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Stanton B Gelvin - One of the best experts on this subject based on the ideXlab platform.
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editorial Agrobacterium biology and its application to transgenic plant production
Frontiers in Plant Science, 2015Co-Authors: Hauhsuan Hwang, Stanton B Gelvin, Erhmin LaiAbstract:The extraordinary Agrobacterium research story started from the search for the causative agent of crown gall disease more than 100 years ago. Agrobacterium tumefaciens was first isolated from grapevine galls in 1897 and later isolated from Paris daisy in 1907 (Cavara, 1897a,b; Smith and Townsend, 1907). The Agrobacterium infection mechanism involves processing and transfer of a specific DNA fragment (the transferred-DNA, T-DNA) from a bacterial tumor-inducing (Ti) plasmid. Transfer to the plant occurs via a type IV secretion system (T4SS), after which T-DNA is integrated into the plant host genome (Gelvin, 2010; Lacroix and Citovsky, 2013). This interkingdom DNA transfer leads to overproduction of the plant hormones auxin and cytokinin, resulting in tumors. The interkingdom DNA transfer ability of Agrobacterium and the possibility to replace the oncogenes in the T-DNA with genes of interest has made Agrobacterium-mediated transformation the most popular technique to generate transgenic plants. This Research Topic provides a collection of reviews and original research articles on Agrobacterium genes involved in bacterial physiology/virulence and plant genes involved in transformation and defense against Agrobacterium. A review by Kado (2014) provides a historical overview of how A. tumefaciens was first established as the cause of crown gall disease. In this review, Kado highlights key early plant pathology and milestone molecular biology studies leading to the conclusion that the expression of oncogenes in native T-DNA is the cause of tumor growth in plants. With the solid foundation of these pioneering discoveries, A. tumefaciens evolved from a phytopathogen to a powerful genetic transformation tool for plant biology and biotechnology research. The first complete genome sequence of an Agrobacterium species (A. tumefaciens C58) was completed in 2001 (Goodner et al., 2001; Wood et al., 2001). The 5.67-megabase genome of this strain carries one circular chromosome, one linear chromosome, and two megaplasmids: the Ti plasmid pTiC58 and a second plasmid, pAtC58. In the review by Platt et al. (2014), the properties, ecology, evolution, and complex interactions of these two A. tumefaciens megaplasmids are discussed. The costs and benefits to A. tumefaciens strains carrying the Ti plasmid and/or the pAtC58 plasmid are discussed and presented from an ecological and evolutionary perspective. Modeling predictions are presented for the relative cost and benefits to A. tumefaciens strains harboring the Ti and/or the pAtC58 plasmids determined by environmental resources. Conjugation and amplification of the Ti plasmid are regulated by the TraI/TraR quorum-sensing (QS) system and conjugal opines. Lang and Faure (2014) review current knowledge of the genetic networks and molecular basis of the A. tumefaciens quorum sensing system. These authors also discuss the biological and ecological impact of the QS system on Ti plasmid conjugation, copy number, and interactions between Agrobacterium and host plants. During the initial interaction between Agrobacterium and plant cells, bacteria sense various plant-derived signals in the rhizosphere with the help of Ti plasmid-encoded virulence gene (vir gene) and chromosomal virulence gene (chv gene) products. The current knowledge of how A. tumefaciens senses and reacts to different plant-derived signals are summarized in the review article by Subramoni et al. (2014), which also discusses the mechanisms of how the plant hormones auxin, salicylic acid, and ethylene, affect bacterial virulence. Finally, this review discusses the complexity and intricacy of Agrobacterium signaling pathways and the underlying regulatory mechanisms during the initial host cell recognition to maximize subsequent successful infection. In the original research article by Lin et al. (2014), the mechanistic regulation of the membrane sensor VirA protein is further dissected. VirA histidine kinase and the cytoplasmic response regulator VirG protein together play a central role in regulating vir gene expression in response to phenolics. Based on a homology model of the VirA linker region, various mutant and chimeric VirA proteins were generated and examined for their ability to induce VirB promoter activity. The ability of VirA to sense and respond to three separate input signals, phenolics, sugars, and environmental pH, plays a significant role in securing successful infection. Agrobacterium attachment to plant cells is an important early step in crown gall disease progression. Motile bacteria swim toward host cells and then physically interact with host cells to form aggregates and establish a multicellular bacterial community known as a