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differentiating indigenous soybean bradyrhizobium and rhizobium spp. of indian soils.soybean is extensively cultivated worldwide and is the largest source of biologically fixed nitrogen among legumes. it is nodulated by both slow and fast growing rhizobia. indigenous soybean rhizobia in vertisols of central india were assessed for utilization of 35 carbon sources and intrinsic resistance to 19 antibiotics. there was greater utilization of trehalose and raffinose by fast growers (87 and 73 % by fast vs. 35 and 30 % by slow growers); but slow growers had higher ability to utilize ...201325320421
the thuefgkab operon of rhizobia and agrobacterium tumefaciens codes for transport of trehalose, maltitol, and isomers of sucrose and their assimilation through the formation of their 3-keto derivatives.the thu operon (thuefgkab) in sinorhizobium meliloti codes for transport and utilization functions of the disaccharide trehalose. sequenced genomes of members of the rhizobiaceae reveal that some rhizobia and agrobacterium possess the entire thu operon in similar organizations and that mesorhizobium loti maff303099 lacks the transport (thuefgk) genes. in this study, we show that this operon is dedicated to the transport and assimilation of maltitol and isomers of sucrose (leucrose, palatinose, a ...201323772075
a genome-wide survey of highly expressed non-coding rnas and biological validation of selected candidates in agrobacterium tumefaciens.agrobacterium tumefaciens is a plant pathogen that has the natural ability of delivering and integrating a piece of its own dna into plant genome. although bacterial non-coding rnas (ncrnas) have been shown to regulate various biological processes including virulence, we have limited knowledge of how agrobacterium ncrnas regulate this unique inter-kingdom gene transfer. using whole transcriptome sequencing and an ncrna search algorithm developed for this work, we identified 475 highly expressed ...201323950988
genetic analysis of agrobacterium tumefaciens unipolar polysaccharide production reveals complex integrated control of the motile-to-sessile switch.many bacteria colonize surfaces and transition to a sessile mode of growth. the plant pathogen agrobacterium tumefaciens produces a unipolar polysaccharide (upp) adhesin at single cell poles that contact surfaces. here we report that elevated levels of the intracellular signal cyclic diguanosine monophosphate (c-di-gmp) lead to surface-contact-independent upp production and a red colony phenotype due to production of upp and the exopolysaccharide cellulose, when a. tumefaciens is incubated with ...201323829710
large deletions in the patc58 megaplasmid of agrobacterium tumefaciens can confer reduced carriage cost and increased expression of virulence genes.the accessory plasmid patc58 of the common laboratory strain of agrobacterium tumefaciens confers numerous catabolic functions and has been proposed to play a role in virulence. genomic sequencing of evolved laboratory strains of a. tumefaciens revealed the presence of multiple deletion events in the at plasmid, with reductions in plasmid size ranging from 25% to 30% (115-194 kb). flanking both ends of the sites of these deletions is a short-nucleotide repeat sequence that is in a single copy in ...201323783172
dynamic ftsa and ftsz localization and outer membrane alterations during polar growth and cell division in agrobacterium tumefaciens.growth and cell division in rod-shaped bacteria have been primarily studied in species that grow predominantly by peptidoglycan (pg) synthesis along the length of the cell. rhizobiales species, however, predominantly grow by pg synthesis at a single pole. here we characterize the dynamic localization of several agrobacterium tumefaciens components during the cell cycle. first, the lipophilic dye fm 4-64 predominantly stains the outer membranes of old poles versus growing poles. in cells about to ...201323674672
visualization of vire2 protein translocation by the agrobacterium type iv secretion system into host cells.type iv secretion systems (t4ss) can mediate the translocation of bacterial virulence proteins into host cells. the plant pathogen agrobacterium tumefaciens uses a t4ss to deliver a vird2-single stranded dna complex as well as the virulence proteins vird5, vire2, vire3, and virf into host cells so that these become genetically transformed. besides plant cells, yeast and fungi can efficiently be transformed by agrobacterium. translocation of virulence proteins by the t4ss has so far only been sho ...201324376037
generating phenotypic diversity in a fungal biocatalyst to investigate alcohol stress tolerance encountered during microbial cellulosic biofuel production.consolidated bioprocessing (cbp) of lignocellulosic biomass offers an alternative route to renewable energy. the crop pathogen fusarium oxysporum is a promising fungal biocatalyst because of its broad host range and innate ability to co-saccharify and ferment lignocellulose to bioethanol. a major challenge for cellulolytic cbp-enabling microbes is alcohol inhibition. this research tested the hypothesis that agrobacterium tumefaciens--mediated transformation (atmt) could be exploited as a tool to ...201324147009
coral-associated bacteria, quorum sensing disrupters, and the regulation of biofouling.marine biofouling, the settlement of microorganisms and macroorganisms on structures submerged in seawater, although economically detrimental, is a successful strategy for survival in hostile environments, where coordinated bacterial communities establish biofilms via the regulation of quorum sensing (qs) communication systems. the inhibition of qs activity among bacteria isolated from different coral species was investigated to gain further insight into its potency in the attenuation, or even t ...201323777289
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
at a supra-physiological concentration, human sexual hormones act as quorum-sensing inhibitors.n-acylhomoserine lactone (ahl)-mediated quorum-sensing (qs) regulates virulence functions in plant and animal pathogens such as agrobacterium tumefaciens and pseudomonas aeruginosa. a chemolibrary of more than 3500 compounds was screened using two bacterial ahl-biosensors to identify qs-inhibitors (qsis). the purity and structure of 15 qsis selected through this screening were verified using hplc ms/ms tools and their activity tested on the a. tumefaciens and p. aeruginosa bacterial models. the ...201324376718
evaluation of a new high-throughput method for identifying quorum quenching bacteria.quorum sensing (qs) is a population-dependent mechanism for bacteria to synchronize social behaviors such as secretion of virulence factors. the enzymatic interruption of qs, termed quorum quenching (qq), has been suggested as a promising alternative anti-virulence approach. in order to efficiently identify qq bacteria, we developed a simple, sensitive and high-throughput method based on the biosensor agrobacterium tumefaciens a136. this method effectively eliminates false positives caused by in ...201324121744
