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[On hybrid embryo culture in vitro of Syringa L].

Syringa L. is the famous ornamental shrub in China, but its embryo always dies before seed mature during cross breeding, and hence, the breeding work is very difficult. The main object of this study is using embryo culture in vitro to get the seedling directly and to improve the succeed rate of cross breeding. The factors that influenced the embryo culture were researched in detail. The results showed that the optimal medium for embryo culture was Monnier, and the second was MS or LS, which meant that the embryo of Syringa needed abundant macroelements and microelements, especially Ca2+ and K+. The optimal sugar concentration was 50 g.L-1. At this level, the sugar could offer enough nutrition and high osmotic pressure for embryo. When the embryo age was 50-60 days, the culture was easy to be succeed. At this time, the cotyledon in ovule began to form, or organ began to differentiation, so, the embryo was very easy to germinate, and the seedling was very easy to form. Proper coconut milk, glutamic acid or glutamine, and activated charcoal could improve the germination and growth of the embryo. When the BA of low concentration (0.01 mg.L-1) was joined in the medium, the germination rate could be improved. The best NAA concentration was 0.01 mg.L-1.

Amino Acids↗

[Syringa species: molecular marking of species and cultivars].

RAPD analysis was carried out with 22 accessions of the genus Syringa, including six species, one interspecific hybrid, and 15 cultivars. In total, 500 polymorphic fragments were detected; species-specific and cultivar-specific markers were identified. For the first time, genetic polymorphism and genome similarity coefficients were estimated and phylogenetic relationships were established for the genus Syringa.

DNA, Plant↗

[Analysis of the coding region for signal peptide-containing proteins in Pseudomonas syringae pv. tomato genome].

The completed 5 615 ORFs of chromosome sequence of Pseudomonas syringae pv. tomato were analyzed by SignalP 3.0. The result revealed that 679 ORFs consisted of putative secretary proteins with signal peptides. 107 ORFs of signal peptides have been named. The length of most signal peptides was different from 19 amino acids to 31 amino acids, and the most dominant one was 23 amino acids in length. The size of most ORFs varied between 101 ~ 400 amino acids. The frequency of particular amino acids in signal peptides by statistical approach was 48.54% for hydrophobic, 18.67% for uncharged, 24.54% for negative and 8.00% for positive amino acids. The most frequent amino acid was alanine, and the least was isoleucine. Three classes of signal peptides were found in the genome of P. syringae pv. tomato: 501 ORFs belong to secretary signal peptides, 36 ORFs belong to twin-arginine signal peptides, and 15 ORFs belong to lipoprotein signal peptides. Type IV pilin signal peptide and bacteriocins and pheronoes signal peptide was not found in the genome.

Alanine↗

Characterization of two epiphytic bacteria from soybean leaves with antagonistic activities against Pseudomonas syringae pv. glycinea.

The strains 48b/90 and 22d/93 are naturally occurring ephiphytes which were isolated from soybean leaves. On the basis of pheno- and genotypic characteristics 48b/90 was identified as Erwinia herbicola and 22d/93 as Pseudomonas syringae. These two isolates produced biological active substances against different indicator organisms. The E. herbicola strain showed clear antagonistic properties against Escherichia coli and Pseudomonas syringae pv. glycinea, but not against Geotrichum candidum. 22d/93 was active against P. glycinea and G. candidum, but not against E. coli. Strain 48b/90 produced at least two different inhibitors: an antibiotic substance and an inhibitor of the alginate synthesis. Strain 22d/93 produced at least three different compounds inhibitory to P. glycinea and one to G. candidum. Their activities against the bacterial blight pathogen, P. glycinea, can be observed in planta, too. Under the influences of the antagonists the pathogen multiplied at lower rates and to lower stationary phase population levels. The development of bacterial blight symptoms was suppressed.

Anti-Bacterial Agents↗

Differential effects of growth temperature on ice nuclei active at different temperatures that are produced by cells of Pseudomonas syringae.

