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quorum-quenching limits quorum-sensing exploitation by signal-negative invaders.some bacteria produce and perceive quorum-sensing (qs) signals that coordinate several behaviours, including the costly processes that are exoenzyme production and plasmid transfer. in the case of plasmid transfer, the emergence of qs signal-altered invaders and their policing are poorly documented. in agrobacterium tumefaciens, the virulence ti-plasmid encodes both synthesis and sensing of qs-signals, which promote its transfer from a donor to a recipient cell. here, we reported that qs-altered ...201728054641
characteristics of n-acylhomoserine lactones produced by hafnia alvei h4 isolated from spoiled instant sea cucumber.this study aimed to identify n-acylhomoserine lactone (ahl) produced by hafnia alvei h4, which was isolated from spoiled instant sea cucumber, and to investigate the effect of ahls on biofilm formation. two biosensor strains, chromobacterium violaceum cv026 and agrobacterium tumefaciens kyc55, were used to detect the quorum sensing (qs) activity of h. alvei h4 and to confirm the existence of ahl-mediated qs system. thin layer chromatography (tlc) and high resolution triple quadrupole liquid chro ...201728379194
growth kinetics and scale-up of agrobacterium tumefaciens.production of recombinant proteins in plants through agrobacterium-mediated transient expression is a promising method of producing human therapeutic proteins, vaccines, and commercial enzymes. this process has been shown to be viable at a large scale and involves growing large quantities of wild-type plants and infiltrating the leaf tissue with a suspension of agrobacterium tumefaciens bearing the genes of interest. this study examined one of the steps in this process that had not yet been opti ...201728357545
transgenic pigeonpea events expressing cry1ac and cry2aa exhibit resistance to helicoverpa armigera.independent transgenic pigeonpea events were developed using two cry genes. transgenic cry2aa-pigeonpea was established for the first time. selected transgenic events demonstrated 100% mortality of helicoverpa armigera in successive generations. lepidopteran insect helicoverpa armigera is the major yield constraint of food legume pigeonpea. the present study was aimed to develop h. armigera-resistant transgenic pigeonpea, selected on the basis of transgene expression and phenotyping. agrobacteri ...201728352969
agrobacterium delivers vire2 protein into host cells via clathrin-mediated endocytosis.agrobacterium tumefaciens can cause crown gall tumors on a wide range of host plants. as a natural genetic engineer, the bacterium can transfer both single-stranded dna (ssdna) [transferred dna (t-dna)] molecules and bacterial virulence proteins into various recipient cells. among agrobacterium-delivered proteins, vire2 is an ssdna binding protein that is involved in various steps of the transformation process. however, it is not clear how plant cells receive the t-dna or protein molecules. usin ...201728345032
is there any crosstalk between the chemotaxis and virulence induction signaling in agrobacterium tumefaciens?agrobacterium tumefaciens, a soil-born phytopathogenic bacterium, is well known as a nature's engineer due to its ability to genetically transform the host by transferring a dna fragment (called t-dna) from its ti plasmid to host-cell genome. to combat the harsh soil environment and seek the appropriate host, a. tumefaciens can sense and be attracted by a large number of chemical compounds released by wounded host. as a member of α-proteobacterium, a. tumefaciens has a chemotaxis system differen ...201728342941
characterization of a novel nicotine degradation gene cluster ndp in sphingomonas melonis ty and its evolutionary analysis.sphingomonas melonis ty utilizes nicotine as a sole source of carbon, nitrogen, and energy through a variant of the pyridine and pyrrolidine pathways (vpp). a 31-kb novel nicotine-degrading gene cluster, ndp, in strain ty exhibited a different genetic organization with the vpp cluster in strains ochrobactrum rhizosphaerae sjy1 and agrobacterium tumefaciens s33. genes in vpp were separated by a 20-kb interval sequence, while genes in ndp were localized together. half of the homolog genes were in ...201728337179
the effect of tobramycin incorporated with bismuth-ethanedithiol loaded on niosomes on the quorum sensing and biofilm formation of pseudomonas aeruginosa.pseudomonas aeruginosa is an opportunistic pathogen, enable of causing infections especially in immunocompromised patients. recently many isolates developed multiple drug resistance, resulting in treatment failure in serious infections. in this study, the effect of tobramycin incorporated with bismuth-ethanedithiol loaded on niosomes on the quorum sensing and biofilm production by p. aeruginosa was evaluated. thin layer hydration method with cholesterol (30%), span 40 and tween 40 were used to m ...201728323149
characterization of vdasp f2 secretory factor from verticillium dahliae by a fast and easy gene knockout system.the vascular wilt fungus verticillium dahliae produces persistent resting structures known as microsclerotia, which enable long-term survival of this plant pathogen in soil. the completed genome sequence of v. dahliae has facilitated large-scale investigations of individual gene functions using gene-disruption strategies based on agrobacterium tumefaciens-mediated transformation (atmt). however, the construction of gene-deletion vectors and screening of deletion mutants have remained challenging ...201728291379
optimization of agrobacterium-mediated transformation in soybean.high transformation efficiency is a prerequisite for study of gene function and molecular breeding. agrobacterium tumefaciens-mediated transformation is a preferred method in many plants. however, the transformation efficiency in soybean is still low. the objective of this study is to optimize agrobacterium-mediated transformation in soybean by improving the infection efficiency of agrobacterium and regeneration efficiency of explants. firstly, four factors affecting agrobacterium infection effi ...201728286512
mature luffa leaves (luffa cylindrica l.) as a tool for gene expression analysis by agroinfiltration.we exploited the potential of cucurbits for ectopic gene expression. agroinfiltration is a simple and commonly used method to obtain transient expression of foreign genes in plants. in contrast to in vitro transformation techniques, agroinfiltration can be used for genetic modification of mature plant tissues. although the cucurbits are commonly used as model plants for molecular biology and biotechnology studies, to date there are no literature sources on the possibility of transient gene expre ...201728270826
arsenite oxidation regulator aior regulates bacterial chemotaxis towards arsenite in agrobacterium tumefaciens gw4.some arsenite [as(iii)]-oxidizing bacteria exhibit positive chemotaxis towards as(iii), however, the related as(iii) chemoreceptor and regulatory mechanism remain unknown. the as(iii)-oxidizing bacterium agrobacterium tumefaciens gw4 displays positive chemotaxis towards 0.5-2 mm as(iii). genomic analyses revealed a putative chemoreceptor-encoding gene, mcp, located in the arsenic gene island and having a predicted promoter binding site for the as(iii) oxidation regulator aior. expression of mcp ...201728256605
