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high frequency agrobacterium tumefaciens-mediated plant transformation induced by ammonium nitrate.success in plant genetic transformation depends on the efficiency of explant regeneration and transgene integration. whereas the former one depends on explant totipotency, the latter depends on the activity of host dna repair and chromatin organisation factors. we analyzed whether factors that result in an increase in recombination frequency can also increase transformation efficiency. here, we report that a threefold increase in the concentration of nh(4)no(3) in the growth medium results in mo ...200919221758
paenibacillus polymyxa antagonizes oomycete plant pathogens phytophthora palmivora and pythium aphanidermatum.to find sustainable alternatives to the application of synthetic chemicals for oomycete pathogen suppression.200919226403
transformation of nasturtium officinale, barbarea verna and arabis caucasica for hairy roots and glucosinolate-myrosinase system production.hairy roots of nasturtium officinale, barbarea verna and arabis caucasica with active glucosinolate-myrosinase system were obtained after transformation with agrobacterium rhizogenes. hairy roots of n. officinale produced phenylalanine-derived gluconasturtiin and glucotropaeolin (max. 24 and 7 mg g(-1) dw). b. verna and a. caucasica hairy roots produced gluconasturtiin (max. 41 mg g(-1) dw) and methionine-derived glucoiberverin (max. 32 mg g(-1) dw), respectively. treatment of the roots with ami ...200919229477
genetic evidence for cheb- and cher-dependent chemotaxis system in a. tumefaciens toward acetosyringone.agrobacterium tumefaciens has homologues of chemotaxis genes of the alpha-subgroup of proteobacteria, which include orf1, orf2, chey1, chea, cher, cheb, chey2, orf9, and orf10 and organized in one operon [wright et al. a chemotaxis cluster from agrobacterium tumefaciens. gene 1998;220:83-9]. two mutants of a. tumefaciens were created by in-frame deletions of cheb and cher using suicide vector, pk18mobsacb. the chemosensory behaviour of mutants studied on swarm plates and in blind-well chemotaxis ...200919231145
anthraquinones from natural and transformed roots of plocama pendula.five new and 15 known anthraquinones have been isolated from plocama pendula root cultures transformed by agrobacterium rhizogenes. the new anthraquinones were named balonone, balonone methyl ether, and plocamanones a, b, and c. moreover, lucidin dimethyl ether was obtained for the first time as a natural compound. on the other hand, a study of p. pendula natural roots afforded another two new anthraquinones, i.e., plocamanone d and chionone. the anthraquinone isozyganein dimethyl ether has been ...200919235160
[effects of natural and hybrid lectins on the legume-rhizobium interactions].the effects of hybrid lectins--full-sized pea pisum sativum lectin (psl) with the carbohydrate-binding region of white melilot melilotus albus lectin or wild licorice astragalus glycyphyllos lectin substituted for the corresponding psl region (psl/mal and psl/agl, correspondingly)--on the legume-rhizobium symbiosis were studied. the treatment of the rhizobium leguminosarum bv. viciae in the alfalfa (medicago sativa) rhizosphere with psl induced formation of uninfected pseudonodules on its roots, ...200919235514
expression of a transgene exchanged by the recombinase-mediated cassette exchange (rmce) method in plants.we developed a site-directed integration (sdi) system for agrobacterium-mediated transformation to precisely integrate a single copy of a desired gene into a predefined target locus by recombinase-mediated cassette exchange (rmce). we produced site-specific transgenic tobacco plants from four target lines and examined expression of the transgene in t1 site-specific transgenic tobacco plants, which were obtained by backcrossing. we found that site-specific transgenic plants from the same target l ...200919241079
white clover nodulation index in heavy metal contaminated soils- a potential bioindicator.the morphological effects of heavy metal stress on the nodulation ability of rhizobium spp. and growth of white clover (trifolium repens l.) were studied in the laboratory under controlled conditions. fourteen topsoils were collected from an area with elevated metal concentrations (cd, zn, and pb). white clover was cultivated using a specialized "rhizotron" method to observe the development of root and nodule characteristics. results show effects of increasing heavy metal concentrations on nodul ...200919244489
bioactive metabolites produced by chaetomium globosum, an endophytic fungus isolated from ginkgo biloba.a novel cytotoxic chlorinated azaphilone derivative named chaetomugilin d (1), together with three known metabolites, chaetomugilin a (2), chaetoglobosins a (3) and c (4), has been isolated by a bioassay-guided fractionation from the etoac extract of the cultures of chaetomium globosum, an endophytic fungus found in the leaves of ginkgo biloba. structure of 1 was established by analyses of spectroscopic methods, including 2d-nmr experiments (cosy, noesy, hmqc, and hmbc). compounds 1-4 displayed ...200919246197
genome sequences of three agrobacterium biovars help elucidate the evolution of multichromosome genomes in bacteria.the family rhizobiaceae contains plant-associated bacteria with critical roles in ecology and agriculture. within this family, many rhizobium and sinorhizobium strains are nitrogen-fixing plant mutualists, while many strains designated as agrobacterium are plant pathogens. these contrasting lifestyles are primarily dependent on the transmissible plasmids each strain harbors. members of the rhizobiaceae also have diverse genome architectures that include single chromosomes, multiple chromosomes, ...200919251847
interactions between brucella suis virb8 and its homolog traj from the plasmid psb102 underline the dynamic nature of type iv secretion systems.the proteinvirb8 plays a critical role in the assembly and function of the agrobacterium tumefaciens virb type iv secretion system (t4ss). the structure of the periplasmic domain of both a. tumefaciens and brucella suis virb8 has been determined, and site-directed mutagenesis has revealed amino acids involved in the dimerization of virb8 and interactions with virb4 and virb10. we have shown previously that traj, the virb8 homologue from psb102, and the chimeric protein trajb8, encompassing the c ...200919251859
assembly of agrobacterium phytochromes agp1 and agp2 with doubly locked bilin chromophores.the natural chromophore of most bacterial and fungal phytochromes is biliverdin (bv), which is incorporated in a covalent manner into the protein. upon photoconversion between the red light-absorbing form pr and the far-red light-absorbing form pfr, the stereochemistry of the chromophore around the c15 methine bridge changes from z anti to e anti. recombinant phytochromes agp1 and agp2 from agrobacterium tumefaciens were assembled with a set of synthetic chromophores, including 2,18-et-bv, 3,18- ...200919253981
transient expression of minimum linear gene cassettes in onion epidermal cells via direct transformation.a new method without any special devices for direct transformation of linear gene cassettes was developed. its feasibility was verified through 5'-fluorescent dye (fluorescein isothiocyanate, fitc)-labeled fluorescent tracing and transient expression of a gus reporter gene. minimal linear gene cassettes, containing necessary regulation elements and a gus reporter gene, was prepared by polymerase chain reaction and dissolved in transformation buffer solution to 100 ng/ml. the basic transformation ...200919255730
