| [subcellular localization and resistance to gibberella fujikuroi of atelhyprp2 in transgenic tobacco]. | the subcellular localization and the resistance to fungal pathogen gibberella fujikuroi of the protein encoded by arabidopsis atelhyprp2 (earli1-like hybrid proline-rich protein 2, at4g12500) were investigated using transgenic tobacco plants. the coding sequence of atelhyprp2 was amplified from genomic dna of col-0 ecotype. after restriction digestion, the pcr fragment was ligated into pcambia1302 to produce a fusion expression vector, pcambia1302-atelhyprp2-gfp. then the recombinant plasmid was ... | 2014 | 25007583 |
| ghh2a12, a replication-dependent histone h2a gene from gossypium hirsutum, is negatively involved in the development of cotton fiber cells. | ghh2a12 was preferentially expressed at the initiation and early elongation stage of cotton fiber development, and overexpression of ghh2a12 caused retardation of fiber initiation and produced shorter fibers. histone h2a is a component of eukaryotic chromatin whose function has not been studied in cotton. we have isolated an h2a gene encoding 156 amino acids, named ghh2a12. like other plant histone h2as, ghh2a12 contains a typical spkk motif in the carboxy-terminal and a plant-unique peptide-bin ... | 2014 | 25001001 |
| bidirectional promoter trapping t-dna for insertional mutagenesis in verticillium dahliae. | transfer dna (t-dna)-based random insertional mutagenesis is a universal forward genetic approach for gene identification and cloning in many phytopathogenic fungi. in a large number of randomly selected transformants, screening for mutants with a specific phenotype is laborious, especially for pathogenicity-defective mutants. to accelerate mutant screening and gene identification, a bidirectional promoter-trapping ti binary vector, 1300-bisgfp-hyg, was constructed and deployed in this study. mo ... | 2014 | 24993736 |
| agrobacterium tumefaciens-mediated genetic transformation of haptophytes (isochrysis species). | isochrysis galbana and isochrysis sp. are economically important microalgae from the division of haptophytes. here, we report agrobacterium-mediated stable dna transfer into their nuclear genomes. initial studies were performed to standardize co-cultivation media and determine the sensitivity of the microalgae to selective agents. up to 1 mg/ml of the antibiotic hygromycin did not inhibit growth, whereas both the haptophytes bleached in artificial seawater (asw) medium containing micromolar conc ... | 2014 | 24993358 |
| integrative gene transfer in the truffle tuber borchii by agrobacterium tumefaciens-mediated transformation. | agrobacterium tumefaciens-mediated transformation is a powerful tool for reverse genetics and functional genomic analysis in a wide variety of plants and fungi. tuber spp. are ecologically important and gastronomically prized fungi ("truffles") with a cryptic life cycle, a subterranean habitat and a symbiotic, but also facultative saprophytic lifestyle. the genome of a representative member of this group of fungi has recently been sequenced. however, because of their poor genetic tractability, i ... | 2014 | 24949275 |
| xyloglucan endotransglucosylase-hydrolase17 interacts with xyloglucan endotransglucosylase-hydrolase31 to confer xyloglucan endotransglucosylase action and affect aluminum sensitivity in arabidopsis. | previously, we reported that although the arabidopsis (arabidopsis thaliana) xyloglucan endotransglucosylase-hydrolase31 (xth31) has predominately xyloglucan endohydrolase activity in vitro, loss of xth31 results in remarkably reduced in vivo xyloglucan endotransglucosylase (xet) action and enhanced al resistance. here, we report that xth17, predicted to have xet activity, binds xth31 in yeast (saccharomyces cerevisiae) two-hybrid and coimmunoprecipitations assays and that this interaction may b ... | 2014 | 24948835 |
| a putative soybean gmssos1 confers enhanced salt tolerance to transgenic arabidopsis sos1-1 mutant. | the cdna of gmssos1, a putative plasma membrane na(+)/h(+) antiporter gene isolated from glycine max, glycine soja, and their hybrid, was constructed into plant expression vector pcambia 1300 and then transformed with agrobacterium tumefaciens under the control of camv 35s promoter to arabidopsis thaliana wild-type (wt) and mutant (atsos1-1) plants. by hygromycin resistance detection and pcr analysis, transgenic plants (wt35s:gmssos1 and atsos1-1 35s:gmssos1) were obtained. seed germination, see ... | 2015 | 24934653 |
| an efficient agrobacterium-mediated transformation system for poplar. | poplar is a model system for the regeneration and genetic transformation of woody plants. to shorten the time required for studies of transgenic poplar, efforts have been made to optimize transformation methods that use agrobacterium tumefaciens. in this study, an agrobacterium infective suspension was treated at 4 °c for at least 10 h before infecting explants. by transforming the populus hybrid clone "nanlin895" (populus deltoides×p. euramericana) with agrobacterium harboring the pbi121:carnac ... | 2014 | 24933641 |
| a golden gate modular cloning toolbox for plants. | plant synthetic biology requires robust and efficient methods for assembling multigene constructs. golden gate cloning provides a precision module-based cloning technique for facile assembly of multiple genes in one construct. we present here a versatile resource for plant biologists comprising a set of cloning vectors and 96 standardized parts to enable golden gate construction of multigene constructs for plant transformation. parts include promoters, untranslated sequences, reporters, antigeni ... | 2014 | 24933124 |
| functional characterization of 5-enopyruvylshikimate-3-phosphate synthase from alkaliphilus metalliredigens in transgenic arabidopsis. | although a large number of aroa enzymes (epsps: 5-enopyruvylshikimate-3-phosphate synthase) have been identified, cloned, and tested for glyphosate resistance, only two aroa variants, derived from agrobacterium tumefaciens strain cp4 and zea mays, have been utilized to produce the commercial glyphosate-resistant crops. here, we have used a pcr-based twostep dna synthesis method to synthesize an aroa gene (aroaa. metalliredigens) from alkaliphilus metalliredigens, encoding a new epsps. furthermor ... | 2014 | 24931502 |
| enzymatic properties and substrate specificity of a bacterial phosphatidylcholine synthase. | phosphatidylcholine (pc) is a rare membrane lipid in bacteria, but is crucial for virulence of the plant pathogen agrobacterium tumefaciens and various other pathogens. agrobacterium tumefaciens uses two independent pc biosynthesis pathways. one is dependent on the integral membrane protein pc synthase (pcs), which catalyzes the conversion of cytidine diphosphate-diacylglycerol (cdp-dag) and choline to pc, thereby releasing a cytidine monophosphate (cmp). here, we show that pcs consists of eight ... | 2014 | 24931117 |