abiotic and biotic factors responsible for antimonite oxidation in agrobacterium tumefaciens gw4.antimonite [sb(iii)]-oxidizing bacteria can transform the toxic sb(iii) into the less toxic antimonate [sb(v)]. recently, the cytoplasmic sb(iii)-oxidase anoa and the periplasmic arsenite [as(iii)] oxidase aioab were shown to responsible for bacterial sb(iii) oxidation, however, disruption of each gene only partially decreased sb(iii) oxidation efficiency. this study showed that in agrobacterium tumefaciens gw4, sb(iii) induced cellular h2o2 content and h2o2 degradation gene kata. gene knock-out ...201728252030
cascade of chromosomal rearrangements caused by a heterogeneous t-dna integration supports the double-stranded break repair model for t-dna integration.transferred dna (t-dna) from agrobacterium tumefaciens can be integrated into the plant genome. the double-stranded break repair (dsbr) pathway is a major model for t-dna integration. from this model, we expect that two ends of a t-dna molecule would invade into a single dna double-stranded break (dsb) or independent dsbs in the plant genome. we call the later phenomenon a heterogeneous t-dna integration, which has never been observed. in this work, we demonstrated it in an arabidopsis t-dna ins ...201728244154
agrobacterium-delivered virulence protein vire2 is trafficked inside host cells via a myosin xi-k-powered er/actin network.agrobacterium tumefaciens causes crown gall tumors on various plants by delivering transferred dna (t-dna) and virulence proteins into host plant cells. under laboratory conditions, the bacterium is widely used as a vector to genetically modify a wide range of organisms, including plants, yeasts, fungi, and algae. various studies suggest that t-dna is protected inside host cells by vire2, one of the virulence proteins. however, it is not clear how agrobacterium-delivered factors are trafficked t ...201728242680
effects upon metabolic pathways and energy production by sb(iii) and as(iii)/sb(iii)-oxidase gene aioa in agrobacterium tumefaciens gw4.agrobacterium tumefaciens gw4 is a heterotrophic arsenite [as(iii)]/antimonite [sb(iii)]-oxidizing strain. the as(iii) oxidase aioab is responsible for as(iii) oxidation in the periplasm and it is also involved in sb(iii) oxidation in agrobacterium tumefaciens 5a. in addition, sb(iii) oxidase anoa and cellular h2o2 are also responsible for sb(iii) oxidation in strain gw4. however, the deletion of aioa increased the sb(iii) oxidation efficiency in strain gw4. in the present study, we found that t ...201728241045
diversity of nodular bacteria of scorpiurus muricatus in western algeria and their impact on plant growth.a total of 51 bacterial strains were isolated from root nodules of scorpiurus muricatus sampled from six regions of western algeria. strains diversity was assessed by rep-pcr amplification fingerprinting which had allowed grouping the isolates into 28 different clusters. partial nucleotide sequencing of 16srrna gene and blast analysis revealed that root nodules of scorpiurus muricatus were colonized by different species close to rhizobium vignae, rhizobium radiobacter, rhizobium leguminosarum, p ...201728235183
the widely used nicotiana benthamiana 16c line has an unusual t-dna integration pattern including a transposon sequence.nicotiana benthamiana is employed around the world for many types of research and one transgenic line has been used more extensively than any other. this line, 16c, expresses the aequorea victoria green fluorescent protein (gfp), highly and constitutively, and has been a major resource for visualising the mobility and actions of small rnas. insights into the mechanisms studied at a molecular level in n. benthamiana 16c are likely to be deeper and more accurate with a greater knowledge of the gfp ...201728231340
animal component-free agrobacterium tumefaciens cultivation media for better gmp-compliance increases biomass yield and pharmaceutical protein expression in nicotiana benthamiana.transient expression systems allow the rapid production of recombinant proteins in plants. such systems can be scaled up to several hundred kilograms of biomass, making them suitable for the production of pharmaceutical proteins required at short notice, such as emergency vaccines. however, large-scale transient expression requires the production of recombinant agrobacterium tumefaciens strains with the capacity for efficient gene transfer to plant cells. the complex media often used for the cul ...201728221723
an agrobacterium tumefaciens strain with gamma-aminobutyric acid transaminase activity shows an enhanced genetic transformation ability in plants.agrobacterium tumefaciens has the unique ability to mediate inter-kingdom dna transfer, and for this reason, it has been utilized for plant genetic engineering. to increase the transformation frequency in plant genetic engineering, we focused on gamma-aminobutyric acid (gaba), which is a negative factor in the agrobacterium-plant interaction. recent studies have shown contradictory results regarding the effects of gaba on vir gene expression, leading to the speculation that gaba inhibits t-dna t ...201728220841
production of taxadiene by engineering of mevalonate pathway in escherichia coli and endophytic fungus alternaria alternata tpf6.taxol (paclitaxel) is a diterpenoid compound with significant and extensive applications in the treatment of cancer. the production of taxol and relevant intermediates by engineered microbes is an attractive alternative to the semichemical synthesis of taxol. in this study, based on a previously developed platform, the authors first established taxadiene production in mutant e. coli t2 and t4 by engineering of the mevalonate (mva) pathway. the authors then developed an agrobacterium tumefaciens- ...201728217906
selection of transformation-efficient barley genotypes based on tfa (transformation amenability) haplotype and higher resolution mapping of the tfa loci.the genetic substitution of transformation amenability alleles from 'golden promise' can facilitate the development of transformation-efficient lines from recalcitrant barley cultivars. barley (hordeum vulgare) cv. 'golden promise' is one of the most useful and well-studied cultivars for genetic manipulation. in a previous report, we identified several transformation amenability (tfa) loci responsible for agrobacterium-mediated transformation using the f2 generation of immature embryos, derived ...201728204911
membrane remodeling by a bacterial phospholipid-methylating enzyme.membrane deformation by proteins is a universal phenomenon that has been studied extensively in eukaryotes but much less in prokaryotes. in this study, we discovered a membrane-deforming activity of the phospholipid n-methyltransferase pmta from the plant-pathogenic bacterium agrobacterium tumefaciens pmta catalyzes the successive three-step n-methylation of phosphatidylethanolamine to phosphatidylcholine. here, we defined the lipid and protein requirements for the membrane-remodeling activity o ...201728196959
development of a colourimetric assay for glycosynthases.the synthesis of glycosidic structures by catalysis via glycosynthases has gained much interest due to the potential high product yields and specificity of the enzymes. nevertheless, the characterisation and implementation of new glycosynthases is greatly hampered by the lack of high-throughput methods for reaction analysis and screening of potential glycosynthase variants. fluoride detection, via silyl ether chemosensors, has recently shown high potential for the identification of glycosynthase ...201728193496
