Molecular and genealogical analysis of grain dormancy in Japanese wheat varieties,with specific focus on MOTHER OF FT AND TFL1 on chromosome 3A

In the wheat (Triticum aestivum L.) cultivar ‘Zenkoujikomugi’, a single nucleotide polymorphism (SNP) in the promoter of MOTHER OF FT AND TFL1 on chromosome 3A (MFT-3A) causes an increase in the level of gene expression, resulting in strong grain dormancy. We used a DNA marker to detect the ‘Zenkoujikomugi’-type (Zen-type) SNP and examined the genotype of MFT-3A in Japanese wheat varieties, and we found that 169 of 324 varieties carry the Zen-type SNP. In Japanese commercial varieties, the frequency of the Zen-type SNP was remarkably high in the southern part of Japan, but low in the northern part. To examine the relationship between MFT-3A genotype and grain dormancy, we performed a germination assay in three wheat-growing seasons. On average, the varieties carrying the Zen-type SNP showed stronger grain dormancy than the varieties carrying the non-Zen-type SNP. Among commercial cultivars, ‘Iwainodaichi’ (Kyushu), ‘Junreikomugi’ (Kinki-Chugoku-Shikoku), ‘Kinuhime’ (Kanto-Tokai), ‘Nebarigoshi’ (Tohoku-Hokuriku), and ‘Kitamoe’ (Hokkaido) showed the strongest grain dormancy in each geographical group, and all these varieties, except for ‘Kitamoe’, were found to carry the Zen-type SNP. In recent years, the number of varieties carrying the Zen-type SNP has increased in the Tohoku-Hokuriku region, but not in the Hokkaido region.     

Naturally occurring antifungal agents against Zygosaccharomyces bailii and their synergism

Polygodial was found to exhibit a fungicidal activity against a food spoilage yeast, Zygosaccharomyces bailii, with the minimum fungicidal concentration (MFC) of 50 microg/mL (0.17 mM). The time-kill curve study showed that polygodial was fungicidal at any growth stage. The primary action of polygodial comes from its ability to disrupt the native membrane-associated function of integral proteins as nonionic surface active agents (surfactants) followed by a decrease in plasma membrane fluidity. The fungicidal activity of polygodial was increased 128-fold in combination with a sublethal amount (equivalent of 1/2 MFC) of anethole and vice versa relative to the fungicidal activity of anethole. The fungicidal activity of sorbic acid was enhanced 512-fold in combination with 1/2 MFC of polygodial. Conversely, the fungicidal activity of polygodial was enhanced 128-fold in combination with 1/2 MFC of sorbic acid.     

Studies on absorption and hydrolysis of ethyl alpha-D-glucoside in rat intestine

Ethyl alpha-D-glucoside (alpha-EG) is normally contained in Sake, which has been taken by Japanese people since ancient times. In this study, the intestinal absorption of alpha-EG was investigated using rat everted intestinal sac. Furthermore, the alpha-EG hydrolytic activity in rat intestine was compared with disaccharides hydrolytic activities, and the effects of alpha-EG on disaccharides hydrolysis were examined using crude enzyme preparation from rat intestinal acetone powder. Glucose liberated from alpha-EG was detected in a serosal solution of everted rat intestinal sac, but it was only less than 4% of absorbed intact alpha-EG. alpha-EG absorption into small intestinal tissue was reduced by elimination of sodium ion from the mucosal solution or under the presence of phlorizin. The hydrolytic activity for alpha-EG was detected in crude enzyme preparation from rat intestinal acetone powder, but it showed a low value as compared to those for disaccharides. alpha-EG showed mixed type inhibition for maltose and sucrose hydrolysis, but inhibitory concentrations of alpha-EG required for 50% inhibition for the maltose and sucrose hydrolysis were higher than those of arabinose and acarbose. In conclusion, a small amount of alpha-EG was hydrolyzed and most of it was absorbed via SGLT1 as an intact form in the rat small intestine, and the inhibitory effect of alpha-EG on disaccharides hydrolysis was weak.     

Purification and characterization of a corrinoid compound from Chlorella tablets as an algal health food

Vitamin B(12) content of an algal health food, Chlorella tablets (Chlorella sp.), was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor-chemiluminescence methods. The values of 200.9-211.6 microg/100 g dry weight determined by the chemiluminescence method were similar to the values (201.3-285.7 microg/100 g dry weight) determined by the microbiological method. A corrinoid compound was purified to homogeneity from the Chlorella tablets and characterized. The purified corrinoid compound was identified as vitamin B12, on the basis of silica gel 60 TLC, C18 reversed-phase HPLC, 1H NMR spectroscopy, and UV-Vis spectroscopy.     

