Localization and responsiveness of a cowpea apyrase VsNTPase1 to phytopathogenic microorganisms

Apyrases (NTPases) are associated with both compatible and incompatible interactions between plants and microorganisms. Previously we reported that the ATPase activities of cell-wall-bound apyrases of several leguminous plants, such as pea, cowpea, soybean, and kidney bean, were enhanced by a glycoprotein elicitor and were inhibited in a species-specific manner by mucin-type glycopeptide suppressors secreted from a pea pathogenic fungus, Mycosphaerella pinodes. In this study, we isolated two apyrase genes, VsNTPase1 and VsNTPase2, from a cDNA library of Vigna sinensis Endl. cv. Sanjakusasage. Based on phylogenetic analysis, VsNTPase1 may belong to a group that responds to environmental stimuli. In a transient assay using DNA bombardment, a fusion protein of green fluorescent protein (GFP) and the N-terminal putative signal sequence of VsNTPase1 was distributed in the nucleus, cytoplasm (cytoskeletal structure), and cell wall. On the other hand, a fusion protein of GFP and the N-terminal putative VsNTPase2-signal sequence was localized in the cytoplasm, especially in small particles (perhaps mitochondria). A recombinant VsNTPase1 expressed in Spodoptera frugiperda 21 cells responded directly to signal molecules from several phytopathogenic microorganisms. Here, we discuss the role of apyrases in recognizing and responding to exogenous signals. The nucleotide sequences of VsNTPase1 and VsNTPase2 in this article have been submitted to DDBJ as accession numbers AB196769 and AB196770, respectively.     

Searching by Differential Display for New Genes Implicated in Induced Disease Resistance of Cucumber Treated with Acibenzolar-S-methyl

Acibenzolar-S-methyl (ASM) is a chemical activator of systemic disease resistance in plants. In this study, we used differential display to identify ASM-inducible defense response genes involved in induced disease resistance. As a result, we cloned three ASM-inducible genes from cucumber, encoding a chitinase, a putative protein disulfide isomerase and a putative mitochondrial-protein-like protein. Expression of these genes was induced within 24 hr after treatment of cucumber leaves with ASM. These results suggest that differential display is a useful tool for understanding the mode of action of ASM and defense responses. Received 6 September 2000/ Accepted in revised form 11 April 2001     

Limitations in the exploitation of N-phenylcarbamates and N-phenylformamidoximes to control benzimidazole-resistant Venturia nashicola on Japanese pear

The efficacy of N -phenylcarbamates and N -phenylformamidoximes against benzimidazole-resistant strains of Venturia nashicola was tested experimentally in Japanese pear orchards. Both compounds gave effective control when applied in an orchard where highly benzimidazole-resistant strains were predominant. However, unsatisfactory control was observed in an orchard where the fungal population was dominated by fungal strains with intermediate or weak benzimidazole resistance. Strains resistant to N -phenylcarbamates and N -phenylformamidoximes were widely distributed in Japanese pear orchards, suggesting that effective use of these compounds to control benzimidazole-resistant strains of V. nashicola would not be practicable. Increased sensitivity to N -phenylformamidoximes in highly benzimidazole-resistant isolates was shown to be controlled by a single chromosomal gene, but progenies that were highly resistant to both a benzimidazole fungicide and a N -phenylformamidoximes appeared in crosses between parents resistant to each fungicide alone. Similar strains were also found in pear orchards, and this'double resistance'was shown to be heritable.     

Iron deficiency causes zinc excess in Zea mays

Iron deficiency stress causes a severe reduction in plant growth. Although Fe deficiency causes an imbalance in divalent heavy metal nutrients, the mechanisms underlying the growth reduction caused by this imbalance remain unclear. We investigated Zn uptake and accumulation in maize under Fe-deficient conditions. Under Fe-deficient conditions, Zn uptake was 15-fold higher and Zn accumulation was 16-fold higher than that under normal nutrient conditions. The Zn content of maize leaves under Fe-deficient conditions was >0.4 mg g⁻¹ dry weight, which was higher than the content of plants grown in a nutrient solution containing 50 µM ZnCl₂. Plant growth under conditions of both Fe and Zn deficiency was significantly higher than that under only Fe-deficient conditions. Moreover, Fe deficiency increased the thiol content of the plant. These results indicate that Fe deficiency causes excess uptake and accumulation of Zn, and that the stress resulting from the Zn overload accelerates growth reduction in maize.     

Isolation of C11 Cyclopentenones from Two Didemnid Species,Lissoclinum sp. and Diplosoma sp.

A series of new C₁₁ cyclopentenones 1–7 was isolated, together with four known metabolites 9/10, 12 and 13, from the extract of the didemnid ascidian Lissoclinum sp. The other didemnid ascidian Diplosoma sp. contained didemnenones 1, 2 and 5, and five known metabolites 8–12. The structures of 1–7 were elucidated by spectroscopic analyses. Cytotoxicity of the isolated compounds was evaluated against three human cancer cell lines (HCT116, A431 and A549).     

Preference for safflower oil in rats exposed to a cold environment under free-feeding conditions

There are several benefits to a high-fat diet for animals exposed to cold, including improved tolerance to severe cold conditions and increased survival rates in cold environments. It is therefore of interest to examine whether animals exposed to cold will selectively consume lipids. We examined the intake of safflower oil (SO) by rats exposed to cold (4 +/- 2 degrees C) under a feeding condition in which the rats were given free access to SO. Rats exposed to cold consumed more SO than those housed at 25 +/- 2 degrees C. This finding suggests that rats prefer SO in a cold environment. There was no significant difference in the ratio of calories of SO ingested to that of matter (standard laboratory chow plus SO) ingested between rats exposed to cold and those at 25 +/- 2 degrees C. The high SO intake also affected cold tolerance and metabolite kinetics in the rats. Factors that affected the SO intake of rats exposed to cold are also discussed.     

Rhabdomyosarcoma in a ferret (Mustela putorius furo)

A 5-year-old spayed male ferret showed a subcutaneous mass in the right lateral thoracic wall. Microscopic examination revealed that the neoplasm had proliferated in the subcutis with infiltration into the surrounding tissues. A packed bundle of large polymorphic neoplastic cells, containing abundant eosinophilic cytoplasm and a round to ovoid, occasionally bizarre nucleus, were arranged interwoven. The neoplasm had metastasized to the right axillary lymph node. The neoplastic cells were intensively positive for vimentin, desmin and myoglobin. Skeletal muscle type creatine phosphokinase-positive granules were detected in the cytoplasm. Ultrastructurally, various amounts of disorganized myofibrils with focal density resembling the Z-band were shown in the cytoplasm of the neoplastic cells. The neoplasia was diagnosed as rhabdomyosarcoma.     

Congenital myopathy in Japanese Black calves

Two Japanese Black bull calves from a dam showed muscular weakness and became recumbent after birth. At necropsy, skeletal muscles, including face, neck, body and proximal and distal forelimb and hindlimb were extremely pale in color and edematous. Histopathological examination of skeletal muscles revealed degenerative changes as follows: replacement of muscle with fat, variation in muscle fiber diameter, internal nuclei, central core-like structures, and vacuolar and hyaline degeneration of muscle fibers. Ultrastructurally, the lesions were characterized by focal myofibrillar disorganization with streaming or irregularity of Z bands. The present cases seem to be classified as congenital myopathy based on pathological alterations and age of onset.