rubberwood processing companies and statistics from estate rubber plantations and wood retailing companies. The result was that the end use of rubberwood in Cambodia was the sawntimber products, being exported to Vietnam and China. The yield rate from rubberwood stumpage into sawn timber was between 11.6 and 14.9%, closely matching the rates of Malaysia and Thailand. The average production of rubberwood roundwood from 1996 to 2011 was 254,697 m³/year, of which sawn timber as well as residuals in secondary processing was 84,559 and 170,138 m³/year, respectively. Although the timber production of Cambodia has declined sharply due to the strict restriction of natural forest logging, rubberwood could explain more than 60% of the domestic industrial roundwood production from 2000. We suggest the necessity of monitoring the expansion of rubber plantations in new rubber producing countries and linking the rubberwood production with the regional wood resource management.     

Variation and Segregation for Rachis Fragility in Spelt Wheat, Triticum Spelta L.

Spelt wheat, Triticum spelta L., has been proved to be rich-sources of useful genes for tolerance to biotic and abiotic stress, and grain quality. But this crop plant has some undesirable traits including glume tenacity and brittle rachis. Free-threshing and reduced fragility of rachis are very important traits for cultivation. The objectives in the present study were to investigate genetic variation of rachis fragility in a wide range of spelt accessions, to examine its genetic segregation pattern, and to clarify if rachis fragility is associated with dosage of chromosome 5A in aneuploid lines of bread wheat. The results demonstrated that spelt germplasm contains a wide range of rachis toughness, and thus selection of spelt wheat with desirable characteristics combined with an appropriate level of tough rachis would be possible. Spike morphology in the F₂ plants was segregated into the three types, square-headed, speltoid, and compactoid. The F₂ plants with compactoid spikes had the most brittle rachis, followed by the speltoid and square-headed spike F₂ plants. Rachis fragility in bread wheat also had genetic variation and was associated with dosage of chromosome 5A.     

Biological dinitrogen fixation and soil microbial biomass carbon as influenced by free-air carbon dioxide enrichment (FACE) at three levels of nitrogen fertilization in a paddy field

An experiment was conducted in an Andosol paddy field in Shizukuishi (Iwate Prefecture, Japan) to determine the effects of free-air CO₂ enrichment (FACE) on biological N₂-fixation activity and soil microbial biomass C at three levels of N application. Rice (Oryza sativa L. cv. Akitakomachi) plants were grown under ambient CO₂ or FACE (ambient +200 µmol mol^-1 CO₂) conditions throughout the growing season with each treatment having four replicated plots. Three levels of N fertilizer (high, standard and low; 15, 9 and 4 g N m^-2, respectively) were applied to examine the effect of different N availability under both CO₂ conditions. Soil samples were collected at four different times from upper and lower soil layers (0-1-cm and 1-10-cm soil depths, respectively) and analysed for biological N₂-fixation (BNF) activity and microbial biomass C (MBC) by the acetylene reduction and chloroform fumigation-extraction methods, respectively. The amounts of chlorophyll-type compounds (Chls), an index of algal growth, and soil available C were also determined. Compared to the ambient CO₂ treatment, the FACE treatment had significantly higher BNF activity in both the upper and lower soil layers at ripening only in low-N soil and at harvest at all three levels of N fertilization rates. MBC was significantly increased by FACE in both the upper and lower soil layers from the middle to later period of the growing season compared to the ambient CO₂ treatment. The FACE treatment increased the Chls in the upper soil layers at ripening only in low-N soil and at harvest at all three levels of N fertilization rates. The amount of soil available C was not significantly different between FACE and ambient CO₂ treatments in both the upper and lower soil layers throughout the cropping season. From these results it can be concluded that the FACE treatment had a significantly positive influence on BNF activity, MBC and Chls at different levels of N fertilization rates in paddy field during the cropping season.     

Displacement above the hypocenter of the 2011 Tohoku-Oki earthquake

The moment magnitude (M(w)) = 9.0 2011 Tohoku-Oki mega-thrust earthquake occurred off the coast of northeastern Japan. Combining Global Positioning System (GPS) and acoustic data, we detected very large sea-floor movements associated with this event directly above the focal region. An area with more than 20 meters of horizontal displacement, that is, four times larger than those detected on land, stretches several tens of kilometers long along the trench; the largest amount reaches about 24 meters toward east-southeast just above the hypocenter. Furthermore, nearly 3 meters of vertical uplift occurred, contrary to observed terrestrial subsidence.