Review: Plant Binary Vectors of Ti Plasmid in <i>Agrobacterium tumefaciens</i> with a Broad Host-Range Replicon of pRK2, pRi, pSa or pVS1 — Oak Academic Publishing
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Review: Plant Binary Vectors of Ti Plasmid in <i>Agrobacterium tumefaciens</i> with a Broad Host-Range Replicon of pRK2, pRi, pSa or pVS1
Department of Plant Pathology and Crop Physiology, Louisiana State University and LSU AgCenter, Baton Rouge, USA
1 Department of Plant Pathology and Crop Physiology, Louisiana State University and LSU AgCenter, Baton Rouge, USA
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 supe rior 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).
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