cloning and characterization of sidhn, a novel dehydrin gene from saussurea involucrata kar. et kir. that enhances cold and drought tolerance in tobacco.saussurea involucrata kar. et kir. is a hardy, dicotyledonous plant featured by strong tolerance to severe stress conditions. in a previous study, we created a cdna library to characterize genetic factors associated with the cold acclimation response of s. involucrata. the full-length cdna of one dehydrin-like gene, sidhn, was obtained by rt-pcr and the sidhn gene was further characterized in this study. the full-length sidhn cdna consists of 703 base pairs (bp) with a 333bp open reading frame e ...201728167030
re-sequencing transgenic plants revealed rearrangements at t-dna inserts, and integration of a short t-dna fragment, but no increase of small mutations elsewhere.transformation resulted in deletions and translocations at t-dna inserts, but not in genome-wide small mutations. a tiny t-dna splinter was detected that probably would remain undetected by conventional techniques. we investigated to which extent agrobacterium tumefaciens-mediated transformation is mutagenic, on top of inserting t-dna. to prevent mutations due to in vitro propagation, we applied floral dip transformation of arabidopsis thaliana. we re-sequenced the genomes of five primary transf ...201728155116
overexpression of the ascorbate peroxidase gene from eggplant and sponge gourd enhances flood tolerance in transgenic arabidopsis.previously, we found that the flood resistance of eggplant (solanum melongena) and sponge gourd (luffa cylindrica) enhanced ascorbate peroxidase (apx) activity under flooding, and consequently, both the smapx and lcapx genes were cloned. in this study, the smapx and lcapx genes were transferred under a ubiquitin promoter to arabidopsis (at) via agrobacterium tumefaciens. the expression and amount of apx and apx activities of the smapx and lcapx transgenic lines were significantly higher than tho ...201728150172
the signal peptide-like segment of hpaxm is required for its association to the cell wall in transgenic tobacco plants.harpins, encoded by hrp (hypersensitive response and pathogenicity) genes of gram-negative plant pathogens, are elicitors of hypersensitive response (hr). hpaxm is a novel harpin-like protein described from cotton leaf blight bacteria, xanthomonas citri subsp. malvacearum-a synonym of x. campestris pv. malvacearum (smith 1901-1978). a putative signal peptide (1-mnslntqiganssfl-15) of hpaxm was predicted in the nitroxyl-terminal (n-terminal)by signalp (signalp 3.0 server). here, we explored the f ...201728141855
agrobacteria enhance plant defense against root-knot nematodes on tomato.the increased incidence of the crown gall disease caused by agrobacterium tumefaciens has long been associated with activities of root-knot nematodes (meloidogyne spp.). pot experiments on tomato were designed to assess plant vitality, nematode reproduction, and crown gall incidence in combined infection with agrobacterium and meloidogyne spp. on tomato roots. results suggest that tomato plants infected with pathogenic a. tumefaciens 2 days before the nematodes show enhanced plant defense agains ...201728134593
agrobacterium-mediated transformation of leaf base segments.agrobacterium-mediated transformation has become a routine method of genetic engineering of cereals, gradually replacing the biolistic protocols. simple integration patterns of transgenic loci, decent transformation efficiency, and technical simplicity are the main advantages offered by this method. here we present a detailed protocol for the production of transgenic oat plants by agrobacterium-mediated transformation of leaf base segments. the use of leaf explants as target tissues for transfor ...201728132145
an oxidoreductase aioe is responsible for bacterial arsenite oxidation and resistance.previously, we found that arsenite (as(iii)) oxidation could improve the generation of atp/nadh to support the growth of agrobacterium tumefaciens gw4. in this study, we found that aioe is induced by as(iii) and located in the arsenic island near the as(iii) oxidase genes aioba and co-transcripted with the arsenic resistant genes arsr1-arsc1-arsc2-acr3-1. aioe belongs to trka family corresponding the electron transport function with the generation of nadh and h(+). an aioe in-frame deletion stra ...201728128323
t-6b allocates more assimilation product for oil synthesis and less for polysaccharide synthesis during the seed development of arabidopsis thaliana.as an agrobacterium tumefaciens t-dna oncogene, t-6b induces the development of tumors and the enation syndrome in vegetative tissues of transgenic plants. most of these effects are related to increases in soluble sugar contents. to verify the potential roles of t-6b in the distribution of carbon in developing seeds, not in vegetative tissues, we fused an endosperm-specific promoter to the t-6b gene for expression in transgenic arabidopsis thaliana plants.201728127400
construction of agrobacterium tumefaciens-mediated tomato black ring virus infectious cdna clones.tomato black ring virus (tbrv, genus nepovirus) infects a wide range of economically important plants such as tomato, potato, tobacco and cucumber. here, a successful construction of infectious full-length cdna clones of the tbrv genomic rnas (rna1 and rna2) is reported for the first time. the engineered constructs consisting of pcr-amplified dnas were cloned into binary vector pjl89 immediately downstream of a double cauliflower mosaic virus (camv) 35s promoter, and upstream of the hepatitis de ...201728109755
highly efficient transformation system for malassezia furfur and malassezia pachydermatis using agrobacterium tumefaciens-mediated transformation.malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. the absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. here we describe, a highly efficient agrobacterium-mediated genetic transformation system for malassezia furfur and m. pachydermatis. a binary t-dna vector with the hygromycin b phosphotransferase (hpt) selection marke ...201728064034
expression of talaromyces thermophilus lipase gene in trichoderma reesei by homologous recombination at the cbh1 locus.cbh1 (cellobiohydrolase) comprises the majority of secreted proteins by trichoderma reesei. for expression of talaromyces thermophilus lipase gene in t. reesei, a self-designed cbh1 promoter was applied to drive the lipase gene expression cassette which was bracketed by flanking sequences of cbh1 gene for homologous recombination. protoplast and agrobacterium-mediated plasmid transformations were performed and compared, resultantly, transformation mediated by agrobacterium was overall proved to ...201728039549
multifaceted roles of the ras guanine-nucleotide exchange factor chrgf in development, pathogenesis, and stress responses of colletotrichum higginsianum.the infection process of colletotrichum higginsianum, which causes a disease of crucifers, involves several key steps: conidial germination, appressorial formation, appressorial penetration, and invasive growth in host tissues. in this study, the chrgf gene encoding a ras guanine-nucleotide exchange factor protein was identified by screening t-dna insertion mutants generated from agrobacterium tumefaciens-mediated transformation that were unable to cause disease on the host arabidopsis thaliana. ...201728026997
agrobacterium tumefaciens-mediated transformation as an efficient tool for insertional mutagenesis of cercospora zeae-maydis.an efficient agrobacterium tumefaciens-mediated transformation (atmt) approach was developed for the plant pathogenic fungus, cercospora zeae-maydis, which is the causative agent of gray leaf spot in maize. the transformation was evaluated with five parameters to test the efficiencies of transformation. results showed that spore germination time, co-cultivation temperature and time were the significant influencing factors in all parameters. randomly selected transformants were confirmed and the ...201727965011
