Effective method for the isolation of non-nodulating mutants of<i>Rhizobium trifolii</i>

Zurkowski, W; Lorkiewicz, Z

doi:10.1017/s0016672300018814

Cited by 60 publications

(24 citation statements)

References 5 publications

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“…In the fast-growing species of rhizobia, genes determining the early steps of nodule development (nod genes) were located on large indigenous plasmids (2,11,22,23,26,38,47).…”

mentioning

confidence: 99%

Two gene clusters of Rhizobium meliloti code for early essential nodulation functions and a third influences nodulation efficiency

Putnoky¹,

Kondorosi²

1986

J Bacteriol

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A pLAFR1 cosmid clone (pPP346) carrying the nodulation region of the symbiotic plasmid pRme4lb was isolated from a gene library of Rhizobium meliloti 41 by direct complementation of a Nod-deletion mutant of R. meliloti. Agrobacterium tumefaciens and Rhizobium species containing pPP346 were able to form ineffective nodules on alfalfa. The 24-kilobase insert in pPP346 carries both the common nodulation genes and genes involved in host specificity of nodulation. It was shown that these two regions are essential and sufficient to determine the early events in nodulation. A new DNA region influencing the kinetics and efficiency of nodulation was also localized on the symbiotic megaplasmid at the right side of the nif genes.Rhizobia induce the formation of nitrogen-fixing nodules on the roots of leguminous plants. In the fast-growing species of rhizobia, genes determining the early steps of nodule development (nod genes) were located on large indigenous plasmids (2,11,22,23,26,38,47).In Rhizobium meliloti a very high molecular weight plasmid (megaplasmid) confers the ability to nodulate the host plant alfalfa (Medicago sativa) (2, 38). When this megaplasmid was mobilized into other Rhizobium species or Agrobacterium tumefaciens, the transconjugants induced nodules on alfalfa, indicating that the early steps of nodulation and host range specificity are coded by this symbiotic plasmid (pSym) (31, 44). These nodules, however, developed only to a certain stage: there was no initiation of infection thread formation, and bacteria were not released into the inner part of the nodule (44, 46).The nod genes are closely linked to the nif genes in R. meliloti (2, 38). In strain 1021, nod mutations were mapped 25 kilobases (kb) away from the nifKDH genes (33). In strain 41, R-prime plasmids carrying the essential nod genes and the nif structural genes were isolated (1), and the physical map of this region was constructed (29). The smallest nod-nif R-prime carried an insert of 90 kb. nod mutations were localized in two clusters within this region. On an 8.5-kb EcoRI fragment, nodulation genes conserved in a wide range of rhizobia (common nod genes) (29) and consisting of four genes (nodABC and nodD) were identified (41, 43; M. Gottfert, B. Horvath, E. Kondorosi, P. Putnoky, F. Rodriguez-Quiniones, and A. Kondorosi, J. Mol. Biol., in press, manuscript in preparation). A similar arrangement of the nodABC and nodD genes was found in strain 1021 (14,25). Genes determining host range specificity (hsn genes) for alfalfa were localized on a 6.8-kb EcoRI fragment about 6 kb away from the 8.5-kb common nod fragment (29). The 6.8-kb fragment carries four hsn genes (

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“…In the fast-growing species of rhizobia, genes determining the early steps of nodule development (nod genes) were located on large indigenous plasmids (2,11,22,23,26,38,47).…”

mentioning

confidence: 99%

Two gene clusters of Rhizobium meliloti code for early essential nodulation functions and a third influences nodulation efficiency

Putnoky¹,

Kondorosi²

1986

J Bacteriol

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“…The strain B-W was subjected to plasmid curing (Zurkowski and Lorkiewcz, 1978) at 43°C to check the loss of 2, 3-dihydroxybenzoic acid (DHB) production in the glucose and MnSO4 containing medium at 28°C and antibiotic resistance. Plasmid was not detected in the cured strain B-W. Curing at 40 or 42°C of strain B-W was not successful.…”

Section: Resultsmentioning

confidence: 99%

“…The siderophore production medium for strain B-W and cured strain was a chemically defined one Curing of plasmid from Acinetobacter sp. B-W Plasmid curing was done by the heat treatment procedure of Zurkowski and Lorkiewcz (1978). For heat curing, overnight nutrient broth cultures were inoculated to approximately 10 6 cells/ml into NB medium.…”

Section: Microorganisms Media and Culture Conditionsmentioning

confidence: 99%

Effect of plasmid curing on the 2, 3-dihydroxybenzoic acid production and antibiotic resistance of Acinetobacter sp. B-W

Kim¹,

Kim²,

Yang³

2016

The Korean Journal of Microbiology

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Acinetobacter sp. B-W producing siderophore, 2, 3-dihydroxybenzoic acid (DHB) was analyzed for plasmid content. Strain B-W harbored plasmid of 20 kb in size. Growth at 43°C was effective in producing mutant cured of plasmid of strain B-W. This mutant lost the ability to produce 2, 3-DHB. Formation of siderophore halos on the chrome azurol S (CAS) agar medium was not detected by cured strain B-W. pHs of supernatants of wild type strain B-W and cured mutant grown in glucose and MnSO4 containing medium at 28°C for 3 days were 4.5 and 8.5, respectively. Antibiotic resistance against ampicillin, actinomycin D, bacitracin, lincomycin, and vancomycin was lost in cured mutant. Plasmid curing of strain B-W resulted in drastic reduction of minimal inhibitory concentration (MIC) of several antibiotics. E. coli DH5α was transformed with plasmid isolated from strain B-W. The transformant E. coli DH5α harbored a plasmid of the same molecular size as that of the donor plasmid. Transformant E. coli DH5α produced 2, 3-DHB and contained antibiotic resistant ability. Thus a single plasmid of 20 kb seemed to be involved in 2, 3-DHB production. Genes encoding resistance to antibiotics were also supposed to be located on this plasmid.

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“…The derivative of R. leguminosarum strain A171 that has been cured of its smallest plasmid fails to nodulate peas (Casse et al, 1979). Similarly, heat-treated R. trifolii strains were enriched for nonnodulating strains (Zurkowski and Lorkiewicz, 1978). In R. leguminosarum strain 300 the mutant strain 6007, which carries a deletion in the 205 Md plasmid, is Nod and is restored to Nod Fix by the introduction of derivatives of the plasmid pRL1JI , but not of the apparently related bacteriocinogenic plasmids pRL3JI and pRL4JI.…”

Section: Riiizouiui Plasmids (I) Physical Studiesmentioning

confidence: 99%

“…Zurkowski and Lorkiewicz (1978) obtained non-nodulating mutants of R. trifolii after incubation for seven days at 35°C. Similarly Casse eta!.…”

Section: Riiizouiui Plasmids (I) Physical Studiesmentioning

confidence: 99%

The genetic analysis of Rhizobium in relation to symbiotic nitrogen fixation

1980

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Effective method for the isolation of non-nodulating mutants ofRhizobium trifolii

Cited by 60 publications

References 5 publications

Two gene clusters of Rhizobium meliloti code for early essential nodulation functions and a third influences nodulation efficiency

Two gene clusters of Rhizobium meliloti code for early essential nodulation functions and a third influences nodulation efficiency

Effect of plasmid curing on the 2, 3-dihydroxybenzoic acid production and antibiotic resistance of Acinetobacter sp. B-W

The genetic analysis of Rhizobium in relation to symbiotic nitrogen fixation

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