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Scientific Reports Jul 2019Ustilago maydis is a biotrophic pathogen and well-established genetic model to understand the molecular basis of biotrophic interactions. U. maydis suppresses plant...
Ustilago maydis is a biotrophic pathogen and well-established genetic model to understand the molecular basis of biotrophic interactions. U. maydis suppresses plant defense and induces tumors on all aerial parts of its host plant maize. In a previous study we found that U. maydis induced leaf tumor formation builds on two major processes: the induction of hypertrophy in the mesophyll and the induction of cell division (hyperplasia) in the bundle sheath. In this study we analyzed the cell-type specific transcriptome of maize leaves 4 days post infection. This analysis allowed identification of key features underlying the hypertrophic and hyperplasic cell identities derived from mesophyll and bundle sheath cells, respectively. We examined the differentially expressed (DE) genes with particular focus on maize cell cycle genes and found that three A-type cyclins, one B-, D- and T-type are upregulated in the hyperplasic tumorous cells, in which the U. maydis effector protein See1 promotes cell division. Additionally, most of the proteins involved in the formation of the pre-replication complex (pre-RC, that assure that each daughter cell receives identic DNA copies), the transcription factors E2F and DPa as well as several D-type cyclins are deregulated in the hypertrophic cells.
Topics: Cell Division; Cell Enlargement; Gene Expression Regulation, Plant; Host-Pathogen Interactions; Plant Diseases; Plant Leaves; Plant Tumors; Transcriptional Activation; Transcriptome; Ustilago; Zea mays
PubMed: 31308451
DOI: 10.1038/s41598-019-46734-3 -
Plant Physiology Jul 2012Upon Agrobacterium tumefaciens infection of a host plant, Tumor morphology root (Tmr) a bacterial adenosine phosphate-isopentenyltransferase (IPT), creates a metabolic...
Upon Agrobacterium tumefaciens infection of a host plant, Tumor morphology root (Tmr) a bacterial adenosine phosphate-isopentenyltransferase (IPT), creates a metabolic bypass in the plastid for direct synthesis of trans-zeatin (tZ) with 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate as the prenyl donor. To understand the biological importance of Tmr function for gall formation, we compared Tmr and Trans-zeatin secretion (Tzs) another agrobacterial IPT that functions within the bacterial cell. Although there is no significant difference in their substrate specificities in vitro, ectopic overexpression of Tzs in Arabidopsis (Arabidopsis thaliana) resulted in the accumulation of comparable amounts of tZ- and N⁶-(Δ²-isopentenyl)adenine (iP)-type cytokinins, whereas overexpression of Tmr resulted exclusively in the accumulation of tZ-type cytokinins. Ectopic expression of Tzs in plant cells yields only small amounts of the polypeptide in plastid-enriched fractions. Obligatory localization of Tzs into Arabidopsis plastid stroma by translational fusions with ferredoxin transit peptide (TP-Tzs) increased the accumulation of both tZ- and iP-type cytokinins. Replacement of tmr on the Ti plasmid with tzs, TP-tzs, or an Arabidopsis plastidic IPT induced the formation of smaller galls than wild-type A. tumefaciens, and they were accompanied by the accumulation of iP-type cytokinins. Tmr is thus specialized for plastid localization and preferential usage of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate in vivo and is important for efficient gall formation.