biofilm. Various genetic and environmental factors that affect Agrobacterium attachment and biofilm formation are reviewed in the article by Heindl et al. (2014). The functions of different types of exopolysaccharides that constitute the biofilm and underlying mechanisms involving how the second messenger cyclic-di-GMP, the ChvG/ChvI system, phosphorus levels, and oxygen tension influence bacterial attachment and virulence are also summarized. In the review article by Matthysse (2014), early studies and current knowledge of the mechanisms of polar and lateral bacterial attachment are summarized. These two mechanisms both contribute to bacterial attachment. When the environmental calcium and phosphate levels and pH values are low, polar attachment predominates. In addition, the phospholipids (PLs), phosphatidylcholine (PC), and phosphate-free lipid ornithine lipids (OLs) contribute to Agrobacterium virulence. In the review by Aktas et al. (2014), the biosynthetic pathways and the physiological roles of these membrane lipids are summarized. The typical eukaryotic membrane lipid PC is not frequently found in bacteria, but it constitutes almost 22% of the Agrobacterium membrane lipid. Interestingly, PCs and OLs may play opposite roles in Agrobacterium virulence. The reduction of tumor formation in a PC-deficient Agrobacterium mutant may result from impaired vir gene expressions controlled by VirA/VirG. The absence of OLs in A. tumefaciens may decrease host defense responses and therefore cause earlier and larger tumor formation. Plant cells have a variety of receptors that recognize so-called microbe- or pathogen-associated molecular patterns (MAMPs or PAMPs), and subsequently activate plant defense responses, a process known as Pattern-recognition receptor-Triggered Immunity (PTI) (Boller and Felix, 2009; Boller and He, 2009). Agrobacteriium may utilize effectors to hijack plant systems and evade plant defense responses. Pitzschke (2013) reviews strategies used by Agrobacterium to turn plant defense responses to its own advantage. Infected plant cells initiate a mitogen-activated protein kinase signaling cascade that causes VIP1 (Agrobacterium VirE2-interacting protein 1) phosphorylation and translocation into the plant nucleus to induce defense gene expression. On the other hand, Agrobacterium may hijack VIP1 to help T-DNA enter the plant nucleus. Based on the current knowledge of plant defense responses against Agrobacterium infection, Pitzschke (2013) discusses several biotechnological approaches to increase transformation efficiency. In another review by Gohlke and Deeken (2014), early plant responses to Agrobacterium, including various defense responses, hypersensitive responses, and phytohormone level alterations are discussed. The alterations in plant morphology, nutrient translocation, and metabolism caused by crown gall tumor formation are also reviewed. The authors summarize important genomic, epigenomic, transcriptomic, and metabolomic studies that reveal epigenetic changes associated with T-DNA integration and gall development. Subsequently, Hwang et al. (2015) review important pathogenic elicitors, host cell receptor molecules, and their downstream signal transduction pathways in host plants during the PAMP-triggered immune response. They highlight recent discoveries linking plant immunity to endomembrane trafficking and actin dynamic changes. Effects of both the host physiology, including hormone levels, circadian clock, developmental stages, and environmental factors, including light exposure lengths and temperature, on plant defense responses and bacterial virulence are reviewed and discussed. In nature, evidence of ancient horizontal gene transfers (HGT) from Agrobacterium to plants has been observed in the genera Nicotiana and Linaria. Sequences homologous to mikimopine-type Agrobacterium rhizogenes pRiA4 T-DNA were first discovered in the genome of untransformed tree tobacco, Nicotiana glauca, and named “cellular T-DNA” (cT-DNA; White et al., 1983). Matveeva and Lutova (2014) review cT-DNA organization, distribution, expression regulation, and a possible correlation with genetic tumor formation in Nicotiana species. They also review recent findings of cT-DNA in the genomes of Linaria species and in other dicotyledonous families. The authors suggest that plants maintaining cT-DNA in their genomes may potentially benefit microorganisms in the rhizosphere by secreting opines in the root zone. They also propose that footprints of ancient pRi T-DNA insertions in the plant genome may provide selective advantage to these plants. With this Research Topic we provide a platform for scientists to share their understanding of Agrobacterium biology and how Agrobacterium transforms plants. These contributions demonstrate how a highly active research community in plant and microbial sciences can elucidate important pathogenesis questions. Future research on Agrobacteium will continue to advance our understanding of plant-pathogen interactions, and provide new insights useful for plant genetic engineering.