| construction of a standard reference plasmid containing seven target genes for the detection of transgenic cotton. | insect resistance and herbicide tolerance are the dominant traits of commercialized transgenic cotton. in this study, we constructed a general standard reference plasmid for transgenic cotton detection. target genes, including the cowpea trypsin gene cpti, the insect resistance gene cry1ab/1ac, the herbicide tolerance gene cp4-epsps, the agrobacterium tumefaciens nopaline synthase (nos) terminator that exists in transgenic cotton and part of the endogenous cotton sadi gene were amplified from pl ... | 2014 | 24929128 |
| evolution of enzymatic activities in the enolase superfamily: galactarate dehydratase iii from agrobacterium tumefaciens c58. | the genome of agrobacterium tumefaciens c58 encodes 12 members of the enolase superfamily (ens), eight of which are members of the mandelate racemase (mr) subgroup and, therefore, likely to be acid sugar dehydratases. using a library of 77 acid sugars for high-throughput screening, one protein (uniprot entry a9cg74; locus tag atu4196) showed activity with both m-galactarate and d-galacturonate. two families of galactarate dehydratases had been discovered previously in the ens, galrd/talrd [yew, ... | 2014 | 24926996 |
| stable expression of mtld gene imparts multiple stress tolerance in finger millet. | finger millet is susceptible to abiotic stresses, especially drought and salinity stress, in the field during seed germination and early stages of seedling development. therefore developing stress tolerant finger millet plants combating drought, salinity and associated oxidative stress in these two growth stages is important. cellular protection through osmotic adjustment and efficient free radical scavenging ability during abiotic stress are important components of stress tolerance mechanisms i ... | 2014 | 24922513 |
| gene targeting using the agrobacterium tumefaciens-mediated crispr-cas system in rice. | the type ii clustered, regularly interspaced, short palindromic repeat (crispr)/ crispr-associated protein 9 (cas9) system is a novel molecular tool for site-specific genome modification. the crispr-cas9 system was recently introduced into plants by transient or stable transformation. | 2014 | 24920971 |
| protozoan alkbh8 oxygenases display both dna repair and trna modification activities. | the alkbh family of fe(ii) and 2-oxoglutarate dependent oxygenases comprises enzymes that display sequence homology to alkb from e. coli, a dna repair enzyme that uses an oxidative mechanism to dealkylate methyl and etheno adducts on the nucleobases. humans have nine different alkbh proteins, alkbh1-8 and fto. mammalian and plant alkbh8 are trna hydroxylases targeting 5-methoxycarbonylmethyl-modified uridine (mcm5u) at the wobble position of trnagly(ucc). in contrast, the genomes of some bacteri ... | 2014 | 24914785 |
| optimization of genetic transformation of artemisia annua l. using agrobacterium for artemisinin production. | artemisinin, a sesquiterpene lactone endoperoxide isolated from the medicinal plant artemisia annua l., is a choice and effective drug for malaria treatment. due to the low yield of artemisinin in plants, there is a need to enhance the production of artemisinin from a. annua and biotechnological technique may be one of the methods that can be used for the purpose. | 2014 | 24914301 |
| mutational analysis of the ve1 immune receptor that mediates verticillium resistance in tomato. | pathogenic verticillium species are economically important plant pathogens that cause vascular wilt diseases in hundreds of plant species. the ve1 gene of tomato confers resistance against race 1 strains of verticillium dahliae and v. albo-atrum. ve1 encodes an extracellular leucine-rich repeat (elrr) receptor-like protein (rlp) that serves as a cell surface receptor for recognition of the recently identified secreted verticillium effector ave1. to investigate recognition of ave1 by ve1, alanine ... | 2014 | 24911915 |
| construction and functional analysis of trichoderma harzianum mutants that modulate maize resistance to the pathogen curvularia lunata. | agrobacterium tumefaciens-mediated transformation (atmt) was used to generate an insertional mutant library of the mycelial fungus trichoderma harzianum. from a total of 450 mutants, six mutants that showed significant influence on maize resistance to c. lunata were analyzed in detail. maize coated with these mutants was more susceptible to c. lunata compared with those coated with a wild-type (wt) strain. similar to other fungal atmt libraries, all six mutants were single copy integrations, whi ... | 2014 | 24901960 |
| psim24 is a novel versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants. | we have constructed a small and highly efficient binary ti vector psim24 for plant transformation with maximum efficacy. in the psim24 vector, the size of the backbone of the early binary vector pkylxm24 (genbank accession no. hm036220; a derivative of pkylx71) was reduced from 12.8 kb to 7.1 kb. the binary vector psim24 is composed of the following genetic elements: left and right t-dna borders, a modified full-length transcript promoter (m24) of mirabilis mosaic virus with duplicated enhancer ... | 2014 | 24897541 |
| functional characterization of pmt2, encoding a protein-o-mannosyltransferase, in the human pathogen cryptococcus neoformans. | diazobenzoic acid b (dbb), also known as diazonium blue b or fast blue b, can be used to distinguish basidiomycetous yeasts from ascomycetes. this chemical has long been used for the taxonomic study of yeast species at the phylum level, but the mechanism underlying the dbb staining remains unknown. to identify molecular targets of dbb staining, we isolated agrobacterium tumefaciens-mediated insertional mutants of cryptococcus neoformans, a basidiomycetous pathogenic yeast, which were negative to ... | 2014 | 24892554 |
| hemoglobin uptake by paracoccidioides spp. is receptor-mediated. | iron is essential for the proliferation of fungal pathogens during infection. the availability of iron is limited due to its association with host proteins. fungal pathogens have evolved different mechanisms to acquire iron from host; however, little is known regarding how paracoccidioides species incorporate and metabolize this ion. in this work, host iron sources that are used by paracoccidioides spp. were investigated. robust fungal growth in the presence of the iron-containing molecules hemi ... | 2014 | 24831516 |
| a betasatellite-dependent begomovirus infects ornamental rose: characterization of begomovirus infecting rose in pakistan. | the begomovirus genus of the family geminiviridae comprises the largest group of geminiviruses. the list of begomoviruses is continuously increasing as a result of improvement in the methods for identification. ornamental rose plants (rosa chinensis) with highly stunted growth and leaf curling were found in faisalabad, pakistan. plants were analyzed for begomovirus infection, through rolling circle amplification and pcr methods. based on complete genome sequence homologies with other begomovirus ... | 2014 | 24781196 |