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
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
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
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
recent advances in the understanding of agrobacterium rhizogenes-derived genes and their effects on stress resistance and plant metabolism.it is commonly accepted that the plant pathogens agrobacterium rhizogenes and agrobacterium tumefaciens, acting via their t-dna oncogenes, disturb hormone metabolism or hormone perception pathways in plants, thereby attaining their aim of successful pathogenesis. in this work, we summarize recent data on the a. rhizogenes rolc and rolb oncogenes in comparison to the a. tumefaciens 6b oncogene with respect to their effects on the physiology of transformed cells. the newly discovered functions of ...201323576052
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
selection of a suitable disc bioassay for the screening of anti-tumor molecules.the crown gall induced in potato discs by agrobacterium tumefaciens is becoming largely utilised in screening anti-tumor agents. the present work is showing that beet discs are more adequate for the anti-tumor screening test. in fact, maximal tumor induction was observed on beet discs (87.5%), followed by carrot discs (75%) and potato discs (68.5%). beet discs present the most sensibility to crown gall disease with a fast expression of symptoms and more visible galls without any staining need. t ...201324711759
[establishment of transformation system of lonicera macranthoides mediated by agrobacterium tumefaciens].based on the system of regeneration,the genetic transformation system of lonicera macranthoides was established.201325090668
retracted article: agrobacterium tumefaciens-mediated transformation and efficient regeneration of a timber yielding plant dalbergia sissoo roxb. 201324431531
transformation of morinda citrifolia via simple mature seed imbibition method.morinda citrifolia, is a valuable medicinal plant with a wide range of therapeutic properties and extensive transformation study on this plant has yet been known. present study was conducted to establish a simple and reliable transformation protocol for m. citrifolia utilising agrobacterium tumefaciens via direct seed exposure. in this study, the seeds were processed by tips clipping and dried and subsequently incubated in inoculation medium. four different parameters during the incubation such ...201324517006
[isolation and characterization of petroleum catabolic broad-host-range plasmids from shen-fu wastewater irrigation zone].based on triparental mating, we isolated a total of eight broad host range (bhr) petroleum hydrocarbon catabolic plasmids from the soils, sediments, and wastewater samples in the shen-fu irrigation zone. the antibiotic resistance of the plasmids was tested, and then, the plasmids were transferred to escherichia coli ec100. the plasmids carrying no antibiotic resistance were tagged by minitn5 transposon consisting of antibiotic resistant genes. the pcr-based incompatibility test revealed that the ...201324564162
[genetic transformation of the fungus gliocladium sp. mediated by agrobacterium tumefaciens].to construct a transformation system in gliocladium sp. 6.102, a cordyceps-colonizing fungus producing a variety of epipolythiodioxopiperazine (etp) compounds with drug potentials.201324617266
genetic transformation of major cereal crops.of the more than 50,000 edible plant species in the world, at least 10,000 species are cereal grains. three major cereal crops, rice (oryza sativa), maize (zea mays), and wheat (triticum sp.), provide two-thirds of the world's food energy intake. although crop yields have improved tremendously thanks to technological advances in the past 50 years, population increases and climate changes continue to threaten the sustainability of current crop productions. whereas conventional and marker-assisted ...201324166432
higher plant transformation: principles and molecular tools.in higher plants, genetic transformation, which is part of the toolbox for the study of living organisms, had been reported only 30 years ago, boosting basic plant biology research, generating superior crops, and leading to the new discipline of plant biotechnology. here, we review its principles and the corresponding molecular tools. in vitro regeneration, through somatic embryogenesis or organogenesis, is discussed because they are prerequisites for the subsequent agrobacterium tumefaciens-med ...201324166431
the roles of bacterial and host plant factors in agrobacterium-mediated genetic transformation.the genetic transformation of plants mediated by agrobacterium tumefaciens represents an essential tool for both fundamental and applied research in plant biology. for a successful infection, culminating in the integration of its transferred dna (t-dna) into the host genome, agrobacterium relies on multiple interactions with host-plant factors. extensive studies have unraveled many of such interactions at all major steps of the infection process: activation of the bacterial virulence genes, cell ...201324166430
from the tumor-inducing principle to plant biotechnology and its importance for society.this dialogue was held between the guest editors of the special issue on "plant transgenesis" of the int. j. dev. biol. and marc van montagu. research in the group of marc van montagu and jeff schell in the 1970s was essential to reveal how the phytopathogenic bacterium agrobacterium tumefaciens transfers dna to host plants to cause crown gall disease. knowledge of the molecular mechanism underlying gene transfer, subsequently led to the development of plant transgene technology, an indispensabl ...201324166428
fundamental discoveries and simple recombination between circular plasmid dnas led to widespread use of agrobacterium tumefaciens as a generalized vector for plant genetic engineering.fundamental research aimed to determine the limits of the agrobacterium transfer dna (t-dna) element that stably inserted into plant nuclear dna to cause crown gall tumor formation. the t-dna borders were discovered to be exceedingly precise, revealing that t-dna insertion into the plant genome was reproducible and exact. deletion of the internal regions of the t-dna, to remove the tumor forming genes, while retaining the t-dna borders, resulted again in efficient dna transfer to plant cells, bu ...201324166427
thirty years of transgenic research in plants.in 1983, the first transgenic tissues and plants were generated by means of disarmed agrobacterium tumefaciens strains, in which the oncogenes had been replaced by antibiotic resistance markers. hence, this special issue of the international journal of developmental biology celebrates 30 years of transgenic research in plants! eminent scientists working in the field of plant transformation or plant biotechnology have contributed to this publication and reviewed the state of the art of their part ...201324166426
