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the effect of a unique halide-stabilizing residue on the catalytic properties of haloalkane dehalogenase data from agrobacterium tumefaciens c58.haloalkane dehalogenases catalyze the hydrolysis of carbon-halogen bonds in various chlorinated, brominated and iodinated compounds. these enzymes have a conserved pair of halide-stabilizing residues that are important in substrate binding and stabilization of the transition state and the halide ion product via hydrogen bonding. in all previously known haloalkane dehalogenases, these residues are either a pair of tryptophans or a tryptophan-asparagine pair. the newly-isolated haloalkane dehaloge ...201323490078
effects of bacteria on cadmium bioaccumulation in the cadmium hyperaccumulator plant beta vulgaris var. cicla l.to investigate the effects of two cadmium-tolerant bacteria, staphylococcus pasteuri (s. pasteuri x1) and agrobacterium tumefaciens (a. tumefaciens x2), on cadmium uptake by the cadmium hyperaccumulator plant beta vulgaris var. cicla l., a pot experiment with artificially contaminated soil was conducted. the results demonstrated that both cadmium-tolerant bacteria enhanced the dry weight of beta vulgaris var. cicla l. the total dry weights of plants in the control ck20, s. pasteuri x1 and a. tum ...201323488173
an accurate method for estimation of the intracellular aqueous volume of escherichia coli cells.the intracellular aqueous volumes (v(in)) of microorganisms are fundamental data that can be used for estimating absolute cellular enzyme and metabolite concentrations. because traditional methods are time-consuming and costly, the v(in) data have been largely estimated ambiguously. here we developed an nad(+) concentration-dependent method and demonstrated its usefulness for accurate estimation of the v(in) value of escherichia coli cells. the v(in) value of e. coli bl21(de3) cells was determin ...201323481146
agrobacterium tumefaciens-mediated transformation as a tool for insertional mutagenesis in thermophilic fungus thermomyces lanuginosus.to establish a stable transformation system of the thermophilic fungus thermomyces lanuginosus for its insertional mutagenesis.201223457794
enhanced tolerance to drought and salt stresses in transgenic faba bean (vicia faba l.) plants by heterologous expression of the pr10a gene from potato.we report for the first time that expression of potato pr10a gene in faba bean causes enhanced tolerance to drought and salinity. grain legumes such as soybean (glycine max l. merrill), pea (pisum sativum l.) and faba bean (vicia faba l.) are staple sources of protein for human and animal nutrition. among grain legumes, faba bean is particularly sensitive to abiotic stress (in particular osmotic stress due to lack of water or enhanced soil salinity) and often suffers from severe yield losses. ma ...201323455709
exploiting plant virus-derived components to achieve in planta expression and for templates for synthetic biology applications.this review discusses the varying roles that have been played by many plant-viral regulatory sequences and proteins in the creation of plant-based expression systems and virus particles for use in nanotechnology. essentially, there are two ways of expressing an exogenous protein: the creation of transgenic plants possessing a stably integrated gene construction, or the transient expression of the desired gene following the infiltration of the gene construct. both depend on disarmed strains of ag ...201323452220
in vivo analysis of dna binding and ligand interaction of blcr, an iclr-type repressor from agrobacterium tumefaciens.agrobacterium tumefaciens blcr represses transcription of the blcabc operon, which is involved in metabolism of γ-butyrolactone, and this repression is alleviated by succinate semialdehyde (ssa). blcr exists as a homodimer, and the blcabc promoter dna contains two blcr-binding sites (ir1 and ir2) that correspond to two blcr dimers. in this study, we established an in vivo system to examine the ssa-responsive control of blcr transcriptional regulation. the endogenous blcr, encoded in the patc58 p ...201323449918
anti-quorum sensing potential of crude kigelia africana fruit extracts.the increasing incidence of multidrug-resistant pathogens has stimulated the search for novel anti-virulence compounds. although many phytochemicals show promising antimicrobial activity, their power lies in their anti-virulence properties. thus the quorum sensing (qs) inhibitory activity of four crude kigelia africana fruit extracts was assessed qualitatively and quantitatively using the chromobacterium violaceum and agrobacterium tumefaciens biosensor systems. inhibition of qs-controlled viola ...201323447012
coordination of division and development influences complex multicellular behavior in agrobacterium tumefaciens.the α-proteobacterium agrobacterium tumefaciens has proteins homologous to known regulators that govern cell division and development in caulobacter crescentus, many of which are also conserved among diverse α-proteobacteria. in light of recent work demonstrating similarity between the division cycle of c. crescentus and that of a. tumefaciens, the functional conservation for this presumptive control pathway was examined. in c. crescentus the ctra response regulator serves as the master regulato ...201323437210
optimization of agrobacterium mediated genetic transformation of cotyledonary node explants of vigna radiata.a reproducible and highly efficient protocol for genetic transformation mediated by agrobacterium has been established for greengram (vigna radiata l. wilczek). double cotyledonary node (dcn) explants were inoculated with agrobacterium tumefaciens strain lba 4404 harboring a binary vector pcambia 2301 containing neomycin phosphotransferase (npt ii) gene as selectable marker, β-glucuronidase (gus) as a reporter (uida) gene and annexin 1 bj gene. important parameters like optical density of agroba ...201223420384
stable transformation of suspension-cultured glycyrrhiza inflata batalin cells with agrobacterium tumefaciens.a protocol for the efficient genetic transformation of licorice (glycyrrhiza inflata batalin) cells in suspension culture using agrobacterium tumefaciens-mediated t-dna delivery is described. g. inflata cells in suspension culture were infected with a. tumefaciens strain lba4404 harbouring the binary vector pcambia1303, which contains the beta-glucuronidase (gus) reporter gene and a hygromycin resistance gene (hpt ii), respectively, under the transcriptional control of the camv35s promoter. opti ...201223413755
soybean genetic transformation: a valuable tool for the functional study of genes and the production of agronomically improved plants.transgenic plants represent an invaluable tool for molecular, genetic, biochemical and physiological studies by gene overexpression or silencing, transposon-based mutagenesis, protein sub-cellular localization and/or promoter characterization as well as a breakthrough for breeding programs, allowing the production of novel and genetically diverse genotypes. however, the stable transformation of soybean cannot yet be considered to be routine because it depends on the ability to combine efficient ...201223412849
dna methylation mediated control of gene expression is critical for development of crown gall tumors.crown gall tumors develop after integration of the t-dna of virulent agrobacterium tumefaciens strains into the plant genome. expression of the t-dna-encoded oncogenes triggers proliferation and differentiation of transformed plant cells. crown gall development is known to be accompanied by global changes in transcription, metabolite levels, and physiological processes. high levels of abscisic acid (aba) in crown galls regulate expression of drought stress responsive genes and mediate drought st ...201323408907
