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regeneration and agrobacterium-mediated transformation of the apomictic species eulaliopsis binata.protocols for regeneration and agrobacterium-mediated transformation of the apomictic species eulaliopsis binata were developed. initially, seeds of four genotypes of e. binata were incubated on a callus induction murashige and skoog (ms) basal medium supplemented with three concentrations of 2,4-dichlorophenoxyacetic acid (2,4-d). it was found that 36.2 % of explants developed highly friable callus on medium containing 3.0 mg l(-1) 2,4-d. based on frequency of callus induction, the genotype nei ...201323873641
ectopic localization of auxin and cytokinin in tobacco seedlings by the plant-oncogenic ak-6b gene of agrobacterium tumefaciens ake10.the oncogenic 6b gene of agrobacterium tumefaciens induces a number of morphological and metabolic alterations in plants. although molecular functions associated with the 6b genes have been proposed, including auxin transport, sugar transport, transcriptional regulation, and mirna metabolism, so far an unequivocal conclusion has not been obtained. we investigated the association between auxin accumulation and tumor development of the tobacco seedlings expressing the ak-6b gene under the control ...201323873395
the glyceraldehyde-3-phosphate dehydrogenase promoter of the food yeast candida utilis strain nrrl y-660 is functional in agrobacterium tumefaciens.the glyceraldehyde-3-phosphate dehydrogenase promoter of the food yeast candida utilis strain nrrl y-660 was cloned to create a novel integrative vector for agrobacterium tumefaciens-mediated transformation. the new binary vector harbors β-glucuronidase activity as reporter and kanamicin/geneticin resistance as selection marker. recombinant clones of a. tumefaciens show kanamycin resistance and high β-glucuronidase activity under the control of the c. utilis promoter. this finding can be explain ...201323873160
transformation of ltp gene into brassica napus to enhance its resistance to sclerotinia sclerotiorum.rapeseed (brassica napus l.) is one of the most important economic crops worldwide, and sclerotinia sclerotiorum is the most dangerous disease that affects its yield greatly. lipid transfer protein (ltp) has broad-spectrum anti-bacterial and fungal activities. in this study, b. napus was transformed using agrobacterium tumefaciens harboring the plasmid-containing ltp gene to study its possible capability of increasing plant's resistance. first, we optimized the petiole genetic transformation sys ...201323866620
systematic dissection of the agrobacterium type vi secretion system reveals machinery and secreted components for subcomplex formation.the type vi secretion system (t6ss) is widely distributed in pathogenic proteobacteria. sequence and structural analysis of t6ss reveals a resemblance to the t4 bacteriophage tail, in which an outer sheath structure contracts an internal tube for injecting nucleic acid into bacterial cells. however, the molecular details of how this phage tail-like t6ss structure is assembled in vivo and executed for exoprotein or effector secretion remain largely unknown. here, we used a systematic approach to ...201323861778
plant biotechnology for food security and bioeconomy.this year is a special year for plant biotechnology. it was 30 years ago, on january 18 1983, one of the most important dates in the history of plant biotechnology, that three independent groups described agrobacterium tumefaciens-mediated genetic transformation at the miami winter symposium, leading to the production of normal, fertile transgenic plants (bevan et al. in nature 304:184-187, 1983; fraley et al. in proc natl acad sci usa 80:4803-4807, 1983; herrera-estrella et al. in embo j 2:987- ...201323860797
identification of host proteins modulated by the virulence factor ac2 of tomato chlorotic mottle virus in nicotiana benthamiana.tomato, one of the most important crops cultivated worldwide, has been severely affected by begomoviruses such as the tomato chlorotic mottle virus (tocmov). virulence factor ac2 is considered crucial for a successful virus-plant interaction and is known to act as a transcriptional activator and in some begomoviruses to function as an rna silencing suppressor factor. however, the exact functions of the ac2 protein of the begomovirus tocmov are not yet established. the aim of the present study wa ...201323533094
yeast competence for exogenous dna uptake: towards understanding its genetic component.the demonstration of spontaneous yeast competence shows that artificial transformation rests on naturally occurring cellular processes. such natural competence is either biologically mediated or environmentally induced. for instance, wild yeast might be transformed through conjugation by cell-to-cell contact mediated either by escherichia coli or agrobacterium tumefaciens. moreover, natural competence can be enhanced by mechanical and physiological mechanisms. on the other hand, artificial yeast ...201323529158