different and new nod factors produced by rhizobium tropici ciat899 following na+ stress.the root nodule bacterium rhizobium tropici strain ciat899 is highly stress resistant. it grows under acid conditions, in large amounts of salt, and at high osmotic pressure. an earlier study reported a substantial qualitative and quantitative effect of acid stress on the biosynthesis of nod factors. the aim of the present work was to investigate the effect of high salt (nacl) concentrations, another common stress factor, on nod factor production. for this purpose, thin-layer chromatography, hpl ...200919260963
variability in the level of rna silencing suppression caused by triple gene block protein 1 (tgbp1) from various potexviruses during infection.rna silencing is an important defence mechanism against virus infection, and many plant viruses encode rna silencing suppressors as a counter defence. in this study, we analysed the rna silencing suppression ability of multiple virus species of the genus potexvirus. nicotiana benthamiana plants exhibiting rna silencing of a green fluorescent protein (gfp) transgene showed reversal of gfp fluorescence when systemically infected with potexviruses. however, the degree of gfp fluorescence varied amo ...200919264652
changes in motility of the rhizobacterium azospirillum brasilense in the presence of plant lectins.the plant-beneficial bacterium azospirillum brasilense can swim in liquids and swarm or migrate with the formation of microcolonies in soft media. to get closer to understanding the influence of natural environments on a. brasilense motility, we studied the individual and social movement of the bacterium in the presence of various plant lectins. the lectins with specificity for n-acetyl-beta-d-glucosamine oligomers (wheat germ, solanum tuberosum and ulex europeus agglutinins) decreased a. brasil ...200917317126
novel scar primers for specific and sensitive detection of agrobacterium vitis strains.an agrobacterium vitis-specific dna fragment (pavs3) was generated from pcr polymorphic bands amplified by primer urp 2r. a. vitis specificity of this fragment was confirmed by southern hybridization with genomic dna from different agrobacterium species. sequence-characterized amplified region (scar) markers were developed for a. vitis specific detection, using 24-mer oligonucleotide primers designed from the flanking ends of the 670bp insert in pavs3. the scar primers amplified target sequences ...200917467252
changes in azospirillum brasilense motility and the effect of wheat seedling exudates.the rhizobacterium azospirillum brasilense sp245 swims, swarms (swa(+) phenotype) or, very rarely, migrates with the formation of granular macrocolonies (gri(+) phenotype). our aims were (i) to identify sp245 mutants that swarm faster than the parent strain or differ from it in the mode of spreading and (ii) to compare the mutants' responses to wheat seedling exudates. in isotropic liquid media, the swimming speeds of all motile a. brasilense strains were not influenced by the exudates. however, ...200917707621
optimization of agrobacterium-mediated transformation conditions in mature embryos of elite wheat.immature embryos have been used frequently as target tissues in the genetical transformation of wheat. however, obtaining a large number of high quality immature embryos throughout the year is a laborious and delicate process, because of the need to cultivate the plants under controlled conditions. to circumvent this, we have employed mature embryos rather than immature ones as starter explants for agrobacterium-mediated transformation of an elite wheat (triticum aestivum l.) cultivar em12. the ...200917906943
variation of 16s-23s internally transcribed spacer sequence and intervening sequence in rdna among the three major agrobacterium species.we analyzed 16s-23s internally transcribed spacer (its) and neighboring sequences among 37 strains belonging to the three major pathogenic agrobacterium species, in order to know variation in each species and to develop a simple discrimination method. number of its size variation was 9, 4, and 7 in agrobacterium tumefaciens, agrobacterium vitis, and agrobacterium rhizogenes, respectively. the its sequence of most strains in each species was distinguishable from that of the other two species. the ...200917997085
bacterial population dynamics in a reverse-osmosis water purification system determined by fluorescent staining and pcr-denaturing gradient gel electrophoresis.the bacterial population dynamics in an industrial scale reverse-osmosis (ro) water purification system were analyzed by fluorescent staining methods and denaturing gradient gel electrophoresis (dgge). bacterial numbers increased with storage in a tank, and bacterial diversity changed during the water purification process. a dna sequence-based analysis of the major bands on the dgge gel revealed that simonsiella sp. (betaproteobacteria) was abundant in the source water (activated sludge-treated ...200921566369
agrobacterium tumefaciens and a. rhizogenes use different proteins to transport bacterial dna into the plant cell nucleus.agrobacterium tumefaciens and a. rhizogenes transport single-stranded dna (ssdna; t-strands) and virulence proteins into plant cells through a type iv secretion system. dna transfer initiates when vird2 nicks border sequences in the tumour-inducing plasmid, attaches to the 5' end, and pilots t-strands into plant cells. agrobacterium tumefaciens translocates ssdna-binding protein vire2 into plant cells where it targets t-strands into the nucleus. some a. rhizogenes strains lack vire2 but transfer ...200921255274
bacillus-based biocontrol of fusarium disease on tomato cultures in burundi.the biocontrol potential of a plant growth-promoting rhizobacterium (pgpr) bacillus subtilis s499 on tomato was studied in open field sites in low attitude area of the plain of imbo in burundi. application of the pgpr strain on seed before sowing have significantly increased growth and fruit yield of tomato plants in addition to its remarkable control of a local important disease caused by a fungus type-fusarium.. this pathogen causing large tosses in burundi tomato plantings is closely related ...200920222546
natural abundances of 15n and 13c in leaves of some n2-fixing and non-n2-fixing trees and shrubs in syria.a survey study was conducted on man-made plantations located at two different areas in the arid region of syria to determine the variations in natural abundances of the (15)n and (13)c isotopes in leaves of several woody legume and non-legume species, and to better understand the consequence of such variations on nitrogen fixation and carbon assimilation. in the first study area (non-saline soil), the delta(15)n values in four legume species (acacia cyanophylla,-1.73 per thousand acacia farnesia ...200920183233
agrobacterium rhizogenes-mediated transformation of hypericum tomentosum l and hypericum tetrapterum fries.this is the first evidence on successful agrobacterium rhizogenes-mediated genetic transformation of two species from the genus hypericum, h. tomentosum l. and h. tetrapterum fries. hairy root cultures were induced from root segments of both hypericum species by two agropine wild-type strains of a. rhizogenes, atcc 15834 and a4. the transgenic character of the hairy root cultures was proved by pcr amplification of the rolabcd genes. in some h. tetrapterum transgenic lines aux genes were detected ...200920158159