establishment of an agroinoculation system for broad bean wilt virus 2.we determined the complete nucleotide sequence of a broad bean wilt virus 2 (bbwv-2) isolate from gentian in japan. the full-length rna1 and rna2 sequences, excluding poly(a) tails, were 5955 and 3600 nucleotides long, respectively. analysis indicated that, in contrast to other bbwv-2 isolates, the 5' end of both rna1 and rna2 starts with a guu sequence. we successfully inoculated nicotiana benthamiana with bbwv-2 by infiltrating a mixed suspension of two agrobacterium tumefaciens clones carryin ...201323404460
[scopolamine and hyoscyamine synthesis in hair roots culture of datura metel].to establish the hair roots culture system of datura metel and study the hair roots growth and biosynthesis of scopolamine and hyoscyamine in hair roots culturing system.201223397717
preparation of d-gulose from disaccharide lactitol using microbial and chemical methods.when an m31 strain of agrobacterium tumefaciens was grown in a mineral salt medium at 30 °c containing 1.0% lactitol as sole carbon source, a keto-sugar was efficiently accumulated in the supernatant. this oxidation from lactitol to the keto-sugar was caused by m31 cells grown with medium containing a disaccharide unit, including sucrose, lactitol, lactose, maltose, or maltitol, suggesting that the enzyme is inducible. m31 also demonstrated good growth characteristics in tryptic soy broth (tsb) ...201323391912
profligate biotin synthesis in α-proteobacteria - a developing or degenerating regulatory system?biotin (vitamin h) is a key enzyme cofactor required in all three domains of life. although this cofactor was discovered over 70 years ago and has long been recognized as an essential nutrient for animals, our knowledge of the strategies bacteria use to sense biotin demand is very limited. the paradigm mechanism is that of escherichia coli in which bira protein, the prototypical bi-functional biotin protein ligase, both covalently attaches biotin to the acceptor proteins of central metabolism an ...201323387333
efficient gene knockout in the maize pathogen setosphaeria turcica using agrobacterium tumefaciens-mediated transformation.setosphaeria turcica, a hemibiotrophic pathogenic dothideomycete, is the causal agent of northern leaf blight of maize, which periodically causes significant yield losses worldwide. to explore molecular mechanisms of fungal pathogenicity and virulence to the host, an efficient targeted gene knockout transformation system using agrobacterium tumefaciens was established with field collected strains. the starting materials, incubation time, induction medium type, agrobacterium cell density, and met ...201323384859
agroinoculation of beet necrotic yellow vein virus cdna clones results in plant systemic infection and efficient polymyxa betae transmission.agroinoculation is a quick and easy method for the infection of plants with viruses. this method involves the infiltration of tissue with a suspension of agrobacterium tumefaciens carrying binary plasmids harbouring full-length cdna copies of viral genome components. when transferred into host cells, transcription of the cdna produces rna copies of the viral genome that initiate infection. we produced full-length cdna corresponding to beet necrotic yellow vein virus (bnyvv) rnas and derived repl ...201323384276
[construction and pathogenicity tests of a mutated specific fragment scf73 in verticillium dahliae].to identify preliminarily the specific fragment scf73's function in verticillium dahlia virulence.201223383504
plant tumour biocontrol agent employs a trna-dependent mechanism to inhibit leucyl-trna synthetase.leucyl-trna synthetases (leurss) have an essential role in translation and are promising targets for antibiotic development. agrocin 84 is a leurs inhibitor produced by the biocontrol agent agrobacterium radiobacter k84 that targets pathogenic strains of a. tumefaciens, the causative agent of plant tumours. agrocin 84 acts as a molecular trojan horse and is processed inside the pathogen into a toxic moiety (tm84). here we show using crystal structure, thermodynamic and kinetic analyses, that thi ...201323361008
unexpected phytostimulatory behavior for escherichia coli and agrobacterium tumefaciens model strains.plant-beneficial effects of bacteria are often underestimated, especially for well-studied strains associated with pathogenicity or originating from other environments. we assessed the impact of seed inoculation with the emblematic bacterial models agrobacterium tumefaciens c58 (plasmid-cured) or escherichia coli k-12 on maize seedlings in nonsterile soil. compared with the noninoculated control, root biomass (with a. tumefaciens or e. coli) and shoot biomass (with a. tumefaciens) were enhanced ...201323360460
tsw gene-based resistance is triggered by a functional rna silencing suppressor protein of the tomato spotted wilt virus.as a result of contradictory reports, the avirulence (avr) determinant that triggers tsw gene-based resistance in capsicum annuum against the tomato spotted wilt virus (tswv) is still unresolved. here, the n and nss genes of resistance-inducing (ri) and resistance-breaking (rb) isolates were cloned and transiently expressed in resistant capsicum plants to determine the identity of the avr protein. it was shown that the nss(ri) protein triggered a hypersensitive response (hr) in tsw-containing ca ...201323360130
type iv secretion machinery: molecular architecture and function.bacteria have evolved several secretion machineries to bring about transport of various virulence factors, nutrients, nucleic acids and cell-surface appendages that are essential for their pathogenesis. t4s (type iv secretion) systems are versatile secretion systems found in various gram-negative and gram-positive bacteria and in few archaea. they are large multisubunit translocons secreting a diverse array of substrates varying in size and nature from monomeric proteins to nucleoprotein complex ...201323356253
expression and large-scale production of human tissue plasminogen activator (t-pa) in transgenic tobacco plants using different signal peptides.an attempt was made to assess the expression level and targeting of a human protein entitled recombinant tissue plasminogen activator (rt-pa) through accumulation in three cellular compartments including the endoplasmic reticulum and cytosolic and apoplastic spaces in transgenic tobacco plants. in this context, three chimeric constructs pbi-sp-tpa, pbi-tpa-kdel, and pbi-ext-tpa were employed and transferred into the tobacco plants through a popular transformation-based system called agrobacteriu ...201323354501
isolation and characterization of quorum-sensing signalling molecules in pseudomonas aeruginosa isolates recovered from nosocomial infections.pseudomonas aeruginosa is one of the most common pathogens in nosocomial infections. many studies have documented the role of quorum-sensing (qs) systems in antibiotic tolerance of p. aeruginosa. n-acyl homoserine lactones (ahls) serve as qs signalling molecules and can be a target for modulating bacterial pathogenicity. in this study, nosocomial isolates of p. aeruginosa were characterized for the presence of different types of qs signalling molecules. ahls were solvent extracted and quantified ...201323331337