Topics: Agrobacterium tumefaciens; Alkyl and Aryl Transferases; Arabidopsis; Biological Transport; Cytokinins; Hydroxylation; Organophosphates; Plant Roots; Plant Tumors; Plants, Genetically Modified; Plastids; Prenylation; Subcellular Fractions; Substrate Specificity; Zeatin
PubMed: 22589470
DOI: 10.1104/pp.112.198572 -
Annual Review of Phytopathology 2009Pantoea agglomerans, a widespread epiphyte and commensal bacterium, has evolved into an Hrp-dependent and host-specific tumorigenic pathogen by acquiring a plasmid... (Review)
Review
Pantoea agglomerans, a widespread epiphyte and commensal bacterium, has evolved into an Hrp-dependent and host-specific tumorigenic pathogen by acquiring a plasmid containing a pathogenicity island (PAI). The PAI was evolved on an iteron plasmid of the IncN family, which is distributed among genetically diverse populations of P. agglomerans. The structure of the PAI supports the premise of a recently evolved pathogen. This review offers insight into a unique model for emergence of new bacterial pathogens. It illustrates how horizontal gene transfer was the major driving force in the creation of the PAI, although a pathoadaptive mechanism might also be involved. It describes the crucial function of plant-produced indole-3-acetic acid (IAA) and cytokinines (CK) in gall initiation as opposed to the significant but secondary role of pathogen-secreted phytohormones. It also unveils the role of type III effectors in determination of host specificity and evolution of the pathogen into pathovars. Finally, it describes how interactions between the quorum sensing system, hrp regulatory genes, and bacterially secreted IAA or CKs affect gall formation and epiphytic fitness.
Topics: Biological Evolution; Gene Transfer, Horizontal; Genomic Islands; Host-Pathogen Interactions; Pantoea; Plant Tumors
PubMed: 19400643
DOI: 10.1146/annurev-phyto-080508-081803 -
NatureAgrobacterium tumefaciens is the causative agent of crown gall, a plant tumour that can arise on most species of dicotyledonous plants. The tumour-inducing capacity of...
Agrobacterium tumefaciens is the causative agent of crown gall, a plant tumour that can arise on most species of dicotyledonous plants. The tumour-inducing capacity of the bacterium requires the presence of a large plasmid, designated the Ti plasmid, which itself contains two regions essential for tumour formation-the T(umour)-region and the Vir(ulence)-region. The T-region is transferred to plant cells by an unknown mechanism, and becomes stably integrated into the plant genome. The Vir-region has been identified by transposon mutagenesis, but the DNA of this region has never been detected in tumour lines. However, trans-complementation of Vir mutants indicates that genes of the Vir-region are functional in the bacterium. Moreover, the Vir- and T-regions can be physically separated in A. tumefaciens without loss of tumour-inducing capacity. Seven loci, designated virA-F and virO (refs 17, 20-22), have been identified in the Vir-region of the octopine Ti plasmid, but their functions are unknown. As virC mutants in the octopine-type plasmid pTiB6 are invariably avirulent in tests on various plant species, this gene seems to be essential for virulence and we are studying it in detail. We report here that the promoter of virC shows no detectable activity in A. tumefaciens and Escherichia coli K-12 grown in standard medium, but that its activity is induced by a plant product.
Topics: Gene Expression Regulation; Genes, Bacterial; Plant Tumors; Plasmids; Promoter Regions, Genetic; Rhizobium
PubMed: 6504164
DOI: 10.1038/312564a0 -
Journal of Bacteriology Aug 1969The ability of 31 strains of Agrobacterium to initiate the production of a tumor growth factor (TGF) which is associated with crown-gall tumors on primary pinto bean...