npr1 protein regulates pathogenic and symbiotic interactions between rhizobium and legumes and non-legumes.legumes are unique in their ability to establish symbiotic interaction with rhizobacteria from rhizobium genus, which provide them with available nitrogen. nodulation factors (nfs) produced by rhizobium initiate legume root hair deformation and curling that entrap the bacteria, and allow it to grow inside the plant. in contrast, legumes and non-legumes activate defense responses when inoculated with pathogenic bacteria. one major defense pathway is mediated by salicylic acid (sa). sa is sensed a ...200920027302
transformation of the plant kalanchoë daigremontiana using agrobacterium tumefaciens.kalanchoë daigremontiana can be stably transformed using the agrobacterium tumefaciens-mediated t-dna transfer method, as described here. sterilized plant tissue is cocultivated with an a. tumefaciens suspension, transformants are selected and the shoots are grown in rooting medium and then in soil. plant phenotypes can be examined approximately 3 mo after transfer of plants to soil.200920147048
unveiling the intergenic transcriptome across rhizobium etli's lifecycle. 200920420221
[agrobacterium-mediated transformation of flax with a mutant tubulin gene responsible for resistance to dinitroaniline herbicides].agrobacterium tumefaciens was used to transform fiber flax with the pbituba8 plasmid carrying the mutant alpha-tubulin gene imparting resistance to dinitroaniline herbicides and the nptii selective marker gene imparting resistance to canamycin. the transformants were selected in parallel on media containing canamycin and trifluralin (a dinitroaniline herbicide). the transgenic nature of the resultant regenerants resistant to dinitroaniline herbicides was confirmed by means of southern blotting a ...200919947549
[toxicological evaluation of biopreparations on the basis of nitrogen-fixing bacteria].a comparative analysis of results of toxicological research of microbiological preparations on the basis of different species of nitrogen-fixing microorganisms of azotobacter, agrobacterium, azospirillum general and pathogenic properties of strains-producers has been carried out. a possibility to improve methodical principles of toxicological estimation and hygienic regulation of associative nitrogen-fixing microorganisms-producers and preparations on their basis in the industrial objects and en ...200920455429
t-dna insertion, plasmid rescue and integration analysis in the model mycorrhizal fungus laccaria bicolor.ectomycorrhiza is a mutualistic symbiosis formed between fine roots of trees and the mycelium of soil fungi. this symbiosis plays a key role in forest ecosystems for the mineral nutrition of trees and the biology of the fungal communities associated. the characterization of genes involved in developmental and metabolic processes is important to understand the complex interactions that control the ectomycorrhizal symbiosis. agrobacterium-mediated gene transfer (amt) in fungi is currently opening ...200821261845
endophytic bacteria in the rice plant.endophytic bacteria are defined as bacteria detected inside surface-sterilized plants or extracted from inside plants and having no visibly harmful effects on the plants. various kinds of endophytic bacteria, such as pantoea, methylobacterium, azospirillum, herbaspirillum, burkholderia and rhizobium etc., have been found inside rice plants. this minireview summarizes and discusses recent studies of endophytic bacteria residing in rice plants, focusing on flora, origin, movement, and interaction ...200821558696
impact of organic fertilizers with and without chemical fertilizers on soil chemical properties and the establishment of nitrogen-fixing bacteria in the rhizosphere.effects of organic fertilizers with and without the application of chemical fertilizers for seven years as part of a wheat-pearl millet cropping sequence on soil chemical properties and the establishment of nitrogen fixing bacteria in the rhizosphere were examined. the application of farmyard manure, poultry manure, and sugarcane filter cake alone or in combination with chemical fertilizers improved the soil organic c, total n, p, and k status. larger populations of azotobacter chroococcum and r ...200821558724
high-efficiency agrobacterium-mediated transformation of brachypodium distachyon inbred line bd21-3.brachypodium distachyon (brachypodium) is a small grass with biological attributes (rapid generation time, small genome, diploid accessions, small stature and simple growth requirements) that make it suitable for use as a model system. in addition, a growing list of genomic resources have been developed or are currently under development including: cdna libraries, bac libraries, est sequences, bac end sequences, a physical map, genetic markers, a linkage map and, most importantly, the complete g ...200817999063
over-expression of a gibberellin 2-oxidase gene from phaseolus coccineus l. enhances gibberellin inactivation and induces dwarfism in solanum species.gibberellins (gas) are endogenous hormones that play a predominant role in regulating plant stature by increasing cell division and elongation in stem internodes. the product of the ga 2-oxidase gene from phaseolus coccineus (pcga2ox1) inactivates c(19)-gas, including the bioactive gas ga(1 )and ga(4), by 2beta-hydroxylation, reducing the availability of these gas in plants. the pcga2ox1 gene was introduced into solanum melanocerasum and s. nigrum (solanaceae) by agrobacterium-mediated transform ...200817999064
structural characterizations of lipids a by ms/ms of doubly charged ions on a hybrid linear ion trap/orbitrap mass spectrometer.here, a new 'one pot' and fast approach is described, based on electrospray ionization (esi) of negative ions by using a hybrid linear ion trap/orbitrap mass spectrometer (ltq/orbitrap) for ms and ms/ms analysis. by this method the distribution of the primary and secondary acyl residues of the intact lipid a is inferred by analysis of the esi spectra measured in positive and negative mode. the analysis of these data allows an unequivocal assignment of the fatty acid distribution. this methodolog ...200817975853
crystal structure of pyridoxamine-pyruvate aminotransferase from mesorhizobium loti maff303099.pyridoxamine-pyruvate aminotransferase (ppat; ec 2.6.1.30) is a pyridoxal 5'-phosphate-independent aminotransferase and catalyzes reversible transamination between pyridoxamine and pyruvate to form pyridoxal and l-alanine. the crystal structure of ppat from mesorhizobium loti has been solved in space group p4(3)2(1)2 and was refined to an r factor of 15.6% (r(free) = 20.6%) at 2.0 a resolution. in addition, the structures of ppat in complexes with pyridoxamine, pyridoxal, and pyridoxyl-l-alanine ...200817989071
phage specificity and lipopolysaccarides of stem- and root-nodulating bacteria (azorhizobium caulinodans, sinorhizobium spp., and rhizobium spp.) of sesbania spp.phage susceptibility pattern and its correlation with lipopolysaccharide (lps) and plasmid profiles may help in understanding the phenotypic and genotypic diversity among highly promiscuous group of rhizobia nodulating sesbania spp.; 43 phages were from two stem-nodulating bacteria of s. rostrata and 16 phages were from root-nodulating bacteria of s. sesban, s. aegyptica and s. rostrata. phage susceptibility pattern of 38 sesbania nodulating bacteria was correlated with their lps rather than pla ...200817989956