The temperature at which ice-nucleating bacteria are grown causes differences of 100- to 10,000-fold in the fraction of cells that nucleate ice at a given temperature (ice nucleation frequency). Ice nucleation frequencies of cells of Pseudomonas syringae grown at temperatures that ranged from 9 to 33 degrees C were examined in order to more accurately characterize physiological effects on ice nuclei active at temperatures of from about -2 to -10 degrees C, the temperature range for this phenotype. Large differences in ice nucleation frequency occurred at all but the lowest assay temperatures in cells of P. syringae grown in the temperature range of 15 to 33 degrees C. These differences in ice nucleation frequency may be attributed, at least in part, to post-translational factors. Because other studies have indicated that ice nuclei active at the lowest assay temperatures may reflect the amount of ice nucleation protein produced, while higher nucleation temperatures reflect aggregates of this ice nucleation protein, data was normalized to the frequency of ice nuclei active at the lowest ice nucleation temperatures (which also correspond to the most abundant nuclei). This was done in order to develop a baseline of comparison for cells grown at different temperatures that more clearly shows possible post-translational effects such as aggregation of the nucleation protein. After this normalization was performed, and in contrast to the results noted above, the number of ice nuclei in cells grown at 9, 15, and 20 degrees C that were active at different assay temperatures was very similar. Differences in ice nucleation frequency that occurred over all assay temperatures in cells grown between 9 and 20 degrees C may be attributed to differences in the total number of nuclei present in the population of cells. The large effects of growth temperature on nucleation frequency have important implications for estimating numbers of ice nucleating bacteria in environmental samples by determining the number of bacterial ice nuclei in such samples.

Bacterial Outer Membrane Proteins↗

Molecular characterization and hrp dependence of the avirulence gene avrPto from Pseudomonas syringae pv. tomato [corrected].

The avrPto avirulence gene from Pseudomonas syringae pv. tomato (Pst) race 0 governs race-specific resistance to bacterial speck disease in tomato cultivars containing the Pto resistance gene. The avrPto gene encodes 0.7 and 0.75 kb mRNAs whose predicted translation product is a mostly hydrophilic 164 amino acid protein of 18.3 kD a that reveals no homology to protein sequences in GenBank or EMBL databases. Highest expression of avrPto in cell culture is observed in minimal media containing sugars and sugar alcohols as carbon sources and lowest expression in minimal media containing tricarboxylic acid intermediates and in complex media. Expression of avrPto in planta is induced within 1 h following infection of both resistant and susceptible tomato plants by Pst, and increases over the first 6 h. Transcription of avrPto requires the hrpSR pathogenicity functions, but is independent of other Pst hrp genes. A region of the avrPto promoter shows homology to hrp box sequences upstream of other P. syringae genes that require the hrpSR locus for expression, and both avirulence activity and avrPto mRNA accumulation are abolished by deletions extending into this region. The avrPto transcription start site maps 31 nucleotides downstream of the hrp box motif.

Amino Acid Sequence↗

Homology of the root adhesin of Pseudomonas fluorescens OE 28.3 with porin F of P. aeruginosa and P. syringae.

The gene encoding the root adhesin from the outer membrane of Pseudomonas fluorescens OE 28.3 was isolated from a genomic lambda EMBL3 library and sequenced. The deduced protein (32104 daltons) displayed strong homology with the amino- and carboxyterminal parts of porin F (OprF) from P. aeruginosa and P. syringae. Significant homology was also found within the C-terminal domain of the OmpA proteins from Enterobacteria and major outer membrane proteins from Neisseria species. However, a cysteine-rich domain present in the OprFs of P. aeruginosa and P. syringae is absent from the adhesin of P. fluorescens. Instead, it contains a shorter sequence with eight alternating proline residues.

Adhesins, Bacterial↗

Ice crystallization by Pseudomonas syringae.

Several bacterial species can serve as biological ice nuclei. The best characterized of these is Pseudomonas syringae, a widely distributed bacterial epiphyte of plants. These biological ice nuclei find various applications in different fields, but an optimized production method was required in order to obtain the highly active cells which may be exploited as ice nucleators. The results presented here show that P. syringae cells reduce supercooling of liquid or solid media and enhance ice crystal formation at sub-zero temperatures, thus leading to a remarkable control of the crystallization phenomenon and a potential for energy savings. Our discussion focuses on recent and future applications of these ice nucleators in freezing operations, spray-ice technology and biotechnological processes.

Biotechnology↗

The release of alginate lyase from growing Pseudomonas syringae pathovar phaseolicola.

Pseudomonas syringae pathovar phaseolicola, which produces alginate during stationary growth phase, displayed elevated extracellular alginate lyase activity during both mid-exponential and late-stationary growth phases of batch growth. Intracellular activity remained below 22% of the total activity during exponential growth, suggesting that alginate lyase has an extracellular function for this organism. Extracellular enzyme activity in continuous cultures, grown in either nutrient broth or glucose-simple salts medium, peaked at 60% of the washout rate, although nutrient broth-grown cultures displayed more than twice the activity per gram of cell mass. These results imply that growth rate, nutritional composition, or both initiate a release of alginate lyase from viable P. syringae pv. phaseolicola, which could modify its entrapping biofilm.