| expression, characterization, and improvement of a newly cloned halohydrin dehalogenase from agrobacterium tumefaciens and its application in production of epichlorohydrin. | a gene encoding halohydrin dehalogenase (hhdh) from agrobacterium tumefaciens cctcc m 87071 was cloned and expressed in escherichia coli. to increase activity and stability of hhdh, 14 amino acid residues around the active site and substrate-binding pocket based on the structural analysis and molecular docking were selected as targets for site-directed mutagenesis. the studies showed that the mutant hhdh (mut-hhdh) enzyme had a more accessible substrate-binding pocket than the wild-type hhdh (wt ... | 2014 | 24777710 |
| thc6 protein, isolated from trichoderma harzianum, can induce maize defense response against curvularia lunata. | mutant t66 was isolated from 450 mutants (constructed with agrobacterium tumefaciens-mediated transformation method) of trichoderma harzianum. maize seeds coated with t66 were more susceptible to curvularia lunata when compared with those coated with wild-type (wt) strain. the disease index of maize treated with t66 and wt were 62.5 and 42.1%, respectively. further research showed t-dna has inserted into the orf of one gene, which resulted in the functional difference between wt and t66. the gen ... | 2015 | 24771614 |
| crystal structure of the novel haloalkane dehalogenase data from agrobacterium tumefaciens c58 reveals a special halide-stabilizing pair and enantioselectivity mechanism. | a novel haloalkane dehalogenase data from agrobacterium tumefaciens c58 belongs to the hld-ii subfamily and hydrolyzes brominated and iodinated compounds, leading to the generation of the corresponding alcohol, a halide ion, and a proton. because data possesses a unique asn-tyr pair instead of the asn-trp pair conserved among the subfamily members, which was proposed to keep the released halide ion stable, the structural basis for its reaction mechanism should be elucidated. here, we determined ... | 2014 | 24770384 |
| draft genome sequence of umbilicaria muehlenbergii kolrilf000956, a lichen-forming fungus amenable to genetic manipulation. | umbilicaria muehlenbergii strain kolrilf000956 is amenable to agrobacterium tumefaciens-mediated transformation (atmt), making it the only known genetically tractable lichen-forming fungus to date. we report another advancement in lichen genetics, a draft genome assembly for u. muehlenbergii with a size of 34,812,353 bp and a gc content of 47.12%, consisting of seven scaffolds. | 2014 | 24762942 |
| the detection of t-nos, a genetic element present in gmos, by cross-priming isothermal amplification with real-time fluorescence. | an isothermal cross-priming amplification (cpa) assay for agrobacterium tumefaciens nopaline synthase terminator (t-nos) was established and investigated in this work. a set of six specific primers, recognizing eight distinct regions on the t-nos sequence, was designed. the cpa assay was performed at a constant temperature, 63 °c, and detected by real-time fluorescence. the results indicated that real-time fluorescent cpa had high specificity, and the limit of detection was 1.06 × 10(3) copies o ... | 2014 | 24748469 |
| identification of effective pb resistant bacteria isolated from lens culinaris growing in lead contaminated soils. | soil bacteria are a new phytoremediation system for the removal of heavy metals from soils. in this study, fifteen soil bacteria were isolated from root nodules of lentil growing in heavy metals contaminated soils, particularly by lead. molecular characterization of the collection showed a large diversity, including agrobacterium tumefaciens, rahnella aquatilis, pseudomonas, and rhizobium sp. these soil bacteria had a wide range of tolerance to heavy metals. among them, strains of a. tumefaciens ... | 2015 | 24740715 |
| the myosin motor domain-containing chitin synthase pdchsvii is required for development, cell wall integrity and virulence in the citrus postharvest pathogen penicillium digitatum. | chitin is an essential component of the fungal cell wall and a potential target in the development of new antifungal compounds, due to its presence in fungi and not in plants or vertebrates. chitin synthase genes (chs) constitute a complex family in filamentous fungi and are involved in fungal development, morphogenesis, pathogenesis and virulence. in this study, additional chs genes in the citrus postharvest pathogen penicillium digitatum have been identified. comparative analyses included each ... | 2014 | 24727399 |
| 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 ... | 2013 | 24711759 |
| a simple and reliable multi-gene transformation method for switchgrass. | a simple and reliable agrobacterium -mediated transformation method was developed for switchgrass. using this method, many transgenic plants carrying multiple genes-of-interest could be produced without untransformed escape. switchgrass (panicum virgatum l.) is a promising biomass crop for bioenergy. to obtain transgenic switchgrass plants carrying a multi-gene trait in a simple manner, an agrobacterium-mediated transformation method was established by constructing a gateway-based binary vector, ... | 2014 | 24700247 |
| tawrky68 responses to biotic stresses are revealed by the orthologous genes from major cereals. | wrky transcription factors have been extensively characterized in the past 20 years, but in wheat, studies on wrky genes and their function are lagging behind many other species. to explore the function of wheat wrky genes, we identified a tawrky68 gene from a common wheat cultivar. it encodes a protein comprising 313 amino acids which harbors 19 conserved motifs or active sites. gene expression patterns were determined by analyzing microarray data of tawrky68 in wheat and of orthologous genes f ... | 2014 | 24688294 |
| identification of the bona fide dhdps from a common plant pathogen. | agrobacterium tumefaciens is a gram-negative soil-borne bacterium that causes crown gall disease in many economically important crops. the absence of a suitable chemical treatment means there is a need to discover new anti-crown gall agents and also characterize bona fide drug targets. one such target is dihydrodipicolinate synthase (dhdps), a homo-tetrameric enzyme that catalyzes the committed step in the metabolic pathway yielding meso-diaminopimelate and lysine. interestingly, there are 10 pu ... | 2014 | 24677246 |