The ability of 31 strains of Agrobacterium to initiate the production of a tumor growth factor (TGF) which is associated with crown-gall tumors on primary pinto bean leaves was determined. Extracts from bean leaves inoculated with these bacteria were tested and they showed that 16 of the 19 strains that induced tumors on the leaves also initiated TGF production. The three strains for which no TGF was detected were of low infectivity and included two strains of A. tumefaciens and a strain of A. rhizogenes. Five of the 12 strains that did not induce pinto bean leaf tumors were found to initiate TGF production. Representatives of A. tumefaciens, A. rhizogenes, and A. radiobacter among these 12 strains were present in both categories. Mixed inocula composed of one of the three infectious TGF-negative strains and one of the five nontumorigenic TGF-positive strains resulted in increased growth of tumors induced by the former. These growth changes were not correlated with changes in tumor number. The ability of different strains to show these tumor growth complementation effects corresponded fully with their ability to initiate TGF, as determined by the assay of leaf extracts. The nontumorigenic TGF-positive strains also promoted the growth of tumors initiated by low concentrations of strain B6. These complementation effects were due, therefore, to the same TGF found in extracts of B6 inoculated leaves and of leaves inoculated with most tumorigenic as well as many nontumorigenic strains of Agrobacterium. Heat-inactivated cells of strain B6 failed to initiate sufficient TGF to be detected in extracts, and heat-inactivated cells of several strains failed to show tumor growth complementation, indicating bacterial viability to be one prerequisite for TGF initiation. Heat inactivated cells also inhibited TGF production by viable cells, similar to their ability to inhibit tumor initiation. Consequently, bacteria capable of attaching to the A. tumefaciens infection site may initiate one of four patterns of events: (i) TGF production only, (ii) tumor induction only, (iii) both, or (iv) neither. Suggestive evidence for a second tumor-associated growth factor is presented.
Topics: Plant Growth Regulators; Plant Tumors; Rhizobium
PubMed: 5808077
DOI: 10.1128/jb.99.2.496-502.1969 -
Progress in Experimental Tumor Research 1972
Review
Topics: Amino Acids; Cell Division; Cell Membrane Permeability; Cell Transformation, Neoplastic; Plant Growth Regulators; Plant Tumors; Purines; RNA, Transfer; Rhizobium
PubMed: 4552169
DOI: 10.1159/000392509 -
Yi Chuan = Hereditas Nov 2011In recent years, the functional mechanisms of the oncogenens from Agrobacterium in plants were received more and more attentions. 6b genes, derived from the T-DNA... (Review)
Review
In recent years, the functional mechanisms of the oncogenens from Agrobacterium in plants were received more and more attentions. 6b genes, derived from the T-DNA fragment, are vital carcinogenesis factors of plants and belong to rolB genes family. In plants, 6b genes can affect phytohormone levels and carbohydrate contents, and can also cause accumulation of secondary metabolites, as well as change the relative genes expression. The specific mechanisms behind these impacts remain to be researched in-depth. In this paper, the function, structure, activity, and acting mode of the 6b genes were summarized, which provide a theoretical foundation for further study and application of these functional genes.
Topics: Agrobacterium; Bacterial Proteins; DNA, Bacterial; Plant Development; Plant Tumors; Plants
PubMed: 22120076
DOI: 10.3724/sp.j.1005.2011.01212 -
Nature Communications Oct 2016Antibiotic-producing microbes evolved self-resistance mechanisms to avoid suicide. The biocontrol Agrobacterium radiobacter K84 secretes the Trojan Horse antibiotic...
Antibiotic-producing microbes evolved self-resistance mechanisms to avoid suicide. The biocontrol Agrobacterium radiobacter K84 secretes the Trojan Horse antibiotic agrocin 84 that is selectively transported into the plant pathogen A. tumefaciens and processed into the toxin TM84. We previously showed that TM84 employs a unique tRNA-dependent mechanism to inhibit leucyl-tRNA synthetase (LeuRS), while the TM84-producer prevents self-poisoning by expressing a resistant LeuRS AgnB2. We now identify a mechanism by which the antibiotic-producing microbe resists its own toxin. Using a combination of structural, biochemical and biophysical approaches, we show that AgnB2 evolved structural changes so as to resist the antibiotic by eliminating the tRNA-dependence of TM84 binding. Mutagenesis of key resistance determinants results in mutants adopting an antibiotic-sensitive phenotype. This study illuminates the evolution of resistance in self-immunity genes and provides mechanistic insights into a fascinating tRNA-dependent antibiotic with applications for the development of anti-infectives and the prevention of biocontrol emasculation.