Plant Diseases↗

The rpoS gene in Pseudomonas syringae is important in surviving exposure to the near-UV in sunlight.

The near-UV component of sunlight decreased culturability of the leaf epiphyte and plant pathogen Pseudomonas syringae. Exposure of the wild-type cells for 4 h to UV-A and UV-B in sunlight was ten fold more detrimental than exposure to sunlight with just UV-A. Sensitivity to UV-A especially increased in a mutant of P. syringae lacking the global regulatory sigma factor, RpoS. No RpoS-mutant cells were culturable after 4 h of exposure to near-UV sunlight. These findings suggest that both UV-A and UV-B wavelengths cause damage to the bacterial cell and that the RpoS protein regulates protective measures for the leaf-associated pseudomonad.

Bacterial Proteins↗

Mechanism of action of Pseudomonas syringae phytotoxin, syringomycin. Interaction with the plasma membrane of wild-type and respiratory-deficient strains of Saccharomyces cerevisiae.

The effects of the phytotoxin, syringomycin, produced by Pseudomonas syringae pv. syringae, were examined on cells of a wild-type and a respiratory-deficient (rho0) mutant of Saccharomyces cerevisiae. The growth of both strains in liquid culture was inhibited by 0.5 micrograms syringomycin per ml and higher. Uptake rates of tetraphenylphosphonium and dimethyloxazolidine ions in cell suspensions of both strains increased when 1.5 micrograms per ml syringomycin was added. These responses were kinetically and quantitatively similar in the two strains and indicated increases in electrical potential (cell interior negative) and pH differences (cell interior alkaline) across the plasma membrane. Glucose (0.1 M) enhanced the effect on the electrical potential, was required for the pH changes, and increased the cellular ATP levels. These results show that the effects of syringomycin are energy-dependent and are due to alterations of plasma membrane and not to mitochondrial function.

Adenosine Triphosphate↗

Molecular cloning in Escherichia coli, expression, and nucleotide sequence of the gene for the ethylene-forming enzyme of Pseudomonas syringae pv. phaseolicola PK2.

The gene for the ethylene-forming enzyme of Pseudomonas syringae pv. phaseolicola PK2 was found to be encoded by an indigenous plasmid, designated pPSP1. The gene for the ethylene-forming enzyme was cloned and expressed in Escherichia coli JM109. Nucleotide sequence analysis of the clone revealed an open reading frame that encodes 350 amino acids (mol. wt. 39,444). In a comparison with other proteins, the homology score for the entire amino-acid sequence of the ethylene-forming enzyme of Pseudomonas syringae versus ethylene-forming enzymes from plants and 2-oxoglutarate-dependent dioxygenases was low. However, functionally significant regions are conserved.

Amino Acid Sequence↗

Conservation of plasmids among plant-pathogenic Pseudomonas syringae isolates of diverse origins.

Thirty isolates of Pseudomonas syringae pv. tabaci, pv. angulata (pathogens on tobacco), pv. coronafaciens, and pv. striafaciens (pathogens on oats) were examined for plasmid DNAs. The strains were obtained from plants throughout the world, some over 50 years ago. Of the 22 tobacco pathogens, 16 contain predominantly one type of plasmid, the pJP27.00 type. The remaining six tobacco-specific strains do not harbor detectable plasmids. The oat pathogens contain one, two, or three plasmids. DNA homology studies indicate that the plasmid DNAs are highly conserved. More importantly, the plasmids harbored by strains isolated from one host plant are conserved most stringently; e.g., the plasmids from the tobacco pathogens are, with one exception, indistinguishable by restriction endonuclease digestion and Southern hybridization. There is also extensive homology among plasmids indigenous to the oat-specific P. syringae pv. coronafaciens and pv. striafaciens strains.

Base Sequence↗

Sequence of a gene encoding periplasmic Pseudomonas syringae ankyrin.

A gene encoding ankyrin (Ank) was isolated from a genomic library of the plant pathogen Pseudomonas syringae pathovar syringae strain 61 (Pss61). The gene encodes an 183-amino-acid (aa) polypeptide which has homology to the 33-aa repeat domain of mammalian Ank and Ank homologs from other bacteria, animals and plants.

