Evaluation of seed yield and seed yield components in red–yellow (Pisum fulvum) and Ethiopian (Pisum abyssinicum) peas

Red–yellow (Pisum fulvum Sibth. et Sm.) and Ethiopian (Pisum abyssinicum A. Br.) peas have become of increasing interest to breeders in the last decade, as they have been found to be partially or completely tolerant to various biotic stresses, such as to attack by pea weevil, mildew blight or rust. A trial was carried out at the Institute of Field and Vegetable Crops at Rimski ?an?evi from 2005 to 2007, with 13 accessions each of red–yellow and Ethiopian peas sown at 100 viable seeds m^?2, in plots of 0.5 m². Seed yield per plant in red–yellow pea was significantly and positively correlated with seed number per plant (r = 0.881**), pod number per plant (r = 0.839**) and number of fertile nodes (r = 0.820**). The highest positive correlation among the agronomic characteristics in Ethiopian pea was between number of fertile nodes and number of pods (r = 0.937**). Seed yield in Ethiopian pea was highly significantly correlated with number of seeds (r = 0.807**), pods (r = 0.692*), and fertile nodes (r = 0.638*). The results suggest that plant morphology of the progenies between red–yellow or Ethiopian peas and grain-type common pea could not differ significantly from that of individual parents. That means that it could be possible to develop hybrid lines that could keep the desirable morphological traits of grain-type common pea, such as lodging-tolerance and high seed yields, and to make an introgression of a specific resistance from wild pea taxa, especially by back-crosses with the former. However, one must always be aware of unpredicted outcomes as a result of rather different genetic basis of individual seed yield components.     

Effects of astaxanthin‐enriched yeast on mucosal IgA induction in the jejunum and ileum of weanling mice

The present study was conducted to clarify the effects of astaxanthin‐enriched yeast on the concentration of immunoglobulin A (IgA), the numbers of IgA antibody‐secreting cells (ASC) and the messenger RNA (mRNA) expression of IgA C‐region in the jejunum and ileum of weanling mice. Weanling mice were fed rodent feed or astaxanthin‐enriched yeast‐supplemented rodent feed for 7, 14 or 21 days. Supplemental astaxanthin‐enriched yeast increased the numbers of IgA ASC in the jejunum and ileum after 7, 14 and 21 days of treatment. Supplemental astaxanthin‐enriched yeast increased IgA concentrations in the jejunum after 21 days of treatment, but IgA concentrations in the ileum were not affected by the treatment. The mRNA expressions of IgA C‐region in the jejunum after 14 and 21 days of treatment and the ileum after 14 days of treatment were enhanced by supplementation of astaxanthin‐enriched yeast. These results indicate that supplementation of astaxanthin‐enriched yeast is effective to enhance the numbers of IgA ASC in the jejunum and ileum and IgA concentrations in the ileum of weanling mice.     

Effects of high potassium chloride supplementation on water intake and bodyweight gains in pregnant and lactating mice

Thirty‐one ICR pregnant mice were assigned to a control or a potassium chloride (KCl) diet group to clarify the effects of KCl supplementation on water intake, bodyweight gains and serum components in pregnant and lactating mice, and 5% KCl was supplemented in KCl diets from 6.5 days post coitus to 1 or 14 days after parturition. Feed intake was not affected by treatment, but supplemental KCl decreased bodyweight gains of lactating mice and their neonatal mice. Water intake and urine volume of KCl supplemented mice were significantly higher than those of control mice during pregnancy and supplemental KCl decreased serum urea N in pregnant mice. Supplemental KCl increased water intake drastically in lactating mice immediately after parturition and increased serum K at 14 days after parturition. Histological alteration using hematoxylin–eosin was not found in the kidney of each mouse at 1 or 14 days after parturition. These results indicate that high KCl supplementation accelerates water intake in lactating mice and prevents bodyweight gains of maternal and neonatal mice during lactation.     

Wood friction characteristics during exposure to high pressure: influence of wood/metal tool surface finishing conditions

Friction that arises during processing for the deformation of wood under relatively high pressure levels (ca. >1 MPa) is an important factor to be taken into account when wood is processed. However, few studies on such friction have been published. The present study was undertaken to evaluate the influence of surface finishing conditions on the nominal friction coefficient (μ) of the wood and metal tool surfaces. Sticking friction was likely to arise on a relatively coarse metal surface, and the type of metal tool surface finishing was found to have large impact on the friction mechanism. The friction characteristics during exposure to high pressure seem to be affected not only by the interface contact characteristics, but also by the deformation characteristics of wood during compressive load or measurement. The value of μ on water-saturated wood was equal or higher than that on dry wood, which suggests that the contact characteristics between these two types of wood are significantly different. The water content in wood was shown to affect both the interface contact and deformation characteristics of wood. The value of μ was not significantly affected by the wood surface finishing conditions; however, changes in μ during sliding differed slightly, depending on the finishing conditions.     