transcriptome profiling and functional analysis of agrobacterium tumefaciens reveals a general conserved response to acidic conditions (ph 5.5) and a complex acid-mediated signaling involved in agrobacterium-plant interactions.agrobacterium tumefaciens transferred dna (t-dna) transfer requires that the virulence genes (vir regulon) on the tumor-inducing (ti) plasmid be induced by plant phenolic signals in an acidic environment. using transcriptome analysis, we found that these acidic conditions elicit two distinct responses: (i) a general and conserved response through which agrobacterium modulates gene expression patterns to adapt to environmental acidification and (ii) a highly specialized acid-mediated signaling re ...200817993523
the effect of co-cultivation and selection parameters on agrobacterium-mediated transformation of chinese soybean varieties.in the present study, an efficient agrobacterium-mediated gene transformation system was developed for soybean [glycine max (l.) merrill] based on the examinations of several factors affecting plant transformation efficiency. increased transformation efficiencies were obtained when the soybean cotyledonary node were inoculated with the agrobacterium inoculum added with 0.02% (v/v) surfactant (silwet l-77). the applications of silwet l-77 (0.02%) during infection and l-cysteine (600 mg l(-1)) dur ...200818004571
agrobacterium-mediated transformation of the temperate grass brachypodium distachyon (genotype bd21) for t-dna insertional mutagenesis.brachypodium distachyon is a promising model system for the structural and functional genomics of temperate grasses because of its physical, genetic and genome attributes. the sequencing of the inbred line bd21 (http://www.brachypodium.org) started in 2007. however, a transformation method remains to be developed for the community standard line bd21. in this article, a facile, efficient and rapid transformation system for bd21 is described using agrobacterium-mediated transformation of compact e ...200818004984
signal molecules in the peanut-bradyrhizobia interaction.main nodulation signal molecules in the peanut-bradyrhizobia interaction were examined. flavonoids exuded by arachis hypogaea l. cultivar tegua were genistein, daidzein and chrysin, the latest being released in lower quantities. thin layer chromatography analysis from genistein-induced bacterial cultures of three peanut bradyrhizobia resulted in an identical nod factor pattern, suggesting low variability in genes involved in the synthesis of these molecules. structural study of nod factor by mas ...200818030448
toxic effects of arsenic on sinorhizobium-medicago sativa symbiotic interaction.recently, the rhizobium-legume symbiotic interaction has been proposed as an interesting tool in bioremediation. however, little is known about the effect of most common contaminants on this process. the phytotoxic effects of arsenic on nodulation of medicago sativa have been examined in vitro using the highly arsenic resistant and symbiotically effective sinorhizobium sp. strain ma11. the bacteria were able to grow on plates containing as concentrations as high as 10 mm. nevertheless, as little ...200818039553
the hypothetical protein atu4866 from agrobacterium tumefaciens adopts a streptavidin-like fold.atu4866 is a 79-residue conserved hypothetical protein of unknown function from agrobacterium tumefaciens. protein sequence alignments show that it shares > or =60% sequence identity with 20 other hypothetical proteins of bacterial origin. however, the structures and functions of these proteins remain unknown so far. to gain insight into the function of this family of proteins, we have determined the structure of atu4866 as a target of a structural genomics project using solution nmr spectroscop ...200818042676
methanolysis of ethyl esters of n-acetyl amino acids catalyzed by cyclosophoraoses isolated from rhizobium meliloti.methanolysis of four ethyl esters, n-acetyl-l-phenylalanine ethyl ester, n-acetyl-l-tyrosine ethyl ester, n-acetyl-l-tryptophan ethyl ester, and ethyl phenylacetate was catalyzed by a mixture of microbial cyclooligosaccharides termed cyclosophoraoses isolated from rhizobium meliloti. cyclosophoraoses [cyclic-(1-->2)-beta-d-glucans, collectively 'cys'] are a mixture of large-ring molecules consisting of various numbers of glucose residues (17-27) linked by beta-(1-->2)-glycosidic bonds. cys as a ...200818045579
using 23s rdna to identify contaminations of escherichia coli in agrobacterium tumefaciens cultures. 200817945175
improvement of agrobacterium-mediated transformation in hi-ii maize (zea mays) using standard binary vectors.high-frequency transformation of maize (zea mays l.) using standard binary vectors is advantageous for functional genomics and other genetic engineering studies. recent advances in agrobacterium tumefaciens-mediated transformation of maize have made it possible for the public to transform maize using standard binary vectors without a need of the superbinary vector. while maize hi-ii has been a preferred maize genotype to use in various maize transformation efforts, there is still potential and n ...200817938932
application of physical and genetic map of rhizobium leguminosarum bv. trifolii ta1 to comparison of three closely related rhizobial genomes.a combined physical and genetic map of rhizobium leguminosarum biovar trifolii ta1 (rtta1) genome was constructed and used in comparison of chromosomal organization with the closely related r. leguminosarum bv. viciae 3841 (rlv) and rhizobium etli cnf42 (rhe). this approach allowed evaluation of chromosome and genome plasticity and provided important insights into r. leguminosarum lineage diversity. mssi, smii, paci, and i-ceui restriction endonucleases were chosen for the analysis, generating f ...200817960422
identification and functional analysis of the fusaricidin biosynthetic gene of paenibacillus polymyxa e681.fusaricidin, a peptide antibiotic consisting of six amino acids, has been identified as a potential antifungal agent from paenibacillus polymyxa. here, we report the complete sequence of the fusaricidin synthetase gene (fusa) identified from the genome sequence of a rhizobacterium, p. polymyxa e681. the gene encodes a polypeptide consisting of six modules in a single open-reading frame. interestingly, module six of fusa does not contain an epimerization domain, which suggests that the sixth amin ...200817980146
phytoremediation of coal mine spoil dump through integrated biotechnological approach.field experiment was conducted on mine spoil dump on an area of 10 ha, to restore the fertility and productivity of the coal mine spoil dump using integrated biotechnological approach. the approach involves use of effluent treatment plant sludge (etp sludge), as an organic amendment, biofertilizers and mycorrihzal fungi along with suitable plant species. the results of the study indicated that amendment with effluent treatment plant sludge (etp sludge), @ 50 ton/ha improved the physico-chemical ...200817980580
protein-protein interactions within type iii secretion system-dependent pili of rhizobium sp. strain ngr234.pili synthesized by the type iii secretion system of rhizobium species strain ngr234 are essential for protein secretion and thus for efficient symbiosis with many legumes. isolation and partial purification of these pili showed that they are composed of at least three proteins, nopa, nopb, and nopx. using biochemical assays, we show here that these proteins interact directly with one another.200817981961