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Transgenic Arabidopsis plants expressing Agrobacterium tumefaciens VirD2 protein are less susceptible to Agrobacterium transformation.
Molecular plant pathology, 2006Co-Authors: Hauhsuan Hwang, Kirankumar S Mysore, Stanton B GelvinAbstract:Agrobacterium tumefaciens causes crown gall disease on many plant species and can result in considerable economic losses. Here we report a new strategy to control crown gall disease by over-expressing Agrobacterium tumefaciens VirD2 protein in plants. Transgenic Arabidopsis plants over-expressing virD2 from constitutive or wound-inducible promoters are less susceptible to Agrobacterium-mediated transformation. Additionally, the transient introduction of an A. tumefaciens virD2 gene in tobacco BY-2 cells reduces subsequent Agrobacterium-mediated transformation.
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transfer of t dna and vir proteins to plant cells by Agrobacterium tumefaciens induces expression of host genes involved in mediating transformation and suppresses host defense gene expression
Plant Journal, 2003Co-Authors: Hongmei Jiang, R W Doerge, Stanton B GelvinAbstract:Agrobacterium tumefaciens is a plant pathogen that incites crown gall tumors by transferring to and expressing a portion of a resident plasmid in plant cells. Currently, little is known about the host response to Agrobacterium infection. Using suppressive subtractive hybridization and DNA macroarrays, we identified numerous plant genes that are differentially expressed during early stages of Agrobacterium-mediated transformation. Expression profiling indicates that Agrobacterium infection induces plant genes necessary for the transformation process while simultaneously repressing host defense response genes, thus indicating successful utilization of existing host cellular machinery for genetic transformation purposes. A comparison of plant responses to different strains of Agrobacterium indicates that transfer of both T-DNA and Vir proteins modulates the expression of host genes during the transformation process.
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Agrobacterium mediated plant transformation the biology behind the gene jockeying tool
Microbiology and Molecular Biology Reviews, 2003Co-Authors: Stanton B GelvinAbstract:Agrobacterium tumefaciens and related Agrobacterium species have been known as plant pathogens since the beginning of the 20th century. However, only in the past two decades has the ability of Agrobacterium to transfer DNA to plant cells been harnessed for the purposes of plant genetic engineering. Since the initial reports in the early 1980s using Agrobacterium to generate transgenic plants, scientists have attempted to improve this “natural genetic engineer” for biotechnology purposes. Some of these modifications have resulted in extending the host range of the bacterium to economically important crop species. However, in most instances, major improvements involved alterations in plant tissue culture transformation and regeneration conditions rather than manipulation of bacterial or host genes. Agrobacterium-mediated plant transformation is a highly complex and evolved process involving genetic determinants of both the bacterium and the host plant cell. In this article, I review some of the basic biology concerned with Agrobacterium-mediated genetic transformation. Knowledge of fundamental biological principles embracing both the host and the pathogen have been and will continue to be key to extending the utility of Agrobacterium for genetic engineering purposes.
