Influences of soil acidity and variety of plant on the populations of Rhizobium trifolii found in nodules of Trifolium subterraneum L. Public Deposited

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  • Complementary methods of strain identification were sought to delineate the composition of the population of Rhizobium trifolii found in nodules of Trifolium subterraneum L. when exposed to an Abiqua silty clay loam (Cumulic Ultic Haploxeroll) containing a resident population of R. trifolii. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE) was used in a microslab system to elucidate the protein profile patterns of the isolates. Antisera were raised to four isolates which showed distinctly different protein profile patterns. Serological tests showed that the majority of nodule isolates could be placed into four serogroups, isolates within two of the groups (16 and 36), were antigenically identical whereas isolates in the other two groups (6 and 27) were antigenically heterogeneous. SAS PAGE revealed that antigenically identical isolates could be subdivided further. Soil acidity affected which members of the indigenous population nodulated T. subterraneum L. cv. Mt. Barker. Representatives of serogroup 6 occupied the greatest percentage of the nodules formed on plants grown at low pH and were a minor nodule occupant at the higher pH. In contrast serogroup 36 was virtually absent in nodules formed at low pH and was a dominant serogroup at the higher pH. Despite the isolates within serogroups 6 and 36 being antigenically identical separation of cellular proteins by SDS-PAGE and determination of symbiotic effectiveness revealed that serogroups 6 and 36 were composed minimally of eight and twelve different strains. The cultivar of T. subterraneum L. was found to impact upon the composition of the nodule occupants. Between 68 and 90% of nodule occupants formed on cvs. Mt. Barker, Nangeela and Howard were placed into the four identifiable serogroups. Only in the case of cv. Woogenellup were the majority of isolates unidentifiable with the antisera at our disposal. In the unlimed soil different serogroups dominated each cultivar. In limed soil serogroup 27 was the dominant identifiable serogroup on all cultivars. Gel-immune-diffusion revealed three serotypes within serogroup 27; serotype 27-A was dominant in nodules on cv. Nangeela alone, whereas serotype 27-B was dominant on both cvs. Mt. Barker and Woogenellup. Separation of cellular proteins by SDS-PAGE revealed four and five different strains within serotypes 27-A and 27-B respectively. Only one strain of serotype 27-B could be considered a common nodule occupant on both cultivars Mt. Barker and Woogenellup. Two of three strains of serotype 27-B were found on cv. Mt. Barker grown in both soil treatments whereas only one of four strains of serotype 27-A was found on cv. Nangeela grown in both soil treatments.
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