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  • 1
    ISSN: 1432-0789
    Keywords: Earthworms ; Rhizobium ; Colonization ; Medicago sativa ; Apporectodea trapezoides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A greenhouse study was performed to examine the ability of the earthworms Microscolex dubius and Aporrectodea trapezoides to transport Rhizobium meliloti L5-30R through soil. When R. meliloti L5-30R was inoculated into either ezi-mulch (a pelleted formulation of cereal-pea straw), oat hay, pea hay, or sheep dung and placed on the soil surface together with either A. trapezoides or M. dubius, 〉104 colony-forming units (CFU) of R. meliloti L5-30R g-1 soil were detected at 90 mm soil depth after 18 days. In the absence of earthworms, R. meliloti L5-30R was not detected at 90 mm soil depth after this time. In a second experiment using ezi-mulch as the inoculant material and in the presence of A. trapezoides (in a number equivalent to 471 or 785 m-2), ca. 103 CFU of R. meliloti L5-30R per 10 mm of alfalfa root were detected at 0–30, 30–60, and 60–90 mm soil depth after 18 days, while 〈3 CFU were detected per 10 mm of root in the absence of A. trapezoides. In a third experiment in which R. meliloti L5-30R was distributed evenly through soil at the start of the experiment, A. trapezoides (in a number equivalent to 157, 471, or 785 m-2) significantly decreased the survival of L5-30R in soil after 40 days of incubation at 15°C, but not after 20 days. The decrease in survival of R. meliloti L5-30R was correlated with the density of A. trapezoides. These results demonstrate that A. trapezoides can increase root colonization of alfalfa by R. meliloti L5-30R, but may also reduce the ability of R. meliloti L5-30R to survive in soil.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Agrobacterium ; Crown gall ; Biological control ; Agrocin 84 ; Agrocin plasmid transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Agrobacterium radiobacter strain K84 is used commercially for the biological control of crown gall. It contains the conjugative plasmid pAgK84, which encodes the synthesis of agrocin 84, an antibiotic that inhibits many pathogenic agrobacteria. A breakdown of control is threatened by the transfer of pAgK84 to pathogens, which then become resistant to agrocin 84. A mutant of pAgK84 with a 5.9-kb deletion overlapping the transfer (Tra) region was constructed using recombinant DNA techniques. The BamHI fragment B1 which covers most of the Tra region was cloned in pBR325 and its internal EcoRI fragments D1 and H, which overlap the Tra region, were removed, leaving 3.7 kb and 0.5 kb of pAgK84 on either side of the deletion. The latter was increased to 3.3 kb by adding EcoRI fragment D2 from a BamHI fragment C clone. The modified pBR325 clone was mobilized into Agrobacterium strain NT1 harbouring pAgK84 with a Tn5 insertion just outside the Tra region but covered by the deletion. A Tra+ cointegrate was formed between the Tn5-insertion derivative and the pBR325-based deletion construct by homologous recombination. The cointegrate was transferred by conjugation to a derivative of strain K84 lacking pAgK84, in which a second recombination event generated a stable deletion-mutant by deletion-marker exchange. The resultant new strain of A. radiobacter, designated K1026, shows normal agrocin 84 production. Mating experiments show that the mutant plasmid, designated pAgK1026, is incapable of conjugal transfer at a detectable frequency.
    Type of Medium: Electronic Resource
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