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A comparison of the fluorescent ELISA and antibiotic resistance identification techniques for use in ecological experiments with Rhizobium trifolii
Author(s) -
RENWICK ANNABEL,
JONES D. GARETH
Publication year - 1985
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1985.tb01448.x
Subject(s) - rhizobia , rhizobium , strain (injury) , soil water , biology , soil bacteria , bacteria , fluorescence , trifolium repens , peat , agronomy , horticulture , symbiosis , ecology , genetics , physics , quantum mechanics , anatomy
The fluorescent ELISA technique for the identification of bacteria was compared with antibiotic resistant mutants as marker systems for use with Rhizobium trifolii in root nodules and in soil. With an effective(CP3B) and an ineffective (R4) strain as a mixed 1:1 inoculum, there was a highly significant correlation ( P < 0.001) between the two techniques when the plants were grown at pH 5.5 when the majority of nodules were inhabited by the effective strain. At pH 6.5, where the ineffective strain predominated in the nodules, there was no correlation. The reason was that 85% of R4 nodules had volumes less than 0.1 mm 3 with bacterial numbers obviously below the necessary threshold for detection using the serological method. Both methods were efficient at enumerating rhizobia from soils although the recovery rate from a brown earth soil was significantly higher than from a peat soil. Fluorescent ELISA was able to detect rhizobia at 8.0 times 10 5 cells/ml soil suspension (1 g soil to 10 ml water) in the brown earth soil and at 2.0 times 10 5 cells/ml in the peat soil. The results are discussed in terms of the limitations of both techniques in ecological studies.