enhancement of heterologous gene expression in flammulina velutipes using polycistronic vectors containing a viral 2a cleavage sequence.agrobacterium tumefaciens-mediated transformation for edible mushrooms has been previously established. however, the enhancement of heterologous protein production and the expression of multi-target genes remains a challenge. in this study, heterologous protein expression in the enoki mushroom flammulina velutipes was notably enhanced using 2a peptide-mediated cleavage to co-express multiple copies of single gene. the polycistronic expression vectors were constructed by connecting multi copies o ...201323516605
synthesis and structure-activity relationship of n-(cinnamyl) chitosan analogs as antimicrobial agents.the current study focuses on the preparation of new n-(cinnamyl) chitosan derivatives as antimicrobial agents against nine types of crop-threatening pathogens. chitosan was reacted with a set of aromatic cinnamaldehyde analogs by reductive amination involving formation of the corresponding imines, followed by reduction with sodium borohydride to produce n-(cinnamyl) chitosan derivatives. the structural characterization was confirmed by (1)h and (13)c nmr spectroscopy and the degrees of substitut ...201323511055
transformation of apple (malus × domestica) using mutants of apple acetolactate synthase as a selectable marker and analysis of the t-dna integration sites.apple acetolactate synthase mutants were generated by site-specific mutagenesis and successfully used as selection marker in tobacco and apple transformation. t-dna/apple genome junctions were analysed using genome-walking pcr and sequencing. an agrobacterium-mediated genetic transformation system was developed for apple (malus × domestica), using mutants of apple acetolactate synthase (als) as a selectable marker. four apple als mutants were generated by site-specific mutagenesis and subsequent ...201323494389
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
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
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
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
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
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
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
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
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
better rooting procedure to enhance survival rate of field grown malaysian eksotika papaya transformed with 1-aminocyclopropane-1-carboxylic acid oxidase gene.a high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. we report in this paper an improved rooting method for the production of transformed malaysian eksotika papaya with high survival rate when transferred to the field. shoots were regenerated from embryogenic calli transformed with antisense and rnai constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (aco) genes using the agrobacterium tumefa ...201325969786
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
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
agrobacterium tumefaciens-mediated in planta seed transformation strategy in sugarcane.an efficient, reproducible and genotype-independent in planta transformation has been standardized for sugarcane using seed as explant. transgenic sugarcane production through agrobacterium infection followed by in vitro regeneration is a time-consuming process and highly genotype dependent. to obtain more number of transformed sugarcane plants in a relatively short duration, sugarcane seeds were infected with agrobacterium tumefaciens eha 105 harboring pcambia 1304-bar and transformed plants we ...201323749098
overexpression of the glutamine synthetase gene modulates oxidative stress response in rice after exposure to cadmium stress.overexpression of osgs gene modulates oxidative stress response in rice after exposure to cadmium stress. our results describe the features of transformants with enhanced tolerance to cd and abiotic stresses. glutamine synthetase (gs) (ec 6.3.1.2) is an enzyme that plays an essential role in the metabolism of nitrogen by catalyzing the condensation of glutamate and ammonia to form glutamine. exposure of plants to cadmium (cd) has been reported to decrease gs activity in maize, pea, bean, and ric ...201323743654
n-acylhomoserine lactone-degrading bacteria isolated from hatchery bivalve larval cultures.quorum sensing (qs) systems, which depend on n-acylhomoserine lactone (ahl) signal molecules, mediate the production of virulence factors in many pathogenic microorganisms. one hundred and forty-six bacterial strains, isolated from a bivalve hatchery, were screened for their capacity to degrade five synthetic ahls [n-butyryl-dl-homoserine lactone (c4-hsl), n-hexanoyl-dl-homoserine lactone (c6-hsl), n-octanoyl-dl-homoserine lactone (c8-hsl), n-decanoyl-dl-homoserine lactone (c10-hsl) and n-dodeca ...201323743010