an r2r3 myb transcription factor determines red petal colour in an actinidia (kiwifruit) hybrid population.red colour in kiwifruit results from the presence of anthocyanin pigments. their expression, however, is complex, and varies among genotypes, species, tissues and environments. an understanding of the biosynthesis, physiology and genetics of the anthocyanins involved, and the control of their expression in different tissues, is required. a complex, the mbw complex, consisting of r2r3-myb and bhlh transcription factors together with a wd-repeat protein, activates anthocyanin 3-o-galactosyltransfe ...201323324587
ellagic acid derivatives from terminalia chebula retz. downregulate the expression of quorum sensing genes to attenuate pseudomonas aeruginosa pao1 virulence.burgeoning antibiotic resistance in pseudomonas aeruginosa has necessitated the development of anti pathogenic agents that can quench acylhomoserine lactone (ahl) mediated qs with least risk of resistance. this study explores the anti quorum sensing potential of t. chebula retz. and identification of probable compounds(s) showing anti qs activity and the mechanism of attenuation of p. aeruginosa pao1 virulence factors.201323320085
enhanced withanolide production by overexpression of squalene synthase in withania somnifera.withania somnifera commonly known as ashwagandha, is held in high repute in traditional indian medicine, largely due to the presence of steroidal lactone phytocompounds collectively known as withanolides, such as withanolide a, withaferin a and withanone. these withanolides have diverse pharmacological properties and are prospective high-value drug candidates. to meet the ever-increasing demands of these compounds, plant cell technology offers a viable alternative. in this study, a key enzyme in ...201323313565
[transgenic plant regeneration of tobacco (nicotiana tabacum) haboring mammalian cyp2e1 gene].cyp2e1 enzyme encoded by cyp2e1 gene plays an important role in metabolism of heterogeneous organics in mammalian liver cells. the transgenic plant with cyp2e1 can metabolize various low molecular weight organic pollutants. however, it is unclear the mechanism of expression control of cyp2e1 in transgenic plant. in this study, plasmid psld50-6 with cyp2e1 and pkh200 with gus as control were transformed into agrobacterium tumefaciens gv3101 separately. then, the cyp2e1 or gus genes were transferr ...201223311134
optimization of factors influencing microinjection method for agrobacterium tumefaciens-mediated transformation of tomato.a simple and efficient protocol for agrobacterium-mediated genetic transformation of tomato was developed using combination of non-tissue culture and micropropagation systems. initially, esam region of 1-day-old germinated tomato seeds were microinjected for one to five times with agrobacterium inoculums (od(600) = 0.2-1.0). the germinated seeds were cocultivated in the ms medium fortified with (0-200 mm) acetosyringone and minimal concentrations of (0-20 mg l(-1)) kanamycin, and the antibiotic ...201323306888
agrobacterium tumefaciens-mediated transformation of the soybean pathogen phomopsis longicolla.to facilitate functional genomics in the soybean pathogen phomopsis longicolla, we developed a robust agrobacterium tumefaciens-mediated transformation system that yielded 150-250 transformants per 1×10(6) conidia of p. longicolla. this first report of p. longicolla transformation provides a useful tool for insertional mutagenesis in an increasingly important pathogen of soybean.201323305924
isolation of bacterial type iv machine subassemblies.the bacterial type iv secretion systems (t4sss) deliver dna and protein substrates to bacterial and eukaryotic target cells generally by a mechanism requiring direct contact between donor and target cells. recent advances in defining the architectures of t4sss have been made through isolation of machine subassemblies for further biochemical and ultrastructural analysis. here, we describe a protocol for isolation and characterization of virb protein complexes from the paradigmatic virb/vird4 t4ss ...201323299736
purification and biochemical characterization of a highly thermostable bacteriocin isolated from brevibacillus brevis strain gm100.a bacteriocin-producing (11,000 au ml(-1)) strain was isolated from the rhizosphere of healthy algerian plants ononis angustissima lam., and identified as brevibacillus brevis strain gm100. the bacteriocin, called bac-gm100, was purified to homogeneity from the culture supernatant, and, based on maldi-tof/ms analysis, was a monomer protein with a molecular mass of 4375.66 da. the 21 n-terminal residues of bac-gm100 displayed 65% homology with thurincin h from bacillus thuringiensis. bac-gm100 wa ...201323291759
rhizobium radiobacter endophthalmitis following intravitreal ranibizumab injection.we present the first reported case of acute endophthalmitis due to rhizobium radiobacter after an intravitreal injection of ranibizumab for neovascular age-related macular degeneration.201223275789
transcription factor atbzip60 regulates expression of ca(2+)-dependent protein kinase genes in transgenic cells.the arabidopsis thaliana bzip60 (atbzip60) transcription factor regulates stress signaling. however, its molecular mechanism remains to be elucidated. in this investigation, cell suspension cultures of two different plant species rice (oryza sativa l.) and white pine (pinus strobes l.) were transformed using agrobacterium tumefaciens strain lba4404 harboring pbi-atzip60. integration of the atbzip60 gene into the genome of rice and white pine has been confirmed by polymerase chain reaction (pcr), ...201223275191
agrobacterium tumefaciens recognizes its host environment using chve to bind diverse plant sugars as virulence signals.agrobacterium tumefaciens is a broad host range plant pathogen that combinatorially recognizes diverse host molecules including phenolics, low ph, and aldose monosaccharides to activate its pathogenic pathways. chromosomal virulence gene e (chve) encodes a periplasmic-binding protein that binds several neutral sugars and sugar acids, and subsequently interacts with the vira/virg regulatory system to stimulate virulence (vir) gene expression. here, a combination of genetics, x-ray crystallography ...201223267119
agrobacterium tumefaciens-mediated transformation of penicillium expansum pe-12 and its application in molecular breeding.lipase produced by penicillium expansum is widely used in laundry detergent and leather industry; however, the absence of an efficient transformation technology sets a major obstacle for further enhancement of its lipase productivity through advanced gene engineering. in this work, agrobacterium tumefaciens-mediated transformation (atmt) was investigated for p. expansum pe-12 transformation, using hygromycin phosphotransferase (hph) as a selectable marker gene. as a result, we revealed that the ...201223265791
assignment of pterin deaminase activity to an enzyme of unknown function guided by homology modeling and docking.of the over 22 million protein sequences in the nonredundant trembl database, fewer than 1% have experimentally confirmed functions. structure-based methods have been used to predict enzyme activities from experimentally determined structures; however, for the vast majority of proteins, no such structures are available. here, homology models of a functionally uncharacterized amidohydrolase from agrobacterium radiobacter k84 (arad3529) were computed on the basis of a remote template structure. th ...201323256477