targeted gene deletion in aspergillus fumigatus using microbial machinery and a recyclable marker.the emerging invasive fungal pathogen aspergillus fumigatus causes very serious infections among immunocompromised patient populations. while the genome of this pathogen has been sequenced, a major barrier to better understanding the complex biology of this eukaryotic organism is a lack of tools for efficient genetic manipulation. to improve upon this, we have generated a new gene deletion system for a. fumigatus using yeast recombinational cloning and agrobacterium tumefaciens mediated transfor ...201324161898
treatment of rhizobium radiobacter bacteremia in a critically ill trauma patient.to report the first case of rhizobium radiobacter bacteremia in a critically ill trauma patient.201324259594
cytokinins secreted by agrobacterium promote transformation by repressing a plant myb transcription factor.agrobacterium-mediated transformation is the most widely used technique for generating transgenic plants. however, many crops remain recalcitrant. we found that an arabidopsis myb family transcription factor (mtf1) inhibited plant transformation susceptibility. mutating mtf1 increased attachment of several agrobacterium strains to roots and increased both stable and transient transformation in both susceptible and transformation-resistant arabidopsis ecotypes. cytokinins from agrobacterium tumef ...201324255177
agrobacterium tumefaciens-mediated transformation of the lichen fungus, umbilicaria muehlenbergii.transformation-mediated mutagenesis in both targeted and random manners has been widely applied to decipher gene function in diverse fungi. however, a transformation system has not yet been established for lichen fungi, severely limiting our ability to study their biology and mechanism underpinning symbiosis via gene manipulation. here, we report the first successful transformation of the lichen fungus, umbilicaria muehlenbergii, via the use of agrobacterium tumefaciens. we generated a total of ...201324386304
agrobacterium tumefaciens-mediated transformation of valsa mali: an efficient tool for random insertion mutagenesis.valsa mali is a causal agent of apple and pear trees canker disease, which is a destructive disease that causes serious economic losses in eastern asia, especially in china. the lack of an efficient transformation system for valsa mali retards its investigation, which poses difficulties to control the disease. in this research, a transformation system for this pathogen was established for the first time using a. tumefaciens-mediated transformation (atmt), with the optimal transformation conditio ...201324381526
detection of genetically modified tomato using pcr coupled with muparaflo microfluidics microarrays.genetically modified (gm) tomatoes have been approved for commercialization in many countries since the first gm tomato flavr savr was permitted for planting in 1994. to meet the requirement of the gm tomatoes labeling policy, in this study we firstly set up the conventional pcr and multiplex pcr detection system for screening the universal elements transformed into tomato, such as cauliflower mosaic virus 35s (camv 35s) promoter, nopaline synthase (nos) terminator of agrobacterium tumefaciens, ...201324266223
generation of doubled haploid transgenic wheat lines by microspore transformation.microspores can be induced to develop homozygous doubled haploid plants in a single generation. in the present experiments androgenic microspores of wheat have been genetically transformed and developed into mature homozygous transgenic plants. two different transformation techniques were investigated, one employing electroporation and the other co-cultivation with agrobacterium tumefaciens. different tissue culture and transfection conditions were tested on nine different wheat cultivars using ...201324260351
overexpression of cytokinin dehydrogenase genes in barley (hordeum vulgare cv. golden promise) fundamentally affects morphology and fertility.barley is one of the most important cereal crops grown worldwide. it has numerous applications, but its utility could potentially be extended by genetically manipulating its hormonal balances. to explore some of this potential we identified gene families of cytokinin dehydrogenases (ckx) and isopentenyl transferases, enzymes that respectively irreversibly degrade and synthesize cytokinin (ck) plant hormones, in the raw sequenced barley genome. we then examined their spatial and temporal expressi ...201324260147
global analysis of differentially expressed genes and proteins in the wheat callus infected by agrobacterium tumefaciens.agrobacterium-mediated plant transformation is an extremely complex and evolved process involving genetic determinants of both the bacteria and the host plant cells. however, the mechanism of the determinants remains obscure, especially in some cereal crops such as wheat, which is recalcitrant for agrobacterium-mediated transformation. in this study, differentially expressed genes (degs) and differentially expressed proteins (deps) were analyzed in wheat callus cells co-cultured with agrobacteri ...201324278131
β-1,3-glucanase class iii promotes spread of pvy(ntn) and improves in planta protein production.glucanases are enzymes regulating the size exclusion limit and permeability of plasmodesmata and play a role in biotic stress. in plant genomes, they are encoded as relatively large gene families divided into four classes. most studies of plant virus interactions have focused on glucanases from classes i and ii. in our study, we have evaluated the role of the β-1,3-glucanase class iii (glu-iii) gene in the potato-potato virus y(ntn) (pvy(ntn)) interaction and implemented the findings to plant bi ...201324273610
agrobacterium-mediated genetic transformation using cotyledons in japanese pear (pyrus pyrifolia).genetic transformation was successfully established producing both transformed adventitious shoots and calli in japanese pear (pyrus pyrifolia nakai) by using cotyledons as explants. cotyledons of five cultivars were co-cultivated with agrobacterium tumefaciens strain lba4404 carrying the pbin19-sgfp, which contained a green fluorescent protein gene and the neomycin phosphotransferase gene. in order to increase transformation efficiency, sonication and ethylenedioxybis (ethylamine)-n,n,n',n'-tet ...201324273422
unlocking the potential of tropical root crop biotechnology in east africa by establishing a genetic transformation platform for local farmer-preferred cassava cultivars.cassava genetic transformation capacity is still mostly restricted to advanced laboratories in the usa, europe and china; and its implementation and maintenance in african laboratories has remained scarce. the impact of transgenic technologies for genetic improvement of cassava will depend largely on the transfer of such capabilities to researchers in africa, where cassava has an important socioeconomic niche. a major constraint to the development of genetic transformation technologies for cassa ...201324400011
