Characterization of genes for novel thaumatin-like proteins in Cryptomeria japonica

Thaumatin-like proteins (TLPs) are induced by a variety of phytopathogens in many plants and several TLPs are allergenic. Previously, we isolated three TLP-encoding cDNAs (Cry j 3.1, Cry j 3.2 and Cry j 3.3) from a cDNA library derived from the pollen of Cryptomeria japonica D. Don. Here, we describe three new TLP cDNAs (Cry j 3.4, Cry j 3.5 and Cry j 3.6). We compared the sequences, the genetic map location and the expression patterns of the Cry j 3 genes. The amino acid sequence predicted from Cry j 3.5 exhibits only limited similarity to those predicted from the other Cry j 3 genes. Linkage analysis showed that the Cry j 3.1 to Cry j 3.4 genes are located in the same linkage group, but Cry j 3.5 is located in a different group. Organ-specificity and induction by stresses and plant hormones differed among the Cry j 3 mRNAs. In pollen grains, the Cry j 3.5 mRNA expression level was higher than that of the other Cry j 3 genes. Exposure to UV-B and salt stress induced expression of Cry j 3.1. The ethylene-releasing compound ethephon strongly induced expression of Cry j 3.4. Salt stress and salicylic acid also induced expression of Cry j 3.4. Abscisic acid weakly induced expression of Cry j 3.5. Arachidonic acid strongly induced expression of Cry j 3.4 and Cry j 3.6, and weakly induced that of Cry j 3.3, whereas expression of Cry j 3.1 and Cry j 3.5 was unaffected. These results suggest that the roles of TLPs and the cascades that regulate their expression differ among the members of the TLP family in C. japonica.     

Partial characterization and ontogenetic development of pancreatic digestive enzymes in Japanese eel Anguilla japonica larvae

The pancreatic digestive enzymes, trypsin, chymotrypsin, lipase and amylase were partially characterized, and changes in their activities were examined during the initial ontogeny of Japanese eel Anguilla japonica larvae from 5 to 34 days post-hatching (dph). The pH optima of the eel larval enzymes were narrower than those other fish species; trypsin activity was highest at pH 9, chymotrypsin and amylase activities were highest at pH 7 and 8, and lipase activity was highest at pH 8 and 9. In an analysis of thermal profiles, the larval pancreatic enzymes had a high optimal temperature and high thermal stability, which are typical of fish from the tropics. At 12 and 13 dph, lipase activity and gene expression levels of trypsin (-a and -b), lipase and amylase decreased markedly, suggesting a marked change in larval metabolism at that time. These data could be useful in the development of artificial larval diets in Japanese eel.     

Integrating remote sensing and GIS for prediction of rice protein contents

In this study, protein content (PC) of brown rice before harvest was established by remote sensing (RS) and analyzed to select the key management factors that cause variation of PC using a GIS database. The possibility of finding out the key management factors using GreenNDVI was tested by combining RS and a GIS database. The study site was located at Yagi basin (Japan) and PC for seven districts (85 fields) in 2006 and nine districts (73 fields) in 2007 was investigated by a rice grain taste analyzer. There was spatial variability between districts and temporal variability within the same fields. PC was predicted by the average of GreenNDVI at sampling points (Point GreenNDVI) and in the field (Field GreenNDVI). The accuracy of the Point GreenNDVI model (r ² > 0.424, RMSE < 0.256%) was better than for the Field GreenNDVI model (r ² > 0.250, RMSE < 0.298%). A general-purpose model (r ² = 0.392, RMSE = 0.255%) was established using 2 years data. In the GIS database, PC was separated into two parts to compare the difference in PC between the upper (mean + 0.5SD) and lower (mean − 0.5SD) parts. Differences in PC were significant depending on the effective cumulative temperature (ECT) from transplanting to harvest (Factor 4) in 2007 but not in 2006. Because of the difference in ECT depending on vegetation term (from transplanting to sampling), PC was separated into two groups based on the mean value of ECT as the upper (UMECT) and lower (LMECT) groups. In 2007, there were significant differences in PC at LMECT group between upper and lower parts depending on the ECT from transplanting to last top-dressing (Factor 2), the amount of nitrogen fertilizer at top-dressing (Factor 3) and Factor 4. When the farmers would have changed their field management, it would have been possible to decrease protein contents. Using the combination of RS and GIS in 2006, it was possible to select the key management factor by the difference in the Field GreenNDVI.     