genetic diversity and symbiotic evolution of rhizobia from root nodules of coronilla varia.ninety symbiotic rhizobial isolates from root nodules of coronilla varia growing in the shaanxi province of china were characterized. combined with the results of rflp patterns, six genotypes were defined among the rhizobial strains and they were divided into three genomic genera. these included mesorhizobium sp., m. alhagi, m. amorphae, m. metallidurans/m. gobiense as the dominant group (86.7%), and rhizobium yanglingense and agrobacterium tumefaciens as the minor groups, according to analysis ...201323245852
reconciliation of sequence data and updated annotation of the genome of agrobacterium tumefaciens c58, and distribution of a linear chromosome in the genus agrobacterium.two groups independently sequenced the agrobacterium tumefaciens c58 genome in 2001. we report here consolidation of these sequences, updated annotation, and additional analysis of the evolutionary history of the linear chromosome, which is apparently limited to the biovar i group of agrobacterium.201323241979
the calcineurin-responsive transcription factor crz1 is required for conidation, full virulence and dmi resistance in penicillium digitatum.the ca(2+)/calmodulin-dependent pathway is a critical signal transduction pathway that mediates various cellular responses in multiple organisms. calcineurin acts through zinc finger transcription factor crz1 in fungi. in the present study, we identified an ortholog of crz1, pdcrz1, in penicillium digitatum, the most important pathogen of postharvest citrus. to determine the function(s) of pdcrz1, a gene-disruption mutant was generated using an agrobacterium tumefaciens-mediated transformation ( ...201323238263
canthaxanthin production with modified mucor circinelloides strains.canthaxanthin is a natural diketo derivative of β-carotene primarily used by the food and feed industries. mucor circinelloides is a β-carotene-accumulating zygomycete fungus and one of the model organisms to study the carotenoid biosynthesis in fungi. in this study, the β-carotene ketolase gene (crtw) of the marine bacterium paracoccus sp. n81106 fused with fungal promoter and terminator regions was integrated into the m. circinelloides genome to construct stable canthaxanthin-producing strains ...201323224586
functional annotation and three-dimensional structure of an incorrectly annotated dihydroorotase from cog3964 in the amidohydrolase superfamily.the substrate specificities of two incorrectly annotated enzymes belonging to cog3964 from the amidohydrolase superfamily were determined. this group of enzymes are currently misannotated as either dihydroorotases or adenine deaminases. atu3266 from agrobacterium tumefaciens c58 and oant2987 from ochrobactrum anthropi atcc 49188 were found to catalyze the hydrolysis of acetyl-(r)-mandelate and similar esters with values of k(cat)/k(m) that exceed 10(5) m(-1) s(-1). these enzymes do not catalyze ...201223214420
a really useful pathogen, agrobacterium tumefaciens.bacteria of the genus agrobacterium are very useful and unusual plant pathogens. through a rare inter-kingdom dna transfer, the bacteria move some of their genes into their host's genome, thereby inducing the host cells to proliferate and produce opines, nutrients sources for the pathogen. agrobacterium's ability to transfer dna makes can be adapted to introduce other genes, such as those encoding useful traits, into plant genomes. the development of agrobacterium as a tool to transform plants i ...201223213133
transformation of miniature potted rose (rosa hybrida cv. linda) with p( sag12 )-ipt gene delays leaf senescence and enhances resistance to exogenous ethylene.key message : the p ( sag12 ) -ipt gene was transferred to miniature rose, as the first woody species, resulting in increased ethylene resistance due to specific up-regulation of the ipt gene under senescence promoting conditions. transgenic plants of rosa hybrida 'linda' were obtained via transformation with agrobacterium tumefaciens strain harboring the binary vector psg529(+) containing the p( sag12 )-ipt construct. a. tumefaciens strains agl1, gv3850 and lba4404 (containing p(35s)-intgus gen ...201323207761
long chain n-acyl homoserine lactone production by enterobacter sp. isolated from human tongue surfaces.we report the isolation of n-acyl homoserine lactone-producing enterobacter sp. isolate t1-1 from the posterior dorsal surfaces of the tongue of a healthy individual. spent supernatants extract from enterobacter sp. isolate t1-1 activated the biosensor agrobacterium tumefaciens ntl4(pzlr4), suggesting production of long chain ahls by these isolates. high resolution mass spectrometry analysis of these extracts confirmed that enterobacter sp. isolate t1-1 produced a long chain n-acyl homoserine la ...201223202161
a septation related gene acseph in acremonium chrysogenum is involved in the cellular differentiation and cephalosporin production.t-dna inserted mutants of acremonium chrysogenum were constructed by agrobacterium tumefaciens-mediated transformation (atmt). one mutant 1223 which grew slowly was selected. tail-pcr and sequence analysis indicated that a putative septation protein encoding gene acseph was partially deleted in this mutant. acseph contains nine introns, and its deduced protein acseph has a conserved serine/threonine protein kinase catalytic (s_tkc) domain at its n-terminal region. acseph shows high similarity wi ...201323201539
protective efficacy against chlamydophila psittaci by oral immunization based on transgenic rice expressing momp in mice.avian chlamydiosis is caused by chlamydophila psittaci (cp. psittaci) and major outer membrane protein (momp) of cp. psittaci is an excellent vaccine candidate. in this study, the momp gene was expressed in rice callus by the agrobacterium tumefaciens vector. the production of protein in transgenic rice seeds was confirmed and quantified by western-blot and elisa, the results demonstrating that the antigen was expressed stably. the transgenic rice seeds expressing the momp protein were administe ...201323196208
in planta expression or delivery of potato aphid macrosiphum euphorbiae effectors me10 and me23 enhances aphid fecundity.the interactions between aphids and their host plants seem to be analogous to those of plant-microbial pathogens. unlike microbial pathogen effectors, little is known about aphid effectors and their ability to interfere with host immunity. to date, only three functional aphid effectors have been reported. to identify potato aphid (macrosiphum euphorbiae) effectors, we developed a salivary gland transcriptome using illumina technology. we generated 85 million illumina reads from salivary glands a ...201323194342
[synthesis and antimicrobial ivestigation of 8-quinolinols glucosaminides].in the phase transfer system solid calcium carbonate--acetonitrile per acetylated o-beta-d-glucosaminides of 8-quinolinol, 2-methyl-8-quinolinol and 5-chloro-8-quinolinol were synthesized. zemplen deacetylation was led to corresponding trioles. obtained o-beta-d-glucosaminides were identified with 1h-nmr spectroscopy. antimicrobial activity of deacetylated forms has been studied in a test of inhibition of bioluminescence of marine luminescent bacteria vibrio fischeri f1 and method of the serial ...201323189563
the 2.5 å structure of the enterococcus conjugation protein tram resembles virb8 type iv secretion proteins.conjugative plasmid transfer is the most important means of spreading antibiotic resistance and virulence genes among bacteria and therefore presents a serious threat to human health. the process requires direct cell-cell contact made possible by a multiprotein complex that spans cellular membranes and serves as a channel for macromolecular secretion. thus far, well studied conjugative type iv secretion systems (t4ss) are of gram-negative (g-) origin. although many medically relevant pathogens ( ...201223188825