indica rice cultivar irga 424, transformed with cry genes of b. thuringiensis, provided high resistance against spodoptera frugiperda (lepidoptera: noctuidae).plant expression of the entomopathogenic bacteria bacillus thuringiensis cry gene has reduced the damage created by insect pests in several economically important cultures. for this study, we have conducted genetic transformation of the indica rice "irga 424", via agrobacterium tumefaciens, using the b. thuringiensis cry1aa and cry1b genes, with the objective of obtaining rice plants resistant to the insect pests from this culture. the gene constructions harbor the promoters maize proteinase inh ...201324498761
structural insight into l-ribulose 3-epimerase from mesorhizobium loti.l-ribulose 3-epimerase (l-re) from mesorhizobium loti has been identified as the first ketose 3-epimerase that shows the highest observed activity towards ketopentoses. in the present study, the crystal structure of the enzyme was determined to 2.7 å resolution. the asymmetric unit contained two homotetramers with the monomer folded into an (α/β)8-barrel carrying four additional short α-helices. the overall structure of m. loti l-re showed significant similarity to the structures of ketose 3-epi ...201324311575
agrobacterium-mediated transformation of the white-rot fungus physisporinus vitreus.the biotechnologically important white-rot fungus physisporinus vitreus was co-cultivated with agrobacterium tumefaciens agl-1 carrying plasmids with nourseothricin resistance as the selectable marker gene and red fluorescence protein as a visual marker. mitotically stable transformed isolates were obtained showing red fluorescence protein activity.201324050950
vea and laea transcriptional factors regulate ochratoxin a biosynthesis in aspergillus carbonarius.ochratoxin a (ota) is a mycotoxin with nephrotoxic, teratogenic and immunotoxic properties which represents a serious risk for human and animal health. aspergillus carbonarius is considered the main ota-producing species in grapes and products such as raisins, wine or juices, although it has also been isolated from coffee, cocoa and cereals. till now not much information is available about regulatory mechanisms of ota production by a. carbonarius. a better understanding of how environmental fact ...201324041999
tropaeolum tops tobacco - simple and efficient transgene expression in the order brassicales.transient expression systems are valuable tools in molecular biology. agrobacterial infiltration of leaves is well-established in tobacco, but has led to limited success in the model plant arabidopsis thaliana. an efficient expression system combining the advantages of arabidopsis (well-characterised) and the simplicity of leaf infiltration is desirable. here, i describe agrobacterium tumefaciens-mediated transformation of tropaeolummajus (nasturtium, order brassicales) as a remarkably simple, c ...201324039923
new bacteriocin from bacillus clausii straingm17: purification, characterization, and biological activity.a bacteriocin-producing strain (9,000 au/ml) was isolated from the rhizosphere of algerian healthy plants ononis angustissima lam. and identified as bacillus clausii strain gm17. the bacteriocin, called bac-gm17, was purified from the culture supernatant after heat treatment, ammonium sulfate precipitation, sephadex g-50 chromatography and mono q fast-performance liquid chromatography (fplc). based on matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis, the puri ...201324037515
high-level extracellular production of d-psicose-3-epimerase with recombinant escherichia coli by a two-stage glycerol feeding approach.the aim of this study is to achieve high-level extracellular production of d-psicose-3-epimerase (dpe) with recombinant escherichia coli. high-level production of dpe is one of the key factors in d-psicose production. in the present study, the gene aal45544.1 from agrobacterium tumefaciens str. c58 was modified by artificial synthesis for overexpression in e. coli. the total dpe activity reached 3.96 u ml(-1) after optimization of the media composition, induction temperature, and concentration o ...201324037040
indole inhibits bacterial quorum sensing signal transmission by interfering with quorum sensing regulator folding.quorum sensing (qs)-dependent biofilm formation and motility were controlled by aqsr in acinetobacter oleivorans dr1. qs-controlled phenotypes appeared to be inhibited by indole and the aqsr mutant had the same phenotypes. we demonstrated that the turnover rate of aqsr became more rapid without the n-acylhomoserine lactone (ahl) signal, and that indole could increase the expression of many protease and chaperone proteins. the addition of exogenous indole decreased the expression of two aqsr-targ ...201324025605
construction of an engineering strain producing high yields of α-transglucosidase via agrobacterium tumefaciens-mediated transformation of asperillus niger.in this study, agrobacterium tumefaciens-mediated transformation (atmt) was used in breeding industrial strains for the purpose of improving α-transglucosidase production. firstly, an efficient atmt system for asperillus niger was established by optimization of several influencing factors, in which transformation efficiency was improved up to 14-fold compared with the initial conditions. furthermore, binary vector pbi-glu containing an α-transglucosidase expression cassette was constructed and t ...201324018680
[analytical methods for control of foodstuffs made from bioengineered plants].foodstuffs made by modern biotechnology are requiring for special control. analytical methods used for these purposes are being constantly perfected. when choosing a strategy for the analysis, several factors have to be assessed: specificity, sensitivity, practically of the method and time efficiency. to date, the gmo testing methods are mainly based on the inserted dna sequences and newly produced proteins in gmos. protein detection methods are based mainly on elisa. the specific detection of a ...201324006752
increased 1-aminocyclopropane-1-carboxylate deaminase activity enhances agrobacterium tumefaciens-mediated gene delivery into plant cells.agrobacterium-mediated transformation is a useful tool for the genetic modification in plants, although its efficiency is low for several plant species. agrobacterium-mediated transformation has three major steps in laboratory-controlled experiments: the delivery of t-dna into plant cells, the selection of transformed plant cells, and the regeneration of whole plants from the selected cells. each of these steps must be optimized to improve the efficiency of agrobacterium-mediated plant transform ...201324000136