| genome sequence of the octopine-type agrobacterium tumefaciens strain ach5. | we have sequenced the complete genome of the plant pathogen agrobacterium tumefaciens strain lba4213, a derivative of the wild-type strain a. tumefaciens ach5 and the ancestor of a. tumefaciens strain lba4404 used in genetic engineering. the genome consists of a circular chromosome and a linear chromosome, as well as a megaplasmid and a tumor-inducing plasmid. | 2014 | 24675863 |
| involvement of the acr3 and dcta anti-porters in arsenite oxidation in agrobacterium tumefaciens 5a. | microbial arsenite (asiii) oxidation forms a critical piece of the arsenic cycle in nature, though our understanding of how and why microorganisms oxidize asiii remains rudimentary. our model organism agrobacterium tumefaciens 5a contains two distinct ars operons (ars1 and ars2) that are similar in their coding region content. the ars1 operon is located nearby the aio operon that is essential for asiii oxidation. the asiii/h(+) anti-porters encoded by acr3-1 and acr3-2 are required for maximal a ... | 2015 | 24674103 |
| fate of arsenate following arsenite oxidation in agrobacterium tumefaciens gw4. | the fate of arsenate (as(v) ) generated by microbial arsenite (as(iii) ) oxidation is poorly understood. agrobacterium tumefaciens wild-type strain (gw4) was studied to determine how the cell copes with as(v) generated in batch culture. gw4 grown heterotrophically with mannitol used as(iii) as a supplemental energy supply as reflected by enhanced growth and increased cellular levels of nadh and atp. under low phosphate (pi) conditions and presence of as(iii) oxidation, up to ∼ 50% of the resulti ... | 2015 | 24673976 |
| antimicrobial activities of novel mannosyl lipids isolated from the biocontrol fungus simplicillium lamellicola bcp against phytopathogenic bacteria. | the antagonistic fungus simplicillium lamellicola bcp has been developed as a microbial biopesticide that effectively controls the development of various plant diseases caused by both pathogenic bacteria and pathogenic fungi. antibacterial bioassay-directed fractionation was used to isolate mannosyl lipids from s. lamellicola bcp, and the structures of these compounds were elucidated using spectral analysis and chemical degradation. three novel mannosyl lipids were characterized and identified a ... | 2014 | 24660753 |
| expression of a fungal ferulic acid esterase in alfalfa modifies cell wall digestibility. | alfalfa (medicago sativa) is an important forage crop in north america owing to its high biomass production, perennial nature and ability to fix nitrogen. feruloyl esterase (ec 3.1.1.73) hydrolyzes ester linkages in plant cell walls and has the potential to further improve alfalfa as biomass for biofuel production. | 2014 | 24650274 |
| cloning, transformation and expression of proinsulin gene in tomato (lycopersicum esculentum mill.). | plants are among promising and suitable platform systems for production of recombinant biopharmaceutical proteins due to several features such as safety, no need for fermentation, inexpensive investment, and fast and easy scale-up. human insulin is one of the most widely used medicines in the world. up to now different expression systems including escherichia coli, yeast and cho have been exploited for producing recombinant human insulin and a variety of different recombinant insulin are extensi ... | 2014 | 24644433 |
| virus-induced gene silencing and agrobacterium tumefaciens-mediated transient expression in nicotiana tabacum. | virus-induced gene silencing (vigs) is a rapid method for transient silencing of plant genes. in this chapter, we describe the methodology for tobacco rattle virus (trv)-based vigs in nicotiana tabacum. in combination with subsequent co-expression of the tomato immune receptor ve1 and the corresponding verticillium effector ave1 through agrobacterium tumefaciens-mediated transient transformation (agroinfiltration), we established a rapid system for assessing the requirement of candidate plant ge ... | 2014 | 24643561 |
| structural basis for high substrate-binding affinity and enantioselectivity of 3-quinuclidinone reductase atqr. | (r)-3-quinuclidinol, a useful compound for the synthesis of various pharmaceuticals, can be enantioselectively produced from 3-quinuclidinone by 3-quinuclidinone reductase. recently, a novel nadh-dependent 3-quinuclidionone reductase (atqr) was isolated from agrobacterium tumefaciens, and showed much higher substrate-binding affinity (>100 fold) than the reported 3-quinuclidionone reductase (rrqr) from rhodotorula rubra. here, we report the crystal structure of atqr at 1.72 å. three nadh-bound p ... | 2014 | 24642255 |
| agrobacterium tumefaciens-mediated transformation of lasiodiplodia theobromae, the causal agent of gummosis in cashew nut plants. | lasiodiplodia theobromae is a major pathogen of many different crop cultures, including cashew nut plants. this paper describes an efficient agrobacterium tumefaciens-mediated transformation (atmt) system for the successful delivery of t-dna, transferring the genes of green fluorescent protein (gfp) and hygromycin b phosphotransferase (hph) to l. theobromae. when the fungal pycnidiospores were co-cultured with a. tumefaciens harboring the binary vector with hph-gfp gene, hygromycin-resistant fun ... | 2014 | 24634294 |
| dimerization of vird2 binding protein is essential for agrobacterium induced tumor formation in plants. | the type iv secretion system (t4ss) is the only bacterial secretion system known to translocate both dna and protein substrates. the virb/d4 system from agrobacterium tumefaciens is a typical t4ss. it facilitates the bacteria to translocate the vird2-t-dna complex to the host cell cytoplasm. in addition to protein-dna complexes, the virb/d4 system is also involved in the translocation of several effector proteins, including vire2, vire3 and virf into the host cell cytoplasm. these effector prote ... | 2014 | 24626239 |
| [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. | 2013 | 24617266 |
| attachment of agrobacterium tumefaciens to leaf tissue in response to infiltration conditions. | transient expression of recombinant proteins in plant tissues following agrobacterium-mediated gene transfer is a promising technique for rapid protein production. however, transformation rates and transient expression levels can be sub-optimal depending on process conditions. attachment of agrobacterium tumefaciens to plant cells is an early, critical step in the gene transfer pathway. bacterial attachment levels and patterns may influence transformation and, by extension, transient expression. ... | 2014 | 24616410 |
| an efficient agrobacterium-mediated transformation method for the edible mushroom hypsizygus marmoreus. | hypsizygus marmoreus is one of the major edible mushrooms in east asia. as no efficient transformation method, the molecular and genetics studies were hindered. the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene of h. marmoreus was isolated and its promoter was used to drive the hygromycin b phosphotransferase (hph) and enhanced green fluorescent protein (egfp) in h. marmoreus. agrobacterium tumefaciens-mediated transformation (atmt) was successfully applied in h. marmoreus. the transformat ... | 2014 | 24612605 |
