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  • 1
    ISSN: 1432-0789
    Keywords: Nodulation ; Competitiveness ; Rhizobium sp. (Cicer) ; lacZ fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We constructed lacZ fusions in Rhizobium sp. (Cicer) by random Tn5-lacZ mutagenesis. The lacZ+ fusants formed blue colonies on a Rhizobial minimal medium containing 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal). Rhizobium sp. (Cicer) fusant HSL-2 was identified in nodules and soil in a mixed population on the basis of the lacZ+ phenotype. Nodule occupancy of inoculated Rhizobium sp. (Cicer) HSL-2 (lacZ+) was assessed by directly streaking the nodule sap on X-gal plates. This method revealed differences between rhizobia carrying identical antibiotic markers. The rhizobial population in soil was estimated by direct plate counts using a medium containing X-gal. Introduction of lacZ into the Rhizobium sp. thus provided a simple and direct method for identifying strains from nodules and soil.
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  • 2
    ISSN: 1432-0789
    Keywords: Bradyrhizobium spp. ; Rhizobium spp. ; Cajanus cajan ; Pigeonpea ; Acetylene reduction assay (ARA) ; Curing ; Nodulation ; Plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Fifty-six isolates of Rhizobium and Bradyrhizobium spp. (Cajanus) were studied for their plasmid profile and N2-fixation efficacy. One to three plasmids were reproducibly detected in all the Rhizobium spp. strains but no plasmid was detected in the Bradyrhizobium spp. strains. Rhizobium sp. strain P-1 was mutagenized by Tn5 and three nod− and six nod+fix− were screened for symbiotic parameters. Neomycin-sensitive mutants were isolated by elevated temperatrue (40°C) from tranconjugants carrying Tn5 insertions. The high temperature “cured” these mutants from the single large plasmid present in the parent strain P-1. All these cured mutants were nod−, indicating that the genes for nodulation were present on this plasmid, which is readily cured at a high temperature (40°C). The high temperature in the semi-arid zones of Haryana could be responsible for the low nodulation of pigeonpea because the plasmid carrying the nodulation genes is cured at 40°–45°C giving rise to non-nodulating mutants.
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