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NewAgrobacterium helper plasmids for gene transfer to plants
Transgenic Research, 1993Co-Authors: Elizabeth E Hood, Stanton B Gelvin, Leo Sjoerd Melchers, Andre HoekemaAbstract:We describe the construction of new helper Ti plasmids for Agrobacterium -mediated plant transformation. These plasmids are derived from three different Agrobacterium tumefaciens Ti plasmids, the octopine plasmid pTiB6, the nopaline plasmid pTiC58, and the L,L-succinamopine plasmid pTiBo542. The T-DNA regions of these plasmids were deleted using site-directed mutagenesis to yield replicons carrying the vir genes that will complement binary vectors in trans . Data are included that demonstrate strain utility. The advantages of Agrobacterium strains harbouring these ‘disamed’ Ti plasmids for plant transformation via Agrobacterium are discussed.
Xavier Nesme - One of the best experts on this subject based on the ideXlab platform.
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Agrobacterium rosae sp. nov., isolated from galls on different agricultural crops
Systematic and Applied Microbiology, 2018Co-Authors: Nemanja Kuzmanović, Joanna Pu\l Awska, Kornelia Smalla, Xavier NesmeAbstract:The plant tumorigenic strain NCPPB 1650T isolated from Rosa\texttimeshybrida, and four nonpathogenic strains isolated from tumors on grapevine (strain 384), raspberry (strain 839) and blueberry (strains B20.3 and B25.3) were characterized by using polyphasic taxonomic methods. Based on 16S rRNA gene phylogeny, strains were clustered within the genus Agrobacterium. Furthermore, multilocus sequence analysis (MLSA) based on the partial sequences of atpD, recA and rpoB housekeeping genes indicated that five strains studied form a novel Agrobacterium species. Their closest relatives were Agrobacterium sp. R89-1, Agrobacterium rubi and Agrobacterium skierniewicense. Authenticity of the novel species was confirmed by average nucleotide identity (ANI) and in silico DNADNA hybridization (DDH) comparisons between strains NCPPB 1650T and B20.3, and their closest relatives, since obtained values were considerably below the proposed thresholds for the species delineation. Whole-genome-based phylogeny further supported distinctiveness of the novel species, that forms together with A. rubi, A. skierniewicense and Agrobacterium sp. R89-1 a well-delineated sub-clade of Agrobacterium spp. named "rubi\textquotedblright. As for other species of the genus Agrobacterium, the major fatty acid of the strains studied was 18:1 w7c (73.4278.12%). The five strains studied were phenotypically distinguishable from other species of the genus Agrobacterium. Overall, polyphasic characterization showed that the five strains studied represent a novel species of the genus Agrobacterium, for which the name Agrobacterium rosae sp. nov. is proposed. The type strain of A. rosae is NCPPB 1650T (=DSM 30203T=LMG 230T=CFBP 4470T=IAM 13558T=JCM 20915T).
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Phylogeny of the Rhizobium-Allorhizobium-Agrobacterium clade supports the delineation of Neorhizobium gen. nov.
