Method for rapid detection of the PvCesA3 gene allele conferring resistance to mandipropamid, a carboxylic acid amide fungicide, in Plasmopara viticola populations

BACKGROUND: The occurrence of carboxylic acid amide (CAA)‐fungicide‐resistant Plasmopara viticola populations is becoming a serious problem in the control of grapevine downy mildew worldwide. RESULTS: The authors have developed a method, which utilises PCR‐RFLP, for the rapid detection of resistance to the CAA fungicide mandipropamid in P. viticola populations. With this method, a glycine‐to‐serine substitution at codon 1105 of the cellulose synthase gene PvCesA3 of CAA‐fungicide‐resistant P. viticola was easily detected, although no resistant P. viticola was detected from 398 isolates in Japan. CONCLUSION: It is proposed that the PCR‐RFLP method is a reliable tool for the rapid detection of CAA‐fungicide‐resistant P. viticola isolates. Only 4 h was required from the sampling of symptoms to the phenotyping of fungicide resistance. Copyright © 2011 Society of Chemical Industry     

Spore-breaking capabilities of collembolans and their feeding habitat within sporocarps

To examine the relationship between the spore-breaking capability of collembolans and their localization within fungal sporocarps we investigated the proportion of broken spores in the feces of four collembolan species of the genus Hypogastrura using the spores of Hypsizygus marmoreus and analyzed the spatial distribution of the collembolan species within sporocarps as an index of their feeding habits. The collembolan species with a higher capability to break spores were observed more frequently from the spore-bearing surfaces of sporocarps. The collembolan species with a lower capability to break spores were found more frequently from the non-spore-bearing interiors of the sporocarps. These results suggest that the relative value of spores in the diet varied with the capability of collembolans to break spores.     

Relationship between the glass transition of soy protein and molecular structure

The change in molecular structure of the soy protein samples as a result of the microbial transglutaminase treatment was studied using solid-state (13)C NMR spectroscopy and circular dichroism (CD), and the relation to the glass transition temperature (T(g)) was examined. From NMR measurements, the structure of the local region of the C(alpha) methine was observed to change, and the region had relatively high mobility. From CD measurements, the structural change seemed to be caused by the change in the secondary structure (disintegration of the beta-structure). By comparison with the T(g) of another protein, the state of the secondary structure of a protein was suggested to be a key in determining its T(g).     

Relationship between antibacterial activity of (+)-catechin derivatives and their interaction with a model membrane

(+)-Catechin derivatives with different alkyl chain lengths were synthesized from (+)-catechin and various straight chain alkylaldehydes in the presence of methyl mercaptan, and their antibacterial activities against Gram-positive bacteria were evaluated. The antibacterial activity increased markedly with elongation of the alkyl chain lengths of the derivatives and reached a maximum at a chain of four to seven carbons. Subsequently, interaction of the (+)-catechin derivatives with a model membrane using liposome was investigated. The derivatives with a chain of three carbons or more were found to have very strong affinity for the membrane. The injury action of the derivatives against the membrane was examined with liposome in which calcein was enclosed as a fluorescent indicator. The leakage was observed in the derivatives with chain lengths of four carbons or more. Particularly the derivatives with chains longer than five carbons are considered to destroy the liposome membrane judging from the degree of the fluorescent leakage. These results implied that the lipophilicity and disrupting ability of the (+)-catechin derivatives to the liposome membrane participate in their antibacterial activity.     

Determination of tri-n-butyltin and di-n-butyltin compounds in fish by gas chromatography with flame photometric detection

An analytical method is described for the simultaneous quantitative determination of tri-n-butyltin and di-n-butyltin compounds in fish. The sample was extracted with 0.5N HCl-methanol, and the methanol solution was extracted with hexane. The extract was purified by gel permeation chromatography and treated with Grignard reagent to yield the methyl derivatives, which were determined by gas chromatography with flame photometric detection operated in the tin mode (610 nm). Recoveries of tri-n-butyltin chloride (Bu3SnCl) and di-n-butyltin dichloride (Bu2SnCl2) spiked to fish at the levels of 0.2 and 1.0 ppm ranged from 80 to 105%. Detection limits were 0.02 micrograms/g for both compounds. Tri-n-butyltin compounds equivalent to Bu3SnCl levels of 0.07-2.0 ppm and di-n-butyltin compounds equivalent to Bu2SnCl2 levels of 0.02-0.11 ppm were found in reared yellowtails, and these values showed good agreement with the results from gas chromatographic-mass spectrometric analysis.     

