Insecticidal activity of Sikimi extract and its modulation of the GABAA receptor channel

Sikimi plant (also known as Japanese star anise), Illicium anisatium, is toxic to mammals. Extracts of Sikimi were studied for their insecticidal activity against the larvae of mosquito, Culex quinquefasciatus, and for their mechanism of action on ion channels. Crude methanol extract and its ethyl acetate-soluble fractions were insecticidally active, with EC₅₀ values of 63·0 μg ml^-1 and 43·7 μg ml^-1, respectively. The ethyl acetate-soluble fraction was perfused through the bathing solution and the current induced by a brief (10 ms) application of GABA by pressure ejection through pipette electrode was recorded by the whole-cell patch clamp technique. The extract suppressed GABA-induced currents irreversibly with an EC₅₀ value of 0·42 μg ml^-1. The time constant of current fitted to the single exponential function was shortened by the ethyl acetate-soluble fraction at concentrations ranging from 0·1 μg ml^-1 to 10 μg ml^-1 in a concentration-dependent manner. It was concluded that Sikimi extracts decreased the affinity of GABA for its binding site on the GABA receptor, thereby suppressing GABA-induced currents. © 1998 SCI     

Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals and prevalence of lukM-lukF-PV genes by bacteriophages in bovine isolates

Leukotoxin family genes in Staphylococcus aureus isolated from domestic animals were examined by polymerase chain reaction. LukS and lukF genes were detected in all 48 avian and 72 porcine isolates of S. aureus. LukE and lukD genes, located in a putative staphylococcal pathogenicity island (Sapln3/Saplm3), were recognized in 44 (91.7%) of 48 avian isolates, but these genes were not detected in porcine isolates. In 297 bovine isolates collected from mastitic cow's milk and bulk milk from dairy farms in two regions, lukM and lukF-PV(P83) genes in addition to lukS-lukF and lukE-lukD genes were detected in 100 (62.5%) of the 160 isolates from Ishikawa and in118 (86.1%) of the 137 isolates from Hokkaido. When the lysogeny of S. aureus bovine isolates was examined by treatment with mitomycin C, clearing of the culture due to cell lysis was observed in 34 (91.9%) of 37 lukM-lukF-PV(P83) genes--positive isolates. In addition, we isolated a novel lukM-lukF-PV(P83)-carrying (designated phiLukM), and revealed that the lukM-lukF-PV(P83) genes were located very close to an amidase gene on the temperate phage genomes. These results suggest horizontal transmission of lukM-lukF-PV(P83) genes by temperate bacteriophages in S. aureus of bovine origin.     

Flowering Date1, a major photoperiod sensitivity gene in adzuki bean,is a soybean floral repressor E1 ortholog

Adzuki bean is an important legume crop originating in temperate regions, with photoperiod in sensitivity being a key factor in its latitudinal adaptation. The Flowering Date1 (FD1) gene has a large effect on the photoperiodic response of flowering time, but the molecular basis for the effect of this locus is undetermined. The present study delimited the FD1 locus to a 17.1 kb sequence, containing a single gene, an E1 ortholog (VaE1). A comparison between Vigna angularis ‘Shumari’ (photoperiod insensitive) and ‘Acc2265’ (photoperiod sensitive) identified 29 insertions/deletions and 178 SNPs upstream of VaE1 in the FD1 locus. VaE1 expression in ‘Acc2265’ was greater under long-day than short-day conditions, whereas VaE1 expression in ‘Shumari’ was lower regardless of day length. These findings suggested that responsible gene of FD1 is a VaE1, which acts as a floral repressor by being upregulated in response to long-day conditions. The inability to upregulate VaE1 under long-day conditions was linked to its ability to flower under these conditions. These results provide greater understanding of the molecular control of a flowering date and clues enabling the breeding of adzuki bean at higher latitudes.     