agrobacterium tumefaciens c58 uses actr and fnrn to control nirk and nor expression.agrobacterium tumefaciens can grow anaerobically via denitrification. to learn more about how cells regulate production of nitrite and nitric oxide, experiments were carried out to identify proteins involved in regulating expression and activity of nitrite and nitric oxide reductase. transcription of nnrr, required for expression of these two reductases, was found to be under control of fnrn. insertional inactivation of the response regulator actr significantly reduced nirk expression and nir ac ...200817981975
phylogenetic diversity based on rrs, atpd, reca genes and 16s-23s intergenic sequence analyses of rhizobial strains isolated from vicia faba and pisum sativum in peru.in this study 17 isolates from effective nodules of vicia faba and pisum sativum var. macrocarpum growing in different soils from peru were isolated and characterized. the isolates, presenting 11 different rapd profiles, were distributed in three groups on the basis of their 16s-rflp patterns. the 16s rrna gene sequences of strains from 16s-rflp groups i, ii and iii were closely related (identities higher than 99.5%) to rhizobium leguminosarum bv. trifolii dsm 30141 (=atcc 14480), r. leguminosar ...200817985116
resistance to grapevine leafroll associated virus-2 is conferred by post-transcriptional gene silencing in transgenic nicotiana benthamiana.grapevine leafroll-associated virus-2 (glrav-2) is an important component of the leafroll disease complex in grapevine. we have previously sequenced the glrav-2 genome and identified the coat protein (cp) gene. the objective of this study is to test the concept of pathogen-derived resistance against a closterovirus associated with grapevine leafroll disease. because glrav-2 is capable of infecting nicotiana benthamiana, we decided to test the concept on this herbaceous host. thirty-seven t(0) tr ...200817912600
generation of transgenic plants of a potential oilseed crop camelina sativa by agrobacterium-mediated transformation.camelina sativa is an alternative oilseed crop that can be used as a potential low-cost biofuel crop or a source of health promoting omega-3 fatty acids. currently, the fatty acid composition of camelina does not uniquely fit any particular uses, thus limit its commercial value and large-scale production. in order to improve oil quality and other agronomic characters, we have developed an efficient and simple in planta method to generate transgenic camelina plants. the method included agrobacter ...200817899095
efficient generation of transgenic barley: the way forward to modulate plant-microbe interactions.stable genetic transformation represents the gold standard approach to the detailed elucidation of plant gene functions. this is particularly relevant in barley, an important experimental model widely employed in applied molecular, genetic and cell biological research, and biotechnology. presented are details of the establishment of a protocol for agrobacterium-mediated gene transfer to immature embryos, which enables the highly efficient generation of transgenic barley. advancements were achiev ...200817905476
overexpression of the arabidopsis gai gene in apple significantly reduces plant size.genetic engineering is an attractive method to obtain dwarf plants in order to eliminate the extensive use of growth retardants in horticultural crop production. in this study, we evaluated the potential of using the arabidopsis gai (gibberellic acid insensitive) gene to dwarf apple trees. the gai gene under 35s promoter was introduced in the apple rootstock a2 and the cultivars gravenstein and mcintosh through agrobacterium-mediated transformation. one transgenic clone was recovered for gravens ...200817932677
overexpression of hva1 gene from barley generates tolerance to salinity and water stress in transgenic mulberry (morus indica).late embryogenesis abundant (lea) proteins are members of a large group of hydrophilic proteins found primarily in plants. the barley hva1 gene encodes a group 3 lea protein and is induced by aba and water deficit conditions. we report here the over expression of hva1 in mulberry under a constitutive promoter via agrobacterium-mediated transformation. molecular analysis of the transgenic plants revealed the stable integration and expression of the transgene in the transformants. transgenic plant ...200817932781
genetic transformation of nerica, interspecific hybrid rice between oryza glaberrima and o. sativa, mediated by agrobacterium tumefaciens.we developed an efficient gene transfer method mediated by agrobacterium tumefaciens for introgression of new rice for africa (nerica) cultivars, which are derivatives of interspecific hybrids between oryza glaberrima steud. and o. sativa l. freshly isolated immature embryos were inoculated with a. tumefaciens lba4404 that harbored binary vector pbig-ubi::gus or pig121hm, which each carried a hygromycin-resistance gene and a gus gene. growth medium supplemented with 500 mg/l cefotaxime and 20 mg ...200817934736
production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration.genetic transformation of mature trees is difficult because adult tissues are recalcitrant to agrobacterium tumefaciens infection and transformation and because transgenic mature events are less competent for regeneration. we have shown that reinvigoration allows manipulation of the vegetative phase to increase the potential for transformation and regeneration without loss of competence for flowering and fruiting. to produce transgenic plants from clementine mandarin (citrus clementina hort. ex ...200817938114
development of an in planta method for transformation of alfalfa (medicago sativa).conventional methods in transforming alfalfa (medicago sativa) require multiple tissue culture manipulations that are time-consuming and expensive, while applicable only to a few highly regenerable genotypes. here, we describe a simple in planta method that makes it possible to transform a commercial variety without employing selectable marker genes. basically, young seedlings are cut at the apical node, cold-treated, and vigorously vortexed in an agrobacterium suspension also containing sand. a ...200817851774
structural biology of membrane-intrinsic beta-barrel enzymes: sentinels of the bacterial outer membrane.the outer membranes of gram-negative bacteria are replete with integral membrane proteins that exhibit antiparallel beta-barrel structures, but very few of these proteins function as enzymes. in escherichia coli, only three beta-barrel enzymes are known to exist in the outer membrane; these are the phospholipase ompla, the protease ompt, and the phospholipidcolon, two colonslipid a palmitoyltransferase pagp, all of which have been characterized at the structural level. structural details have al ...200817880914
metabolic engineering of novel ketocarotenoid production in carrot plants.carotenoids constitute a vast group of pigments that are ubiquitous throughout nature. carrot (daucus carota l.) roots provide an important source of dietary beta-carotene (provitamin a), alpha-carotene and lutein. ketocarotenoids, such as canthaxanthin and astaxanthin, are produced by some algae and cyanobacteria but are rare in plants. ketocarotenoids are strong antioxidants that are chemically synthesized and used as dietary supplements and pigments in the aquaculture and neutraceutical indus ...200817682834