demonstration of crispr/cas9/sgrna-mediated targeted gene modification in arabidopsis, tobacco, sorghum and rice.the type ii crispr/cas system from streptococcus pyogenes and its simplified derivative, the cas9/single guide rna (sgrna) system, have emerged as potent new tools for targeted gene knockout in bacteria, yeast, fruit fly, zebrafish and human cells. here, we describe adaptations of these systems leading to successful expression of the cas9/sgrna system in two dicot plant species, arabidopsis and tobacco, and two monocot crop species, rice and sorghum. agrobacterium tumefaciens was used for delive ...201323999092
transient gene expression in medicago truncatula leaves via agroinfiltration.transient expression is a powerful method for the functional characterization of genes. in this chapter, we outline a protocol for the transient expression of constructs in medicago truncatula leaves using agrobacterium tumefaciens infiltration. using quantitative real-time pcr we demonstrate that the infiltration of a construct containing the legume anthocyanin production 1 (lap1) transcription factor results in the strong upregulation of key biosynthetic genes and the accumulation of anthocyan ...201323996318
stable transformation of medicago truncatula cv. jemalong for gene analysis using agrobacterium tumefaciens.medicago truncatula is a model legume that has all the genomic resources to be suitable as a model for functional genomics. transformation to produce transgenic plants is part of this toolkit, enabling a spectrum of approaches to study gene function: including knockdown, overexpression, reporter genes for gene expression, and proteins tagged with fluorescent proteins such as gfp. a special genetic line is necessary for transformation and jemalong 2ha derived from cv. jemalong is used in the meth ...201323996317
efficient gene targeting in penicillium chrysogenum using novel agrobacterium-mediated transformation approaches.the industrial production of β-lactam antibiotics by penicillium chrysogenum has increased tremendously over the last decades, however, further optimization via classical strain and process improvement has reached its limits. the availability of the genome sequence provides new opportunities for directed strain improvement, but this requires the establishment of an efficient gene targeting (gt) system. recently, mutations affecting the non-homologous end joining (nhej) pathway were shown to incr ...201323994321
molecular tools for functional genomics in filamentous fungi: recent advances and new strategies.advances in genetic transformation techniques have made important contributions to molecular genetics. various molecular tools and strategies have been developed for functional genomic analysis of filamentous fungi since the first dna transformation was successfully achieved in neurospora crassa in 1973. increasing amounts of genomic data regarding filamentous fungi are continuously reported and large-scale functional studies have become common in a wide range of fungal species. in this review, ...201323988676
antitumor, antioxidant and antimicrobial studies of substituted pyridylguanidines.a series of n-pivaloyl-n'-(alkyl/aryl)-n''-pyridylguanidine of general formula c₄h₉conhc(nr¹r²)npy have been synthesized and characterized using elemental analysis, ft-ir, multinuclear nmr spectroscopy, and in the case of compounds 7 and 11, by single crystal x-ray diffraction (xrd). the synthesized guanidines were tested for antitumor activities against potato tumor, and showed excellent inhibition against agrobacterium tumefaciens (at10)-induced tumor. the antioxidant and antimicrobial activit ...201323985956
genetic transformation in citrus.citrus is one of the world's important fruit crops. recently, citrus molecular genetics and biotechnology work have been accelerated in the world. genetic transformation, a biotechnological tool, allows the release of improved cultivars with desirable characteristics in a shorter period of time and therefore may be useful in citrus breeding programs. citrus transformation has now been achieved in a number of laboratories by various methods. agrobacterium tumefaciens is used mainly in citrus tran ...201323983635
live-cell imaging of microtubules and microtubule-associated proteins in arabidopsis thaliana.microtubules and microtubule-associated proteins (maps) play fundamental roles in plant growth and morphogenesis. the ability to observe microtubules and maps in living cells using fluorescent protein fusions has propelled plant scientists forward and given them the opportunity to answer longstanding biological questions. in combination with the genetic resources available in the model plant arabidopsis thaliana, our mechanistic understanding of how the microtubule cytoskeleton affects plant lif ...201323973076
functional analysis of autophagy genes via agrobacterium-mediated transformation in the vascular wilt fungus verticillium dahliae.autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in foliar infection by many plant pathogenic fungi. however, the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood. here, we report the establishment of an agrobacterium tumefaciens-mediated transformation (atmt) system and its application to investigate two autophag ...201323969251
genome sequence of the quorum-quenching agrobacterium tumefaciens strain wrt31.agrobacterium tumefaciens strain wrt31 is a quorum-sensing signal-degrading bacterium that has been isolated from the rhizosphere of tobacco plants. this strain belongs to a. tumefaciens genomovar g1, is avirulent on various putative host plants, devoid of ti plasmid, and contains the blcc gene encoding a gamma-butyrolactonase.201323969055
genetic circuits that govern bisexual and unisexual reproduction in cryptococcus neoformans.cryptococcus neoformans is a human fungal pathogen with a defined sexual cycle. nutrient-limiting conditions and pheromones induce a dimorphic transition from unicellular yeast to multicellular hyphae and the production of infectious spores. sexual reproduction involves cells of either opposite (bisexual) or one (unisexual) mating type. bisexual and unisexual reproduction are governed by shared components of the conserved pheromone-sensing cpk1 mapk signal transduction cascade and by mat2, the m ...201323966871
optimization, validation, and application of a real-time pcr protocol for quantification of viable bacterial cells in municipal sewage sludge and biosolids using reporter genes and escherichia coli.biosolids result from treatment of sewage sludge to meet jurisdictional standards, including pathogen reduction. once government regulations are met, materials can be applied to agricultural lands. culture-based methods are used to enumerate pathogen indicator microorganisms but may underestimate cell densities, which is partly due to bacteria existing in a viable but non-culturable physiological state. viable indicators can also be quantified by realtime polymerase chain reaction (qpcr) used wi ...201323958912