penicillin v acylases from gram-negative bacteria degrade n-acylhomoserine lactones and attenuate virulence in pseudomonas aeruginosa.virulence pathways in gram-negative pathogenic bacteria are regulated by quorum sensing mechanisms, through the production and sensing of n-acylhomoserine lactone (ahl) signal molecules. enzymatic degradation of ahls leading to attenuation of virulence (quorum quenching) could pave the way for the development of new antibacterials. penicillin v acylases (pvas) belong to the ntn hydrolase superfamily, together with ahl acylases. pvas are exploited widely in the pharmaceutical industry, but their ...201727933456
type iv effector proteins involved in the medicago-sinorhizobium symbiosis.in this study, we investigated genetic elements of the type iv secretion system (t4ss) found in sinorhizobium spp. and the role they play in symbiosis. sinorhizobium meliloti and s. medicae each contain a putative t4ss similar to that used by agrobacterium tumefaciens during pathogenesis. the cre reporter assay for translocation system was used to validate potential effector proteins. both s. meliloti and s. medicae contained the effector protein tfea, which was translocated into the host plant. ...201727918247
transcriptional activation of virulence genes of rhizobium etli.recently, rhizobium etli, in addition to agrobacterium spp., has emerged as a prokaryotic species whose genome encodes a functional machinery for dna transfer to plant cells. to understand this r. etli-mediated genetic transformation, it would be useful to define how its vir genes respond to the host plants. here, we explored the transcriptional activation of the vir genes contained on the r. etli p42a plasmid. using a reporter construct harboring lacz under the control of the r. etli vire promo ...201728069822
the arginine pairs and c-termini of the sso7c4 from sulfolobus solfataricus participate in binding and bending dna.the sso7c4 from sulfolobus solfataricus forms a dimer, which is believed to function as a chromosomal protein involved in genomic dna compaction and gene regulation. here, we present the crystal structure of wild-type sso7c4 at a high resolution of 1.63 å, showing that the two basic c-termini are disordered. based on the fluorescence polarization (fp) binding assay, two arginine pairs, r11/r22' and r11'/r22, on the top surface participate in binding dna. as shown in electron microscopy (em) imag ...201728068385
rhizobium radiobacter in pulmonary abscess associated with postgripal necrotizing pneumonia. 201727897436
fitness costs restrict niche expansion by generalist niche-constructing pathogens.we investigated the molecular and ecological mechanisms involved in niche expansion, or generalism, versus specialization in sympatric plant pathogens. nopaline-type and octopine-type agrobacterium tumefaciens engineer distinct niches in their plant hosts that provide different nutrients: nopaline or octopine, respectively. previous studies revealed that nopaline-type pathogens may expand their niche to also assimilate octopine in the presence of nopaline, but consequences of this phenomenon on ...201727801902
metabolic response of agrobacterium tumefaciens 5a to arsenite.wide-spread abundance in soil and water, coupled with high toxicity have put arsenic at the top of the list of environmental contaminants. early studies demonstrated that both concentration and the valence state of inorganic arsenic (arsenite, as(iii) vs. arsenate as(v)) can be modulated by microbes. using genetics, transcriptomic and proteomic techniques, microbe-arsenic detoxification, respiratory as(v) reduction and as(iii) oxidation have since been examined. the effect of arsenic exposure on ...201727871140
arabidopsis nahg plants as a suitable and efficient system for transient expression using agrobacterium tumefaciens. 201727867105
transformation of the cucurbit powdery mildew pathogen podosphaera xanthii by agrobacterium tumefaciens.the obligate biotrophic fungal pathogen podosphaera xanthii is the main causal agent of powdery mildew in cucurbit crops all over the world. a major limitation of molecular studies of powdery mildew fungi (erysiphales) is their genetic intractability. in this work, we describe a robust method based on the promiscuous transformation ability of agrobacterium tumefaciens for reliable transformation of p. xanthii. the a. tumefaciens-mediated transformation (atmt) system yielded transformants of p. x ...201727864969
dna labelling of varieties covered by patent protection: a new solution for managing intellectual property rights in the seed industry.plant breeders' rights are undergoing dramatic changes due to changes in patent rights in terms of plant variety rights protection. although differences in the interpretation of »breeder's exemption«, termed research exemption in the 1991 upov, did exist in the past in some countries, allowing breeders to use protected varieties as parents in the creation of new varieties of plants, current developments brought about by patenting conventionally bred varieties with the european patent office (suc ...201727567633
testing transgenic aspen plants with bar gene for herbicide resistance under semi-natural conditions.obtaining herbicide resistant plants is an important task in the genetic engineering of forest trees. transgenic european aspen plants (populus tremula l.) expressing the bar gene for phosphinothricin resistance have been produced using agrobacterium tumefaciens-mediated transformation. successful genetic transformation was confirmed by pcr analysis for thirteen lines derived from two elite genotypes. in 2014-2015, six lines were evaluated for resistance to herbicide treatment under semi-natural ...201727437143
low-affinity iron transport protein uvt3277 is important for pathogenesis in the rice false smut fungus ustilaginoidea virens.ustilaginoidea virens is the causal agent of rice false smut disease resulting in quantitative and qualitative losses in rice. to gain insights into the pathogenic mechanisms of u. virens, we established a t-dna insertion mutant library of u. virens through agrobacterium tumefaciens-mediated transformation and selected an enhanced pathogenicity mutant (i.e., b3277). we analyzed the biological characteristics of the wild-type p1 and b3277. the growth rate and sporulation of b3277 were decreased c ...201727306226
function analysis of caffeoyl-coa o-methyltransferase for biosynthesis of lignin and phenolic acid in salvia miltiorrhiza.in this study, we cloned a full-length cdna and the genomic dna sequence of smccoaomt (genbank id jq007585) from salvia miltiorrhiza. the 744-bp open-reading frame encodes a protein of 247 amino acids that shares 95 % similarity with one in vitis vinifera. real-time quantitative pcr analysis revealed that smccoaomt is most highly expressed in the stems and can be induced by methyl jasmonate (meja) and xc-1 treatment. to evaluate its function in vivo, we generated rna interference transgenic plan ...201727613617
lipopeptides from a novel bacillus methylotrophicus 39b strain suppress agrobacterium crown gall tumours on tomato plants.this study aims to characterise the antibacterial activity of a novel bacillus methylotrophicus strain named 39b against tumourigenic agrobacterium tumefaciens c58 and b6 strains. it also aims to identify the compound that is responsible for its activity and to evaluate its efficiency to control crown gall disease in tomato plants.201727252109