the helicobacter pylori protein cagm is located in the transmembrane channel that is required for caga translocation.helicobacter pylori (h. pylori) is a human gastric pathogen that colonizes the stomach in more than 50 % of the world's human population. infection with this bacterium can induce several gastric diseases ranging from gastritis to peptic ulcer and gastric cancer. virulent h. pylori isolates harboring the cag pathogenicity island (cag pai), which encodes a type iv secretion system (t4ss), form a pilus for the injection of its major virulence protein caga into gastric cells. several cag pai genes h ...201323736225
botrytis cinerea chitin synthase bcchsvi is required for normal growth and pathogenicity.fungal chitin synthase of classes v and vi (or vii), which contain an additional n-terminal myosin motor domain, have been shown to play important roles in pathogenesis. to study the function of bcchsvi in botrytis cinerea, bcchs6 gene was disrupted through agrobacterium tumefaciens-mediated transformation. the bcchs6 disruption mutant exhibited a 45.5 % increasing in its chitin content when compared with wild strain. the qrt-pcr analysis revealed that in bcchs6 mutant the expression of bcchs6 w ...201323722656
l-glutamine and l-glutamic acid facilitate successful agrobacterium infection of recalcitrant tea cultivars.the first step in agrobacterium tumefaciens infection of plants is the establishment of cell-cell contact between the two partners. however, failure to establish such contact makes many plants and explants recalcitrant to a. tumefaciens infection. tea is one such example where even the popular inducer, acetosyringone failed to facilitate a. tumefaciens infection due to the presence of high amounts of bactericidal/bacteriostatic polyphenols. quinones are formed as a result of polyphenols oxidatio ...201323712792
in planta fitness-cost of the atu4232-regulon encoding for a selective gaba-binding sensor in agrobacterium.gaba (gamma-aminobutyric acid) mediates cell-to-cell communication in eukaryotes and interspecies communication in host-microbe interactions. agrobacterium tumefaciens induces the development of plant tumor in which gaba accumulates. two periplasmic binding proteins atu2422 and atu4243 and their appropriate abc-transporter are involved in the binding and importation of gaba. the structure of the selective gaba-binding atu4243 reveals a gaba conformation similar to a proposed model of gaba bound ...201323710277
investigation of the fusarium virguliforme fvtox1 mutants revealed that the fvtox1 toxin is involved in foliar sudden death syndrome development in soybean.the soil borne fungus, fusarium virguliforme, causes sudden death syndrome (sds) in soybean, which is a serious foliar and root rot disease. the pathogen has never been isolated from the diseased foliar tissues; phytotoxins produced by the pathogen are believed to cause foliar sds symptoms. one of these toxins, a 13.5-kda acidic protein named fvtox1, has been hypothesized to interfere with photosynthesis in infected soybean plants and cause foliar sds. the objective of this study is to determine ...201323702608
implantable vascular access port-associated bloodstream infection caused by rhizobium radiobacter: a case report.rhizobium radiobacter is an uncommon opportunistic pathogen present in soil. it has been particularly associated with indwelling intravascular devices in immunocompromised patients. reported herein is a case of r. radiobacter bloodstream infection associated with an implantable vascular access port, which was easily controlled with antimicrobial treatment and did not require removal of the intravascular device, in a child diagnosed with acute lymphoblastic leukemia. also included is a review of ...201323692845
a flexible c-terminal linker is required for proper ftsz assembly in vitro and cytokinetic ring formation in vivo.assembly of the cytoskeletal protein ftsz into a ring-like structure is required for bacterial cell division. structurally, ftsz consists of four domains: the globular n-terminal core, a flexible linker, 8-9 conserved residues implicated in interactions with modulatory proteins, and a highly variable set of 4-10 residues at its very c terminus. largely ignored and distinguished by lack of primary sequence conservation, the linker is presumed to be intrinsically disordered. here we employ genetic ...201323692518
fast induction of biosynthetic polysaccharide genes lpxa, lpxe, and rkpi of rhizobium sp. strain prf 81 by common bean seed exudates is indicative of a key role in symbiosis.rhizobial surface polysaccharides (sps) are, together with nodulation (nod) factors, recognized as key molecules for establishment of rhizobia-legume symbiosis. in rhizobium tropici, an important nitrogen-fixing symbiont of common bean (phaseolus vulgaris l.), molecular structures and symbiotic roles of the sps are poorly understood. in this study, rhizobium sp. strain prf 81 genes, belonging to the r. tropici group, were investigated: lpxa and lpxe, involved in biosynthesis and modification of ...201323652766