Effects of β‐carotene‐enriched dry carrots on β‐carotene status and colostral immunoglobulin in β‐carotene‐deficient Japanese Black cows

Data from 18 β‐carotene‐deficient Japanese Black cows were collected to clarify the effects of feeding β‐carotene‐enriched dry carrots on β‐carotene status and colostral immunoglobulin (Ig) in cows. Cows were assigned to control or carrot groups from 3 weeks before the expected calving date to parturition, and supplemental β‐carotene from dry carrots was 138 mg/day in the carrot group. Plasma β‐carotene concentrations in the control and carrot groups at parturition were 95 and 120 μg/dL, and feeding dry carrots slightly improved plasma β‐carotene at parturition. Feeding dry carrots increased colostral IgA concentrations in cows and tended to increase colostral IgG₁, but colostral IgM, IgG₂, β‐carotene and vitamin A were not affected by the treatment. Feeding dry carrots had no effects on plasma IgG₁, IgA and IgM concentrations in cows, but plasma IgG₁ concentrations decreased rapidly from 3 weeks before the expected calving date to parturition. These results indicate that feeding β‐carotene‐enriched dry carrots is effective to enhance colostral IgA and IgG₁ concentrations in β‐carotene‐deficient cows.     

Allelopathic potential and an allelopathic substance in mango leaves

The present study was aimed to determine the allelopathic potential of mango (Mangifera indica L.) leaves and to identify allelopathic substances in the leaves. The aqueous methanol extracts of mango leaves inhibited seedling growth of garden cress, radish, rapeseed, foxtail fescue and crabgrass, and the inhibitory effects increased with the increasing extract concentration, suggesting that mango leaves may contain allelopathic substances. The extract was then purified by several chromatographic runs through a bioassay-directed fractionation, and an growth inhibitory substance was isolated and identified by spectral data as methyl-3,4,5-trihydroxybenzoate (methyl gallate). Methyl gallate at the concentration of 10 mM inhibited 98.6% and 99.8% of root and shoot growth of garden cress relative to the control, respectively, and 94.4% and 49.3% of those of foxtail fescue, respectively. The concentrations of methyl gallate required for 50% growth inhibition (I₅₀) on garden cress roots and shoots were 3.9 and 3.3 mM, and those on foxtail fescue roots and shoots were 1.5 and 9.5 mM, respectively. It is the first report of an allelopathic substance in mango leaves and allelopathic activity of methyl gallate. Therefore, the mango leaves may have potential as a soil additive material for the weed management options.     

Mapping of QTLs controlling epicotyl length in adzuki bean (Vigna angularis)

Epicotyl length (ECL) of adzuki bean (Vigna angularis) affects the efficiency of mechanized weeding and harvest. The present study investigated the genetic factors controlling ECL. An F₂ population derived from a cross between the breeding line ‘Tokei1121’ (T1121, long epicotyls) and the cultivar ‘Erimo167’ (common epicotyls) was phenotyped for ECL and genotyped using simple sequence repeats (SSRs) and single-nucleotide polymorphism (SNP) markers. A molecular linkage map was generated and fifty-two segregating markers, including 27 SSRs and 25 SNPs, were located on seven linkage groups (LGs) at a LOD threshold value of 3.0. Four quantitative trait loci (QTLs) for ECL, with LOD scores of 4.0, 3.4, 4.8 and 6.4, were identified on LGs 2, 4, 7 and 10, respectively; together, these four QTLs accounted for 49.3% of the phenotypic variance. The segregation patterns observed in F₅ residual heterozygous lines at qECL10 revealed that a single recessive gene derived from T1121 contributed to the longer ECL phenotype. Using five insertion and deletion markers, this gene was fine mapped to a ~255 kb region near the end of LG10. These findings will facilitate marker-assisted selection for breeding in the adzuki bean and contribute to an understanding of the mechanisms associated with epicotyl elongation.     