Systematic and Applied Microbiology, 2014Co-Authors: Seyed Abdollah Mousavi, Céline Lavire, Xavier Nesme, Niklas Wahlberg, Ludovic Vial, Lars Paulin, Philippe De Lajudie, Kristina LindströmAbstract:The genera Agrobacterium, Allorhizobium, and Rhizobium belong to the family Rhizobiaceae. However, the placement of a phytopathogenic group of bacteria, the genus Agrobacterium, among the nitrogen-fixing bacteria and the unclear position of Rhizobium galegae have caused controversy in previous taxonomic studies. To resolve uncertainties in the taxonomy and nomenclature within this family, the phylogenetic relationships of generic members of Rhizobiaceae were studied, but with particular emphasis on the taxa included in Agrobacterium and the "R. galegae complex" (R. galegae and related taxa), using multilocus sequence analysis (MLSA) of six protein-coding housekeeping genes among 114 rhizobial and agrobacterial taxa. The results showed that R. galegae, R. vignae, R. huautlense, and R. alkalisoli formed a separate clade that clearly represented a new genus, for which the name Neorhizobium is proposed. Agrobacterium was shown to represent a separate cluster of mainly pathogenic taxa of the family Rhizobiaceae. A. vitis grouped with Allorhizobium, distinct from Agrobacterium, and should be reclassified as Allorhizobium vitis, whereas Rhizobium rhizogenes was considered to be the proper name for former Agrobacterium rhizogenes. This phylogenetic study further indicated that the taxonomic status of several taxa could be resolved by the creation of more novel genera.
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rapid and efficient identification of Agrobacterium species by reca allele analysis Agrobacterium reca diversity
Microbial Ecology, 2010Co-Authors: Denis Costechareyre, Ali Rhouma, Céline Lavire, Perrine Portier, David Chapulliot, Franck Bertolla, Ali Boubaker, Yves Dessaux, Xavier NesmeAbstract:The analysis of housekeeping recA gene sequences from 138 strains from 13 species or genomic species of Agrobacterium, nine being biovar 1 genomospecies, and the others Agrobacterium larrymoorei, Agrobacterium rubi, Agrobacterium sp. NCPPB 1650, and Agrobacterium vitis and one “former” Agrobacterium species, Rhizobium rhizogenes, led to the identification of 50 different recA alleles and to a clear delineation of the 14 species or genomospecies entirely consistent with that obtained by amplified fragment length polymorphism (AFLP) analysis. The relevance of a recA sequencing approach for epidemiological analyses was next assessed on agrobacterial Tunisian isolates. All Tunisian isolates were found to belong to the Agrobacterium tumefaciens/biovar 1 species complex by both biochemical tests and rrs sequencing. recA sequence analysis further permitted their unambiguous assignment to A. tumefaciens genomospecies G4, G6, G7, and G8 in total agreement with the results of an AFLP-based analysis. At subspecific level, several Tunisian recA alleles were novel, indicating the power and accuracy of recA-based typing for studies of Agrobacterium spp.
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Rapid and Efficient Identification of Agrobacterium Species by recA Allele Analysis
Microbial Ecology, 2010Co-Authors: Denis Costechareyre, Ali Rhouma, Céline Lavire, Perrine Portier, David Chapulliot, Franck Bertolla, Ali Boubaker, Yves Dessaux, Xavier NesmeAbstract:The analysis of housekeeping recA gene sequences from 138 strains from 13 species or genomic species of Agrobacterium, nine being biovar 1 genomospecies, and the others Agrobacterium larrymoorei, Agrobacterium rubi, Agrobacterium sp. NCPPB 1650, and Agrobacterium vitis and one “former” Agrobacterium species, Rhizobium rhizogenes, led to the identification of 50 different recA alleles and to a clear delineation of the 14 species or genomospecies entirely consistent with that obtained by amplified fragment length polymorphism (AFLP) analysis. The relevance of a recA sequencing approach for epidemiological analyses was next assessed on agrobacterial Tunisian isolates. All Tunisian isolates were found to belong to the Agrobacterium tumefaciens/ biovar 1 species complex by both biochemical tests and rrs sequencing. recA sequence analysis further permitted their unambiguous assignment to A. tumefaciens genomospecies G4, G6, G7, and G8 in total agreement with the results of an AFLP-based analysis. At subspecific level, several Tunisian recA alleles were novel, indicating the power and accuracy of recA-based typing for studies of Agrobacterium spp.
Hiroyuki Sawada - One of the best experts on this subject based on the ideXlab platform.