Construction of a linkage map and identification of DNA markers linked to Fom-1, a gene conferring resistance to Fusarium oxysporum f.sp. melonis race 2 in melon

Resistance to Fusarium oxysporum f.sp. melonis race 2 is conferred by a single dominant gene, Fom-1 in melon. Here, we identified DNA markers tightly linked to Fom-1 that could be used for marker assisted selection in breeding programs. First, we developed 125 F₂ plants derived from the cross between melon lines P11 (fom-1fom-1) and MR-1 (Fom-1Fom-1). Using the F₂ population, we constructed a linkage map including 14 SSR markers which had not been mapped previously. Fom-1 was confirmed to be allocated to linkage group 7. Then, we identified four AFLP markers using bulked segregant analysis. The AFLP marker TAG/GCC-470 was completely linked to Fom-1 and other three markers were mapped near Fom-1. TAG/GCC-470 and TCG/GGT-400 were respectively converted to STS and CAPS markers. Usefulness of DNA markers was confirmed in the analysis with several melon cultivars and lines.     

Determination of the rate of 18F-labeled water movement to the leaf and its association with water relations in acid soil-tolerant rice varieties

The visual symptoms of damage by acid sulfate soil look quite similar to those due to soil moisture deficit. The present paper is to confirm if acid soil-tolerance is associated with drought tolerance in terms of leaf water potential, transpiration, and dry matter production. Seedlings of seven acid soil-tolerant (acid-tolerant), and three acid soil-intolerant varieties (acid-intolerant) were subjected to low pH conditions (pH 3.8) for 48 hours. The rate of water uptake was determined continuously by measuring radioactivity in the collar part (target area) of rice leaves exposed to ¹⁸F-labeled water (¹⁸F-water) using a Positron Emitting Tracer Imaging System (PETIS). The PETIS measurement showed that the radioactivity in the target position of leaves of acid-tolerant varieties increased faster than that of the intolerant varieties after the ¹⁸F-water was applied into the vial. In the plants subjected to low pH conditions, the transpiration rate (Tr) and leaf water potential (Ψ) decreased in both acid-tolerant and intolerant varieties. However, the extent of reduction was less in the acid-tolerant varieties than in the intolerant ones. Moreover, the dry matter production rate of the acid-tolerant varieties was significantly higher than that of the acid-intolerant ones in the low pH conditions. This study clearly shows that ¹⁸F-water uptake in a leaf was higher, and water relations were also better maintained in the acid-tolerant varieties compared with intolerant ones, resulting in higher growth rate in the acid-tolerant varieties, when plants were exposed to the acid solution conditions. We conclude that acid soil-tolerance is closely associated with leaf water relations in rice plants.     

Isolation and identification of indigestible pyroglutamyl peptides in an enzymatic hydrolysate of wheat gluten prepared on an industrial scale

An enzymatic hydrolysate of wheat gluten was further digested in vitro with porcine pepsin and pancreatin to obtain an indigestible peptide. Indigestible pyroglutamyl peptide was isolated from the digest by strong cation-exchange, size-exclusion, and reversed-phase chromatographies. The pyroglutamyl peptide was digested with pyroglutamate aminopeptidase, and the digest was reacted with phenyl isothiocyanate. The resultant phenylthiocarbamyl (PTC) peptides were purified by reversed-phase HPLC by using binary gradient elution with ammonium acetate buffer, pH 6.0, and acetonitrile. The PTC peptides were analyzed with an automatic peptide sequencer on the basis of the Edman degradation method with a modified program. Some pyroglutamyl peptides were also analyzed by fast-atom bombardment ionization mass spectrometry without the pyroglutamate amino peptidase digestion. Consequently, pyroGlu-Asn-Pro-Gln, pyroGlu-Gln-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gly-Gln-Gly-Gln, pyroGlu-Gln, pyroGlu-Gln-Pro, pyroGlu-Ile-Pro-Gln, pyroGlu-Ile-Pro, pyroGlu-Gln-Pro-Leu, pyroGlu-Gln-Phe-Pro-Gln, pyroGlu-Ser-Phe-Pro-Gln, pyroGlu-Phe-Pro-Gln, and pyroGlu-Gln-Pro-Pro-Phe-Ser were identified.     