Topics: Agrobacterium tumefaciens; Drug Resistance, Bacterial; Evolution, Molecular; Genes, Bacterial; Kinetics; Leucine-tRNA Ligase; Pest Control, Biological; Plant Tumors; RNA, Transfer
PubMed: 27713402
DOI: 10.1038/ncomms12928 -
Genome Biology and Evolution Jun 2019Plasmids play a crucial role in the ecology of agrobacteria. In this study, we sequenced tumor-inducing (Ti) and opine-catabolic (OC) plasmids in three Rhizobium...
Plasmids play a crucial role in the ecology of agrobacteria. In this study, we sequenced tumor-inducing (Ti) and opine-catabolic (OC) plasmids in three Rhizobium rhizogenes (Agrobacterium biovar 2) strains isolated from the same crown gall tumor on "Colt" cherry rootstock and conducted comparative genomic analyses. Tumorigenic strains C5.7 and C6.5 carry nopaline-type Ti plasmids pTiC5.7/pTiC6.5, whereas the nonpathogenic strain Colt5.8 carries the nopaline-type OC plasmid pOC-Colt5.8. Overall, comparative genomic analysis indicated that pTiC5.7/pTiC6.5 and related Ti plasmids described before (pTiC58 and pTi-SAKURA) originate from a common ancestor, although they have diverged during evolution. On the other hand, plasmid pOC-Colt5.8 was most closely related to the well-known OC plasmid pAtK84b; however, analysis suggested that they had different evolutionary histories and seem to share a more distant common ancestor. Although the reconstruction of the evolutionary history of Ti and OC plasmids is still speculative, we hypothesized that nopaline-type Ti plasmid might originate from the nopaline-type OC plasmid. Our results suggested that OC plasmids are widespread and closely associated with crown gall tumors. Finally, we proposed a thorough scheme for classification of Ti and OC plasmids that is based on separate comparative analysis of each functional element of the plasmid studied.
Topics: Arginine; Conjugation, Genetic; Oxazines; Plant Tumors; Plasmids; Rhizobium; Virulence
PubMed: 31028704
DOI: 10.1093/gbe/evz091 -
The New Phytologist Jan 2018During Agrobacterium (Agrobacterium tumefaciens) infection, the translocated virulence proteins (VirD2, VirE2, VirE3, VirF and VirD5) play crucial roles. It is thought...
During Agrobacterium (Agrobacterium tumefaciens) infection, the translocated virulence proteins (VirD2, VirE2, VirE3, VirF and VirD5) play crucial roles. It is thought that, through protein-protein interactions, Agrobacterium uses and abuses host plant factors and systems to facilitate its infection. Although some molecular functions have been revealed, the roles of VirD5 still need to be further elucidated. Here, plant transformation and tumorigenesis mediated by genetically modified Agrobacterium strains were performed to examine VirD5 roles. In addition, protein-protein interaction-associated molecular and biochemistry technologies were used to reveal and elucidate VirD5 interaction with Arabidopsis VirE2 interacting protein 2 (VIP2). Our results showed that deleting virD5 from Agrobacterium reduced its tumor formation ability and stable transformation efficiency but did not affect the transient transformation efficiency. We also found that VirD5 can interact with Arabidopsis VIP2. Further experiments demonstrated that VirD5 can affect VIP2 binding to cap-binding proteins (CBP20 and CBP80). The tumorigenesis efficiency for cbp80 mutant was not significantly changed, but that for cbp20, cbp20cbp80 mutants were significantly increased. This work demonstrates experimentally that VirD5 is required for efficient Agrobacterium infection and may promote this process by competitive interaction with Arabidopsis VIP2. CBP20 is involved in the Agrobacterium infection process and its effect can be synergistically enhanced by CBP80.
Topics: Agrobacterium tumefaciens; Arabidopsis; Arabidopsis Proteins; Gene Deletion; Plant Tumors; Protein Binding; Protein Transport; Transcription Factors, General; Transformation, Genetic; Virulence Factors
PubMed: 29084344
DOI: 10.1111/nph.14854