targeting fungal genes by diced sirnas: a rapid tool to decipher gene function in aspergillus nidulans.gene silencing triggered by chemically synthesized small interfering rnas (sirnas) has become a powerful tool for deciphering gene function in many eukaryotes. however, prediction and validation of a single sirna duplex specific to a target gene is often ineffective. rna interference (rnai) with synthetic sirna suffers from lower silencing efficacy, off-target effects and is cost-intensive, especially for functional genomic studies. with the explosion of fungal genomic information, there is an i ...201324130711
pigment-protein interactions in phytochromes probed by fluorescence line narrowing spectroscopy.fluorescence line narrowing (fln) spectroscopy was used to study bacteriophytochromes and variants from various species in their red-absorbing pr ground state, including phytochromes agp1 from agrobacterium tumefaciens , drbphp from deinococcus radiodurans , and rpbphp2 and rpbphp3 from rhodopseudomonas palustris . a species-dependent narrowing of the fluorescence emission bands is observed. the results suggest varied pigment-protein interactions, possibly connected to chromophore mobility or ex ...201324205818
concerted transfer of the virulence ti plasmid and companion at plasmid in the agrobacterium tumefaciens-induced plant tumour.the plant pathogen agrobacterium tumefaciens c58 harbours three independent type iv secretion (t4ss) machineries. t4sst-dna promotes the transfer of the t-dna to host plant cells, provoking tumour development and accumulation of opines such as nopaline and agrocinopines. t4sspti and t4sspat control the bacterial conjugation of the ti and at plasmids respectively. expression of t4sspti is controlled by the agrocinopine-responsive transcriptional repressor accr. in this work, we compared the genom ...201324118167
agrobacterium-mediated transformation of promising oil-bearing marine algae parachlorella kessleri.parachlorella kessleri is a unicellular alga which grows in fresh as well as marine water and is commercially important as biomass/lipid feedstock and in bioremediation. the present study describes the successful transformation of marine p. kessleri with the help of agrobacterium tumefaciens. transformed marine p. kessleri was able to tolerate more than 10 mg l(-1) hygromycin concentration. co-cultivation conditions were modulated to allow the simultaneous growth of both marine p. kessleri and a ...201324097049
the involvement of β-amyrin 28-oxidase (cyp716a52v2) in oleanane-type ginsenoside biosynthesis in panax ginseng.panax species are the most popular medicinal herbs. the root of these plants contains pharmacologically active triterpene saponins, also known as ginsenosides, compounds that are divided into dammarane- and oleanane-type triterpenes. two cyp716a subfamily genes (cyp716a47 and cyp716a53v2) were recently characterized, encoding an enzyme catalyzing the hydroxylation of dammarane-type triterpenes in panax ginseng. herein, we report that one cyp716a subfamily gene (cyp716a52v2) isolated from p. gins ...201324092881
genome sequence of the quorum-sensing-signal-producing nonpathogen agrobacterium tumefaciens strain p4.agrobacterium tumefaciens p4 is a quorum-sensing-signal-producing bacterium that has been isolated from the tobacco rhizosphere. this strain belongs to genomospecies 1 of the a. tumefaciens complex; it is avirulent on various putative host plants, devoid of the ti plasmid, and contains a luxi homolog on the at plasmid.201324092790
biological activity of the tzs gene of nopaline agrobacterium tumefaciens gv3101 in plant regeneration and genetic transformation.agrobacterium tumefaciens has been widely used in plant genetic transformation. hormone-encoding genes residing in the t-dna region have been removed, resulting in disarmed agrobacterium strains that are used in various transformation experiments. nopaline agrobacterium strains, however, carry another hormone gene, trans-zeatin synthesizing (tzs), that codes for trans-zeatin in the virulence region of the tumor-inducing plasmids. we investigated the activity and function of the tzs gene of a nop ...201324088018
the transformation of the photo-thermo sensitive genic male-sterile line 261s of rice via an expression vector containing the anti-waxy gene.transgenic photo-thermo sensitive genic male sterility oryza sativa l. cv. "261s" plants with the anti-waxy gene were successfully obtained using an agrobacterium tumefaciens-mediated co-transformation method. marker-free homozygous transgenic lines with the anti-waxy gene were obtained. the setting seed rates of the transgenic plants via self-pollination or via crossing with the restorer line wx99075 rice and the 1000-grain weight of the transgenic plants and the f2 hybrid seeds obtained by cro ...201323853508
assessment of factors influencing the agrobacterium-mediated in planta seed transformation of brinjal (solanum melongena l.).an efficient and reproducible in planta transformation method was developed for brinjal using seed as an explant. the brinjal seeds were infected with agrobacterium tumefaciens eha 105 harbouring pcambia 1301-bar plasmid, and the transformants were selected against basta®. several parameters influencing the in planta seed transformation such as pre-culture duration, acetosyringone concentration, surfactants, duration of sonication, vacuum pressure and vacuum duration have been evaluated. the put ...201323852797
a genetic system for citrus tristeza virus using the non-natural host nicotiana benthamiana: an update.in nature citrus tristeza virus (ctv), genus closterovirus, infects only the phloem cells of species of citrus and related genera. finding that the ctv t36 strain replicated in nicotiana benthamiana (nb) protoplasts and produced normal virions allowed development of the first genetic system based on protoplast transfection with rna transcribed from a full-genome cdna clone, a laborious and uncertain system requiring several months for each experiment. we developed a more efficient system based o ...201323847598
genetic transformation of western clover (trifolium occidentale d. e. coombe.) as a model for functional genomics and transgene introgression in clonal pasture legume species.western clover (trifolium occidentale) is a perennial herb with characteristics compatible for its development as an attractive model species for genomics studies relating to the forage legume, white clover (trifolium repens). its characteristics such as a small diploid genome, self-fertility and ancestral contribution of one of the genomes of t. repens, facilitates its use as a model for genetic analysis of plants transformed with legume or novel genes.201323841995