the methylation of the pcmyb10 promoter is associated with green-skinned sport in max red bartlett pear.varieties of the european pear (pyrus communis) can produce trees with both red- and green-skinned fruits, such as the max red bartlett (mrb) variety, although little is known about the mechanism behind this differential pigmentation. in this study, we investigated the pigmentation of mrb and its green-skinned sport (mrb-g). the results suggest that a reduction in anthocyanin concentration causes the mrb-g sport. transcript levels of pcufgt (for udp-glucose:flavonoid 3-o-glucosyltransferase), th ...201323629835
a putative transmembrane leucine zipper of agrobacterium virb10 is essential for t-pilus biogenesis but not type iv secretion.the agrobacterium tumefaciens virb/vird4 type iv secretion system is composed of a translocation channel and an extracellular t pilus. bitopic virb10, the virb7 lipoprotein, and virb9 interact to form a cell envelope-spanning structural scaffold termed the "core complex" that is required for the assembly of both structures. the related pkm101-encoded core complex is composed of 14 copies each of these virb homologs, and the transmembrane (tm) α helices of virb10-like traf form a 55-å-diameter ri ...201323625845
a rapid assay to quantify the cleavage efficiency of custom-designed nucleases in planta.custom-designed nucleases are a promising technology for genome editing through the catalysis of double-strand dna breaks within target loci and subsequent repair by the host cell, which can result in targeted mutagenesis or gene replacement. implementing this new technology requires a rapid means to determine the cleavage efficiency of these custom-designed proteins in planta. here we present such an assay that is based on cleavage-dependent luciferase gene correction as part of a transient dua ...201323625357
agroinoculation of a full-length cdna clone of cotton leafroll dwarf virus (clrdv) results in systemic infection in cotton and the model plant nicotiana benthamiana.cotton blue disease is the most important viral disease of cotton in the southern part of south america. its etiological agent, cotton leafroll dwarf virus (clrdv), is specifically transmitted to host plants by the aphid vector (aphis gossypii) and any attempt to perform mechanical inoculations of this virus into its host has failed. this limitation has held back the study of this virus and the disease it causes. in this study, a full-length cdna of clrdv was constructed and expressed in vivo un ...201323623981
chloroplast-targeted expression of the codon-optimized truncated cry1ah gene in transgenic tobacco confers a high level of protection against insects.the study of insect-resistant transgenic tobacco provides a good foundation for the further application of the cry1ah gene in other important crops. to improve transgene expression levels and insect resistance, the coding sequence of the novel bacillus thuringiensis insecticidal gene cry1ah (truncated cry1ah) was modified according to the codon bias of the plant by increasing its gc content from the original 37 % to 48, 55, and 63 % (designated m1-cry1ah, m2-cry1ah, and m3-cry1ah, respectively). ...201323620344
high expression of a neutral endo-β-glucanase gene from humicola insolens in trichoderma reesei.the neutral endo-β-glucanase gene cel5a from humicola insolens was cloned and connected with the cellobiohydrolase 1 promoter from trichoderma reesei to construct a recombinant plasmid pcb-heg with the hygromycin b resistance marker. the plasmid was introduced into conidia of t. reesei using the agrobacterium tumefaciens mediated transformation method. eight transformants were obtained on screening plates with sodium carboxymethyl cellulose as the sole carbon source. stable integration of the ce ...201323619969
shape-dependent bactericidal activity of tio2 for the killing of gram-negative bacteria agrobacterium tumefaciens under uv torch irradiation.this paper demonstrated the relative bactericidal activity of photoirradiated (6w-uv torch, λ > 340 nm and intensity = 0.64 mw/cm(2)) p25-tio2 nanoparticles, nanorods, and nanotubes for the killing of gram-negative bacterium agrobacterium tumefaciens lba4404 for the first time. tio2 nanorod (anatase) with length of 70-100 nm and diameter of 10-12 nm, and tio2 nanotube with length of 90-110 nm and diameter of 9-11 nm were prepared from p-25 degussa tio2 (size, 30-50 nm) by hydrothermal method and ...201323608987
booster responses by oral vaccination with transgenic plants against chicken leucocytozoonosis.we developed a transgenic potato (trp/r7) expressing the recombinant r7 (rr7) antigen for use as an oral vaccine to protect against a chicken protozoan disease, chicken leucocytozoonosis. the trp/r7 potato was produced by agrobacterium tumefaciens-mediated transformation and regeneration, and the r7 gene insertion into potato chromosomes was confirmed by genomic polymerase chain reaction and southern hybridization. rr7 antigen expression in trp/r7 potato was also confirmed by sandwich enzyme-lin ...201323607580