Naturally occurring anti-Salmonella agents.

Polygodial and (2E)-hexenal were found to possess antibacterial activity against Salmonella choleraesuis with the minimum bactericidal concentrations (MBC) of 50 microg/mL (0.17 mM) and 100 microg/mL (0.98 mM), respectively. The time kill curve study showed that these two alpha,beta-unsaturated aldehydes were bactericidal against this food-borne bacterium at any stage of growth. However, they showed different effects on the growth of S. choleraesuis. The combination of polygodial and anethole exhibited strong synergism on their bacteriostatic action but only marginal synergism on their bactericidal action.     

Antibacterial activity of akyl gallates against Bacillus subtilis

The antibacterial activity of a series of alkyl gallates (3,4,5-trihydroxybenzoates) against Gram-positive bacteria was tested using a broth dilution method. All of the Gram-positive bacteria tested were susceptible to alkyl gallates, and this activity was found to correlate with the alkyl chain length. The antibacterial activity of alkyl gallates against Bacillus subtilis was a parabolic function of their lipophilicity and maximized with alkyl chain length between C(8) and C(11). Notably, alkyl gallates were found to be bactericidal against B. subtilis ATCC 9372, but this activity was significantly affected by the endospore formation in the culture. The antibacterial activity of alkyl gallates likely comes at least in part from their ability to inhibit the membrane respiratory chain but is not due to the prooxidant action.     

Identification of a novel cytotoxic protein, Cry45Aa, from Bacillus thuringiensis A1470 and its selective cytotoxic activity against various mammalian cell lines

Parasporal inclusion proteins produced by Bacillus thuringiensis strain A1470 exhibit strong cytotoxicity against human leukemic T cells when activated by protease treatment. One of the cytotoxic proteins was separated by anion exchange chromatography and gel filtration chromatography and designated Cry45Aa. Its gene was then expressed in recombinant Escherichia coli, in which the Cry45Aa precursor was accumulated in an inclusion body. It was solubilized in sodium carbonate buffer and processed with proteinase K, and cytotoxic activities of the protein against various mammalian cell lines were evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide assay. The protein exhibited high cytotoxic activity against CACO-2, Sawano, MOLT-4, TCS, and HL60 cells and moderate activity against U-937 DE-4, PC12, and HepG2 cells. On the other hand, the EC50 values against Jurkat, K562, HeLa, A549, Vero, COS-7, NIH3T3, CHO, and four normal tissue cells (human primary hepatocyte cells, UtSMC, MRC-5, and normal T cells) were >2 microg/mL.     

Antifungal mechanism of polygodial

The primary antifungal action of polygodial comes in part from its ability to function as a nonionic surfactant, disrupting the lipid-protein interface of integral proteins and denaturing their conformation. As a result, the antifungal mechanism of this sesquiterpene dialdehyde is associated with the membrane functions or derangement of the membrane. For example, the glucose-induced medium acidification process of Saccharomyces cerevisiae was inhibited by polygodial, presumably caused by inhibition of the plasma membrane H(+)-ATPase. However, the potent antifungal activity of polygodial results from its multiple functions.     

An analysis of wet deposition of sulfate using a trajectory model for East Asia

A long-range transport model for East Asia was developed to estimate the wet deposition of sulfate. The model is a trajectory type which is appropriate for long-term analysis. Trajectories of air masses are calculated by tracing the wind field which changes spatially and temporally. The processes of reactions, rainout removal, intake of sulfate in cloud water into rain water, and dry and wet depositions are considered. It is possible to calculate the concentration of sulfate in precipitation at a receptor by performing material balance in a grid box containing the receptor.The results obtained by the long-range transport model were evaluated through comparison with observation data of acidic deposition. The observation was conducted at 21 stations throughout Japan for one year. The calculated amount of wet deposition of sulfate in Japan was 0.22Tg/y in S equivalent, while the observed amount was 0.29Tg/y. The long-range transport model can predict almost 80% of observed wet deposition. The contributions of domestic anthropogenic sources and volcanic eruption to wet deposition of sulfate in Japan were estimated using the longrange transport model. The ratio of the deposition of sulfate due to Japanese anthropogenic sources to that due to the Asian continental sources was about 1 to 2. Since air stream from the direction of the Asian continent dominates during winter, the contribution of Japan to wet deposition in the region which faces the Sea of Japan amounted to less than 15%. The contribution of the sulfur oxides from volcanoes was about 20%.     

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.