protease inhibitor from insect silk-activities of derivatives expressed in vitro and in transgenic potato.several recombinant derivatives of serine protease inhibitor called silk protease inhibitor 2 (spi2), which is a silk component in galleria mellonella (lepidoptera, insecta), were prepared in the expression vector pichia pastoris. both the native and the recombinant protease inhibitors were highly active against subtilisin and proteinase k. the synthetic spi2 gene with ala codon in the p1 position was fused with mgfp-5 to facilitate detection of the transgene and its protein product. a construct ...201323824530
expression of verocytotoxic escherichia coli antigens in tobacco seeds and evaluation of gut immunity after oral administration in mouse model.verocytotoxic escherichia (e.) coli strains are responsible for swine oedema disease, which is an enterotoxaemia that causes economic losses in the pig industry. the production of a vaccine for oral administration in transgenic seeds could be an efficient system to stimulate local immunity. this study was conducted to transform tobacco plants for the seed-specific expression of antigenic proteins from a porcine verocytotoxic e. coli strain. parameters related to an immunological response and pos ...201323820163
central venous catheter infections in outpatients with pulmonary hypertension treated with continuous iloprost.intravenous prostanoid therapy is one cornerstone of therapy for patients with pre-capillary pulmonary hypertension (ph). long-term central venous catheters expose patients to infectious complications.201323817035
identification of virulence genes in the crucifer anthracnose fungus colletotrichum higginsianum by insertional mutagenesis.to investigate the molecular and genetic mechanisms underlying virulence of colletotrichum higginsianum on arabidopsis thaliana, a t-dna insertion mutant library of c. higginsianum, the causal agent of crucifer anthracnose, was established using agrobacterium tumefaciens-mediated transformation. among 875 transformants tested for virulence on arabidopsis, six mutants with altered virulence, including an appressorial melanin-deficient mutant t734, two mutants defective in penetration, t45 and b30 ...201323806215
the role of fabg3 in fruit ripening and b. cinerea fungal infection of strawberry.in plants, β-glucosidases (bg) have been implicated in developmental and pathogen defense, and are thought to take part in abscisic acid (aba) synthesis via hydrolysis of aba glucose ester to release active aba; however, there is no genetic evidence for the role of bg genes in ripening and biotic/abiotic stress in fruits. to clarify the role of bg genes in fruit, eight fa/fvbg genes encoding β-glucosidase were isolated using information from the genbank strawberry nucleotide database. of the fa/ ...201323802911
taxol production by an endophytic fungus, fusarium redolens, isolated from himalayan yew.different endophytic fungi isolated from himalayan yew plants were tested for their ability to produce taxol. the bapt gene (c-13 phenylpropanoid side chain-coa acetyl transferase) involved in the taxol biosynthetic pathway was used as a molecular marker to screen taxol-producing endophytic fungi. taxol extracted from fungal strain tbpj-b was identified by hplc and ms analysis. strain tbpj-b was identified as fusarium redolens based on the morphology and internal transcribed spacer region of nrd ...201323801250
expression of shigella flexneri ipab gene in tobacco.shigellosis is a leading cause of diarrhea in many developing countries and although the disease can be controlled and managed with antibiotics, the constant emergence of resistant species requiring ever newer antibacterial drugs make development of an effective vaccine necessary. the bacteria are highly contagious and since immunity to shigella is serotype-specific a multi-serotype vaccine is required for adequate protection. proteins encoded by shigella invasion plasmid, which are part of the ...201323799180
improving the glycosyltransferase activity of agrobacterium tumefaciens glycogen synthase by fusion of n-terminal starch binding domains (sbds).glycogen and starch, the major storage carbohydrate in most living organisms, result mainly from the action of starch or glycogen synthases (ss or gs, respectively, ec 2.4.1.21). ssiii from arabidopsis thaliana is an ss isoform with a particular modular organization: the c-terminal highly conserved glycosyltransferase domain is preceded by a unique specific region (ssiii-sd) which contains three in tandem starch binding domains (sbds, named d1, d2 and d3) characteristic of polysaccharide degradi ...201323796574
in vitro antibacterial and antitumor activities of some medicinal plant extracts, growing in turkey.to investigate antibacterial and antitumor activities of 51 different extracts prepared with 3 types of solvents (water, ethanol and methanol) of 16 different plant species (ajuga reptans (a. reptans) l., phlomis pungens (p. pungens) willd., marrubium astracanicum (m. astracanicum) jacq., nepeta nuda (n. nuda) l., stachys annua (s. annua) l., genista lydia (g. lydia) boiss., nuphar lutea (n. lutea) l., nymphaea alba (n. alba) l., vinca minor (v. minor) l., stellaria media (s. media) l., capsella ...201323790332
[infective endocarditis by rhizobium radiobacter. a case report].rhizobium radiobacter is a gram-negative, nitrogen-fixing bacterium, which is found mainly on the ground. it rarely causes infections in humans. it has been associated with bacteremia, secondary to colonization of intravascular catheters, in immunocompromised patients. the aim of this paper was to report the case of an infective endocarditis caused by r. radiobacter, in a 47-year-old male, diagnosed with chronic kidney disease stage 5, on replacement therapy with hemodialysis and who attended th ...201323781714
agrobacterium tumefaciens-mediated genetic transformation of salix matsudana koidz. using mature seeds.an agrobacterium tumefaciens-mediated transformation method was developed for salix matsudana koidz. using mature seeds as starting material. multiple shoots were induced directly from embryonic shoot apices of germinating seeds. although thidiazuron, 6-benzylaminopurine and zeatin induced multiple shoot induction with high frequency, zeatin (4.5 μm) was more effective for elongation of shoots and roots. the binary vector pcambia1303, which contained neomycin phosphotransferase as a selectable m ...201323771952
efficient auto-excision of a selectable marker gene from transgenic citrus by combining the cre/loxp system and ipt selection.a highly efficient cre-mediated deletion system, offering a good alternative for producing marker-free transgenic plants that will relieve public concerns regarding gmos, was first developed in citrus. the presence of marker genes in genetically modified crops raises public concerns regarding their safety. the removal of marker genes can prevent the risk of their flow into the environment and hasten the public's acceptance of transgenic products. in this study, a new construct based on the cre/l ...201323771575