diaporthe endophytica and d. terebinthifolii from medicinal plants for biological control of phyllosticta citricarpa.the citrus industry is severely affected by citrus black spot (cbs), a disease caused by the pathogen phyllosticta citricarpa. this disease causes loss of production, decrease in the market price of the fruit, and reduction in its export to the european union. currently, cbs disease is being treated in orchards with various pesticides and fungicides every year. one alternative to cbs disease control without harming the environment is the use of microorganisms for biological control. diaporthe en ...201727242153
expression of ipt in asakura-sanshoo (zanthoxylum piperitum (l.) dc. f. inerme makino) alters tree architecture, delays leaf senescence, and changes leaf essential oil composition.the ipt gene encodes isopentenyl pyrophosphate transferase, a key enzyme in cytokinin biosynthesis. we introduced ipt under the control of the camv35s promoter into asakura-sanshoo (zanthoxylum piperitum (l.) dc. f. inerme makino) via stable agrobacterium tumefaciens-mediated transformation. three of 3-year-old transgenic asakura-sanshoo lines y5, y16, and y17 were used to evaluate the effects of ipt expression on the morphological characteristics, leaf senescence, and essential oil composition. ...201727182107
rhamnose synthase activity is required for pathogenicity of the vascular wilt fungus verticillium dahliae.the initial interaction of a pathogenic fungus with its host is complex and involves numerous metabolic pathways and regulatory proteins. considerable attention has been devoted to proteins that play a crucial role in these interactions, with an emphasis on so-called effector molecules that are secreted by the invading microbe to establish the symbiosis. however, the contribution of other types of molecules, such as glycans, is less well appreciated. here, we present a random genetic screen that ...201726996832
artificial agrobacterium tumefaciens strains exhibit diverse mechanisms to repress xanthomonas oryzae pv. oryzae-induced hypersensitive response and non-host resistance in nicotiana benthamiana.xanthomonas oryzae pv. oryzae (xoo) rapidly triggers a hypersensitive response (hr) and non-host resistance in its non-host plant nicotiana benthamiana. here, we report that agrobacterium tumefaciens strain gv3101 blocks xoo-induced hr in n. benthamiana when pre-infiltrated or co-infiltrated, but not when post-infiltrated at 4 h after xoo inoculation. this suppression by a. tumefaciens is local and highly efficient to xoo. the hr-inhibiting efficiency of a. tumefaciens is strain dependent. strai ...201727061769
improvement in agrobacterium-mediated transformation of chickpea (cicer arietinum l.) by the inhibition of polyphenolics released during wounding of cotyledonary node explants.agrobacterium-mediated transformation of chickpea (cicer arietinum l.) has been performed using cotyledonary node explants (cns), which release phenolics upon excision that are detrimental to the viability of agrobacterium tumefaciens and result in low transformation frequency. twelve low molecular weight phenolic compounds and salicylic acid were identified in the exudates released upon excision during the preparation of cotyledonary nodes by reverse phase high-performance liquid chromatography ...201726747430
an efficient protocol for agrobacterium-mediated transformation of the biofuel plant jatropha curcas by optimizing kanamycin concentration and duration of delayed selection.jatropha curcas is considered a potential biodiesel feedstock crop. currently, the value of j. curcas is limited because its seed yield is generally low. transgenic modification is a promising approach to improve the seed yield of j. curcas. although agrobacterium-mediated genetic transformation of j. curcas has been pursued for several years, the transformation efficiency remains unsatisfying. therefore, a highly efficient and simple agrobacterium-mediated genetic transformation method for j. c ...201726640597
peritonitis in a peritoneal dialysis patient due to rhizobium radiobacter and moraxella osleonsis: case report and literature review. 201725520491
in vitro regeneration and agrobacterium tumefaciens-mediated genetic transformation in asakura-sanshoo (zanthoxylum piperitum (l.) dc. f. inerme makino) an important medicinal plant.asakura-sanshoo (zanthoxylum piperitum [l.] dc. f. inerme makino) is an important medicinal plant in east asia. transgenic technique could be applied to improve plant traits and analyze gene function. however, there is no report on regeneration and genetic transformation in asakura-sanshoo.201725829778
processing heterogeneous biomass: overcoming the hurdles in model building.plants have been developed as an alternative platform for the production of biopharmaceutical proteins, culminating recently with the fda approval of the first plant-derived recombinant pharmaceutical enzyme for human use (elelysoô by protalix biotherapeutics). among the many different plant-based technologies that have been proposed, transient expression mediated by agrobacterium tumefaciens has proven to be particularly suitable for the rapid production of vaccines in response to emerging pand ...201722895059
improved vectors for agrobacterium mediated genetic manipulation of hypholoma spp. and other homobasidiomycetes.the basidiomycete fungi hypholoma fasciculare and h. sublateritium are both prolific producers of sesquiterpenes and triterpenes, some of which have relevant pharmaceutical properties. although h. sublateritium has been transformed in the past, the low reported efficiencies highlighted the need for establishing an effective simple transformation system for these valuable species. we have optimized agrobacterium tumefaciens-mediated transformation through testing various parameters in these two h ...201728843436
sit4-associated protein is required for pathogenicity of leptosphaeria maculans on brassica napus.an insertional mutant with reduced pathogenicity on brassica napus was identified in the plant pathogenic fungus leptosphaeria maculans. the transfer-dna molecule from agrobacterium tumefaciens inserted into a gene encoding a protein with similarity to sit4-associated proteins (saps). in contrast to saccharomyces cerevisiae which has four members of the sap family, there is a single copy of the gene in l. maculans. the mutant had normal spore production and spore germination, but altered hyphal ...201728840344
differential fine-tuning of gene expression regulation in coffee leaves by ccdreb1d promoter haplotypes under water deficit.despite the importance of the dreb1d gene (also known as cbf4) in plant responses to water deficit and cold stress, studies analysing its regulation by transgenic approaches are lacking. in the current work, a functional study of three ccdreb1d promoter haplotypes (named hp15, hp16 and hp17) isolated from drought-tolerant and drought-sensitive clones of coffea canephora was carried out in plants of c. arabica stably transformed by agrobacterium tumefaciens by analysing their ability to regulate ...201728830103
identification of factors involved in dimorphism and pathogenicity of zymoseptoria tritici.a forward genetics approach was applied in order to investigate the molecular basis of morphological transition in the wheat pathogenic fungus zymoseptoria tritici. z. tritici is a dimorphic plant pathogen displaying environmentally regulated morphogenetic transition between yeast-like and hyphal growth. considering the infection mode of z. tritici, the switching to hyphal growth is essential for pathogenicity allowing the fungus the host invasion through natural openings like stomata. we exploi ...201728829795