| overexpression of s-adenosyl-l-methionine synthetase increased tomato tolerance to alkali stress through polyamine metabolism. | s-adenosyl-l-methionine (sam) synthetase is the key enzyme involved in the biosynthesis of sam, which serves as a common precursor for polyamines (pas) and ethylene. a sam synthetase cdna (slsams1) was introduced into the tomato genome using the agrobacterium tumefaciens transformation method. transgenic plants overexpressing slsams1 exhibited a significant increase in tolerance to alkali stress and maintained nutrient balance, higher photosynthetic capacity and lower oxidative stress compared w ... | 2014 | 24605920 |
| transgenic tomato line expressing modified bacillus thuringiensis cry1ab gene showing complete resistance to two lepidopteran pests. | the modified truncated bt-cry1ab gene of bacillus thuringiensis has been used for the development and selection of over expressing transgenic events in a commercially important variety of tomato (solanum lycopersicum l.) by agrobacterium-mediated leaf-disc transformation procedure. the integration and inheritance of cry1ab gene in t0 transgenic plants and their progenies were determined by pcr, rt-pcr and southern blot hybridization analysis. the toxin expression was monitored by double antibody ... | 2014 | 24600542 |
| roles of agrobacterium tumefaciens membrane-bound ferritin (mbfa) in iron transport and resistance to iron under acidic conditions. | agrobacterium tumefaciens membrane-bound ferritin (mbfa) is a member of the erythrin (er)-vacuolar iron transport family. the mbfa protein has an er or ferritin-like domain at its n terminus and has been predicted to have five transmembrane segments in its c-terminal region. analysis of protein localization using phoa and lacz reporter proteins supported the view that the n-terminal di-iron site is located in the cytoplasm whilst the c-terminal end faces the periplasm. an a. tumefaciens mbfa mut ... | 2014 | 24600024 |
| a new high-frequency agrobacterium-mediated transformation technique for sesamum indicum l. using de-embryonated cotyledon as explant. | in spite of the economic importance of sesame (sesamum indicum l.) and the recent availability of its genome sequence, a high-frequency transformation protocol is still not available. the only two existing agrobacterium-mediated transformation protocols that are available have poor transformation efficiencies of less than 2%. in the present study, we report a high-frequency, simple, and reproducible transformation protocol for sesame. transformation was done using de-embryonated cotyledons via s ... | 2014 | 24590594 |
| a procedure for the transient expression of genes by agroinfiltration above the permissive threshold to study temperature-sensitive processes in plant-pathogen interactions. | localized expression of genes in plants from t-dnas delivered into plant cells by agrobacterium tumefaciens is an important tool in plant research. the technique, known as agroinfiltration, provides fast, efficient ways to transiently express or silence a desired gene without resorting to the time-consuming, challenging stable transformation of the host, the use of less efficient means of delivery, such as bombardment, or the use of viral vectors, which multiply and spread within the host causin ... | 2014 | 24581046 |
| antibacterial and antitumour activities of some plants grown in turkey. | screening of antibacterial and antitumour activities of 33 different extracts prepared with three types of solvents (water, ethanol and methanol) was conducted. the extracts were obtained from 11 different plant species grown in turkey: eryngium campestre l., alchemilla mollis (buser) rothm., dorycnium pentaphyllum scop., coronilla varia l., onobrychis oxyodonta boiss., fritillaria pontica wahlenb., asarum europaeum l., rhinanthus angustifolius c. c. gmelin, doronicum orientale hoffm., campanula ... | 2014 | 26740759 |
| agrobacterium tumefaciens-mediated transformation of corn (zea mays l.) multiple shoots. | an agrobacterium tumefaciens-mediated corn transformation method based on multiple shoot tissue cultures was developed, which is effective with a variety of corn inbred lines and standard binary vectors. six factors that affected the success of corn transformation were tested, including a. tumefaciens strain, corn genotype, tissue culture growth stage, medium composition, co-culture temperature and surfactant treatment. agropine-type bacteria (eha 101 and agl 1) were eightfold more effective tha ... | 2014 | 26019506 |
| rnai mediated curcin precursor gene silencing in jatropha (jatropha curcas l.). | curcin, a type i ribosomal inhibiting protein-rip, encoded by curcin precursor gene, is a phytotoxin present in jatropha (jatropha curcas l.). here, we report designing of rnai construct for the curcin precursor gene and further its genetic transformation of jatropha to reduce its transcript expression. curcin precursor gene was first cloned from jatropha strain darl-2 and part of the gene sequence was cloned in sense and antisense orientation separated by an intron sequence in plant expression ... | 2014 | 24574003 |
| [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 ... | 2013 | 24564162 |
| improved dominant selection markers and co-culturing conditions for efficient agrobacterium tumefaciens-mediated transformation of ustilago scitaminea. | ustilago scitaminea is the causal agent of sugar-cane smut disease. there is, however, no genetic transformation method for it. here we report the development of an efficient mutagenesis method based on agrobacterium tumefaciens-mediated transformation. to improve transformation efficiency, a range of conditions, including the codon-usage preference of the selection marker gene, promoters and the culture conditions for transformation were optimized. a strong promoter to drive marker gene express ... | 2014 | 24563317 |
| agrobacterium tumefaciens-mediated transformation of the causative agent of valsa canker of apple tree valsa mali var. mali. | valsa mali var. mali (vmm), which is the causative agent of valsa canker of apple tree, causes heavy damage to apple production in eastern asia. in this article, we report agrobacterium tumefaciens-mediated transformation (atmt) of vmm and expression of gfp (green fluorescent protein) in this fungus. the transformation system was optimized to a transformation efficiency of approximately 150 transformants/10(6) conidia, and a library containing over 4,000 transformants was generated. the tested t ... | 2014 | 24554343 |