Pressure-induced denaturation of monomer beta-lactoglobulin is partially irreversible: comparison of monomer form (highly acidic pH) with dimer form (neutral pH)

This study was conducted to assess the effect of high hydrostatic pressure on monomer beta-lactoglobulin (BLg) at acid pH by fluorescence spectroscopy under pressure and by circular dichroism (CD) and (1)H NMR spectroscopies after release of pressure. The intrinsic (tryptophan) fluorescence measurement and the study of 8-anilinonaphthalene-1-sulfonate (ANS) binding to BLg indicated that at pH 2.0 the recovery of center of spectral mass or ANS fluorescence was almost complete upon pressure release. No difference in (1)H NMR spectra was observed between pressurized and unpressurized BLg. In addition, NMR detection of the H/D exchange of aromatic protein indicated that the conformation of the vicinity of tryptophan residues could be refolded almost completely after release of pressure. These results seemingly confirm that the pressure-induced denaturation of BLg at pH 2.0 is reversible. However, cis-parinaric acid binding ability of pressurized BLg was largely lost, although its retinol binding ability was the same as its unpressurized one. Furthermore, CD spectra of the far-UV region and 2D NMR spectra evidently revealed the difference in the conformation of the molecule between unpressurized and pressurized BLg. These results are interpreted as an existence of partially fragile structure in the BLg molecule by high pressure.     

Nitrogen Retention in Japanese Cedar Stands in Northern Honshu, with High Nitrogen Deposition

High nitrogen, especially ammonium, input has been observed in Schichinohe, Aomori Prefecture, northeastern Japan. A monitoring study on precipitation, throughfall, and stream water has been carried out to estimate the stage of nitrogen saturation since 1996. Fifty-two to 70% of nitrogen input in throughfall was retained in forest ecosystems. Nitrate concentration in stream water tended to decrease throughout the study. There was no symptom of nitrogen saturation at Japanese cedar stands in Shichinohe, although high nitrogen input in open bulk has been observed. Ammonium (NH₄ ⁺) was retained in the canopy. The ratio of NH₄ ⁺ input in throughfall to that by open bulk was 0.40 – 0.47. Total inorganic nitrogen input under the canopy amounted 0.68 – 0.72 kmolc ha^?1 yr^?1 (9.6 – 10.0 kg N ha^?1 yr^?1). Our results suggests that atmospheric nitrogen input has benefitted the three growth.     

Muscular architecture and VIP-like immunoreactive nerves in the gastroduodenal junction of the chicken

The muscular architecture and the distribution of nerves with vasoactive intestinal polypeptide-like (VIP-like) immunoreactivity were examined in the gastroduodenal junction of the chicken. Both the outer and the inner muscle layers (tunica muscularis) and the muscularis mucosae were examined. The inner muscle layer was divided into two parts, namely its inner and outer parts. The outer muscle layer was not present in the pyloric part but was found in the proventriculus and the duodenum. The muscle strands of the outer muscle layer ran longitudinally but were absent at the isthmus and the pylorus. Most of the outer part of the inner muscle layer was located in or close to the pyloric part. The muscle strands in the outer part connected the pyloric part with other regions, namely the craniodorsal muscle, the tendinous centres and the isthmus. The inner part of the inner muscle layer was widely distributed in the stomach. In the pyloric part, the muscle strands of this layer followed a circular or spiral course. The muscularis mucosae was not present in the caudodorsal and cranioventral muscles of the gizzard. In addition, VIP-like immunoreactive nerve fibres were predominantly distributed in the inner part of inner muscle layer of the pyloric part. These results suggest that the ventriculopyloric orifice and the pyloric part itself constrict with a sphincter-like action.Abbreviations VIP vasoactive intestinal polypeptide     

Muscle Representation within the Hypoglossal Nucleus of the Chicken Studied by Means of Horseradish Peroxidase

The distribution in the chicken of motoneurons innervating the hyolingual muscles, i.e. the Mm. hyoglossus rostralis (HR), hyoglossus obliquus (HO), ceratoglossus (CG), interceratobranchialis (CB). stylohyoideus (YH), serpihyoideus (PH) and cricohyoideus (CR), and the laryngotracheal muscles, comprising the Mm. tracheolateralis (TL), cleido-hyoideus (CL) and sternotrachealis (ST), was examined by retrograde transport of horseradish peroxidase conjugated with wheat-germ agglutinin. Labelled motoneurons are only found in the hypoglossal nucleus. The rostrocaudal distributions of motoneurons projecting to hyolingual muscles are restricted in the hypoglossal nucleus cranial to the obex, and those projecting to laryngotracheal muscles are distributed in the more caudal part of hypoglossal nucleus. Detailed analysis of the data showed that the most rostrally positioned motoneurons in the hypoglossal nucleus supplied to the PH, followed by the CG, CB. HR, YH, HO, CR, TL, CL and ST in that order, overlapping each other. In the hypoglossal nucleus motoneurons innervating the PH and YH have the smallest perikarya. Of the motoneurons in the hypoglossal nucleus, those supplying the laryngotracheal muscles (CL and TL) have the largest perikarya. Motoneurons innervating the other muscles are intermediate in size.