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A. F. Bent, B. N. Kunkel, D. Dahlbeck, K. L. Brown, R. Schmidt, J. Giraudat, J. Leung and B. J. Staskawicz, “RPS2 of Arabidopsis thaliana: A Leucine-Rich Repeat Class of Plant Disease Resistance Genes,” Science, Vol. 265, No. 5180, 1994, pp. 1856-1860. doi:10.1126/science.8091210

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• TITLE: Review: Plant Binary Vectors of Ti Plasmid in Agrobacterium tumefaciens with a Broad Host-Range Replicon of pRK2, pRi, pSa or pVS1

  AUTHORS: Norimoto Murai

  KEYWORDS: Agrobacterium tumefaciens; Binary Vectors; pRK2; pRi; pSA; pVS1; T-DNA; Ti Plasmid

  JOURNAL NAME: American Journal of Plant Sciences, Vol.4 No.4, April 25, 2013

  ABSTRACT: This review chronicles the development of the plant binary vectors of Ti plasmid in Agrobacterium tumefaciens during the last 30 years. A binary vector strategy was designed in 1983 to separate the T-DNA region in a small plasmid from the virulence genes in avirulent T-DNA-less Ti plasmid. The small plant vectors with the T-DNA region have been simply now called binary Ti vectors. A binary Ti vector consist of a broad host-range replicon for propagation in A. tumeraciens, an antibiotic resistance gene for bacterial selection and the T-DNA region that would be transferred to the plant genome via the bacterial virulence machinery. The T-DNA region delimited by the right and left border sequences contains an antibiotic resistance gene for plant selection, reporter gene, and/or any genes of interest. The ColEI replicon was also added to the plasmid backbone to enhance the propagation in Escherichia coli. A general trend in the binary vector development has been to increase the plasmid stability during a long co-cultivation period of A. tumefaciens with the target host plant tissues. A second trend is to understand the molecular mechanism of broad host-range replication, and to use it to reduce the size of plasmid for ease in cloning and for higher plasmid yield in E. coli. The broad host-range replicon of VS1 was shown to be a choice of replicon over those of pRK2, pRi and pSA because of the superior stability and of small well-defined replicon. Newly developed plant binary vectors pLSU has the small size of plasmid backbone (4566 bp) consisting of VS1 replicon (2654 bp), ColE1 replicon (715 bp), a bacterial kanamycin (999 bp) or tetracycline resistance gene, and the T-DNA region (152 bp).