a novel in vitro transformation of lepidium draba l. using rapid direct shoot regeneration.the present research is carried out to study lepidium draba gene transformation for the first time, using direct shoot explants. as a prerequisite for gene transformation, the regeneration conditions in l. draba were optimized. we achieved an efficient and reproducible protocol for successful direct shoot regeneration without intervening callus formation. the results indicate that l. draba is the insistent species of brassicaceae in direct shoot regeneration. various explants of l. draba were ge ...201728828291
endo-cannabinoids system and the toxicity of cannabinoids with a biotechnological approach.cannabinoids have shown diverse and critical effects on the body systems, which alter the physiological functions. synthetic cannabinoids are comparatively innovative misuse drugs with respect to their nature of synthesis. synthetic cannabinoids therapy in healthy, chain smokers, and alcoholic individuals cause damage to the immune and nervous system, eventually leading to intoxication throughout the body. relevant studies were retrieved using major electronic databases such as pubmed, embase, m ...201728827985
metabolism of carbamazepine in plant roots and endophytic rhizobacteria isolated from phragmites australis.carbamazepine (cbz) is a pharmaceutical frequently categorized as a recalcitrant pollutant in the aquatic environment. endophytic bacteria previously isolated from reed plants have shown the ability to promote growth of their host and to contribute to cbz metabolism. in this work, a horseradish (armoracia rusticana) hairy root (hr) culture has been used as a plant model to study the interactions between roots and endophytic bacteria in response to cbz exposure. hrs could remove up to 5% of the i ...201728823920
metabolic engineering of photorespiration.the introduction of two alternative glycolate catabolic pathways in the chloroplasts of arabidopsis thaliana rendered plants with increased biomass. to introduce these synthetic pathways, the selected genes were stepwise integrated in the nuclear genome of wild-type plants. these plants were transformed by agrobacterium tumefaciens carrying the binary vectors using the floral dip method. selection of transformants was conducted using different selection agents and the expression of the transgene ...201728822131
antagonistic properties of some halophilic thermoactinomycetes isolated from superficial sediment of a solar saltern and production of cyclic antimicrobial peptides by the novel isolate paludifilum halophilum.this study has focused on the isolation of twenty-three halophilic actinomycetes from two ponds of different salinity and the evaluation of their ability to exert an antimicrobial activity against both their competitors and several other pathogens. from the 23 isolates, 18 strains showed antagonistic activity, while 19 showed activities against one or more of the seven pathogen strains tested. six strains exhibited consistent antibacterial activity against gram-negative and gram-positive pathoge ...201728819625
genetic engineering approach using early vinca alkaloid biosynthesis genes led to increased tryptamine and terpenoid indole alkaloids biosynthesis in differentiating cultures of catharanthus roseus.catharanthus roseus today occupies the central position in ongoing metabolic engineering efforts in medicinal plants. the entire multi-step biogenetic pathway of its very expensive anticancerous alkaloids vinblastine and vincristine is fairly very well dissected at biochemical and gene levels except the pathway steps leading to biosynthesis of monomeric alkaloid catharanthine and tabersonine. in order to enhance the plant-based productivity of these pharma molecules for the drug industry, cell a ...201728808798
molecular and phenotypic characterization of transgenic wheat and sorghum events expressing the barley alanine aminotransferase.the expression of a barley alanine aminotransferase gene impacts agronomic outcomes in a c3 crop, wheat. the use of nitrogen-based fertilizers has become one of the major agronomic inputs in crop production systems. strategies to enhance nitrogen assimilation and flux in planta are being pursued through the introduction of novel genetic alleles. here an agrobacterium-mediated approach was employed to introduce the alanine aminotransferase from barley (hordeum vulgare), hvalaat, into wheat (triti ...201728801748
rnai-mediated silencing of map kinase signalling genes (fmk1, hog1, and pbs2) in fusarium oxysporum reduces pathogenesis on tomato plants.fusarium oxysporum is a soil-borne plant fungal pathogen, and causes colossal losses in several crop plants including tomato. effective control measures include the use of harmful fungicides and resistant cultivars, but these methods have shown limited success. conventional methods to validate fungal pathogenic genes are labour intensive. therefore, an alternative strategy is required to efficiently characterize unknown pathogenic genes. rna interference (rnai) has emerged as a potential tool to ...201728800849
a hydroponic co-cultivation system for simultaneous and systematic analysis of plant/microbe molecular interactions and signaling.an experimental design mimicking natural plant-microbe interactions is very important to delineate the complex plant-microbe signaling processes. arabidopsis thaliana-agrobacterium tumefaciens provides an excellent model system to study bacterial pathogenesis and plant interactions. previous studies of plant-agrobacterium interactions have largely relied on plant cell suspension cultures, the artificial wounding of plants, or the artificial induction of microbial virulence factors or plant defen ...201728784965
agrobacterium-mediated co-transformation of multiple genes in metarhizium robertsii.fungi of the metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. to establish a co-transformation system for the transformation of multiple m. robertsii genes using agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. subsequently, in the two vectors containing the nourseothricin and phosphinothricin resist ...201728781540
evaluation of a mdmyb10/gfp43 fusion gene for its suitability to act as reporter gene in promoter studies in fragaria vesca l. 'rügen'.a malus domestica mdmyb10 transcription factor gene was previously used as visible marker for successful plant transformation. we combined the mdmyb10 transcription factor gene with a gfp gene to test its viability as a non-destructive, visual, double reporter system for functional promoter studies in transgenic strawberry plants. the gfp gene was fused to mdmyb10 to provide evidence for promoter activity in red colored cells of transformed plant tissue and to exclude artefacts resulting from st ...201728781398
cap analog and potato virus a hc-pro silencing suppressor improve gfp transient expression using an infectious virus vector in nicotiana benthamiana.transient expression of proteins in plants has become a choice to facilitate recombinant protein production with its fast and easy application. on the other hand, host defensive mechanisms have been reported to reduce the efficiency of transient expression in plants. hence, this study was designed to evaluate the effect of cap analog and potato virus a helper component proteinase (pva hc-pro) on green fluorescent protein (gfp) expression efficiency. n. benthamiana leaves were inoculated with cap ...201728775990