one out of four: hspl but no other small heat shock protein of agrobacterium tumefaciens acts as efficient virulence-promoting virb8 chaperone.alpha-crystallin-type small heat shock proteins (shsps) are ubiquitously distributed in most eukaryotes and prokaryotes. four shsp genes named hspl, hspc, hspat1, and hspat2 were identified in agrobacterium tumefaciens, a plant pathogenic bacterium capable of unique interkingdom dna transfer via type iv secretion system (t4ss). hspl is highly expressed in virulence-induced growth condition and functions as a virb8 chaperone to promote t4ss-mediated dna transfer. here, we used genetic and biochem ...201223185409
use of gfp-tagged strains of penicillium digitatum and penicillium expansum to study host-pathogen interactions in oranges and apples.penicillium digitatum and penicillium expansum are responsible for green and blue molds in citrus and pome fruits, respectively, which result in major monetary losses worldwide. in order to study their infection process in fruits, we successfully introduced a green fluorescent protein (gfp) encoding gene into wild type p. digitatum and p. expansum isolates, using agrobacterium tumefaciens-mediated transformation (atmt), with hygromycin b resistance as the selectable marker. to our knowledge, thi ...201223177056
bacteriophytochrome controls carotenoid-independent response to photodynamic stress in a non-photosynthetic rhizobacterium, azospirillum brasilense sp7.ever since the discovery of the role of bacteriophytochrome (bphp) in inducing carotenoid synthesis in deinococcus radiodurans in response to light the role of bphps in other non-photosynthetic bacteria is not clear yet. azospirillum brasilense, a non-photosynthetic rhizobacterium, harbours a pair of bphps out of which abbphp1 is a homolog of atbphp1 of agrobacterium tumefaciens. by overexpression, purification, biochemical and spectral characterization we have shown that abbphp1 is a photochrom ...201223173079
development of an efficient transformation method by agrobacterium tumefaciens and high throughput spray assay to identify transgenic plants for woodland strawberry (fragaria vesca) using nptii selection.key message : we developed an efficient agrobacterium -mediated transformation method using an ac/ds transposon tagging construct for f. vesca and high throughput paromomycin spray assay to identify its transformants for strawberry functional genomics. genomic resources for rosaceae species are now readily available, including the fragaria vesca genome, est sequences, markers, linkage maps, and physical maps. the rosaceae genomic executive committee has promoted strawberry as a translational gen ...201323160638
the promoter of an a9 homolog from the conifer cryptomeria japonica imparts male strobilus-dominant expression in transgenic trees.key message : gus analysis in cryptomeria japonica revealed that the cjmale1 promoter is activated in the male strobilus of c. japonica. toward the development of male sterile technology for cryptomeria japonica, a male strobilus-dominant promoter of c. japonica was isolated. the cjmale1 gene was isolated from a male strobilus-specific suppression subtractive hybridization (ssh) library, and the promoter was isolated by the tail-pcr method. to characterize the cjmale1 promoter, β-glucuronidase ( ...201323160637
development of insect-resistant transgenic cotton with chimeric tvip3a accumulating in chloroplasts.an optimized vip3a gene, designated as vip3a* was chemically synthesized and a thi1 gene chloroplast transit peptide coding sequence was attached to its 5' end to produce the tvip3a*. vip3a* and tvip3a* genes were transformed into gossypium hirsutum cv. zhongmiansuo35 mediated by agrobacterium tumefaciens. four independent transgenic t1 lines with single-copy insertions and unchanged phenotypes (ctv1 and ctv2 for tvip3a*, and cv1 and cv2 for vip3a*) were selected by polymerase chain reaction (pc ...201323143498
agrobacterium rhizogenes-induced cotton hairy root culture as an alternative tool for cotton functional genomics.although well-accepted as the ultimate method for cotton functional genomics, agrobacterium tumefaciens-mediated cotton transformation is not widely used for functional analyses of cotton genes and their promoters since regeneration of cotton in tissue culture is lengthy and labor intensive. in certain cases, a. rhizogenes-induced hairy root culture has been a suitable molecular tool for functional analyses of genes and promoters for plants that are difficult to regenerate by a. tumefaciens-medi ...201323143493
development and bioassay of transgenic chinese cabbage expressing potato proteinase inhibitor ii gene.lepidopteran larvae are the most injurious pests of chinese cabbage production. we attempted the development of transgenic chinese cabbage expressing the potato proteinase inhibitor ii gene (pinii) and bioassayed the pest-repelling ability of these transgenic plants. cotyledons with petioles from aseptic seedlings were used as explants for agrobacterium-mediated in vitro transformation. agrobacterium tumefaciens c58 contained the binary vector pbbbasta-pinii-bar comprising pinii and bar genes. p ...201223136521
recent advances in soybean transformation and their application to molecular breeding and genomic analysis.herbicide-resistant transgenic soybean plants hold a leading market share in the usa and other countries, but soybean has been regarded as recalcitrant to transformation for many years. the cumulative and, at times, exponential advances in genetic manipulation have made possible further choices for soybean transformation. the most widely and routinely used transformation systems are cotyledonary node-agrobacterium-mediated transformation and somatic embryo-particle-bombardment-mediated transform ...201223136488
tnt1 retrotransposon mutagenesis: a tool for soybean functional genomics.insertional mutagenesis is a powerful tool for determining gene function in both model and crop plant species. tnt1, the transposable element of tobacco (nicotiana tabacum) cell type 1, is a retrotransposon that replicates via an rna copy that is reverse transcribed and integrated elsewhere in the plant genome. based on studies in a variety of plants, tnt1 appears to be inactive in normal plant tissue but can be reactivated by tissue culture. our goal was to evaluate the utility of the tnt1 retr ...201323124322
gateway vectors for transformation of cereals.until now, the availability of vectors for transgenic research in cereal crops has been rather limited. we present a novel collection of agrobacterium tumefaciens binary t-dna vectors compatible with gateway recombinational cloning that facilitate the modular assembly of genes of interest together with new regulatory sequences, such as strong constitutive or endosperm-specific brachypodium distachyon promoters. this resource aims at streamlining the creation of vectors and transgenes designed to ...201323121806
one-step agrobacterium mediated transformation of eight genes essential for rhizobium symbiotic signaling using the novel binary vector system phuge.advancement in plant research is becoming impaired by the fact that the transfer of multiple genes is difficult to achieve. here we present a new binary vector for agrobacterium tumefaciens mediated transformation, phuge-red, in concert with a cloning strategy suited for the transfer of up to nine genes at once. this vector enables modular cloning of large dna fragments by employing gateway technology and contains dsred1 as visual selection marker. furthermore, an r/rs inducible recombination sy ...201223112864