Clinical and histopathological evaluation of Dermatophagoides farinae-induced dermatitis in NC/Nga mice orally administered Bacillus subtilis

Probiotic strains have been reported to have the ability to control allergic and inflammatory diseases. In this study, we studied the inhibitory effect of Bacillus subtilis (natto) (BS) on atopic dermatitis. The effects of continuous oral administration of BS for 4 weeks on the development of atopic dermatitis induced by Dermatophagoides farinae body antigen (DF) in NC/Nga (NC) mice were evaluated using 4 groups of mice: group (Gp) DF, DF(+) with no administration of bacteria (n=3); Gp DF/BS, DF(+) and BS(+) (n=5); and Gp PBS, DF(-) with no administration of bacteria (n=3). The mice were gavaged with 1.2 × 10(17) CFU/head of BS 6 times a week for 4 weeks, and DF was applied twice a week for 4 weeks. Histopathological examination revealed significant differences in auricular thickness between Gp DF (664.4 μm, SD=78.0) and Gp DF/BS (278.7 μm, SD = 88.8; p<0.01). The dorsal skin of Gp DF/BS (316.7 μm, SD=187.4) was significantly thinner than that of Gp DF (503 μm, SD=116.3). These results suggest that continuous oral administration of fermented food-derived bacteria (BS) can be effective in alleviating the development of skin lesions induced by DF in NC mice.     

Characterization of canine BCL6 cDNA and detection systems for its protein expression

BCL6 is known to be a key molecule in germinal center (GC) formation of lymph nodes, and its expression profiles have been implicated in the prognosis of diffuse large B-cell lymphoma in humans. The present study was carried out to characterize canine BCL6 cDNA and to indicate the technical methods for detection of the BCL6 protein in dog tissues. The deduced amino acid sequence of canine BCL6 showed close homology to that of human BCL6 (96.3%), especially in the zinc-finger motifs and POZ (poxvirus and zinc finger) domain with complete identity. Immunoblot analysis of a canine lymph node with an anti-human BCL6 monoclonal antibody revealed a band of 80 kDa. Immunohistochemical staining using the same antibody produced positive reactions in the cells exclusively localized in the GC of a canine lymph node. This study will be useful for the molecular classification of canine B-cell lymphomas with different prognoses.     

Co-situs application of controlled-release fertilizers to alleviate iron chlorosis of paddy rice grown in calcareous soil

Abstract

Co-situs is the placement with one application of a sufficient amount of controlled-release fertilizer for an entire growing season at any site, together with seeds or seedlings, without causing fertilizer salt injury. An experiment was conducted to find an efficient method for ameliorating Fe deficiency in two rice cultivars (cv. Tsukinohikari and cv. Sasanishiki) grown in a calcareous soil (pH 9.2, CaCO₃ 384 g kg^?1), which was poor in organic matter (0.1 g kg^?1) and available Fe (3.0 μg g^?1 soil). The field treatments consisted of co-situs application of the following fertilizers: 1) controlled-release NPK fertilizer (CRF-NPK) containing no micronutrients; 2) controlled-release NPK fertilizer containing micronutrients (CRF-M1); and 3) controlled-release NPK fertilizer containing micronutrients (CRF-M2). The main difference between CRF-M1 and CRF-M2 was that the former had larger granules than the latter. All the fertilizers were placed in contact with the roots of rice seedlings at transplanting time. Plants in the CRF-M1 and CRF-M2 treatments had similar lengths, number of stems, leaf age, and leaf color (SPAR value) during the cultivation period. By contrast, plants from the CRF-NPK treatments grew poorly, showed severe chlorosis symptoms of Fe deficiency, and all died on 30 DAT. Plants of both cultivars accumulated more macroand micronutrients with the CRF-M2 treatment than with the CRF-M1 treatment. The grain yield of cv. Tsukinohikari was 0.0, 1,910, and 2,160 kg ha^?1 for the CRF-NPK, CRF-M1, and CRF-M2 treatments, respectively, and 0.0, 2,490, and 2,860 kg ha^?1 for the same treatments for cv. Nihonbare. Chlorosis due to iron deficiency was successfully ameliorated and world-average grain yields were obtained with the co-sites application of both controlled-release fertilizers.