Ti and Ri Plasmids

Suzuki, K.; Tanaka, Katsuyuki; Yamamoto, Sachiko; Kiyokawa, Kazuya; Moriguchi, Kazuki; Yoshida, Kazuo

doi:10.1007/978-3-540-85467-8_6

Cited by 19 publications

(16 citation statements)

References 67 publications

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“…Most genes required for pathogenicity are located on the plasmids (17,33). Ti plasmids are kept stable at a low copy number equivalent to that of the chromosomal DNA in the bacterial cells (32) due to the repABC locus (16,30,34).…”

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Novel Toxin-Antitoxin System Composed of Serine Protease and AAA-ATPase Homologues Determines the High Level of Stability and Incompatibility of the Tumor-Inducing Plasmid pTiC58

et al. 2009

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Stability of plant tumor-inducing (Ti) plasmids differs among strains. A high level of stability prevents basic and applied studies including the development of useful strains. The nopaline type Ti plasmid pTiC58 significantly reduces the transconjugant efficiency for incoming incompatible plasmids relative to the other type, such as octopine-type plasmids. In this study we identified a region that increases the incompatibility and stability of the plasmid. This region was located on a 4.3-kbp segment about 38 kbp downstream of the replication locus, repABC. We named two open reading frames in the segment, ietA and ietS, both of which were essential for the high level of incompatibility and stability. Plasmid stabilization by ietAS was accomplished by a toxin-antitoxin (TA) mechanism, where IetS is the toxin and IetA is the antitoxin. A database search revealed that putative IetA and IetS proteins are highly similar to AAA-ATPases and subtilisin-like serine proteases, respectively. Amino acid substitution experiments in each of the highly conserved characteristic residues, in both putative enzymes, suggested that the protease activity is essential and that ATP binding activity is important for the operation of the TA system. The ietAS-containing repABC plasmids expelled Ti plasmids even in strains which were tolerant to conventional Ti-curing treatments.

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confidence: 99%

“…Differences in Ti plasmid stability are critical for plasmid engineering and evolution (33). However, little is known about the stability factors of Ti plasmids other than the repABC locus.…”

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confidence: 99%

Novel Toxin-Antitoxin System Composed of Serine Protease and AAA-ATPase Homologues Determines the High Level of Stability and Incompatibility of the Tumor-Inducing Plasmid pTiC58

et al. 2009

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“…Complete nucleotide sequences are available for several different types of Ti and Ri plasmids (26). The difference in the auxiliary vir region affects the host range in part.…”

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confidence: 99%

“…However, the large size of Ti and Ri plasmids, approximately 200 kbp, makes structural analysis and modification difficult. Complete nucleotide sequences of several Ti and Ri plasmids (for example, pTi-SAKURA, pTiC58, and pRi1724) have been reported (9,14,24,26,31). Accumulation of such nucleotide sequence information makes targeted replacement easier than it was previously.…”

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Construction of Disarmed Ti Plasmids Transferable between Escherichia coli and Agrobacterium Species