identifying pathogenicity genes in the rubber tree anthracnose fungus colletotrichum gloeosporioides through random insertional mutagenesis.to gain more insight into the molecular mechanisms of colletotrichum gloeosporioides pathogenesis, agrobacterium tumefaciens-mediated transformation (atmt) was used to identify mutants of c. gloeosporioides impaired in pathogenicity. an atmt library of 4128 c. gloeosporioides transformants was generated. transformants were screened for defects in pathogenicity with a detached copper brown leaf assay. 32 mutants showing reproducible pathogenicity defects were obtained. southern blot analysis show ...201323602122
identification of a genomic region containing a novel promoter resistant to glucose repression and over-expression of β-glucosidase gene in hypocrea orientalis eu7-22.a high concentration of glucose in the medium could greatly inhibit the expression of cellulase in filamentous fungi. the aspartic protease from fungus hypocrea orientalis eu7-22 could efficiently express under both induction condition and glucose repression condition. based on the sequence of structure gene of aspartic protease, the upstream sequence harboring the putative promoter proa for driving the expression of aspartic protease was obtained by genome walking. the upstream sequence contain ...201323594998
effect of plant phenolic compounds on biofilm formation by pseudomonas aeruginosa.in the natural environment, bacteria predominantly exist in matrix-enclosed multicellular communities associated with various surfaces, referred to as biofilms. bacteria in biofilms are extremely resistant to antibacterial agents thus causing serious problems for antimicrobial therapy. in this study, we showed that different plant phenolic compounds, at concentrations that did not or weakly suppressed bacterial growth, increased the capacity of pseudomonas aeruginosa pao1 to form biofilms. biofi ...201323594262
the tzs protein and exogenous cytokinin affect virulence gene expression and bacterial growth of agrobacterium tumefaciens.the soil phytopathogen agrobacterium tumefaciens causes crown gall disease in a wide range of plant species. the neoplastic growth at the infection sites is caused by transferring, integrating, and expressing transfer dna (t-dna) from a. tumefaciens into plant cells. a trans-zeatin synthesizing (tzs) gene is located in the nopaline-type tumor-inducing plasmid and causes trans-zeatin production in a. tumefaciens. similar to known virulence (vir) proteins that are induced by the vir gene inducer a ...201323593941
rapid and accurate species and genomic species identification and exhaustive population diversity assessment of agrobacterium spp. using reca-based pcr.agrobacteria are common soil bacteria that interact with plants as commensals, plant growth promoting rhizobacteria or alternatively as pathogens. indigenous agrobacterial populations are composites, generally with several species and/or genomic species and several strains per species. we thus developed a reca-based pcr approach to accurately identify and specifically detect agrobacteria at various taxonomic levels. specific primers were designed for all species and/or genomic species of agrobac ...201323578959
an experimentally tested scenario for the structural evolution of eukaryotic cys2his2 zinc fingers from eubacterial ros homologs.the exact evolutionary origin of the zinc finger (zf) domain is unknown, as it is still not clear from which organisms it was first derived. however, the unique features of the zf domains have made it very easy for evolution to tinker with them in a number of different manners, including their combination, variation of their number by unequal crossing-over or tandem duplication and tuning of their affinity for specific dna sequence motifs through point substitutions. classical cys2his2 zf domain ...201323576569
an active ac/ds transposon system for activation tagging in tomato cultivar m82 using clonal propagation.tomato (solanum lycopersicum) is a model organism for solanaceae in both molecular and agronomic research. this project utilized agrobacterium tumefaciens transformation and the transposon-tagging construct activator (ac)/dissociator (ds)-atag-bar_gosgfp to produce activation-tagged and knockout mutants in the processing tomato cultivar m82. the construct carried hygromycin resistance (hyg), green fluorescent protein (gfp), and the transposase (tpase) of maize (zea mays) activator major transcri ...201323569107
brachypodium sylvaticum, a model for perennial grasses: transformation and inbred line development.perennial species offer significant advantages as crops including reduced soil erosion, lower energy inputs after the first year, deeper root systems that access more soil moisture, and decreased fertilizer inputs due to the remobilization of nutrients at the end of the growing season. these advantages are particularly relevant for emerging biomass crops and it is projected that perennial grasses will be among the most important dedicated biomass crops. the advantages offered by perennial crops ...201324073248