GtxA from Gallibacterium anatis, a cytolytic RTX-toxin with a novel domain organisation

Gallibacterium anatis is a pathogen in chickens and other avian species where it is a significant cause of salpingitis and peritonitis. We found that bacterial cells and cell-free, filter-sterilised culture supernatant from the haemolytic G. anatis biovar haemolytica were highly cytotoxic towards avian-derived macrophage-like cells (HD11). We obtained the genome sequence of G. anatis 12656-12 and used a rational approach to identify a gene predicted to encode a 2026 amino acid RTX-toxin, which we named GtxA (Gallibacterium toxin). The construction of a gtxA knock-out mutant showed gtxA to be responsible for G. anatis’ haemolytic and leukotoxic activity. In addition, Escherichia coli expressing gtxA and an adjacent acyltransferase, gtxC, became cytolytic. GtxA was expressed during in vitro growth and was localised in the extracellular protein fraction in a growth phase dependent manner. GtxA had an unusual modular structure; the C-terminal 1000 amino acids of GtxA were homologous to the classical pore-forming RTX-toxins in other members of Pasteurellaceae. In contrast, the N-terminal approximately 950 amino acids had few significant matches in sequence databases. Expression of truncated GtxA proteins demonstrated that the C-terminal RTX-domain had a lower haemolytic activity than the full-length toxin, indicating that the N-terminal domain was required for maximal haemolytic activity. Cytotoxicity towards HD11 cells was not detected with the C-terminal alone, suggesting that the N-terminal domain plays a critical role for the leukotoxicity.     

Improved method for in situ biolistic transformation to analyze barley–powdery mildew interactions

Although the use of stable transformants is indispensable to elucidate mechanisms underlying molecular plant–pathogen interactions, this approach remains difficult to apply to crops. Alternatively, biolistic transformation has often been used as a transient expression method in various plants. In this study, we developed a method for in situ biolistic transformation without separating leaves from barley seedlings by using a hand-held particle bombardment system because unwounded leaves are preferable for analyzing interactions between barley and Blumeria graminis f. sp. hordei which requires healthy living cells. As a result, we found that the infection rate in intact leaves was higher than in separated leaves and that the transformation efficiencies in leaves were higher when plants were grown in vermiculite rather than in culture soil. Furthermore, we determined the appropriate inoculation time after bombardment to analyze the incompatible interaction and successfully monitored the gradual occurrence of cell death over time. Our system was suitable for relatively long-term follow-up analysis of the fate of each single cell during plant–pathogen interactions.     

cDNA cloning and expression analysis of two heat shock protein genes,Hsp90 and Hsp60, from a sterile Ulva pertusa (Ulvales,Chlorophyta)

Molecular chaperones are synthesized and accumulated under a variety of unfavorable conditions in all organisms. Heat shock protein 90 (Hsp90) and Hsp60, which are classified into the major classes of molecular chaperones, play important roles in cellular stress responses. In this study, we characterized sterile Ulva pertusa Hsp90 (UpHsp90) and UpHsp60 genes which may be involved in tolerance to thermal and heavy metal stresses in this alga. The UpHsp90 cDNA consisting of 2,118 nucleotides encoded a polypeptide of 705 amino acids (AA). On the other hand, the UpHsp60 cDNA consisting of 1,722 nucleotides encoded a protein whose predicted length was 573 AA. The AA sequence alignment and phylogenetic analyses showed that the UpHsp90 and UpHsp60 proteins were more similar to cytoplasmic Hsp90s and mitochondrial Hsp60s, respectively, than to other types of the respective Hsps. Southern blot analysis indicated that the sterile U. pertusa genome had at least two cytoplasmic Hsp90-encoding genes and two mitochondrial Hsp60-encoding genes. The UpHsp90 and UpHsp60 mRNA levels were significantly affected by diurnal and temperature changes, and slightly affected by exposure to heavy metals. These results suggest that UpHsp90 and UpHsp60 genes play particularly important roles in adaptation to diurnal and temperature changes.     

Purification and functional characterization of complement C3 and a novel zymosan-binding protein in tilapia serum

Zymosan, a yeast cell wall preparation that binds activated forms of complement C3, is a useful model target to activate the complement system. In our trial to analyze C3 diversity in Nile tilapia at the protein level using zymosan, we found that a novel 240-kDa serum protein (ZBP-240) also bound to zymosan in addition to C3-derived fragments. In the present study, we aimed to characterize tilapia C3 and ZBP-240, focusing on their immune-related functions. Four distinct C3 isoforms were purified from tilapia serum and shown to possess an intrachain thioester bond. ZBP-240 was also isolated from tilapia serum and examined for its binding properties to various microbial targets. As a result, ZBP-240 showed a wide spectrum of binding to Gram-positive and Gram-negative bacteria and yeasts. Amino acid sequence analysis of CNBr fragments of ZBP-240 suggested that this is a novel protein with no homologous sequence in protein databases. It was also suggested that the binding of ZBP-240 to microbes largely depends on hydrophobic interactions in a divalent-cation-independent manner, and that there may be a divalent-cation-dependent factor that enhances the binding of ZBP-240 in tilapia serum. Interestingly, ZBP-240 showed opsonic activity for tilapia kidney phagocytes at a level comparable to that of C3, implying that ZBP-240 is a novel teleost opsonic serum protein.     

Enthalpy relaxation behavior of dry wood detected by temperature-modulated differential scanning calorimetry

Enthalpy relaxation of dry wood has been investigated by temperature-modulated differential scanning calorimetry. The reversing and non-reversing heat flow changes revealed that enthalpy relaxation occurred in dry wood, which did not exhibit any clear glass transitions. This enthalpy relaxation behavior seemed to differ significantly from those of previously reported isolated lignins, which implies that the microstructure of dry wood possesses a rigid amorphous state derived from interactions among wood components. The observed enthalpy relaxation is considered to be related to other components besides lignin, and the time-dependent physical properties due to unstable states or physical aging of wood originate not only from lignin but also from other components, such as cellulose and hemicellulose and the interactions between them.