a glimpse into the genetic basis of symbiosis between hydrogenophaga and their helper strains in the biodegradation of 4-aminobenzenesulfonate.we report the whole genome sequences of hydrogenophaga intermedia s1 and agrobacterium radiobacter s2, the first reported bacterial co-culture capable of degrading 4-aminobenzenesulfonate (4-abs), a recalcitrant industrial waste product. to gain insights into the genetic basis for the syntrophic interaction between this symbiotic pair and also another recently reported hydrogenophaga associated co-culture, hydrogenophaga sp. pbc and ralstonia sp. pba, we performed detailed genetic analysis of th ...201728775791
delivery of cytoplasmic and apoplastic effectors from phytophthora infestans haustoria by distinct secretion pathways.the potato blight pathogen phytophthora infestans secretes effector proteins that are delivered inside (cytoplasmic) or can act outside (apoplastic) plant cells to neutralize host immunity. little is known about how and where effectors are secreted during infection, yet such knowledge is essential to understand and combat crop disease. we used transient agrobacterium tumefaciens-mediated in planta expression, transformation of p. infestans with fluorescent protein fusions and confocal microscopy ...201728758684
efficacy of eucalyptus cinerea as a source of bioactive compounds for curative biocontrol of crown gall caused by agrobacterium tumefaciens strain b6.this research investigated the eucalyptus cinerea leaves efficiency in the agrobacterium tumefaciens biocontrol, the causative agent of crown gall. gc-ms analysis of the essential oil (eo) showed that the main components were 1,8-cineole (61%) and camphene (15.13%). thanks to its polyphenols, flavonoids, quinones, terpenoids, alkaloids, and tannins richness, the etoac-f exhibited the most potent antibacterial activity in vitro. indeed, compared to the other fractions, it has the lowest mic and m ...201728758126
production of d-psicose from d-glucose by co-expression of d-psicose 3-epimerase and xylose isomerase.d-psicose has been drawing increasing attention in recent years because of its medical and health applications. the production of d-psicose from d-glucose requires the co-expression and synergistic action of xylose isomerase and d-psicose 3-epimerase. to co-express these genes, vector pet-28a(+)-dual containing two t7 promoters and rbs sites and an multiple cloning sites was constructed using the escherichia coli expression plasmid pet-28a(+). the xylose isomerase gene from e. coli mg1665 and th ...201728756856
transient gus gene expression in date palm fruit using agroinjection transformation technique.transient expression of foreign genes in plant tissue is a valuable tool for testing the efficacy of transformation methods. in this work, we present, for the first time, the utilization of agroinjection as an efficient transformation system for gene delivery in date palm fruit. the research utilized agrobacterium tumefaciens strain lba4404 harboring the binary vector pri201-an-gus carrying the beta-glucuronidase (gus) gene, under the control of a camv 35s and kanamycin (nptii) as an antibiotic ...201728755354
identification of a dimeric kdg aldolase from agrobacterium tumefaciens.agrobacterium tumefaciens is a gram-negative bacterium and causative agent of crown gall disease that infects a variety of economically important plants. the annotated a. tumefaciens genome contains 10 putative dapa genes, which code for dihydrodipicolinate synthase (dhdps). however, we have recently demonstrated that only one of these genes (dapa7) encodes a functional dhdps. the function of the other nine putative dapa genes is yet to be determined. here, we demonstrate using bioinformatics th ...201728748551
t-dna-genome junctions form early after infection and are influenced by the chromatin state of the host genome.agrobacterium tumefaciens mediated t-dna integration is a common tool for plant genome manipulation. however, there is controversy regarding whether t-dna integration is biased towards genes or randomly distributed throughout the genome. in order to address this question, we performed high-throughput mapping of t-dna-genome junctions obtained in the absence of selection at several time points after infection. t-dna-genome junctions were detected as early as 6 hours post-infection. t-dna distribu ...201728742090
generation and screening of t-dna insertion mutants mediated by agrobacterium tumefaciens in the garden asparagus stem blight pathogen phomopsis asparagi.the garden asparagus stem blight caused by filamentous fungus phomopsis asparagi exposes a serious threat on asparagus production globally. however, to present, we understand poorly about the molecular mechanisms of fungal pathogenicity. to facilitate functional genomics research of p. asparagi, here we developed a highly efficient and stable agrobacterium tumefaciens-mediated transformation approach which yielded 150-200 transformants per 1 × 10(6) conidia. our results indicated that 25 °c, ace ...201728730458
impact of paracoccin gene silencing on paracoccidioides brasiliensis virulence.among the endemic deep mycoses in latin america, paracoccidioidomycosis (pcm), caused by thermodimorphic fungi of the paracoccidioides genus, is a major cause of morbidity. disease development and its manifestations are associated with both host and fungal factors. concerning the latter, several recent studies have employed the methodology of gene modulation in p. brasiliensis using antisense rna (asrna) and agrobacterium tumefaciens-mediated transformation (atmt) to identify proteins that influ ...201728720727
generation of stable infectious clones of plant viruses by using rhizobium radiobacter for both cloning and inoculation.a novel rhizobium radiobacter (synonym agrobacterium tumefaciens)-mediated approach was developed to generate stable infectious clones of plant viruses. this method uses r. radiobacter for both cloning and inoculation of infectious clones, bypassing the requirement of cloning in e. coli to avoid the instability. only three steps are included in this method: (i) construct viral genome-encoding plasmids in vitro by one-step gibson assembly; (ii) transform the assembled dna products into r. radioba ...201728715654
melanization and ageing are not drawbacks for successful agro-transformation of dark septate endophytes.dark septate endophytes (dses) are melanin-enriched ascomycetous fungi that are abundant in stressed environments. however, little is known about their physiology and metabolism, and dse genes have not been functionally characterized yet. therefore developing molecular genetic tools to investigate the biological function of genes of interest in dses is of major significance. we investigated agrobacterium tumefaciens-mediated transformation (atmt) efficiency in eight dse strains belonging to cado ...201728705394
overexpression of ginseng ugt72al1 causes organ fusion in the axillary leaf branch of arabidopsis.glycosylation of natural compounds increases the diversity of secondary metabolites. glycosylation steps are implicated not only in plant growth and development, but also in plant defense responses. although the activities of uridine-dependent glycosyltransferases (ugts) have long been recognized, and genes encoding them in several higher plants have been identified, the specific functions of ugts in planta remain largely unknown.201728701886