efficient and rapid agrobacterium-mediated genetic transformation of durum wheat (triticum turgidum l. var. durum) using additional virulence genes.genetic transformation of wheat, using biolistics or agrobacterium, underpins a range of specific research methods for identifying genes and studying their function in planta. transgenic approaches to study and modify traits in durum wheat have lagged behind those for bread wheat. here we report the use of agrobacterium strain agl1, with additional vir genes housed in a helper plasmid, to transform and regenerate the durum wheat variety ofanto. the use of the basic psoup helper plasmid with no a ...200817638109
enrichment, isolation, and characterization of 4-chlorophenol-degrading bacterium rhizobium sp. 4-cp-20.the objectives of this research were to monitor the variations of species in mixed cultures during the enrichment period, isolate species and identify and characterize the pure 4-chlorophenol (4-cp) degrading strains from enriched mixed cultures. strain rhizobium sp. 4-cp-20 was isolated from the acclimated mixed culture. the dgge result indicated that strain rhizobium sp. 4-cp-20 was undetectable at the beginning but detectable after 2 weeks of enrichment. the optimum growth temperatures for rh ...200817636393
in vivo characterization of plant promoter element interaction using synthetic promoters.short directly-repeated (dr) dna enhancer elements of plant viral origin were analyzed for their ability, both individually and in combination, to influence in vivo transcription when inserted upstream from a minimal camv35s promoter. synthetic promoters containing multiple copies and/or combinations of dr cassettes were tested for their effect upon reporter gene (luciferase) expression using an agrobacteria-based leaf-infiltration transient assay and within stably transformed plants (nicotiana ...200817653610
growth and nitrogen fixation in lotus japonicus and medicago truncatula under nacl stress: nodule carbon metabolism.lotus japonicus and medicago truncatula model legumes, which form determined and indeterminate nodules, respectively, provide a convenient system to study plant-rhizobium interaction and to establish differences between the two types of nodules under salt stress conditions. we examined the effects of 25 and 50mm nacl doses on growth and nitrogen fixation parameters, as well as carbohydrate content and carbon metabolism of m. truncatula and l. japonicus nodules. the leghemoglobin (lb) content and ...200817728011
a plant growth promoting rhizobacterium, paenibacillus polymyxa strain gbr-1, suppresses root-knot nematode.exposure of root-knot nematode, meloidogyne incognita to various concentrations (5-100%) of culture filtrate of paenibacillus polymyxa gbr-1 under in vitro conditions significantly reduced egg hatch and caused substantial mortality of its juveniles. the increase in the exposure durations of juveniles to culture filtrate and its concentrations increased the mortality rate. similarly, higher concentrations increased its inhibitory effect on egg hatch. in higher concentrations (25-100%) egg hatch w ...200817706411
inactivation of pea genes by rnai supports the involvement of two similar o-methyltransferases in the biosynthesis of (+)-pisatin and of chiral intermediates with a configuration opposite that found in (+)-pisatin.(+)-pisatin, the major phytoalexin of pea (pisum sativum l.), is believed to be synthesized via two chiral intermediates, (-)-7,2'-dihydroxy-4',5'-methylenedioxyisoflavanone [(-)-sophorol] and (-)-7,2'-dihydroxy-4',5'-methylenedioxyisoflavanol [(-)-dmdi]; both have an opposite c-3 absolute configuration to that found at c-6a in (+)-pisatin. the expression of isoflavone reductase (ifr), which converts 7,2'-dihydroxy-4',5'-methylenedioxyisoflavone (dmd) to (-)-sophorol, sophorol reductase (sor), w ...200817707445
enhanced growth and nodulation of pigeon pea by co-inoculation of bacillus strains with rhizobium spp.endophytic bacteria which are known to reside in plant tissues have often been shown to promote plant growth. present study deals with the isolation of putative endophytes from the surface sterilized root nodules of pigeon pea (cajanus cajan) designated as non-rhizobial (nr) isolates. three of these non-rhizobial isolates called nr2, nr4 and nr6 showed plant growth promotion with respect to increase in plant fresh weight, chlorophyll content, nodule number and nodule fresh weight when co-inocula ...200817826983
defects in rhizobial cyclic glucan and lipopolysaccharide synthesis alter legume gene expression during nodule development.cdna array technology was used to compare transcriptome profiles of lotus japonicus roots inoculated with a mesorhizobium loti wild-type and two mutant strains affected in cyclic beta(1-2) glucan synthesis (cgs) and in lipopolysaccharide synthesis (lpsbeta2). expression of genes associated with the development of a fully functional nodule was significantly affected in plants inoculated with the cgs mutant. array results also revealed that induction of marker genes for nodule development was dela ...200818052882
pyrroloquinoline quinone is a plant growth promotion factor produced by pseudomonas fluorescens b16.pseudomonas fluorescens b16 is a plant growth-promoting rhizobacterium. to determine the factors involved in plant growth promotion by this organism, we mutagenized wild-type strain b16 using omegakm elements and isolated one mutant, k818, which is defective in plant growth promotion, in a rockwool culture system. a cosmid clone, pok40, which complements the mutant k818, was isolated from a genomic library of the parent strain. tn3-gusa mutagenesis of pok40 revealed that the genes responsible fo ...200818055583
binding site determinants for the lysr-type transcriptional regulator pcaq in the legume endosymbiont sinorhizobium meliloti.lysr-type transcriptional regulators represent one of the largest groups of prokaryotic regulators described to date. in the gram-negative legume endosymbiont sinorhizobium meliloti, enzymes involved in the protocatechuate branch of the beta-ketoadipate pathway are encoded within the pcadchgb operon, which is subject to regulation by the lysr-type protein pcaq. in this work, purified pcaq was shown to bind strongly (equilibrium dissociation constant, 0.54 nm) to a region at positions -78 to -45 ...200818055594
importance of co-cultivation medium ph for successful agrobacterium-mediated transformation of lilium x formolongi.an efficient system for agrobacterium-mediated transformation of lilium x formolongi was established by preventing the drastic drop of ph in the co-cultivation medium with mes. meristematic nodular calli were inoculated with an overnight culture of a. tumefaciens strain eha101 containing the plasmid pig121-hm which harbored intron-containing beta-glucuronidase (gus), hygromycin phosphotransferase (hpt), and neomycin phosphotransfease ii (nptii) genes. after three days of co-cultivation on 2 g/l ...200818060407
use of 4-methylindole or 7-methyl-dl-tryptophan in a transformant selection system based on the feedback-insensitive anthranilate synthase alpha-subunit of tobacco (asa2).effective selectable markers are needed for basic research and commercial applications that do not involve antibiotic or herbicide resistance. a novel selection system based on a feedback-insensitive anthranilate synthase alpha-subunit of tobacco (asa2) as selectable marker using either 4-methylindole (4mi) or 7-methyl-dl-tryptophan (7mt) as the selection agent was developed. we found that these two components were able to discriminate better between asa2 expressing and untransformed lines than ...200818060408