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a revision of rhizobium frank 1889 with an emended description of the genus and the inclusion of all species of Agrobacterium conn 1942 and allorhizobium undicola de lajudie et al 1998 as new combinations rhizobium radiobacter r rhizogenes r rubi r u
International Journal of Systematic and Evolutionary Microbiology, 2001Co-Authors: J M Young, L D Kuykendall, Esperanza Martinezromero, A Kerr, Hiroyuki SawadaAbstract:Rhizobium, Agrobacterium and Allorhizobium are genera within the bacterial family Rhizobiaceae, together with Sinorhizobium. The species of Agrobacterium, Agrobacterium tumefaciens (syn. Agrobacterium radiobacter), Agrobacterium rhizogenes, Agrobacterium rubi and Agrobacterium vitis, together with Allorhizobium undicola, form a monophyletic group with all Rhizobium species, based on comparative 16S rDNA analyses. Agrobacterium is an artificial genus comprising plant-pathogenic species. The monophyletic nature of Agrobacterium, Allorhizobium and Rhizobium and their common phenotypic generic circumscription support their amalgamation into a single genus, Rhizobium. Agrobacterium tumefaciens was conserved as the type species of Agrobacterium, but the epithet radiobacter would take precedence as Rhizobium radiobacter in the revised genus. The proposed new combinations are Rhizobium radiobacter, Rhizobium rhizogenes, Rhizobium rubi, Rhizobium undicola and Rhizobium vitis.
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Proposal for Rejection of Agrobacterium tumefaciens and Revised Descriptions for the Genus Agrobacterium and for Agrobacterium radiobacter and Agrobacterium rhizogenes
International journal of systematic bacteriology, 1993Co-Authors: Hiroyuki Sawada, Hiroyuki Ieki, Hiroshi Oyaizu, Satoshi MatsumotoAbstract:The 16S rRNA sequences of seven representative Agrobacterium strains, eight representative Rhizobium strains, and the type strains of Azorhizobium caulinodans and Bradyrhizobium japonicum were determined. These strains included the type strains of Agrobacterium tumefaciens, Agrobacterium rhizogenes, Agrobacterium radiobacter, Agrobacterium vitis, Agrobacterium rubi, Rhizobium fredii, Rhizobium galegae, Rhizobium huakuii, Rhizobium leguminosarum, Rhizobium loti, Rhizobium meliloti, and Rhizobium tropici. A phylogenetic analysis showed that the 15 strains of Agrobacterium and Rhizobium species formed a compact phylogenetic cluster clearly separated from the other members of the alpha subclass of the Proteobacteria. However, Agrobacterium species and Rhizobium species are phylogenetically entwined with one another, and the two genera cannot be separated. In the Agrobacterium species, the strains of biovar 1, biovar 2, Agrobacterium rubi, and Agrobacterium vitis were clearly separated. The two biovars exhibited homogeneity in their phenotypic, chemotaxonomic, and phylogenetic characteristics, and two species should be established for the two biovars. We considered the nomenclature of the two biovars, and revised descriptions of Agrobacterium radiobacter(for the biovar 1 strains) and Agrobacterium rhizogenes(for the biovar 2 strains) are proposed. The name Agrobacterium tumefaciens is rejected because the type strain of this species was assigned to Agrobacterium radiobacter, and consequently the description of the genus Agrobacterium is revised.
Vitaly Citovsky - One of the best experts on this subject based on the ideXlab platform.
