Abstract
The Tn5-sacB-labeled symbiotic megaplasmid pMhHN3015c of Mesorhizobium huakuii HN3015 was, respectively, transferred into M. huakuii HN308SR containing three large plasmids of pMhHN308a, pMhHN308b and pMhHN308c, and 7653R-1SR, a symbiotic plasmid pMh7653Rb deleted mutant from M. huakuii 7653R by tri-parent mating. The stable indigenous plasmid pMhHN308c of HN308SR was cured by the introduction of pMhHN3015c and the transconjugant was named as HN308SRN18. The results implied that pMhHN3015c and pMhHN308c were incompatible and might be ascribed to the same incompatibility group. Furthermore, the results from plasmid curing tests of HN308SRN18 containing pMhHN3015c, pMhHN308b, and pMhHN308a showed that not only was pMhHN3015c deleted, but that pMhHN308a was also cured simultaneously. The plasmid profiles of transconjugant 7653R-1SRN18 showed pMhHN3015c could coexist with pMh7563Ra. The plasmid replication repC-like gene sequences were detected by polymerase chain reaction from 7653R-1SRN18, HN308SRN18 and its plasmid-curing derivatives, but failed to detect from plasmid-curing derivatives of 7653R-1SRN18. The repC gene sequence similarities of strains tested were up to 99%. Results from plant nodulation tests showed that introduction of pMhHN3015c failed to restore the nitrogen fixation ability of HN308SRN18 and 7653R-1SRN18.
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