Amino Acid Sequence↗

Resistance to ultraviolet light in Pseudomonas syringae: sequence and functional analysis of the plasmid-encoded rulAB genes.

The indigenous plasmids, pPSR1 and pPSR5, were each shown to confer resistance to ultraviolet light (UV) in Pseudomonas syringae (Ps) pv. syringae FF5. The UV-resistance (UVR) determinant was subcloned from a cosmid library of pPSR1, and sequence analysis revealed the presence of two ORFs, designated rulAB which are homologous to the Escherichia coli umuDC mutagenic DNA repair systems and other plasmid-encoded UVR operons. Amino acid (aa) alignments indicated that RulAB are most closely related to the RumAB proteins from plasmid R391, sharing 40.5% and 48.6% aa identity with RumA and RumB, respectively. UV sensitivity assays with the cloned rulAB genes indicated that the expression of UVR in Ps required a functional recA gene.

Amino Acid Sequence↗

The causal agent of halo blight in bean, Pseudomonas syringae pv. phaseolicola, attaches to stomata via its pili.

The phytopathogenic pseudomonad Pseudomonas syringae pv. phaseolicola causes halo blight of bean (Phaseolus vulgaris L.). Initiation of infection depends on the ability of the cells to adhere to the target cell surface. P. syringae pv. phaseolicola expresses pili, which are the receptors of the lipid-containing dsRNA bacteriophage phi 6. phi 6-resistant bacterial strains can be divided into different piliation types. It was possible to show that the adhesion of the bacteria onto plant cell surface was dependent on the pili. Non-piliated bacterial stains showed a much lower adherence to the leaf surface than strains expressing phi 6 specific pili. Scanning electron microscopy showed that the piliated bacteria attached to the leaf surface at the site of stomata. Non-piliated bacteria were evenly distributed on the leaf surface. All bacterial strains used in this study were capable of causing halo blight if injected into the plant. If the bacteria were sprayed on the plants, followed by spraying of sterile buffer, only piliated bacteria caused symptoms.

Bacterial Adhesion↗

Structural heterogeneity in the lipopolysaccharides of Pseudomonas syringae with O-polysaccharide chains having different repeating units.

Studies by sugar and methylation analyses, Smith degradation, and 1H and 13C NMR spectroscopy revealed a structural heterogeneity in the O-polysaccharides of Pseudomonas syringae pvs. coronafaciens IMV 9030 and atrofaciens IMV 8281 owing to the presence of different types of repeating units. In strain IMV 9030, the major repeating units are a linear alpha-L-rhamnose trisaccharide and a tetrasaccharide (A, n=0 or 1). A minor repeating unit is a branched pentasaccharide with an alpha-L-rhamnose main chain and a lateral 3-acetamido-3,6-dideoxy-D-galactose (D-Fuc3NAc) residue (B, X=2, n=1). In strain IMV 8281, all repeating units are branched and differ in size and position of substitution of one of the alpha-L-rhamnose residues (tetrasaccharide, B, X=3, n=0; pentasaccharides, B, X=2 or 3, n=1). [structure--see text] Reinvestigation of the structure of the branched O-polysaccharide of P. syringae pv. tomato IPGR 140 showed that, together with the major tetrasaccharide repeating unit (B, X=3, n=0) [Knirel, Y. A., et al. Carbohydr. Res. 1993, 243, 199-204], it has a minor pentasaccharide repeating unit (B, X=3, n=1).

Carbohydrate Conformation↗

Chlorosis-inducing products from Pseudomonas syringae pathovars: new N-coronafacoyl compounds.

Liquid cultures of Pseudomonas syringae pv. tomato strains that also produced the phytotoxin coronatine, were found to have a new chlorosis-inducing activity, not previously described. Bioassay-guided fractionation and HPLC analysis revealed two new peaks that were chlorosis-inducing on leaves of bean plants. Mass spectrometry and NMR analyses of the compounds led to the derivation of their structures as coronafacoyl-L-serine and coronafacoyl-L-threonine, respectively. The amino acid C-2 configurations were determined by GC analysis following hydrolysis with 6M HCl. Both compounds have more polar and more acidic properties than coronatine, norcoronatine, and other known coronafacoyl conjugates with non-polar amino acids. Investigation of Pseudomonas syringae pv. glycinea strains that are known to be prolific producers of the coronatine family of compounds revealed the production of coronafacoylserine and coronafacoylthreonine also.

Journal Article↗