Culture-Independent Detection of Changes in Root-Associated Bacterial Populations of Common Bean (Phaseolus vulgaris L.) Following Nitrogen Depletion |
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Authors: | Schallmach E Minz D Jurkevitch E |
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Institution: | (1) Department of Plant Pathology and Microbiology, Faculty of Agricultural, Food and Environmental Quality Sciences, The-Hebrew University of Jerusalem, P.O.B. 12, 76100 Rehovot, Israel |
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Abstract: | The structure of root-associated bacterial populations in the legume common bean (Phaseolus vulgaris L.), was studied in plants grown under nitrogen sufficiency and under conditions inducing nitrogen deficiency. Similar cell
numbers were obtained in the rhizosphere of nitrogen-amended plants as compared to nitrogen-deficient plants and between various
root parts—tip, elongation and branching zones—using DAPI staining. In contrast, a higher proportion of DAPI-stained cells
from the nitrogen-amended plants hybridized with a fluorescence-labeled EUB338 probe for theBacteria domain than cells originating from nitrogen-deficient plants. Shifts in the percentages of EUB338-reactive cells—as well
as in absolute cell number—hybridizing to fluorescent rRNA-directed probes specific for the α and γProteobacteria and for high GC content gram-positive bacteria in separated root segments were detected between the treatments. No such differences
were found using β and δProteobacteria or rRNA group I pseudomonad targeted probes. Denaturating gradient gel electrophoresis profiles of PCR products obtained
from the same samples and amplified withBacteria-domain targeted primers supported the results obtained with the whole cell hybridizations. The advantages and drawbacks of
the techniques applied are discussed. |
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