| the influence of heat stress on auxin distribution in transgenic b. napus microspores and microspore-derived embryos. | plant embryogenesis is regulated by differential distribution of the plant hormone auxin. however, the cells establishing these gradients during microspore embryogenesis remain to be identified. for the first time, we describe, using the dr5 or dr5rev reporter gene systems, the gfp- and gus-based auxin biosensors to monitor auxin during brassica napus androgenesis at cellular resolution in the initial stages. our study provides evidence that the distribution of auxin changes during embryo develo ... | 2014 | 24553810 |
| functions and regulation of quorum-sensing in agrobacterium tumefaciens. | in agrobacterium tumefaciens, horizontal transfer and vegetative replication of oncogenic ti plasmids involve a cell-to-cell communication process called quorum-sensing (qs). the determinants of the qs-system belong to the luxr/luxi class. the luxi-like protein trai synthesizes n-acyl-homoserine lactone molecules which act as diffusible qs-signals. beyond a threshold concentration, these molecules bind and activate the luxr-like transcriptional regulator trar, thereby initiating the qs-regulator ... | 2014 | 24550924 |
| development of an agrobacterium-mediated stable transformation method for the sensitive plant mimosa pudica. | the sensitive plant mimosa pudica has long attracted the interest of researchers due to its spectacular leaf movements in response to touch or other external stimuli. although various aspects of this seismonastic movement have been elucidated by histological, physiological, biochemical, and behavioral approaches, the lack of reverse genetic tools has hampered the investigation of molecular mechanisms involved in these processes. to overcome this obstacle, we developed an efficient genetic transf ... | 2014 | 24533121 |
| the brucella suis ibpa heat-shock chaperone is not required for virulence or for expression of the virb type iv secretion system virb8 protein. | brucella suis, facultative intracellular bacterial pathogen of mammals, and agrobacterium tumefaciens, a plant pathogen, both use a virb type iv secretion system (t4ss) to translocate effector molecules into host cells. hspl, an α-crystalline-type small heat-shock protein, acts as a chaperone for the agrobacterium virb8 protein, an essential component of the virb system. an agrobacterium mutant lacking hspl is attenuated due to a misfunctional t4ss. we have investigated whether ibpa (bra0051), t ... | 2014 | 24517122 |
| 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 ... | 2013 | 24517006 |
| improvement of a gene targeting system for genetic manipulation in penicillium digitatum. | penicillium digitatum is the most important pathogen of postharvest citrus. gene targeting can be done in p. digitatum using homologous recombination via agrobacterium tumefaciens mediated transformation (atmt), but the frequencies are often very low. in the present study, we replaced the ku80 homolog (a gene of the non-homologous end-joining (nhej) pathway) with the hygromycin resistance cassette (hph) by atmt. no significant change in vegetative growth, conidiation, or pathogenicity was observ ... | 2014 | 24510704 |
| 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 ... | 2013 | 24498761 |
| agrobacterium-mediated genetic transformation and plant regeneration of the hardwood tree species fraxinus profunda. | this transformation and regeneration protocol provides an integral framework for the genetic improvement of fraxinus profunda (pumpkin ash) for future development of plants resistant to the emerald ash borer. using mature hypocotyls as the initial explants, an agrobacterium tumefaciens-mediated genetic transformation system was successfully developed for pumpkin ash (fraxinus profunda). this transformation protocol is an invaluable tool to combat the highly aggressive, non-native emerald ash bor ... | 2014 | 24493252 |
| overexpression of rice nac gene snac1 improves drought and salt tolerance by enhancing root development and reducing transpiration rate in transgenic cotton. | the snac1 gene belongs to the stress-related nac superfamily of transcription factors. it was identified from rice and overexpressed in cotton cultivar yz1 by agrobacterium tumefaciens-mediated transformation. snac1-overexpressing cotton plants showed more vigorous growth, especially in terms of root development, than the wild-type plants in the presence of 250 mm nacl under hydroponic growth conditions. the content of proline was enhanced but the mda content was decreased in the transgenic cott ... | 2014 | 24489802 |
| quorum vs. diffusion sensing: a quantitative analysis of the relevance of absorbing or reflecting boundaries. | the consequences of the boundary conditions (signal reflecting vs. signal adsorbing) on bacterial intercellular communication were addressed by a combined physics and microbiology approach. a predictive biophysical model was devised that considered system size, diffusion from given points, signal molecule decay and boundary properties. the theoretical predictions were tested with two experimental agarose-gel-based set-ups for reflecting or absorbing boundaries. n-acyl homoserine lactone (ahl) co ... | 2014 | 24484313 |
| transient down-regulation of the rna silencing machinery increases efficiency of agrobacterium-mediated transformation of arabidopsis. | agrobacterium tumefaciens is a plant pathogen that is widely used in plant transformation. as the process of transgenesis includes the delivery of single-stranded t-dna molecule, we hypothesized that transformation rate may negatively correlate with the efficiency of the rna-silencing machinery. using mutants compromised in either the transcriptional or post-transcriptional gene-silencing pathways, two inhibitors of stable transformation were revealed-ago2 and nrpd1a. furthermore, an immunopreci ... | 2014 | 24472037 |
| physiological and photosynthetic characteristics of indica hang2 expressing the sugarcane pepc gene. | phosphoenolpyruvate carboxylase (pepc) is known to play a key role in the initial fixation of co2 in c4 photosynthesis. the pepc gene from sugarcane (a c4 plant) was introduced into indica rice (hang2), a process mediated by agrobacterium tumefaciens. integration patterns and copy numbers of the gene was confirmed by dna blot analysis. rt-pcr and western blotting results showed that the pepc gene was expressed at both the mrna and protein levels in the transgenic lines. real-time pcr results ind ... | 2014 | 24469712 |
| enhanced resistance to blast fungus in rice (oryza sativa l.) by expressing the ribosome-inactivating protein α-momorcharin. | rice blast caused by magnaporthe grisea is one of the three major diseases that seriously affect the rice production. alpha-momorcharin (α-mc), a ribosome-inactivating protein (rip) isolated from momordica charantia seeds, has antifungal effects in vitro. in this study, the α-mc gene was constitutively expressed under the control of the 2×35s promoter in transgenic rice (oryza sativa l.) using an agrobacterium tumefaciens-mediated method. the nine transgenic plants were obtained and confirmed by ... | 2014 | 24467890 |