variable internal flexibility characterizes the helical capsid formed by agrobacterium vire2 protein on single-stranded dna.agrobacterium is known for gene transfer to plants. in addition to a linear ssdna oligonucleotide, agrobacterium tumefaciens secretes an abundant ssdna-binding effector, vire2. in many ways vire2 adapts the conjugation mechanism to transform the eukaryotic host. the crystal structure of vire2 shows two compact domains joined by a flexible linker. bound to ssdna, vire2 forms an ordered solenoidal shell, or capsid known as the t-complex. here, we present a three-dimensional reconstruction of the v ...201323769668
novel rhizobium lineages isolated from root nodules of the common bean (phaseolus vulgaris l.) in andean and mesoamerican areas.the taxonomic affiliations of nineteen root-nodule bacteria isolated from the common bean (phaseolus vulgaris l.) in mexico, ecuador and brazil were investigated by analyses of 16s rrna and of four protein-coding housekeeping genes. one strain from mexico could be assigned to rhizobium etli and two from brazil to rhizobium leucaenae, whereas another from mexico corresponded to a recently described bean-nodulating species-level lineage related to r. etli and rhizobium phaseoli. ten strains isolat ...201323764913
response of chickpea genotypes to agrobacterium-mediated delivery of chickpea chlorotic dwarf virus (cpcdv) genome and identification of resistance source.chickpea stunt disease caused by chickpea chlorotic dwarf virus (cpcdv) (genus mastrevirus, family geminiviridae) is the most important biotic stress affecting chickpea crops worldwide. a survey conducted on the incidence of stunt disease clearly revealed high incidence of the disease with severe symptom expression in both indigenous and imported genotypes. to manage the disease in a sustainable way, resistant genotypes need to be bred by adopting objective and precise assessment of the disease ...201323955474
stable transformation of ferns using spores as targets: pteris vittata and ceratopteris thalictroides.ferns (pteridophyta) are very important members of the plant kingdom that lag behind other taxa with regards to our understanding of their genetics, genomics, and molecular biology. we report here, to our knowledge, the first instance of stable transformation of fern with recovery of transgenic sporophytes. spores of the arsenic hyperaccumulating fern pteris vittata and tetraploid 'c-fern express' (ceratopteris thalictroides) were stably transformed by agrobacterium tumefaciens with constructs c ...201323933990
agrobacterium-mediated transformation of guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis.guignardia citricarpa is the causal agent of citrus black spot (cbs), an important disease in citriculture. due to the expressive value of this activity worldwide, especially in brazil, understanding more about the functioning of this fungus is of utmost relevance, making possible the elucidation of its infection mechanisms, and providing tools to control cbs. this work describes for the first time an efficient and successful methodology for genetic transformation of g. citricarpa mycelia, which ...201323931121
purple-leaved ficus lyrata plants produced by overexpressing a grapevine vvmyba1 gene.this study established an efficient method of regenerating plants of ficus lyrata and producing purple-leaved f. lyrata plants through genetic transformation using a vvmyba1 gene of grapevine.201323926030
the selectable antibiotic marker, teta(c), increases pseudomonas aeruginosa susceptibility to the herbicide/superoxide generator, paraquat.the presence of the widely used selectable antibiotic marker, teta(c), unexpectedly increased the sensitivity of pseudomonas aeruginosa pao1 to the superoxide-generating herbicide, paraquat. a dna fragment spanning the first 99 amino acids of teta(c) was sufficient to confer paraquat sensitivity. the teta(c)-induced paraquat sensitive phenotype was observed in other gram-negative bacteria such as agrobacterium tumefaciens, salmonella enterica ser. typhimurium and xanthomonas campestris suggestin ...201323907582
silencing agrobacterium oncogenes in transgenic grapevine results in strain-specific crown gall resistance.grapevine rootstock transformed with an agrobacterium oncogene-silencing transgene was resistant to certain agrobacterium strains but sensitive to others. thus, genetic diversity of agrobacterium oncogenes may limit engineering crown gall resistance.201323903949
identification and characterization of the biochemical function of agrobacterium t-complex-recruiting protein atu5117.atu5117 from agrobacterium tumefaciens is a highly conserved protein with a putative nucleotidyltransferase domain in its n-terminal region and a putative higher eukaryotes and prokaryotes nucleotide-binding domain in its c-terminal region. this protein has been shown to be a t-complex-recruiting protein that can recruit t-complex from the cytosol to the polar virb/d4 type iv secretion system (t4ss). however, the biochemical function of atu5117 is still unknown. here, we show that atu5117 is a ( ...201323902381
norway spruce (picea abies) genetic transformation with modified cry3a gene of bacillus thuringiensis.modified versions of the cry3a gene of bacillus thuringiensis (bt) were transferred into norway spruce (picea abies). both the biolistic approach and agrobacterium tumefaciens mediated procedure were employed for transformation of embryogenic tissue (et) cultures. the latter method proved to be more efficient yielding 70 transgenic embryogenic tissue lines compared with 18 lines obtained by biolistics. the modified cry3a genes were driven by a 35s promoter and the nptii screenable selection mark ...201323888296
construction of a sonchus yellow net virus minireplicon: a step toward reverse genetic analysis of plant negative-strand rna viruses.reverse genetic analyses of negative-strand rna (nsr) viruses have provided enormous advances in our understanding of animal viruses over the past 20 years, but technical difficulties have hampered application to plant nsr viruses. to develop a reverse genetic approach for analysis of plant nsr viruses, we have engineered sonchus yellow net nucleorhabdovirus (synv) minireplicon (mr) reporter cassettes for agrobacterium tumefaciens expression in nicotiana benthamiana leaves. fluorescent reporter ...201323885070
inhibition of pbgp43 expression may suggest that gp43 is a virulence factor in paracoccidioides brasiliensis.glycoprotein gp43 is an immunodominant diagnostic antigen for paracoccidioidomycosis caused by paracoccidioides brasiliensis. it is abundantly secreted in isolates such as pb339. it is structurally related to beta-1,3-exoglucanases, however inactive. its function in fungal biology is unknown, but it elicits humoral, innate and protective cellular immune responses; it binds to extracellular matrix-associated proteins. in this study we applied an antisense rna (arna) technology and agrobacterium t ...201323874627
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