endogenous silencing of puccinia triticina pathogenicity genes through in planta-expressed sequences leads to the suppression of rust diseases on wheat.rust fungi are destructive plant pathogens. the draft genomes of several wheat-infecting species have been released and potential pathogenicity genes identified through comparative analyses to fungal pathogens that are amenable to genetic manipulation. functional gene analysis tools are needed to understand the infection process of these obligate parasites and to confirm whether predicted pathogenicity genes could become targets for disease control. we have modified an agrobacterium tumefaciens- ...201323110316
phosphorus limitation increases attachment in agrobacterium tumefaciens and reveals a conditional functional redundancy in adhesin biosynthesis.bacterial responses to phosphorus limitation, commonly inorganic phosphate (p(i)), are important survival mechanisms in a variety of environments. the two-component sensor kinase phor and its cognate response regulator phob are central to the p(i) limitation response of many bacteria and control the large pho regulon. limitation for p(i) significantly increased attachment and biofilm formation by the plant pathogen agrobacterium tumefaciens, and this was driven by phob. surprisingly, it was also ...201223103488
evaluation of four agrobacterium tumefaciens strains for the genetic transformation of tomato (solanum lycopersicum l.) cultivar micro-tom.key message : agrobacterium tumefaciens strains differ not only in their ability to transform tomato micro-tom, but also in the number of transgene copies that the strains integrate in the genome. the transformation efficiency of tomato (solanum lycopersicum l.) cv. micro-tom with agrobacterium tumefaciens strains agl1, eha105, gv3101, and mp90, harboring the plasmid pbi121 was compared. the presence of the nptii and/or uida transgenes in regenerated t(0) plants was determined by pcr, southern b ...201323099543
the pepper extracellular xyloglucan-specific endo-β-1,4-glucanase inhibitor protein gene, caxegip1, is required for plant cell death and defense responses.plants produce various proteinaceous inhibitors to protect themselves against microbial pathogen attack. a xyloglucan-specific endo-β-1,4-glucanase inhibitor1 gene, caxegip1, was isolated and functionally characterized in pepper (capsicum annuum) plants. caxegip1 was rapidly and strongly induced in pepper leaves infected with avirulent xanthomonas campestris pv vesicatoria, and purified caxegip1 protein significantly inhibited the hydrolytic activity of the glycoside hydrolase74 family xylogluca ...201223093361
improvement of coenzyme q10 production: mutagenesis induced by high hydrostatic pressure treatment and optimization of fermentation conditions.coenzyme q10 (coq10, ubiquinone), a potent antioxidative dietary supplement, was produced by submerged fermentation using agrobacterium tumefaciens instead of chemical synthesis or solvent extraction. agrobacterium tumefaciens 1.2554 was subjected to mutagenesis using a series of treatments including high hydrostatic pressure (hhp) treatment, uv irradiation, and diethyl sulfate (des) treatment to obtain mutant strains showing higher coq10 production than wild-type strains. a mutant strain pk38 w ...201223091351
metabolic modeling of denitrification in agrobacterium tumefaciens: a tool to study inhibiting and activating compounds for the denitrification pathway.a metabolic network model for facultative denitrification was developed based on experimental data obtained with agrobacterium tumefaciens. the model includes kinetic regulation at the enzyme level and transcription regulation at the enzyme synthesis level. the objective of this work was to study the key factors regulating the metabolic response of the denitrification pathway to transition from oxic to anoxic respiration and to find parameter values for the biological processes that were modeled ...201223087683
genetic transformation of european chestnut somatic embryos with a native thaumatin-like protein (cstl1) gene isolated from castanea sativa seeds.the availability of a system for direct transfer of antifungal candidate genes into european chestnut (castanea sativa mill.) would offer an alternative approach to conventional breeding for production of chestnut trees tolerant to ink disease caused by phytophthora spp. for the first time, a chestnut thaumatin-like protein gene (cstl1), isolated from chestnut cotyledons, has been overexpressed in three chestnut somatic embryogenic lines. transformation experiments have been performed using an a ...201223086811
methionine biosynthesis in agrobacterium tumefaciens: study of the first enzyme.here we characterize the first step in methionine biosynthesis in agrobacterium tumefaciens, an α-proteobacterium. we explored the meta gene and its products and found several unique properties. although the gene was annotated as a homoserine transsuccinylase, based upon sequence similarity to characterized homologs in other bacteria, including escherichia coli, the enzyme uses acetyl-coa as a substrate and therefore is functionally a transacetylase. moreover, the protein is thermolabile and the ...201323085540
polar auxin transport is essential for gall formation by pantoea agglomerans on gypsophila.the virulence of the bacterium pantoea agglomerans pv. gypsophilae (pag) on gypsophila paniculata depends on a type iii secretion system (t3ss) and its effectors. the hypothesis that plant-derived indole-3-acetic acid (iaa) plays a major role in gall formation was examined by disrupting basipetal polar auxin transport with the specific inhibitors 2,3,5-triiodobenzoic acid (tiba) and n-1-naphthylphthalamic acid (npa). on inoculation with pag, galls developed in gypsophila stems above but not belo ...201323083316
overexpression of the cytochrome p450 monooxygenase (cyp71av1) and cytochrome p450 reductase (cpr) genes increased artemisinin content in artemisia annua (asteraceae).finding an efficient and affordable treatment against malaria is still a challenge for medicine. artemisinin is an effective anti-malarial drug isolated from artemisia annua. however, the artemisinin content of a. annua is very low. we used transgenic technology to increase the artemisinin content of a. annua by overexpressing cytochrome p450 monooxygenase (cyp71av1) and cytochrome p450 reductase (cpr) genes. cyp71av1 is a key enzyme in the artemisinin biosynthesis pathway, while cpr is a redox ...201223079824
multiplex pcr assay for detection of human interferon alpha2b gene in transgenic plants.during the last decade interferons are regarded as potent candidates for generation of plant-based edible vaccines because of broad spectrum of antiviral activities and adjuvant properties. establishment and certification of numerous interferon producing plant systems requests development of fast and efficient multiplex pcr protocol for the transgene detection in gm plants. here we represent a protocol for simultaneous amplification in one assay of fragments of hifn alpha 2b gene and two control ...201623074956
agrobacterium tumefaciens-mediated transformation of phellodendron amurense rupr. using mature-seed explants.an efficient transformation protocol was developed for agrobacterium-mediated transformation of phellodendron amurense rupr. for using explants from mature seeds. the binary vector pcambia1303, which contained hygromycin phosphotransferase (hptii) as a selectable marker gene and β-glucuronidase (gus) as a reporter gene, was used for transformation studies. different factors that affect survival of transformed buds, namely agrobacterium infection method, bacterial strain, pre-culture duration, ac ...201323065217