cis-prenyltransferase interacts with a nogo-b receptor homolog for dolichol biosynthesis in panax ginseng meyer.prenyltransferases catalyze the sequential addition of isopentenyl diphosphate units to allylic prenyl diphosphate acceptors and are classified as either trans-prenyltransferases (tpts) or cis-prenyltransferases (cpts). the functions of cpts have been well characterized in bacteria, yeast, and mammals compared to plants. the characterization of cpts also has been less studied than tpts. in the present study, molecular cloning and functional characterization of a cpt from a medicinal plant, panax ...201728701884
genomic and transcriptomic analyses of agrobacterium tumefaciens s33 reveal the molecular mechanism of a novel hybrid nicotine-degrading pathway.agrobacterium tumefaciens s33 is able to degrade nicotine via a novel hybrid of the pyridine and pyrrolidine pathways. it can be utilized to remove nicotine from tobacco wastes and transform nicotine into important functionalized pyridine precursors for some valuable drugs and insecticides. however, the molecular mechanism of the hybrid pathway is still not completely clear. here we report the genome analysis of strain s33 and its transcriptomes grown in glucose-ammonium medium and nicotine medi ...201728684751
co-expression of beta-glucosidase and laccase in trichoderma reesei by random insertion with enhanced filter paper activity.trichoderma reesei strain rut-c30 was modified with enhanced beta-glycosidase (bgl) activity to balance the cellulase system and generated laccase (lac) protein for lignin degradation. initially, the binary plasmid p1300-w1 was constructed to express t. reesei bgl2 under the control of promoter p pki and t-nos terminator. random insertion was performed via agrobacterium tumefaciens-mediated transformation. a total of 353 mutants were obtained, and 34ptrb2 was exceptionally stable with increased ...201728676913
high resolution crystal structure of a fluoride-inhibited organophosphate-degrading metallohydrolase.metal ion-dependent, organophosphate-degrading enzymes (op hydrolases) have received increasing attention due to their ability to degrade and thus detoxify commonly used pesticides and nerve agents such as sarin and vx. these enzymes thus garner strong potential as bioremediators. the op hydrolase from agrobacterium radiobacter (opda) is one of the most efficient members of this group of enzymes. previous studies have indicated that the choice of the hydrolysis-initiating nucleophile may depend ...201728673485
a genetic transformation method for cadmium hyperaccumulator sedum plumbizincicola and non-hyperaccumulating ecotype of sedum alfredii.the present study demonstrates the development of an agrobacterium-mediated genetic transformation method for species of the sedum genus, which includes the cd/zn hyperaccumulator sedum plumbizincicola and the non-hyperaccumulating ecotype of s. alfredii. multiple shoots were induced from stem nodes of two sedum plants using murashige and skoog (ms) medium containing 0.1 mg/l cytokinin 6-benzyladenine (6-ba) and 1.0 mg/l auxin 1-naphthaleneacetic acid (naa). the shoot primordia were used as dire ...201728670322
development of a mixed microbial culture for thiocyanate and metal cyanide degradation.the degradation capacity of a mixed culture of agrobacterium tumefaciens suts 1 and pseudomonas monteilii suts 2 for thiocyanate and metal cyanide, in the form of zinc and cadmium, has been determined. the growth of a mixed culture of suts 1 and suts 2 in cyanide complexes and the cyanide removal efficiency of a fixed-film bio-column system were studied. the results showed that the mixed culture of bacteria can survive and grow in broth media containing thiocyanate and metal cyanide complexes wi ...201728664381
production, characterization and structural modification of exopolysaccharide-based bioflocculant by rhizobium radiobacter sz4s7s14 and media optimization.synthesis of the exopolysaccharide-based bioflocculant rhizobium radiobacter sz4s7s14 was researched and medium optimized for enhanced production of bioflocculant. d-mannose and yeast extract were found to be best carbon and nitrogen sources for maximal yield of bioflocculant. the bioflocculant samples obtained in different media with different carbon and nitrogen sources were further analyzed by sem-edx and ft-ir. ft-ir spectroscopy of the bioflocculant samples, obtained in different carbon/nit ...201728664366
trimethylphosphate and dimethylphosphate hydrolysis by binuclear cd(ii) , mn(ii) , and zn(ii) -fe(ii) promiscuous organophosphate-degrading enzyme: reaction mechanisms.organophosphate-degrading enzyme from agrobacterium radiobacter p230 exhibits promiscuity, not only in the reactions it catalyzes, but also in the metals it uses to catalyze those reactions. here, three different binuclear metal centers were studied: di-cd(ii) , di-mn(ii) and zn(ii) -fe(ii) . this enzyme uses these metal centers for hydrolyzing trimethyl- and dimethyl-phosphates. both mechanisms were studied at dft level of theory using a cluster model approach. the ground spin state was determi ...201728661038
multiple t-dna delivery to plants using novel mini binary vectors with compatible replication origins.improved plants are necessary to meet human needs. agrobacterium-mediated transformation is the most common method used to rewire plant capabilities. for plant gene delivery, dna constructs are assembled into binary t-dna vectors that rely on broad host range origins for bacterial replication. here we present plx vectors, a set of mini binary t-dna plasmids suitable for type iis restriction endonuclease- and overlap-based assembly methods. plx vectors include replicons from compatible broad host ...201728657330
gr1-like gene expression in lycium chinense was regulated by cadmium-induced endogenous jasmonic acids accumulation.the g1-like gene from the lycium chinense was cloned and transferred into n. tabacum. evidence showed that endogenous ja accumulation was crucial to lcgr gene expression in cadmium-stressed l. chinense. glutathione reductase (gr) plays a vital role in glutathione-ascorbate metabolism and is a key enzyme in maintaining the redox state in plants. jasmonic acids (ja) are important hormones regulating protective responses against bacteria and mechanic damage in plants. at present, the relationship b ...201728656324
rapid deletion production in fungi via agrobacterium mediated transformation of oscar deletion constructs.precise deletion of gene(s) of interest, while leaving the rest of the genome unchanged, provides the ideal product to determine that particular gene's function in the living organism. in this protocol the oscar method of precise and rapid deletion plasmid construction is described. oscar relies on the cloning system in which a single recombinase reaction is carried out containing the purified pcr-amplified 5' and 3' flanks of the gene of interest and two plasmids, pa-hyg oscar (the marker vecto ...201728654073
agrobacterium tumefaciens-mediated transformation of oleaginous yeast lipomyces species.interest in using renewable sources of carbon, especially lignocellulosic biomass, for the production of hydrocarbon fuels and chemicals has fueled interest in exploring various organisms capable of producing hydrocarbon biofuels and chemicals or their precursors. the oleaginous (oil-producing) yeast lipomyces starkeyi is the subject of active research regarding the production of triacylglycerides as hydrocarbon fuel precursors using a variety of carbohydrate and nutrient sources. the genome of ...201728631219
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