Application of Pregerminated Brown Rice for Breadmaking

Pregerminated brown rice (PGBR) prepared by immersing in water was used for breadmaking, and effects on the dough properties and bread qualities were studied to compare with the ungerminated brown rice (BR). The substitution of BR or PGBR for wheat flour produced smaller specific volume in bread than in the control bread without BR or PGBR along with the increasing amount of substitution. However, the bread samples containing BR or PGBR suppressed staling during storage. The improving effect was especially obvious for substitutions of 10 and 20% PGBR as compared with BR. PGBR made viscous dough and retarded the staleness of bread compared with BR. γ‐Aminobutyric acid and oryzanol did not contain in the final BR and PGBR substituted bread, and phytic acid was decomposed ≈54 and 45% for 30% BR and 30% PGBR substituted breads, respectively. But ferulic acid was quite stable in the final baked product. As a result, substitution of PGBR for wheat flour improved the bread quality.     

Rhizosphere pH Profile of Rice Plant Influenced by Al Treatment

To study the mechanisms of Al tolerance in rice, we focused on the change of rhizosphere pH. The 4-d seedlings were treated with Al solution (0, 10, 50 mM) for 24 h. Then each sample was put on an agarose gel including bromocresol green, so that the color of the gel indicated pH change. During 2-h contact, the pH of rhizosphere was decreased gradually, especially for Al-treated samples, showing the specific pH profiles along the root axis. Pretreatment of sample plants with a decoupling reagent 2,4-dinitrophenol (DNP) or a plasma membrane H⁺-ATPase inhibitor Na₃VO₄ did not decrease rhizosphere pH. Therefore, it was suggested that the H⁺ secretion activity was involved with Al-tolerant mechanisms of rice.     

Hydrothermal Treatment of Date Palm Lignocellulose Residue for Organic Fertilizer Conversion: Effect on Cell Wall and Aerobic Degradation Rate

Date palm (Phoenix roebelenii) woodchips, a residue of palm tree plantations, was subjected to hydrothermal treatment (HTT) at mild reaction conditions (160°C < T < 220°C, 0.6 MPa < P < 2.4 MPa) for 30 min, and the effect of treatments on the cell wall (CW) solubilization and subsequent aerobic degradation rate (as CO production) was tested under controlled composting conditions during 63 days of incubation (38°C). The HTT at 160 and 180°C reaction temperatures notably solubilized hemicellulose, decreasing the fraction of this CW polymer from 34.1% in the untreated material, to 9.5 and 4.6% in the respective residues. However, treatment at 200 and 220°C reaction temperatures rapidly liquefied the lignin, which apparently went into solution with hemicellulose and appeared to stabilize a portion of this polysaccharides against hydrolysis. Consequently, the fraction of hemicellulose in 200 and 220°C – treated residues gradually increased; the respective values were 5.8 and 9.4%. The treatment temperature of 180°C was the most effective HTT temperature for subsequent aerobic degradation by solubilizing the largest portion of hemicellulose within the CW, which had two consequences: 1) it supplied additional readily bioavailable form of carbon, which in turn promoted rapid microbial activities in the early stage of decomposition; and 2) it created pores and cavities within the CW, which permitted rapid bacterial penetration and CW degradation. As a consequence, biodegradation of the residue treated under this reaction temperature proceeded rapidly and stability was reached within 21 days, compared to 63 days of continued degradation for the untreated CW. The enhanced biodegradability was also partially linked to the effect of 180°C treatment temperature on solubilization of amorphous cellulose and partial hydrolysis of lignin. Based on the results, the HTT system can successfully be used as a pretreatment step to accelerate the aerobic digestion rate of date palm residues for the production of organic fertilizers.