agrobacterium may delay plant nonhomologous end-joining dna repair via xrcc4 to favor t-dna integration.agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (t-dna) into the plant genome. several plant factors that play a role in agrobacterium attachment to plant cells and transport of t-dna to the nucleus have been identified, but the t-dna integration step during transformation is poorly understood and has been proposed to occur via nonhomologous end-joining (nhej)-mediated double-str ...201223064322
integrated co-regulation of bacterial arsenic and phosphorus metabolisms.arsenic ranks first on the us environmental protection agency superfund list of hazardous substances. its mobility and toxicity depend upon chemical speciation, which is significantly driven by microbial redox transformations. genome sequence-enabled surveys reveal that in many microorganisms genes essential to arsenite (asiii) oxidation are located immediately adjacent to genes coding for functions associated with phosphorus (pi) acquisition, implying some type of functional importance to the m ...201223057575
dynamic localization of two tobamovirus orf6 proteins involves distinct organellar compartments.orf6 is a small gene that overlaps the movement and coat protein genes of subgroup 1a tobamoviruses. the orf6 protein of tomato mosaic virus (tomv) strain l (l-orf6), interacts in vitro with eukaryotic elongation factor 1α, and mutation of the orf6 gene of tobacco mosaic virus (tmv) strain u1 (u1-orf6) reduces the pathogenicity in vivo of tmv, whereas expression of this gene from two other viruses, tobacco rattle virus (trv) and potato virus x (pvx), increases their pathogenicity. in this work, ...201323052393
structural basis for selective gaba binding in bacterial pathogens.gaba acts as an intercellular signal in eukaryotes and as an interspecies signal in host-microbe interactions. structural characteristics of selective eukaryotic gaba receptors and bacterial gaba sensors are unknown. here, we identified the selective gaba-binding protein, called atu4243, in the plant pathogen agrobacterium tumefaciens. a constructed atu4243 mutant was affected in gaba transport and in expression of the gaba-regulated functions, including aggressiveness on two plant hosts and deg ...201223043322
the significance of early accumulation of nanomolar concentrations of no as an inducer of denitrification.denitrifying bacteria have variable ability to perform efficient and balanced denitrification during oxygen depletion. no is often assumed to exert a positive feedback in the transcription of denitrification genes, because no-dependent activators have been identified. the regulatory network of denitrification is complex, however, and the significance of no signalling needs to be studied in vivo. we utilized acetylene-catalysed no oxidation to scavenge no produced by batch cultures of denitrifyin ...201323035849
generation and characterization of the western regional research center brachypodium t-dna insertional mutant collection.the model grass brachypodium distachyon (brachypodium) is an excellent system for studying the basic biology underlying traits relevant to the use of grasses as food, forage and energy crops. to add to the growing collection of brachypodium resources available to plant scientists, we further optimized our agrobacterium tumefaciens-mediated high-efficiency transformation method and generated 8,491 brachypodium t-dna lines. we used inverse pcr to sequence the dna flanking the insertion sites in th ...201223028431
acid-induced type vi secretion system is regulated by exor-chvg/chvi signaling cascade in agrobacterium tumefaciens.the type vi secretion system (t6ss) is a widespread, versatile protein secretion system in pathogenic proteobacteria. several t6sss are tightly regulated by various regulatory systems at multiple levels. however, the signals and/or regulatory mechanisms of many t6sss remain unexplored. here, we report on an acid-induced regulatory mechanism activating t6ss in agrobacterium tumefaciens, a plant pathogenic bacterium causing crown gall disease in a wide range of plants. we monitored the secretion o ...201223028331
expression, purification, crystallization and x-ray analysis of 3-quinuclidinone reductase from agrobacterium tumefaciens.(r)-3-quinuclidinol is a useful chiral building block for the synthesis of various pharmaceuticals and can be produced from 3-quinuclidinone by asymmetric reduction. a novel 3-quinuclidinone reductase from agrobacterium tumefaciens (atqr) catalyzes the stereospecific reduction of 3-quinuclidinone to (r)-3-quinuclidinol with nadh as a cofactor. recombinant atqr was overexpressed in escherichia coli, purified and crystallized with nadh using the sitting-drop vapour-diffusion method at 293 k. cryst ...201223027756
construction of the intermediate vector pvbg2307 by incorporating vital elements of expression vectors pbi121 and pbi221.molecular chaperones of plasmid pbi121 carrying camv35s promoter and a nucleotide sequence of plasmid pbi221 were inserted into plasmid pcambia2300 to construct an intermediate vector: pvbg2307. this novel vector pvbg2307 contains a greatly expanded multiple cloning site with an adjacent imported camv35s promoter sequence. this vector allows controlled transformation of dna in both escherichia coli and agrobacterium tumefaciens. cloned pg, orf456, ipt genes and e8, a fruiting promoter, were ampl ...201223007987
elicitation of andrographolide in the suspension cultures of andrographis paniculata.andrographis paniculata belonging to the family acanthaceae produces a group of diterpene lactones, one of which is the pharmaceutically important-andrographolide. it is known to possess various important biological properties like anticancer, anti-hiv, anti-inflammatory, etc. this is the first report on the production of andrographolide in the cell suspension cultures of andrographis paniculata by 'elicitation'. elicitation was attempted to enhance the andrographolide content in the suspension ...201223001530
expression of agrobacterium tumefaciens octopine ti-plasmid virb8 gene is regulated by translational coupling.eleven proteins of the agrobacterium tumefaciens virb operon are required for type iv secretion. all octopine ti-plasmid ptia6nc virb proteins, except virb8, could be expressed from a cloned monocistronic gene. accumulation of virb8 required translation of the upstream virb7 gene. analysis of chimeric virb8 genes and a newly constructed virb7 deletion mutant agrobacterium ad1275 showed that translation of virb7, and not the gene product, is required for virb8 accumulation. agrobacterium ad1275 a ...201322990025
theoretical prediction and experimental verification of protein-coding genes in plant pathogen genome agrobacterium tumefaciens strain c58.agrobacterium tumefaciens strain c58 is a gram-negative soil bacterium capable of inducing tumors (crown galls) on many dicotyledonous plants. the genome of a. tumefaciens strain c58 was re-annotated based on the z-curve method. first, all the 'hypothetical genes' were re-identified, and 29 originally annotated 'hypothetical genes' were recognized to be non-coding open reading frames (orfs). theoretical evidence obtained from principal component analysis, clusters of orthologous groups of protei ...201222984411