| overexpression of ptsos2 enhances salt tolerance in transgenic poplars. | protein kinases are major signal transduction factors that have a central role in mediating acclimation to environmental changes in eukaryotic organisms. in this study, we cloned and identified three salt overly sensitive 2 (sos2) genes in the woody plant populus trichocarpa, designated as ptsos2.1, ptsos2.2, and ptsos2.3, which were transformed into hybrid poplar clone t89 (populus tremula× populus tremuloides michx clone t89) mediated by agrobacterium tumefaciens. southern and northern blot an ... | 2014 | 24465084 |
| an efficient tool for random insertional mutagenesis: agrobacterium tumefaciens-mediated transformation of the filamentous fungus aspergillus terreus. | agrobacterium tumefaciens-mediated transformation (atmt) has been widely used in various organisms. in the current study, we developed a simple and efficient system for genetic transformation of the filamentous fungus aspergillus terreus using atmt. the transformation protocol was optimized for certain parameters to rapidly generate a library of transferred dna (t-dna) insertion mutants of a. terreus. the presence of mitotically stable hygromycin resistance gene (hph) integration in the genome w ... | 2014 | 24462974 |
| involvement of rad52 in t-dna circle formation during agrobacterium tumefaciens-mediated transformation of saccharomyces cerevisiae. | agrobacterium tumefaciens cells carrying a tumour inducing plasmid (ti-plasmid) can transfer a defined region of transfer dna (t-dna) to plant cells as well as to yeast. this process of agrobacterium-mediated transformation (amt) eventually results in the incorporation of the t-dna in the genomic dna of the recipient cells. all available evidence indicates that t-strand transfer closely resembles conjugal dna transfer as found between gram-negative bacteria. however, where conjugal plasmid dna t ... | 2014 | 24460832 |
| galactaro δ-lactone isomerase: lactone isomerization by a member of the amidohydrolase superfamily. | agrobacterium tumefaciens strain c58 can utilize d-galacturonate as a sole source of carbon via a pathway in which the first step is oxidation of d-galacturonate to d-galactaro-1,5-lactone. we have identified a novel enzyme, d-galactarolactone isomerase (gli), that catalyzes the isomerizaton of d-galactaro-1,5-lactone to d-galactaro-1,4-lactone. gli, a member of the functionally diverse amidohydrolase superfamily, is a homologue of ligi that catalyzes the hydrolysis of 2-pyrone-4,6-dicarboxylate ... | 2014 | 24450804 |
| detection of an o-methyltransferase synthesising acetosyringone in methyl jasmonate-treated tobacco cell-suspensions cultures. | acetosyringone (3',5'-dimethoxy-4'-hydroxyacetophenone) is a well-known and very effective inducer of the virulence genes of agrobacterium tumefaciens but the precise pathway of its biosynthesis in plants is still unknown. we have used two tobacco cell lines, cultured in suspension and exhibiting different patterns of accumulation of acetosyringone in their culture medium upon treatment with methyl jasmonate, to study different steps of acetosyringone biosynthesis. in the two cell lines studied, ... | 2014 | 24445177 |
| dna replicons for plant genome engineering. | sequence-specific nucleases enable facile editing of higher eukaryotic genomic dna; however, targeted modification of plant genomes remains challenging due to ineffective methods for delivering reagents for genome engineering to plant cells. here, we use geminivirus-based replicons for transient expression of sequence-specific nucleases (zinc-finger nucleases, transcription activator-like effector nucleases, and the clustered, regularly interspaced, short palindromic repeat/cas system) and deliv ... | 2014 | 24443519 |
| retracted article: agrobacterium tumefaciens-mediated transformation and efficient regeneration of a timber yielding plant dalbergia sissoo roxb. | | 2013 | 24431531 |
| agroinfiltration and pvx agroinfection in potato and nicotiana benthamiana. | agroinfiltration and pvx agroinfection are two efficient transient expression assays for functional analysis of candidate genes in plants. the most commonly used agent for agroinfiltration is agrobacterium tumefaciens, a pathogen of many dicot plant species. this implies that agroinfiltration can be applied to many plant species. here, we present our protocols and expected results when applying these methods to the potato (solanum tuberosum), its related wild tuber-bearing solanum species (solan ... | 2014 | 24430891 |
| bcpmi2, isolated from non-heading chinese cabbage encoding phosphomannose isomerase, improves stress tolerance in transgenic tobacco. | phosphomannose isomerase (pmi) is an enzyme that catalyses the first step of the l-galactose pathway for ascorbic acid (asa) biosynthesis in plants. to clarify the physiological roles of pmi in asa biosynthesis, the cdna sequence of pmi was cloned from non-heading chinese cabbage (brassica campestris ssp. chinensis makino) and overexpressed in tobacco transformed with agrobacterium tumefaciens. the asa and soluble sugar contents were lower in 35s::bcpmi2 tobacco than in wild-type tobacco. howeve ... | 2014 | 24430300 |
| 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 ... | 2013 | 24400011 |
| molecular strategies to replace the structural metal site in the prokaryotic zinc finger domain. | the specific arrangement of secondary elements in a local motif often totally relies on the formation of coordination bonds between metal ions and protein ligands. this is typified by the ~30 amino acid eukaryotic zinc finger motif in which a β-sheet and an α-helix are clustered around a zinc ion by various combinations of four ligands. the prokaryotic zinc finger domain (found in the ros protein from agrobacterium tumefaciens) is different from the eukaryotic counterpart as it consists of 58 am ... | 2014 | 24389235 |
| 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 ... | 2013 | 24386304 |
| 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 ... | 2013 | 24381526 |
| siderophore biosynthesis genes of rhizobium sp. isolated from cicer arietinum l. | rhizobium bicc 651, a fast-growing strain isolated from root nodule of chickpea (cicer arietinum l.), produced a catechol siderophore to acquire iron under iron poor condition. a tn5-induced mutant (b153) of the strain, bicc 651 impaired in siderophore biosynthesis was isolated and characterized. the mutant failed to grow on medium supplemented with iron chelator and grew less efficiently in deferrated broth indicating its higher iron requirement. the mutant produced less number of nodules than ... | 2014 | 28324476 |
