A binding protein for fungal signal molecules in the cell wall of Pisum sativum

In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous signal molecules.     

Identification of genes expressed during spore germination of Mycosphaerella pinodes

Mycosphaerella blight, caused by Mycosphaerella pinodes, is one of the major diseases of cultivated pea (Pisum sativum L.). To isolate the genes that are up- and down-regulated during spore germination, suppression subtraction hybridization (SSH) was performed between ungerminated and germinated spores. The 232 and 128 clones from forward and reverse libraries, respectively, were collected, sequenced, and analyzed with a BLASTX homology search. About 95% of the 32 selected clones were expressed during spore germination on a paper sheet and during infection of pea leaves. We discuss the applicability of the SSH libraries for analyzing M. pinodes genes involved in the early stage of infection.     

Computer tomography diagnosis of meningoencephalocele in a calf

A 24-day-old female Holstein calf had a soft, painless fluctuating swelling on the median plane in the frontal region, but did not show any clinical symptoms including neurological signs. Computer tomography (CT) distinctly showed the cyst filled with fluid and part of the encephalon. Hence, this swelling was diagnosed as meningoencephalocele, but not meningocele. The meningoencephalocele was successfully repaired surgically. Meningoencephalocele can thus be easily recognised by CT in a calf.     

Estimation of annual suspended sediment yield from a Japanese cypress (Chamaecyparis obtusa) plantation considering antecedent rainfalls

Japanese cypress (Chamaecyparis obtusa) is one of the most common plantation species in Japan. In most Japanese cypress plantations, closure of the forest canopy hinders understory growth because of insufficient management practices. Thus leaf litter barely covers the soil surface. Such plantations are vulnerable to surface erosion triggered by rainfalls, and could yield large amounts of suspended sediment (SS). However, few studies have investigated the annual SS yield. This study aimed to develop a modified model of SS yields, and to accurately estimate and characterize the annual SS yield from a Japanese cypress plantation. For this, hydrological surveys were conducted for 5 years, and stream water was sampled weekly as well as sequentially at 15–60 min intervals in each of 14 rain events in a small forested watershed of Japanese cypress. The root mean square error (RMSE) and the annual SS yield estimates were obtained using three different equations: the rating curve derived from instantaneous SS concentration versus stream discharge relationship (SSC − Q equation); the rating curve derived from cumulative SS yield versus cumulative specific discharge relationship (∑SS − ∑Q equation); and the multiple regression of cumulative SS yield expressed as functions of cumulative specific discharge and antecedent rainfalls (∑SS − ∑Q × AR equation). The ∑SS − ∑Q × AR equation is a modified model considering the effects of antecedent rain history on the SS yield. The parameters in each equation were determined by a nonlinear least square method. Of the three equations, the RMSE was the highest using the SSC − Q equation. Although the ∑SS − ∑Q equation and the ∑SS − ∑Q × AR equation gave similar RMSE values, the SS yields extrapolated by the ∑SS − ∑Q equation would be overestimated. Thus, the 5-year average of annual SS yield estimated by the ∑SS − ∑Q equation was three times larger than that estimated by the ∑SS − ∑Q × AR equation. These results indicate that the ∑SS − ∑Q × AR equation more accurately estimates the annual SS yield, compared with the other two equations. The results also suggest that antecedent rain history, as well as stream discharge, should be considered for appropriately estimating the annual SS yield. The annual SS yield estimated by the ∑SS − ∑Q × AR equation was the largest among 25 undisturbed forested watersheds, except for two watersheds dominated by sandstone. Our results suggest that the annual SS yield is greater from a Japanese cypress plantation under current management practices than from other undisturbed forests.     

Effects of dietary phytol on tissue accumulation of phytanic acid and pristanic acid and on the tissue lipid profiles in mice

Recent in vitro evidence suggests that the phytol‐derived fatty acids, phytanic acid (PA) and pristanic acid (PrA), are components of animal products with the potential to cause both beneficial and harmful effects on human health. In this study, we investigated the in vivo tissue accumulation of PA and PrA and the changes in tissue lipid profiles, using mice fed a phytol‐containing diet. After 4 weeks of treatment with a diet containing 1.0% phytol, plasma, adipose tissue, liver, and brain were collected and their lipid profiles were biochemically and gas‐chromatographically determined. Dietary phytol caused PA and PrA accumulation in the adipose tissue and liver but not in the brain, and reduced plasma and liver triacylglycerol levels. Phytol intake also decreased the fatty acid concentrations in the adipose tissue, especially polyunsaturated fatty acids such as linoleic acid, but increased the concentrations of these fatty acids in the liver. However, dietary phytol had a low impact on the brain lipid profile. This study suggests that dietary phytol intake caused accumulation of PA and PrA and modified lipid profiles in the adipose tissue and liver, but that the brain is an insusceptible tissue to dietary phytol‐induced changes.     

Use of an automatic cell-counting system with LED illumination for enumeration of marine bacteria

ABSTRACT:   The concentration of aquatic bacteria is basic information required to evaluate the status of environments and to assess bacterial contribution to material cycles. However, the standard direct counting method using epifluorescence microscopy (EFM) is tedious and there is variation in the counts among workers. Here an automatic counting system that consists of Bioplorer (BP) and image analysis has been applied to marine bacteria. BP is composed of a light-emitting diode (LED) illuminant, an optical unit, a driving stage and a charge-coupled device camera. In combination with fluorescent labeling and simplified membrane filtration, bacteria are enumerated automatically. The reproducibility, sensitivity and accuracy of the system were tested for natural marine bacteria, in comparison with EFM and flow cytometry (FCM). The counts obtained by BP showed good correlation with those obtained by EFM and FCM methods. The counts were significantly higher in inshore and oceanic samples, indicating high sensitivity with low background noise. Considering its reproducibility, objectivity, ease of use and compact size, BP can be used as a routine tool for counting aquatic bacteria in substitution for EFM or FCM.