diversity of rhizobial bacteria isolated from nodules of the gypsophyte ononis tridentata l. growing in spanish soils.the aim of this work is to describe the diversity and phylogeny of rhizobial bacteria associated to nodules of ononis tridentata l. in different geographical regions of spain. twenty-two bacterial isolates were characterized using several molecular techniques (16s amplified ribosomal deoxyribonucleic acid restriction analysis, fingerprinting, and sequencing) and phylogenies were inferred from their 16s and nodc gene sequences. phylogenetically, the isolates grouped with the genera rhizobium, mes ...200818060448
the agrobacterium tumefaciens dnak: atpase cycle, oligomeric state and chaperone properties.dnak is a molecular chaperone that promotes cell survival during stress by preventing protein misfolding. the chaperone activity is regulated by nucleotide binding and hydrolysis events in the n-terminal atpase domain, which in turn mediate substrate binding and release in the c-terminal substrate binding domain. in this study we determined that atp hydrolysis was the rate limiting step in the atpase cycle of agrobacterium tumefaciens dnak (agt dnak); however the data suggested that agt dnak had ...200818061511
functional analysis of a novel abc transporter abc4 from magnaporthe grisea.the atp-binding cassette (abc) superfamily of membrane transporters has been implicated to play a role in pathogenesis in various phytopathogenic fungi. in an insertional mutagenesis screen for pathogenicity mutants of magnaporthe grisea obtained via agrobacterium tumefaciens-mediated transformation (atmt), a novel gene belonging to the abc transporter family was identified. the gene abc4 was predicted to be 5045 bp in length coding for a protein of 1654 amino acids. the mutant did not form func ...200818034832
a high-throughput agrobacterium-mediated transformation system for the grass model species brachypodium distachyon l.in the ongoing process of developing brachypodium distachyon as a model plant for temperate cereals and forage grasses, we have developed a high-throughput agrobacterium-mediated transformation system for a diploid accession. embryogenic callus, derived from immature embryos of the accession bdr018, were transformed with agrobacterium tumefaciens strain agl1 carrying two t-dna plasmids, pdm805 and pwbv-ds-ubi-bar-ds. transient and stable transformation efficiencies were optimised by varying the ...200818064538
firefly luciferase complementation imaging assay for protein-protein interactions in plants.the development of sensitive and versatile techniques to detect protein-protein interactions in vivo is important for understanding protein functions. the previously described techniques, fluorescence resonance energy transfer and bimolecular fluorescence complementation, which are used widely for protein-protein interaction studies in plants, require extensive instrumentation. to facilitate protein-protein interaction studies in plants, we adopted the luciferase complementation imaging assay. t ...200818065554
core residue replacements cause coiled-coil orientation switching in vitro and in vivo: structure-function correlations for osmosensory transporter prop.protein prop acts as an osmosensory transporter in diverse bacteria. c-terminal residues 468-497 of escherichia coli prop (propec) form a four-heptad homodimeric alpha-helical coiled coil. arg 488, at a core heptad a position, causes it to assume an antiparallel orientation. arg in the hydrophobic core of coiled coils is destabilizing, but arg 488 forms stabilizing interstrand salt bridges with asp 475 and asp 478. mutation r488i destabilizes the coiled coil and elevates the osmotic pressure at ...200818076193
kalanchoe blossfeldiana plants expressing the arabidopsis etr1-1 allele show reduced ethylene sensitivity.transgenic kalanchoe blossfeldiana poelln. with reduced ethylene sensitivity in flowers was obtained by agrobacterium tumefaciens-mediated transformation using the plasmid pbeo210 containing the mutant ethylene receptor gene etr1-1 from arabidopsis thaliana under the control of the flower-specific fbp1-promoter from petunia. three ethylene-resistent t0 lines, 300, 324 and 331, were selected and analyzed for postharvest-performance and morphological characteristics. line 324 was found to be infer ...200818080125
in crystallo capture of a michaelis complex and product-binding modes of a bacterial phosphotriesterase.the mechanism by which the binuclear metallophosphotriesterases (ptes, e.c. 3.1.8.1) catalyse substrate hydrolysis has been extensively studied. the mu-hydroxo bridge between the metal ions has been proposed to be the initiating nucleophile in the hydrolytic reaction. in contrast, analysis of some biomimetic systems has indicated that mu-hydroxo bridges are often not themselves nucleophiles, but act as general bases for freely exchangeable nucleophilic water molecules. herein, we present crystal ...200818082180
growth of hairy-root cultures in various bioreactors for the production of secondary metabolites.agrobacterium rhizogenes-mediated hairy-root cultures are a very promising alternative to the biotechnological exploitation of plant cell cultures. their characteristic capacity for secondary-metabolite production, inherent genetic stability reflected in stable productivity and the possibility of genetic manipulation to increase biosynthetic capacity have initiated a considerable interest, both as a fundamental research tool and as a source of valuable products. one of the most important limitat ...200818086010
rhizobium common nod genes are required for biofilm formation.in legume nitrogen-fixing symbioses, rhizobial nod genes are obligatory for initiating infection thread formation and root nodule development. here we show that the common nod genes, nodd1abc, whose products synthesize core nod factor, a chitin-like oligomer, are also required for the establishment of the three-dimensional architecture of the biofilm of sinorhizobium meliloti. common nod gene mutants form a biofilm that is a monolayer. moreover, adding nod factor antibody to s. meliloti cells in ...200818086203
molecular characterization of microbial mutations induced by ion beam irradiation.a positive selection system for gene disruption using a sucrose-sensitive transgenic rhizobium was established and used for the molecular characterization of mutations induced by ion beam irradiations. single nucleotide substitutions, insertions, and deletions were found to occur in the sucrose sensitivity gene, sacb, when the reporter line was irradiated with highly accelerated carbon and iron ion beams. in all of the insertion lines, fragments of essentially the same sequence and of approximat ...200818068202
diversity of nifh gene pools in the rhizosphere of two cultivars of sorghum (sorghum bicolor) treated with contrasting levels of nitrogen fertilizer.the diversity of nitrogen-fixing bacteria was assessed in the rhizospheres of two cultivars of sorghum (is 5322-c and ipa 1011) sown in cerrado soil amended with two levels of nitrogen fertilizer (12 and 120 kg ha(-1)). the nifh gene was amplified directly from dna extracted from the rhizospheres, and the pcr products cloned and sequenced. four clone libraries were generated from the nifh fragments and 245 sequences were obtained. most of the clones (57%) were closely related to nifh genes of un ...200818070072
combining x-ray and electron-microscopy data to solve crystal structures.low-resolution electron-microscopy reconstructions can be used as search models in molecular replacement or may be combined with existing monomeric structures in order to produce multimeric models suitable for molecular replacement. the technique is described in the case of viral and subviral particles as well as in the case of oligomeric proteins.200818094469