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the roles of bacterial and host plant factors in Agrobacterium mediated genetic transformation
The International Journal of Developmental Biology, 2013Co-Authors: Benoa T Lacroix, Vitaly CitovskyAbstract:The genetic transformation of plants mediated by Agrobacterium tumefaciens represents an essential tool for both fundamental and applied research in plant biology. For a successful infection, culminating in the integration of its transferred DNA (T-DNA) into the host genome, Agrobacterium relies on multiple interactions with host-plant factors. Extensive studies have unraveled many of such interactions at all major steps of the infection process: activation of the bacterial virulence genes, cell-cell contact and macromolecular translocation from Agrobacterium to host cell cytoplasm, intracellular transit of T-DNA and associated proteins (T-complex) to the host cell nucleus, disassembly of the T-complex, T-DNA integration, and expression of the transferred genes. During all these processes, Agrobacterium has evolved to control and even utilize several pathways of host-plant defense response. Studies of these Agrobacterium-host interactions substantially enhance our understanding of many fundamental cellular biological processes and allow improvements in the use of Agrobacterium as a gene transfer tool for biotechnology.
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Agrobacterium: From Biology to Biotechnology - Agrobacterium : from biology to biotechnology
2008Co-Authors: Tzvi Tzfira, Vitaly CitovskyAbstract:Agrobacterium: A disease-causing bacterium.- A Brief History of Research on Agrobacterium Tumefaciens: 1900-1980s.- Agrobacterium and Plant Biotechnology.- The Agrobacterium Tumefaciens C58 Genome.- Agrobacterium-Taxonomy of Plant-Pathogenic Rhizobium Species.- The Initial Steps in Agrobacterium Tumefaciens Pathogenesis: Chemical Biology of Host Recognition.- Agrobacterium-Host Attachment and Biofilm Formation.- Production of a Mobile T-DNA by Agrobacterium Tumefaciens.- Translocation of Oncogenic T-DNA and Effector Proteins to Plant Cells.- Intracellular Transport of Agrobacterium T-DNA.- Mechanisms of T-DNA integration.- Agrobacterium Tumefaciens-Mediated Transformation: Patterns of T-Dna Integration Into the Host Genome.- Function of Host Proteins in The Agrobacterium-Mediated Plant Transformation Process.- The Oncogenes of Agrobacterium Tumefaciens and Agrobacterium Rhizogenes.- Biology of Crown Gall Tumors.- The Cell-Cell Communication System of Agrobacterium Tumefaciens.- Horizontal Gene Transfer.- Agrobacterium-Mediated Transformation of Non-Plant Organisms.- The Bioethics and Biosafety of Gene Transfer.- Agrobacterium-Mediated Gene Transfer: A Lawyer's Perspective.
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Agrobacterium mediated genetic transformation of plants biology and biotechnology
Current Opinion in Biotechnology, 2006Co-Authors: Tzvi Tzfira, Vitaly CitovskyAbstract:Agrobacterium -mediated genetic transformation is the dominant technology used for the production of genetically modified transgenic plants. Extensive research aimed at understanding and improving the molecular machinery of Agrobacterium responsible for the generation and transport of the bacterial DNA into the host cell has resulted in the establishment of many recombinant Agrobacterium strains, plasmids and technologies currently used for the successful transformation of numerous plant species. Unlike the role of bacterial proteins, the role of host factors in the transformation process has remained obscure for nearly a century of Agrobacterium research, and only recently have we begun to understand how Agrobacterium hijacks host factors and cellular processes during the transformation process. The identification of such factors and studies of these processes hold great promise for the future of plant biotechnology and plant genetic engineering, as they might help in the development of conceptually new techniques and approaches needed today to expand the host range of Agrobacterium and to control the transformation process and its outcome during the production of transgenic plants.
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partners in infection host proteins involved in the transformation of plant cells by Agrobacterium
Trends in Cell Biology, 2002Co-Authors: Tzvi Tzfira, Vitaly CitovskyAbstract:Genetic modification of plant cells by Agrobacterium is the only known natural example of DNA transport between kingdoms. While the bacterial factors involved in Agrobacterium infection have been relatively well characterized, studies of their host cellular partners are just beginning. Here, we describe the plant cell factors that might participate in Agrobacterium-mediated genetic transformation and discuss their possible roles in this process. Because Agrobacterium probably adapts existing cellular processes for its life cycle, identifying the host factors participating in Agrobacterium infection might contribute to a better understanding of such basic biological processes as cell communication, intracellular transport and DNA repair and recombination as well as help expand the host range of Agrobacterium as a genetic engineering tool.