an orange fluorescent protein tagging system for real-time pollen tracking.monitoring gene flow could be important for future transgenic crops, such as those producing plant-made-pharmaceuticals (pmps) in open field production. a nicotiana hybrid (nicotiana. tabacum × nicotiana glauca) shows limited male fertility and could be used as a bioconfined pmp platform. effective assessment of gene flow from these plants is augmented with methods that utilize fluorescent proteins for transgenic pollen identification.201324070251
retraction: agrobacterium tumefaciens-mediated transformation and efficient regeneration of a timber yielding plant dalbergia sissoo roxb. 201324068859
[study on human afgf fusion gene transformation with soybean 24 kda oleosin and expression in safflower].to investigate the expression of acidic fibroblast growth factor (afgf) in transgenic safflower and lay the foundation for the use of the plant bioreactor large-scale production afgf.201324066580
development of transgenic papaya through agrobacterium-mediated transformation.transgenic papaya plants were regenerated from hypocotyls and immature zygotic embryo after cocultivation with agrobacterium tumefaciens lba-4404 carrying a binary plasmid vector system containing neomycin phosphotransferase (nptii) gene as the selectable marker and β-glucuronidase (gus) as the reporter gene. the explants were co-cultivated with agrobacterium tumefaciens on regeneration medium containing 500 mg/l carbenicillin + 200 mg/l cefotaxime for one week. the cocultivated explants were tr ...201324066284
agrobacterium tumefaciens-mediated genetic transformation of the taxol-producing endophytic fungus ozonium sp efy21.an efficient agrobacterium tumefaciens-mediated genetic transformation method was successfully established for a newly isolated taxol-producing fungus, ozonium sp efy21. a specific hygromycin b resistance expression vector, pcambia1304'an7-1, was constructed for fungal transformation. key factors affecting transformation efficiency were thoroughly investigated and optimized. pcr amplification and southern hybridization were used to verify the transformation events. this study should pave the way ...201324065647
trans-kingdom horizontal dna transfer from bacteria to yeast is highly plastic due to natural polymorphisms in auxiliary nonessential recipient genes.with the rapid accumulation of genomic information from various eukaryotes in the last decade, genes proposed to have been derived from recent horizontal gene transfer (hgt) events have been reported even in non-phagotrophic unicellular and multicellular organisms, but the molecular pathways underlying hgt remain to be explained. the development of in vitro hgt detection systems, which permit the molecular and genetic analyses of donor and recipient organisms and quantify hgt, are helpful in ord ...201324058593
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
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
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
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
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
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
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
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
an improved agrobacterium-mediated transformation of recalcitrant indica rice (oryza sativa l.) cultivars.agrobacterium-mediated transformation of indica rice varieties has been quite difficult as these are recalcitrant to in vitro responses. in the present study, we established a high-efficiency agrobacterium tumefaciens-mediated transformation system of rice (oryza sativa l. ssp. indica) cv. ir-64, lalat, and iet-4786. agrobacterium strain eha-101 harboring binary vector pig121-hm, containing a gene encoding for β-glucuronidase (gus) and hygromycin resistance, was used in the transformation experi ...201322878598
efficient genetic transformation of withania coagulans (stocks) dunal mediated by agrobacterium tumefaciens from leaf explants of in vitro multiple shoot culture.an efficient and reproducible agrobacterium-mediated genetic transformation of withania coagulans was achieved using leaf explants of in vitro multiple shoot culture. the agrobacterium strain lba4404 harboring the binary vector pig121hm containing β-glucuronidase gene (gusa) under the control of camv35s promoter was used in the development of transformation protocol. the optimal conditions for the agrobacterium-mediated transformation of w. coagulans were found to be the co-cultivation of leaf e ...201322766977
a novel transcriptional regulator, clbr, controls the cellobiose- and cellulose-responsive induction of cellulase and xylanase genes regulated by two distinct signaling pathways in aspergillus aculeatus.the cellobiose- and cellulose-responsive induction of the fiii-avicelase (cbhi), fii-carboxymethyl cellulase (cmc2), and fia-xylanase (xynia) genes is not regulated by xlnr in aspergillus aculeatus, which suggests that this fungus possesses an unknown cellulase gene-activating pathway. to identify the regulatory factors involved in this pathway, we constructed a random insertional mutagenesis library using agrobacterium tumefaciens-mediated transformation of a. aculeatus ncp2, which harbors a tr ...201322851016