Kiyokawa

Yamamoto

Sakuma

et al. 2009

Appl Environ Microbiol

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Agrobacterium-mediated plant transformation has been used widely, but there are plants that are recalcitrant to this type of transformation. This transformation method uses bacterial strains harboring a modified tumor-inducing (Ti) plasmid that lacks the transfer DNA (T-DNA) region (disarmed Ti plasmid). It is desirable to develop strains that can broaden the host range. A large number of Agrobacterium strains have not been tested yet to determine whether they can be used in transformation. In order to improve the disarming method and to obtain strains disarmed and ready for the plant transformation test, we developed a simple scheme to make certain Ti plasmids disarmed and simultaneously maintainable in Escherichia coli and mobilizable between E. coli and Agrobacterium. To establish the scheme in nopaline-type Ti plasmids, a neighboring segment to the left of the left border sequence, a neighboring segment to the right of the right border sequence of pTi-SAKURA, a cassette harboring the pSC101 replication gene between these two segments, the broad-host-range IncP-type oriT, and the gentamicin resistance gene were inserted into a suicidetype sacB-containing vector. Replacement of T-DNA with the cassette in pTiC58 and pTi-SAKURA occurred at a high frequency and with high accuracy when the tool plasmid was used. We confirmed that there was stable maintenance of the modified Ti plasmids in E. coli strain S17-1pir and conjugal transfer from E. coli to Ti-less Agrobacterium strains and that the reconstituted Agrobacterium strains were competent to transfer DNA into plant cells. As the modified plasmid delivery system was simple and efficient, conversion of strains to the disarmed type was easy and should be applicable in studies to screen for useful strains.Agrobacterium-mediated transformation has been considered the most efficient and reliable method for plant biology and biotechnology. This methodology has been established for many plants, but not for others. One of the major factors affecting its applicability is the limited number of donor Agrobacterium strains, because the method depends exclusively on the host ranges of the strains.Wild-type Agrobacterium strains harboring a tumor-inducing (Ti) plasmid are the causative agent of crown gall tumor disease in dicotyledonous plants (35). The transfer DNA (T-DNA) and virulence gene (vir) regions in the Ti plasmid are essential for tumorigenesis. The vir gene products nick the T-DNA region at its left border (LB) and right border (RB) and then transfer T-DNA into plant cells. T-DNA contains phytohormone synthesis genes, whose expression causes infected plants to suffer from unregulated growth (5, 26). The hairy-root-inducing (Ri) plasmid has a similar T-DNA system.The binary vector system (11) is widely used for Agrobacterium-mediated transformation. Binary vectors are small plasmids with a cloning site and a selectable marker gene between LB and RB (2). To ensure transformation without tumorigenicity, Agrobacterium strains used for transformation contain a modified...

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The Ecology of Agrobacterium vitis and Management of Crown Gall Disease in Vineyards

Kuzmanović

Puławska

Hao

et al. 2018

Current Topics in Microbiology and Immunology

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Agrobacterium vitis is the primary causal agent of grapevine crown gall worldwide. Symptoms of grapevine crown gall disease include tumor formation on the aerial plant parts, whereas both tumorigenic and nontumorigenic strains of A. vitis cause root necrosis. Genetic and genomic analyses indicated that A. vitis is distinguishable from the members of the Agrobacterium genus and its transfer to the genus Allorhizobium was suggested. A. vitis is genetically diverse, with respect to both chromosomal and plasmid DNA. Its pathogenicity is mainly determined by a large conjugal tumor-inducing (Ti) plasmid characterized by a mosaic structure with conserved and variable regions. Traditionally, A. vitis Ti plasmids and host strains were differentiated into octopine/cucumopine, nopaline, and vitopine groups, based on opine markers. However, tumorigenic and nontumorigenic strains of A. vitis may carry other ecologically important plasmids, such as tartrate- and opine-catabolic plasmids. A. vitis colonizes vines endophytically. It is also able to survive epiphytically on grapevine plants and is detected in soil exclusively in association with grapevine plants. Because A. vitis persists systemically in symptomless grapevine plants, it can be efficiently disseminated to distant geographical areas via international trade of propagation material. The use of healthy planting material in areas with no history of the crown gall represents the crucial measure of disease management. Moreover, biological control and production of resistant grape varieties are encouraging as future control measures.

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Ti and Ri Plasmids

Cited by 19 publications

References 67 publications

Novel Toxin-Antitoxin System Composed of Serine Protease and AAA-ATPase Homologues Determines the High Level of Stability and Incompatibility of the Tumor-Inducing Plasmid pTiC58

Novel Toxin-Antitoxin System Composed of Serine Protease and AAA-ATPase Homologues Determines the High Level of Stability and Incompatibility of the Tumor-Inducing Plasmid pTiC58

Construction of Disarmed Ti Plasmids Transferable between Escherichia coli and Agrobacterium Species

The Ecology of Agrobacterium vitis and Management of Crown Gall Disease in Vineyards

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