In
Agrobacterium tumefaciens, the stability of Ti plasmids differs depending on the strain. So far, little is known about genes that cause the difference in stability. The
repABC operon is responsible for replication and incompatibility of Ti plasmids. We constructed recombinant plasmids carrying the
repABC operon and different portions of pTi-
SAKURA
. Cells having the recombinant plasmids that harbored a 2.6-kbp
NheI fragment of pTi-
SAKURA
were found to be transformed via conjugation 100-fold less frequently with a small incompatible
repABC plasmid than cells having the recombinant plasmids lacking the 2.6-kbp
NheI fragment. Since the phenomenon occurred only when the resident and incoming plasmids belonged to the same incompatibility group, it was suggested that the 2.6-kbp
NheI fragment bears the potential enhancing incompatibility. The fragment contained an operon consisting of two open reading frames,
tiorf24 and
tiorf25.
tiorf24 is an orphan gene, whereas
tiorf25 is a homologue of a group of plasmid stability genes. Removal of the 2.6-kbp fragment from the resident pTi-
SAKURA
increased the resident plasmid ejection ratio by the incoming
repABC plasmid, whereas addition of the fragment to pTiC58 decreased the ejection ratio, and the loss ratio during growth at 37°C. These data suggest that
tiorf24 and
tiorf25 are responsible for the stability of pTi-
SAKURA
, and reduce, in the host bacterium, the frequency of ejection of the resident plasmid, presumably through an incompatibility mechanism.
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