Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Antigens and antibodies in the ascitic fluid of mice infected with Fasciola gigantica

Fluke antigens were detected in the ascitic fluid of mice infected with Fasciola gigantica. The precipitating antibodies reactive against the antigens from metacercariae were detected as early as 2 weeks after the infection, and those against the adult and immature worms appeared 3 and 4 weeks later.     

Electroencephalogram of Japanese Black calves affected with cerebrocortical necrosis

Electroencephalogram (EEG) examinations were carried out on three Japanese Black calves (1, 2 and 14 week old) affected with cerebrocortical necrosis (CCN) in order to investigate the relationship between EEG abnormalities and encephalic lesions. Some neurologic signs such as circling, astasia, convulsion and opisthotonus appeared in two of the calves (case 1 and 3). The other calf (case 2), however, showed no neurologic signs. The EEGs obtained from the two calves (case 1 and 2), in the awake state, showed constant high amplitude (50-160 microV) and slow (1-4 Hz) activity (HASA). One of these (case 1) had necrosis mainly in the right hemisphere and showed a markedly asymmetric sharp wave. On the other hand, in case 3, the EEG showed diffuse lowered activity, and almost flat in some leads. A decrease in fast activity observed in common with all three animals. These findings suggest that one of the characteristic EEG patterns of CCN patient may be HASA and decreased fast activity. Histopathologic diagnoses of the calves were CCN in various degrees. The degrees of abnormality of the EEGs seemed to accord with the seriousness of the lesions. Because the EEG obtained from the calf having no obvious neurologic signs showed an abnormal pattern, it was considered that EEG examinations in the early stages of disorders are valuable to diagnose CCN.     

Biosynthesis of a syringyl 8-O-4′ neolignan in Eucommia ulmoides: formation of syringylglycerol-8-O-4′-(sinapyl alcohol) ether from sinapyl alcohol

To investigate the biosynthesis and stereochemistry of syringylglycerol-8-O-4′-(sinapyl alcohol) ether (SGSE), a syringyl 8-O-4′ neolignan, feeding experiments and enzyme assays using Eucommia ulmoides were carried out. Diastereoselective formation of erythro-SGSE was found. When [8-¹⁴C]sinapyl alcohol was administered to excised shoots of E. ulmoides, ¹⁴C was incorporated into free SGSE and SGSE glucosides. In stems, incorporation into (+)-erythro-[¹⁴C]SGSE (0.037%) with 9.1% enantiomeric excess (% e.e.) was found; incorporation into the threo isomer was not detectable. Erythro-[¹⁴C]SGSE glucosides (0.047%) dominated over threo forms (0.007%) with 74.0% diastereomeric excess (% d.e.); both diastereomers were levorotatory with 32.0% e.e. and 18.3% e.e., respectively. In leaves, higher incorporation into (−)-erythro-[¹⁴C]SGSE (0.500%, 15.9% e.e.) than into the threo isomer (0.206%, 7.4% e.e.) was observed (41.6% d.e.). (−)-Erythro-[¹⁴C]SGSE glucosides (1.692%, 25.0% e.e.) were produced at higher rates than threo isomers (0.177%, 16.4% e.e.) with 81.0% d.e. In incubations of a mixture of [8-¹⁴C]sinapyl and [8-¹⁴C]coniferyl alcohols with an insoluble enzyme preparation from stems of E. ulmoides, erythro-SGSE was preferentially produced. The highest % d.e. (82.8) was observed at 60 min with the (+)-erythro isomer (21.4% e.e.) and the (−)-threo form (4.3% e.e.).Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002, and the 47th Lignin Symposium, Fukuoka, October 2002     

Atmospheric lead and cadmium deposition within forests in the Kanto district, Japan

Atmospheric lead and cadmium deposition in bulk precipitation and throughfall was investigated at four forests in the Kanto district, Japan, to assess the impact of human activities on the environmental health of forests. Annual lead and cadmium depositions in bulk precipitation ranged from 8.9 to 25.7 g ha⁻¹ year⁻¹ and from 0.77 to 1.30 g ha⁻¹ year⁻¹, respectively. Lead and cadmium deposition increased in the summer at every forest due to large amounts of rainfall. At one of the forests, the depositions were also high in the winter due to heavy snowfall. These depositions were similar to recent depositions observed at other rural and urban sites in Japan and several forests in Europe and North America after 1990. These results indicate that although anthropogenic lead and cadmium are deposited at these rates over wide areas, depositions are still higher than in remote areas.     

Assessing raindrop impact energy at the forest floor in a mature Japanese cypress plantation using continuous raindrop-sizing instruments

The raindrop size distribution of throughfall and open rainfall was monitored continuously during a rainfall event using laser raindrop-sizing instruments (LD gauges), in order to calculate the raindrop impact energy in a plantation of mature Japanese cypress (Chamaecyparis obtusa), where surface erosion at the forest floor had been a problem. Data from two rainfall events were analyzed. The LD gauges recorded qualitative raindrop size distribution, and the capture rate during each rainfall event was used to manipulate raindrop data quantitatively. Throughfall and open rainfall comparisons revealed several important differences. First, throughfall raindrops were fewer in number and larger in size than open rainfall drops. In one rainfall event, for example, throughfall raindrops were less than one-fifth as frequent as open rainfall raindrops; in addition, the maximum throughfall raindrop diameter was 6.35mm compared to 3.31mm for open rainfall raindrops. Second, throughfall raindrops that were larger than the largest open rainfall raindrops comprised 63.8% of the throughfall precipitation by volume. Third, total raindrop impact energy from throughfall was over twice that of open rainfall. Moreover, comparison of throughfall events implied that throughfall raindrops did not always have a uniform distribution between different events or among different periods of time in one rainfall event, in contrast to findings in previous studies which showed that throughfall raindrops had a uniform size distribution independent of rainfall intensity. It is possible that an abrupt transition of throughfall intensity from low to high changes the distribution of throughfall raindrops.