Organization of the genes encoding chalcone synthase in Pisum sativum

To analyze the regulation of defense-related genes by signal molecules produced by phytopathogens, we isolated genes that encode chalcone synthase (CHS) in Pisum sativum. We have obtained seven independent genomic clones that contain at least seven classes of CHS genes, identified by the hybridization analysis to CHS cDNA and by the restriction mapping analysis. Two of the genomic clones (clone 5 and 6) each contain two CHS genes in a tandem repeat. The nucleotide sequence analysis of CHS genomic clone 5 revealed that PsCHS1 and PsCHS2 were corresponding genes of the CHS cDNA clones, pCC6 and pCC2, respectively, as reported earlier. Both genes are interrupted by a single intron of 88 nucleotides with identical sequences, although exonic sequences and 5-flanking sequences are divergent. Nucleotide sequences of the introns in five other classes of CHS genes showed that three classes had an intron of 87 nt with a striking homology to each other, but that the intron of the other two classes of CHS genes showed heterogeneity both in size and nucleotide sequence. 5-upstream regions of PsCHS1 and PsCHS2 did not show sequence homology except the 31 bp identical sequence that contains the CCTACC motif resembling the box-1 sequence. Both PsCHS1 and PsCHS2 genes are shown to be induced by fungal elicitor by a primer extension analysis and a transient transformation analysis using pea protoplasts prepared from suspension cultured-cells.     

Inhibition of ATPase Activity in Pea Plasma Membranes by Fungal Suppressors from Mycosphaerella pinodes and Their Peptide Moieties

The effects of two suppressors of the defense reactions of hostplants, which had been purified from the pea pathogen Mycosphaerellapinodes, as well as the effects of peptide moieties, on theATPase activity in pea plasma membranes were examined in vitro.One of the suppressors, Supprescin B, inhibited the ATPase activityin a non-competitive manner, but the other suppressor, SupprescinA, did not. Supprescin A was observed to reduce the inhibitoryeffect of Supprescin B. A tripeptide, Ser-Ser-Gly, and a hexapeptide,Ser-Ser-Gly-Asp-Glu-Thr, which were the respective peptide moietiesof Supprescin A and B, inhibited the ATPase activity in a competitivemanner. Supprescin B and fragments of the hexapeptide, suchas Asp-Glu-Thr and Gly- Asp-Glu, inhibited not only the ATPaseactivity but also the acid phosphatase activity of plasma membranesin vitro. These results indicate that the acidic amino-acidresidues of the "Asp-Glu" moiety seem to act as inhibitors ofthe phosphatase activity. Thus, the peptide moiety of SupprescinB consists of at least two functional elements. (Received October 23, 1992; Accepted January 18, 1993)     

Liposomal modification with uronate, which endows liposomes with long circulation in vivo, reduces the uptake of liposomes by J774 cells in vitro.

For overcoming rapid removal of liposomes from the bloodstream, we developed reticuloendothelial system (RES)-avoiding liposomes modified with a uronic acid derivative, palmityl-D-glucuronide (PGlcUA). In this current study, we examined the in vitro interaction of PGlcUA-liposomes with J774 cells derived from mouse macrophages. Liposomal association with J774 cells at 37 degrees C did not increase compared with the binding at 4 degrees C when liposomes were modified with PGlcUA. RES-avoiding ability was not specifically endowed by glucuronate but by uronates in general, since palmityl-D-galacturonide showed a similar effect on liposomal clearance in vivo and liposomal uptake in vitro. These facts indicate that modification of the liposomal surface with uronic acid derivatives endows liposomes with a long circulation time in the bloodstream by reducing their uptake by macrophages.     

A novel ganglioside expressed by mouse hematopoietic cell lines.

Mouse progenitor T cell-derived cell lines were established by fusion of cells of hematopoietic organs such as bone marrow and fetal liver with T lymphoma (BW5147) to determine their characteristic cell-surface components. The hybridomas with the phenotype of Thy-1+, CD3-, CD4-, CD8- and expression of T cell receptor gene mRNA (BM216 and FL339) were selected for progenitor T cell-derived cell lines, and their ganglioside compositions were studied. A ganglioside component with a mobility slightly faster than that of bovine brain GD1a on high-performance thin-layer chromatography was found in the cell extracts of these cell lines as one of the most abundant components and was absent in the extract of the parental cell line (BW5147). The structure of the ganglioside was determined to be: NeuAc alpha-Gal beta-Gal beta-Gal alpha-Gal beta-Glc beta-ceramide. Gangliosides with such a sequence have never been found before, suggesting the possibility that the ganglioside is expressed as a surface marker of the cells in hematopoietic organs committed to a specific cell lineage, presumably to T cell lineage. cells in hematopoietic organs committed to a specific     

Development and sexual maturation of Taenia crassiceps (Cestoda) in the golden hamster

The development of cysticerci of Taenia crassiceps to mature tapeworms in the intestinal tract of cortisone-treated and untreated golden hamsters is described. The development of the worms in untreated hamsters is as follows. Maturation and branching of the uterus and the presence of ova in the uterus were observed on day 15 postinfection (PI). By day 21 PI, fertilization had taken place, as evidenced by the presence of sperm in the seminal receptacle. On day 28 PI, shelled eggs were observed in feces. The rate of development of the worms in cortisone-treated hamsters was similar to that observed in untreated hamsters. More worm recovery and a longer period of worm survival were seen in the cortisone-treated than in the untreated hamsters. Thus, both cortisone-treated and untreated golden hamsters can produce normal gravid individuals of T. crassiceps.     