agrobacteria lacking ornithine lipids induce more rapid tumour formation.ornithine lipids (ols) are phosphorus-free membrane lipids that are widespread among gram-negative bacteria. their basic structure consists of a 3-hydroxy fatty acyl group attached in amide linkage to the α-amino group of ornithine and a second fatty acyl group ester-linked to the 3-hydroxy position of the first fatty acid. it has been shown that ols can be hydroxylated within the amide-linked fatty acyl moiety, the secondary fatty acyl moiety or within the ornithine moiety. these modifications ...201222958119
cloning, expression, purification and crystallization of dihydrodipicolinate synthase from agrobacterium tumefaciens.dihydrodipicolinate synthase (dhdps) catalyzes the first committed step of the lysine-biosynthesis pathway in bacteria, plants and some fungi. this study describes the cloning, expression, purification and crystallization of dhdps (np_354047.1) from the plant pathogen agrobacterium tumefaciens (agt-dhdps). enzyme-kinetics studies demonstrate that agt-dhdps possesses dhdps activity in vitro. crystals of agt-dhdps were grown in the unliganded form and in forms with substrate bound and with substra ...201222949190
predictive models for the accumulation of a fluorescent marker protein in tobacco leaves according to the promoter/5'utr combination.the promoter and 5'-untranslated region (5'utr) play a key role in determining the efficiency of recombinant protein expression in plants. comparative experiments are used to identify suitable elements but these are usually tested in transgenic plants or in transformed protoplasts/suspension cells, so their relevance in whole-plant transient expression systems is unclear given the greater heterogeneity in expression levels among different leaves. furthermore, little is known about the impact of ...201322948957
enhancement of ganoderic acid accumulation by overexpression of an n-terminally truncated 3-hydroxy-3-methylglutaryl coenzyme a reductase gene in the basidiomycete ganoderma lucidum.ganoderic acids produced by ganoderma lucidum, a well-known traditional chinese medicinal mushroom, exhibit antitumor and antimetastasis activities. genetic modification of g. lucidum is difficult but critical for the enhancement of cellular accumulation of ganoderic acids. in this study, a homologous genetic transformation system for g. lucidum was developed for the first time using mutated sdhb, encoding the iron-sulfur protein subunit of succinate dehydrogenase, as a selection marker. the tru ...201222941092
functional genomics tools to decipher the pathogenicity mechanisms of the necrotrophic fungus plectosphaerella cucumerina in arabidopsis thaliana.the analysis of the interaction between arabidopsis thaliana and adapted (pcbmm) and nonadapted (pc2127) isolates of the necrotrophic fungus plectosphaerella cucumerina has contributed to the identification of molecular mechanisms controlling plant resistance to necrotrophs. to characterize the pathogenicity bases of the virulence of necrotrophic fungi in arabidopsis, we developed p. cucumerina functional genomics tools using agrobacterium tumefaciens-mediated transformation. we generated pcbmm- ...201322937870
production of engineered long-life and male sterile pelargonium plants.pelargonium is one of the most popular garden plants in the world. moreover, it has a considerable economic importance in the ornamental plant market. conventional cross-breeding strategies have generated a range of cultivars with excellent traits. however, gene transfer via agrobacterium tumefaciens could be a helpful tool to further improve pelargonium by enabling the introduction of new genes/traits. we report a simple and reliable protocol for the genetic transformation of pelargonium spp. a ...201222935247
agroinoculation of the cloned infectious cdnas of lettuce chlorosis virus results in systemic plant infection and production of whitefly transmissible virions.lettuce chlorosis virus (lcv) is a single stranded, positive strand rna virus that is solely transmitted by specific whitefly vectors (bemisia tabaci biotypes a and b) but not by mechanical leaf-rub inoculation. the roles of viral encoded proteins involved in the infection cycle of lcv have not yet been characterized due to the lack of reverse genetic tools. we present here a report of the successful development of an agrobacterium-mediated inoculation system for the cloned cdna constructs of lc ...201222926259
accumulation of glycolipids and other non-phosphorous lipids in agrobacterium tumefaciens grown under phosphate deprivation.phosphate deficiency is characteristic for many natural habitats, resulting in different physiological responses in plants and bacteria including the replacement of phospholipids by glycolipids and other phosphorous-free lipids. the plant pathogenic bacterium agrobacterium tumefaciens, which is free of glycolipids under full nutrition, harbors an open reading frame (orf) coding for a processive glycosyltransferase (named as pgt). this glycosyltransferase was previously shown to synthesize glucos ...201322923441
the novel solanum tuberosum calcium dependent protein kinase, stcdpk3, is expressed in actively growing organs.calcium-dependent protein kinases (cdpks) are key components of calcium regulated signaling cascades in plants. in this work, isoform stcdpk3 from solanum tuberosum was studied and fully described. stcdpk3 encodes a 63 kda protein with an n-terminal variable domain (ntv), rich in prolines and glutamines, which presents myristoylation and palmitoylation consensus sites and a pest sequence indicative of rapid protein degradation. stcdpk3 gene (circa 11 kb) is localized in chromosome 3, shares the ...201222922879
ehrlichia chaffeensis tandem repeat proteins and ank200 are type 1 secretion system substrates related to the repeats-in-toxin exoprotein family.ehrlichia chaffeensis has type 1 and 4 secretion systems (t1ss and t4ss), but the substrates have not been identified. potential substrates include secreted tandem repeat protein (trp) 47, trp120, and trp32, and the ankyrin repeat protein, ank200, that are involved in molecular host-pathogen interactions including dna binding and a network of protein-protein interactions with host targets associated with signaling, transcriptional regulation, vesicle trafficking, and apoptosis. in this study we ...201122919588
plant innate immunity induced by flagellin suppresses the hypersensitive response in non-host plants elicited by pseudomonas syringae pv. averrhoi.a new pathogen, pseudomonas syringae pv. averrhoi (pav), which causes bacterial spot disease on carambola was identified in taiwan in 1997. many strains of this pathovar have been isolated from different locations and several varieties of hosts. some of these strains, such as hl1, are nonmotile and elicit a strong hypersensitive response (hr) in nonhost tobacco leaves, while other strains, such as pa5, are motile and elicit a weak hr. based on the image from a transmission electron microscope, t ...201222911741
overexpression and protein folding of a chimeric beta-glucosidase constructed from agrobacterium tumefaciens and cellvibrio gilvus.in continuation of our investigation on structure and function relationship of beta-glucosidases from mesophilic and thermophilic bacteria, we constructed a chimeric gene by shuffling 17% length in c terminal region of beta-glucosidase of agrobacterium tumefaciens with the corresponding homologous region of cellvibrio gilvus beta-glucosidase. the chimeric gene was overexpressed in e. coli bl21 (de3) using pet vector. however, nearly all of the beta-glucosidase produced was trapped into inclusion ...200222908413
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