Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Record number 347392
Title Quanitative detection and diversity of the pyrrolnitrin biosynthetic locus in soil under different treatments.
Author(s) Garbeva, P.; Voesenek, K.; Elsas, J.D. van
Source Soil Biology and Biochemistry 36 (2004)9. - ISSN 0038-0717 - p. 1453 - 1463.
DOI https://doi.org/10.1016/j.soilbio.2004.03.009
Department(s) Biointeracties and Plant Health
Publication type Refereed Article in a scientific journal
Publication year 2004
Keyword(s) real-time pcr - pseudomonas-fluorescens - burkholderia-cepacia - biological-control - conventional pcr - selective media - strains - genes - quantification - biocontrol
Abstract The prevalence of antibiotic production loci in soil is a key issue of current research aimed to unravel the mechanisms underlying the suppressiveness of soil to plant pathogens. Pyrrolnitrin (PRN) is a key antibiotic involved in the suppression of a range of phytopathogenic fungi. Therefore, field soils from different agricultural regimes, including permanent grassland, arable land under common agricultural rotation and arable land under maize monoculture, were investigated in respect of the prevalence of pyrrolnitrin biosynthetic loci. Primers for detection of the prnD gene were used for initial PCR/hybridisation-based assessments. By this method, evidence was obtained for the contention that PRN production loci were most prevalent in grasslands, however, robust quantitative data were not achieved. To quantify the prevalence of PRN biosynthetic loci, we designed a TaqMan PCR system based on the prnD gene for the real-time quantitative detection of this production locus in soil. The system was found to be specific for prnD sequences from Pseudomonas, Serratia and Burkholderia species. Using pure culture DNA, the prnD gene was detectable down to a level of 60 fg, or approximately 10 gene copies, per amplification reaction. Application of the system to soil DNA spiked with different levels of the target DNA indicated that, in a soil DNA background, specific amplification could be obtained to about the same level of sensitivity. Field soil samples obtained from the different agricultural regimes were then screened for the prevalence of prnD with the real-time PCR system. The quantitative data obtained suggested a strongly enhanced presence of prnD genes in grassland or grassland-derived plots, as compared to the prevalence of this biosynthetic locus in the arable land plots. The implications of these findings are placed in the context of the suppressiveness of soil to phytopathogens, notably Rhizoctonia solani AG3. (C) 2004 Elsevier Ltd. All rights reserved.
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