| a rapid and inexpensive one-tube genomic dna extraction method from agrobacterium tumefaciens. | many methods have been used to isolate genomic dna, but some of them are time-consuming and costly, especially when extracting a large number of samples. here we described an easy protocol using two simple solutions for dna extraction from a. tumefaciens cells. compared with the standard protocol, this protocol allows rapid dna isolation with comparable yield and purity at negligible cost. following this protocol, we have demonstrated: (1) gdna extraction was achieved within 15 min; (2) this met ... | 2014 | 28324444 |
| two fatty acid desaturases, stearoyl-acyl carrier protein δ9-desaturase6 and fatty acid desaturase3, are involved in drought and hypoxia stress signaling in arabidopsis crown galls. | agrobacterium tumefaciens-derived crown galls of arabidopsis (arabidopsis thaliana) contain elevated levels of unsaturated fatty acids and strongly express two fatty acid desaturase genes, ω3 fatty acid desaturase3 (fad3) and stearoyl-acyl carrier protein δ9-desaturase6 (sad6). the fad3-2 mutant with impaired α-linolenic acid synthesis developed significantly smaller crown galls under normal, but not under high, relative humidity. this strongly suggests that fad3 plays a role in increasing droug ... | 2014 | 24368335 |
| identification of functionally important conserved trans-membrane residues of bacterial pib -type atpases. | powered by atp hydrolysis, p(ib) -atpases drive the energetically uphill transport of transition metals. these high affinity pumps are essential for heavy metal detoxification and delivery of metal cofactors to specific cellular compartments. amino acid sequence alignment of the trans-membrane (tm) helices of p(ib)-atpases reveals a high degree of conservation, with ∼ 60-70 fully conserved positions. of these conserved positions, 6-7 were previously identified to be important for transport. howe ... | 2014 | 24350798 |
| cloning and characterization of a novel dehydrin gene, sidhn2, from saussurea involucrata kar. et kir. | saussurea involucrata kar. et kir. is a hardy dicotyledonous plant capable of tolerating severe abiotic stress conditions. in a previous study, we created a cdna library to determine what factors are associated with the cold acclimation response in s. involucrata. from this, a full-length cdna of a dehydrin-like gene (sidhn2) was obtained by rt-pcr. the sidhn2 gene was characterized in this study. the full-length sidhn2 cdna comprised 693 bp containing an open reading frame of 345 bp specifying ... | 2014 | 24337866 |
| maize (zea mays l.) transformation by agrobacterium tumefaciens infection of pollinated ovules. | a novel transformation system was established for maize using agrobacterium infection of in vitro cultured ovules. the maize ovules were isolated 24h after pollination and infected with agrobacterium. the embryos were isolated from the pollinated ovules 2-3 weeks after agrobacterium infection, regenerated to plantlets and investigated for transgene expression and inheritance. experimental evaluations were focused on the four main aspects. firstly, through the introduction of gus gene for monitor ... | 2014 | 24333124 |
| field performance of transgenic sugarcane produced using agrobacterium and biolistics methods. | future genetic improvement of sugarcane depends, in part, on the ability to produce high-yielding transgenic cultivars with improved traits such as herbicide and insect resistance. here, transgenic sugarcane plants generated by different transformation methods were assessed for field performance over 3 years. agrobacterium-mediated (agro) transgenic events (35) were produced using four different agrobacterium tumefaciens strains, while biolistic (biol) transgenic events (48) were produced using ... | 2014 | 24330327 |
| the fusarium oxysporum effector six6 contributes to virulence and suppresses i-2-mediated cell death. | plant pathogens secrete effectors to manipulate their host and facilitate colonization. fusarium oxysporum f. sp. lycopersici is the causal agent of fusarium wilt disease in tomato. upon infection, f. oxysporum f. sp. lycopersici secretes numerous small proteins into the xylem sap (six proteins). most six proteins are unique to f. oxysporum, but six6 is an exception; a homolog is also present in two colletotrichum spp. six6 expression was found to require living host cells and a knockout of six6 ... | 2014 | 24313955 |
| 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 ... | 2013 | 24311575 |
| generation of selectable marker-free transgenic eggplant resistant to alternaria solani using the r/rs site-specific recombination system. | marker-free transgenic eggplants, exhibiting enhanced resistance to alternaria solani , can be generated on plant growth regulators (pgrs)- and antibiotic-free ms medium employing the multi-auto-transformation (mat) vector, pmat21 - wasabi defensin , wherein isopentenyl transferase ( ipt ) gene is used as a positive selection marker. | 2014 | 24311155 |
| stable transformation and reverse genetic analysis of penium margaritaceum: a platform for studies of charophyte green algae, the immediate ancestors of land plants. | the charophyte green algae (cga, streptophyta, viridiplantae) occupy a key phylogenetic position as the immediate ancestors of land plants but, paradoxically, are less well-studied than the other major plant lineages. this is particularly true in the context of functional genomic studies, where the lack of an efficient protocol for their stable genetic transformation has been a major obstacle. observations of extant cga species suggest the existence of some of the evolutionary adaptations that h ... | 2014 | 24308430 |
| direct visualization of agrobacterium-delivered vire2 in recipient cells. | agrobacterium tumefaciens is a natural genetic engineer widely used to deliver dna into various recipients, including plant, yeast and fungal cells. the bacterium can transfer single-stranded dna molecules (t-dnas) and bacterial virulence proteins, including vire2. however, neither the dna nor the protein molecules have ever been directly visualized after the delivery. in this report, we adopted a split-gfp approach: the small gfp fragment (gfp11) was inserted into vire2 at a permissive site to ... | 2014 | 24299048 |
| genome engineering of agrobacterium tumefaciens using the lambda red recombination system. | agrobacterium tumefaciens has been widely used as a tool for transgenesis in plants. the availability of its genome sequence should facilitate the directed engineering of improved properties; however, the current genome engineering options are laborious. here, we investigated whether the lambda red operon can be applied for recombineering of the a. tumefaciens genome. first, we built an expression plasmid for a. tumefaciens employing a tetracycline-inducible promoter to regulate the red operon. ... | 2014 | 24297480 |
| 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 ... | 2013 | 24278131 |
| 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 ... | 2013 | 24273610 |
| 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 ... | 2013 | 24273422 |