the quorum-sensing system avsr-avsi regulates both long-chain and short-chain acyl-homoserine lactones in agrobacterium vitis e26.agrobacterium vitis strain e26 is a nonpathogenic bacterium isolated from grape crown gall. in this study, the identification of a luxr-luxi type quorum-sensing system in strain e26 is reported. this system is involved in the induction of hypersensitive response on tobacco, but not its biocontrol activity. the deduced components avsi(e26) and avsr(e26) show the greatest similarity to avsi(f2/5) and avsr(f2/5), respectively from a. vitis strain f2/5. the mutant in avsi(e26) abolished the producti ...200817906938
rhau of rhizobium leguminosarum is a rhamnose mutarotase.of the nine genes comprising the l-rhamnose operon of rhizobium leguminosarum, rhau has not been assigned a function. the construction of a delta rhau strain revealed a growth phenotype that was slower than that of the wild-type strain, although the ultimate cell yields were equivalent. the transport of l-rhamnose into the cell and the rate of its phosphorylation were unaffected by the mutation. rhau exhibits weak sequence similarity to the formerly hypothetical protein yiil of escherichia coli ...200818156270
salicylic acid and systemic acquired resistance play a role in attenuating crown gall disease caused by agrobacterium tumefaciens.we investigated the effects of salicylic acid (sa) and systemic acquired resistance (sar) on crown gall disease caused by agrobacterium tumefaciens. nicotiana benthamiana plants treated with sa showed decreased susceptibility to agrobacterium infection. exogenous application of sa to agrobacterium cultures decreased its growth, virulence, and attachment to plant cells. using agrobacterium whole-genome microarrays, we characterized the direct effects of sa on bacterial gene expression and showed ...200818156296
thiamine limitation determines the transition from aerobic to fermentative-like metabolism in rhizobium etli ce3.both thiamine and biotin when added to minimal medium subcultures reversed the fermentative-like metabolism exhibited by rhizobium etli ce3. thiamine auxotrophs lacking thicoge genes were used to investigate the role of thiamine in this medium. a thic1169::minitn5lacz1 thiamine auxotroph subjected to the above subcultures resulted in growth arrest, reduced pyruvate-dehydrogenase activity, and a smaller amount of poly-beta-hydroxybutyrate compared with the ce3 strain. moreover, thic and thieb gen ...200818177306
recent advances in nitrogen-fixing acetic acid bacteria.nitrogen is an essential plant nutrient, widely applied as n-fertilizer to improve yield of agriculturally important crops. an interesting alternative to avoid or reduce the use of n-fertilizers could be the exploitation of plant growth-promoting bacteria (pgpb), capable of enhancing growth and yield of many plant species, several of agronomic and ecological significance. pgpb belong to diverse genera, including azospirillum, azotobacter, herbaspirillum, bacillus, burkholderia, pseudomonas, rhiz ...200818177965
efficient and high-throughput vector construction and agrobacterium-mediated transformation of arabidopsis thaliana suspension-cultured cells for functional genomics.we established a large-scale, high-throughput protocol to construct arabidopsis thaliana suspension-cultured cell lines, each of which carries a single transgene, using agrobacterium-mediated transformation. we took advantage of riken arabidopsis full-length (rafl) cdna clones and the gateway cloning system for high-throughput preparation of binary vectors carrying individual full-length cdna sequences. throughout all cloning steps, multiple-well plates were used to treat 96 samples simultaneous ...200818178967
physiological function of ispe, a plastid mep pathway gene for isoprenoid biosynthesis, in organelle biogenesis and cell morphogenesis in nicotiana benthamiana.isoprenoid biosynthesis in plants occurs by two independent pathways: the cytosolic mevalonate (mva) pathway and the plastidic methylerythritol phosphate (mep) pathway. in this study, we investigated the cellular effects of depletion of ispe, a protein involved in the mep pathway, using virus-induced gene silencing (vigs). the ispe gene is preferentially expressed in young tissues, and induced by light and methyl jasmonate. the gfp fusion protein of ispe was targeted to chloroplasts. reduction o ...200818180879
agrobacterium-mediated transformation and insertional mutagenesis in colletotrichum acutatum for investigating varied pathogenicity lifestyles.colletotrichum acutatum is a cosmopolitan pathogen causing economically important diseases known as anthracnose on a wide range of hosts. this fungus exhibits varied pathogenicity lifestyles and the tools essential to understand the molecular mechanisms are still being developed. the transformation methods currently available for this species for gene discovery and functional analysis involve protoplast transformation and are laborious and inefficient. we have developed a protocol for efficient ...200818183501
rivet-a tool for in vivo analysis of symbiotically relevant gene expression in sinorhizobium meliloti.despite significant advances in the development of sensitive tools for studying genetics and signal exchange in legume-rhizobium symbioses, many uncertainties remain about the in vivo role of bacterial and plant signals in symbiotic gene regulation. in this study, we adapted tnpr recombinase-based in vivo expression technology (rivet) to document gene regulation in sinorhizobium meliloti. the substrate for tnpr, the res1-tet-res1 cassette, is stably inherited when cloned into a neutral site of t ...200818184060
a large scale analysis of protein-protein interactions in the nitrogen-fixing bacterium mesorhizobium loti.global viewing of protein-protein interactions (ppis) is a useful way to assign biological roles to large numbers of proteins predicted by complete genome sequence. here, we systematically analyzed ppis in the nitrogen-fixing soil bacterium mesorhizobium loti using a modified high-throughput yeast two-hybrid system. the aims of this study are primarily on the providing functional clues to m. loti proteins that are relevant to symbiotic nitrogen fixation and conserved in other rhizobium species, ...200818192278
subpicosecond midinfrared spectroscopy of the pfr reaction of phytochrome agp1 from agrobacterium tumefaciens.phytochromes are light-sensing pigments found in plants and bacteria. for the first time, the p(fr) photoreaction of a phytochrome has been subject to ultrafast infrared vibrational spectroscopy. three time constants of 0.3 ps, 1.3 ps, and 4.0 ps were derived from the kinetics of structurally specific marker bands of the biliverdin chromophore of agp1-bv from agrobacterium tumefaciens after excitation at 765 nm. vis-pump-vis-probe experiments yield time constants of 0.44 ps and 3.3 ps for the un ...200818192363
a metagenomic analysis of soil bacteria extends the diversity of quorum-quenching lactonases.a metagenomic library of 10,121 clones, generated from bacteria inhabiting a pasture soil from france, was screened for the presence of fosmids conferring either n-acylhomoserine lactone (nahl) synthesis or nahl degradation ability upon their escherichia coli host. no clone producing nahls was identified whereas one, containing a 31 972 bp insert in fosmid p2h8, allowed nahl degradation. this led to the cloning and identification of a gene, qlca, encoding an nahl-lactonase activity, as judged by ...200818201196
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