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genetic transformation of hela cells by Agrobacterium
Proceedings of the National Academy of Sciences of the United States of America, 2001Co-Authors: Talya Kunik, Tzvi Tzfira, Yoram Kapulnik, Yedidya Gafni, Colin Dingwall, Vitaly CitovskyAbstract:Abstract Agrobacterium tumefaciens is a soil phytopathogen that elicits neoplastic growths on the host plant species. In nature, however, Agrobacterium also may encounter organisms belonging to other kingdoms such as insects and animals that feed on the infected plants. Can Agrobacterium, then, also infect animal cells? Here, we report that Agrobacterium attaches to and genetically transforms several types of human cells. In stably transformed HeLa cells, the integration event occurred at the right border of the tumor-inducing plasmid's transferred-DNA (T-DNA), suggesting bona fide T-DNA transfer and lending support to the notion that Agrobacterium transforms human cells by a mechanism similar to that which it uses for transformation of plants cells. Collectively, our results suggest that Agrobacterium can transport its T-DNA to human cells and integrate it into their genome.
Piotr Sobiczewski - One of the best experts on this subject based on the ideXlab platform.
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Rapid and specific identification of four Agrobacterium species and biovars using multiplex PCR.
Systematic and applied microbiology, 2005Co-Authors: Joanna Puławska, Anne Willems, Piotr SobiczewskiAbstract:On the basis of 23S rRNA gene sequences, 1 universal forward and 4 taxon (species/biovar)-specific reverse primers were designed for multiplex PCR to aid in identification and differentiation of Agrobacterium rubi, Agrobacterium vitis and Agrobacterium biovars 1 and 2. In reactions with DNA of 119 bacterial strains belonging to: Agrobacterium, Allorhizobium, Mesorhizobium, Rhizobium, Sinorhizobium and Phyllobacterium, as well as phytopathogenic bacteria representing various genera, the primers developed for identification of A. vitis, A. rubi or Agrobacterium biovar 1 amplified only DNA of strains belonging to these taxa, producing fragments of the expected sizes: 478, 1006 and 184bp, respectively. However, in the case of the primer developed for identification of Agrobacterium biovar 2, the characteristic 1066bp PCR product was obtained not only with DNA of this biovar, but also with DNA of 3 atypical biovar 1 strains and some rhizobial strains. Differentiation between Agrobacterium biovar 2 and the other strains was possible using the restriction analysis of this product with endonuclease Alw26I. The method developed is an excellent tool for rapid classification of these 4 taxa of Agrobacterium.
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Phylogenetic analysis of 23S rRNA gene sequences of. Agrobacterium, Rhizobium and Sinorhizobium strains.
Systematic and Applied Microbiology, 2000Co-Authors: Joanna Puławska, Anne Willems, Martine Maes, Piotr SobiczewskiAbstract:The phylogenetic relationship among twelve Agrobacterium, four Rhizobium, and two Sinorhizobium strains originating from various host plants and geographical regions was studied by analysis of the 23S rDNA sequences. The study included Agrobacterium strains belonging to biovars 1, 2 (with tumor- or hairy-root inducing and non-pathogenic strains), A. vitis, A. rubi; representative species of the Rhizobium genus: R. galegae, R. leguminosarum and R. tropici and Sinorhizobium meliloti strains. The phylogenetic analysis showed that within Agrobacterium, the biovar designation was reflected in the 23S rDNA similarity and that strains of Agrobacterium and Rhizobium are closely related to each other. The results suggest that the taxonomic definition of Agrobacterium and Rhizobium should be considered for revision and that the Agrobacterium-biovar identity is probably a reliable taxonomic trait.