Isolation and Characterization of Double-Stranded RNAs from the Pea Pathogen, Mycosphaerella pinodes

Studies on the structure of the extrachromosomal nucleic acidsin the plant pathogenic fungus, Mycosphaerella pinodes, OMP-1,isolated from Okayama, Japan, indicate that at least three double-stranded(ds) RNAs are present in the cytoplasm, while a strongly pathogenicisolate from England, MP-3, does not contain any of the dsRNAs.The sizes of these dsRNAs are 6.0 kb (L), 4.0 kb (M), and 2.6kb (S), respectively. Weakly pathogenic mutants selected fromUV-irradiated OMP-1 contain considerably higher copy numberof S than does the parental strain, while one of the mutants,OMP-X62, contains no detectable M. OMP-1 was cured of detectabledsRNAs by treatment with cycloheximide. Curing was accompaniedby an increase in virulence and by partial recovery of a colonymorphology similar to that of MP-3. The presence of these dsRNAsapparently results in a reduction in mycelial growth, in thestimulation of sporulation and possibly in the promotion ofhypovirulence. (Received February 19, 1990; Accepted October 30, 1990)     

Purification and characterization of neutral alpha-mannosidase that is activated by Co2+ from Japanese quail oviduct

An alpha-mannosidase was purified from the magnum section of Japanese quail oviduct by ammonium sulfate precipitation, DEAE-Sephacel chromatography, Sephacryl S-300 chromatography, mannan-Sepharose 4B chromatography, and hydroxyapatite chromatography. The purified alpha-mannosidase (referred to as neutral alpha-mannosidase) showed a single band on polyacrylamide gel with or without sodium dodecyl sulfate. Its molecular weight was found to be 330,000 by gel chromatography. Neutral alpha-mannosidase hydrolyzed p-nitrophenyl alpha-D-mannopyranoside and the pyridylamino derivative of Man alpha 1-6(Man alpha 1-3)Man alpha 1-6(Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4GlcNAc (Km value was 3 mM). Mannosyl alpha 1-2 linkages in the pyridylamino derivative of Man alpha 1-2 Man alpha 1-6(Man alpha 1-2Man alpha 1-3)Man alpha 1-6(Man alpha 1-2Man alpha 1-2Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4GlcNAc were hardly hydrolyzed. Its optimum pH was found to be 7.0. The activity of the enzyme was activated by CO2+, and was potently inhibited by Cu2+, Hg2+, swainsonine, and 1-deoxymannojirimycin.     

Elicitor–like Substances Present in Barley and Wheat Seeds

Abstract Barley and wheat seeds contain elicitor–like substances which induce the production of antifungal substances in the host plants effective against powdery mildew fungi. Seed extract per se does not exhibit direct antimicrobial activity but induces the resistance response on the host. It also induces the systemic resistance in juvenile leaves. Host resistance induced by the treatment with seed extract shows some degree of cultivar specificity.     

Mechanism of the protective immunity against murine typhoid: persistence of salmonella L forms in the liver after immunization with live-cell vaccines

Abstract Live-cell vaccines of Salmonella typhimurium , either a sub-lethal dose of a wild-type (strain LT2) or a high dose of its two-heptose Rd₁ mutant (strain SL1004), induced acquired resistance to murine typhoid, which remained 180 days after immunozation. Growth of S. typhimurium as a bacillary form ceased between days 30 and 60 of immunization, but L forms of this bacterium colonized the liver (the mean number of L forms in the liver: 600 L-forming units) even at 180 days post-immunization. In contrast, a high inoculum of either a Ra mutant (strain TV148) of strain LT2 or S. schottmülleri 8006 sharing the same O antigenic components with those of S. typhimurium induced only a short-lived protection in proportion to the number of L forms in the liver, and the protective immunity was lost before day 180. However, there was no significant difference in the salmonella-specific T-cell responses among groups of immunized mice on day 180 of immunization. A lethal infection with strain LT2 in mice which had been immunized 75 days previously with living cells of strain SL1004 resulted in a rapid clearance of the challenge inoculum, together with a rapid elevation of anti- S. typhimurium antibody responses. Thus, the present data suggest that the long-lived immunity conferred upon live S. typhimurium vaccines is attributable to the colonization of this bacterium in the liver as L forms and the ability to colonize the liver as L forms is independent of the brain length of salmonella O-antigens.     

Pathogenicity, Non-Host Hypersensitivity and Host Defence Non-Permissibility Regulatory Gene hrpX is Highly Conserved in Xanthomonas Pathovars

Xanthomonas campestris pv. campestris and X. oryzae pv, oryzae contain the 1428 base pair hrpX gene whose product is involved in the regulation oi hrp genes required for pathogericity, non-host hypersensitivity and non-permissibility of compatible host defence responses. Previous Southern blot hybridization studies have suggested that hrpX is conserved in several X. campestris pathovars and X. oryzae. strains. We have confirmed and extended these findings using hrpX gene amplification by polymerase chain reaction, coupled with Southern blot hybridization analyses. Sixteen distinct pathovars of X. campestris and 12 strains of X. oryzae pv, oryzae were shown to contain homologs of hrpX which were not apparent in heterologous bacteria such as Agrobacterium tumefaciens, A. rhizogenes, Erwinia carolovora ssp. carotovora, Pseudomonas syringae pv, glycinea. P. syringae pv, labaci , and Escherichia coli. The hrpX gene is therefore highly conserved among Xanthomonas species and its gene product strongly resembles positive regulatory proteins of the AraC protein family,