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
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
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
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
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
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
characterization of glyceraldehyde-3-phosphate dehydrogenase gene rtgpd1 and development of genetic transformation method by dominant selection in oleaginous yeast rhodosporidium toruloides.the oleaginous yeast rhodosporidium toruloides, which belongs to the pucciniomycotina subphylum in the basidiomycota, has attracted strong interest in the biofuel community recently due to its ability to accumulate more than 60% of dry biomass as lipid under high-density fermentation. a 3,543-nucleotide (nt) dna fragment of the glyceraldehyde-3-phosphate dehydrogenase gene (gpd1) was isolated from r. toruloides atcc 10657 and characterized in details. the 1,038-nt mrna derived from seven exons e ...201322722909
the selection of recombinant binary plasmids generated by gateway(®) lr cloning in the escherichia coli strain c2110.gateway(®) cloning is widely used in molecular biology laboratories. various binary vectors used for agrobacterium-mediated plant transformation have been modified as destination vectors that are convenient for the sub-cloning of targeted genes from entry plasmids. however, when the destination and entry plasmids have the same antibiotic resistance genes for bacterial selection, the non-recombinant entry plasmid in the lr reaction mixture can compete with the recombinant destination plasmid duri ...201322555851
diversity of sporadic symbionts and nonsymbiotic endophytic bacteria isolated from nodules of woody, shrub, and food legumes in ethiopia.fifty-five bacterial isolates were obtained from surface-sterilized nodules of woody and shrub legumes growing in ethiopia: crotalaria spp., indigofera spp., and erythrina brucei, and the food legumes soybean and common bean. based on partial 16s rrna gene sequence analysis, the majority of the isolates were identified as gram-negative bacteria belonging to the genera achromobacter, agrobacterium, burkholderia, cronobacter, enterobacter, mesorhizobium, novosphingobium, pantoea, pseudomonas, rahn ...201324196581
rapid regeneration and ploidy stability of 'cv ir36' indica rice (oryza sativa. l) confers efficient protocol for in vitro callus organogenesis and agrobacterium tumefaciens mediated transformation.cereal crops are the major targets for transformation mediated crop improvement and ir36 is an early maturing, high yielding, insect and disease resistant rice variety however, it is abiotic stress sensitive. hence, development of an efficient and reproducible micropropagation system via somatic embryogenesis and agrobacterium tumefaciens mediated transformation is prerequisite to develop abiotic stress tolerant ir36. further, genetic stability of analysis of plantlets through rapd and issr and ...201328510899
highly efficient synthesis of (r)-3-quinuclidinol in a space-time yield of 916 g l(-1) d(-1) using a new bacterial reductase arqr.a new keto reductase (arqr), identified from agrobacterium radiobacter ecu2556, can efficiently reduce 3-quinuclidinone in excellent enantioselectivity and high space-time yield for the synthesis of (r)-3-quinuclidinol, a chiral building block of many antimuscarinic agents. this is the first time that a high yield of (r)-3-quinuclidinol up to 916 g l(-1) d(-1) using a bioreduction approach is reported.201324050153
high frequency regeneration via direct somatic embryogenesis and efficient agrobacterium-mediated genetic transformation of tobacco.a direct somatic embryogenesis protocol was developed for four cultivars of nicotiana species, by using leaf disc as an explant. direct somatic embryogenesis of nicotiana by using bap and iaa has not been investigated so far. this method does not require formation of callus tissues which leads to somaclonal variations. the frequency of somatic embryogenesis was strongly influenced by the plant growth hormones. the somatic embryos developing directly from explant tissue were noticed after 6 d of ...201323518589
hfq influences multiple transport systems and virulence in the plant pathogen agrobacterium tumefaciens.the hfq protein mediates gene regulation by small rnas (srnas) in about 50% of all bacteria. depending on the species, phenotypic defects of an hfq mutant range from mild to severe. here, we document that the purified hfq protein of the plant pathogen and natural genetic engineer agrobacterium tumefaciens binds to the previously described srna abcr1 and its target mrna atu2422, which codes for the substrate binding protein of an abc transporter taking up proline and γ-aminobutyric acid (gaba). s ...201222821981
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
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
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