Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays

Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays

14 páginas, ilustraciones y tablas estadísticas

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Main Authors: Campos-Herrera, R., Johnson, E. G., El-Borai, F. E., Stuart, Robin J., Graham, J. H., Duncan, L. W.
Format: artículo biblioteca
Language:English
Published: Association of Applied Biologists 2011
Subjects:Entomopathogenic nematode, Heterorhabditis, Quantitative real-time PCR, Soil ecology, Species specific primers, Steinernema, TaqMan probe,
Online Access:http://hdl.handle.net/10261/60177
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spelling dig-ica-es-10261-601772020-05-19T11:12:44Z Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays Campos-Herrera, R. Johnson, E. G. El-Borai, F. E. Stuart, Robin J. Graham, J. H. Duncan, L. W. Entomopathogenic nematode Heterorhabditis Quantitative real-time PCR Soil ecology Species specific primers Steinernema TaqMan probe 14 páginas, ilustraciones y tablas estadísticas Quantitative real-time PCR (qPCR) techniques are being increasingly used to provide accurate and reliable methods to identify and quantify cryptic organisms in soil ecology. Entomopathogenic nematode (EPN) diversity in Florida is known to be extensive and our phylogenetic studies of the D2D3 and ITS regions showed the occurrence of an additional species-complex in the Steinernema glaseri- group in widely separated locations of the peninsula. To address ecological studies, we developed and used qPCR assays to detect and quantify six species of EPN that are naturally distributed in Florida citrus orchards (Steinernema diaprepesi, Steinernema riobrave, Heterorhabditis indica, Heterorhabditis zealandica, Heterorhabditis floridensis and an undescribed species in the S. glaseri group) and an exotic species, S. glaseri. Species-specific primers and TaqMan® probes were designed from the ITS rDNA region. No nonspecific amplification was observed in conventional or qPCR when the primers and probes were tested using several populations of each of the Florida species and other exotic EPN species. Standard curves were established using DNA from pure cultures. We optimised a protocol for extracting nematodes and DNA from soil samples that can detect one EPN added to nematode communities recovered by conventional extraction protocols. A survey of an 8-ha orchard in April 2009 compared the EPN spatial patterns derived from qPCR to that obtained by baiting soil samples with Galleria mellonella larvae. The patterns were also compared to those derived from the same site in 2000–01 by repeatedly (12 sampling events) baiting soil in situ with caged larvae of the root weevil Diaprepes abbreviatus. The qPCR assay was more efficient than the Galleria baitingmethod for detecting the EPN species composition in population mixtures.Moreover, the spatial patterns of EPN in this orchardwere remarkably stable over the course of nearly a decade. The pattern of H. zealandica detected at the site 8 years earlier was related to those derived by qPCR (P = 0.002) and from sample baiting (P = 0.02). The spatial pattern of H. indica derived from qPCR, but not that from sample baiting, was also related to the earlier pattern (P = 0.01). The qPCR assay developed here is a fast, affordable and accurate method to detect and quantify these EPN species in soil and offers great potential for studying the ecology of EPN. Peer reviewed 2012-11-14T08:50:12Z 2012-11-14T08:50:12Z 2011 artículo http://purl.org/coar/resource_type/c_6501 Annals of applied biology 158:55-68(2011) http://hdl.handle.net/10261/60177 10.1111/j.1744-7348.2010.00433.x 0003-4746 en http://dx.doi.org/10.1111/j.1744-7348.2010.00433.x none Association of Applied Biologists
institution ICA ES
collection DSpace
country España
countrycode ES
component Bibliográfico
access En linea
databasecode dig-ica-es
tag biblioteca
region Europa del Sur
libraryname Biblioteca del ICA España
language English
topic Entomopathogenic nematode
Heterorhabditis
Quantitative real-time PCR
Soil ecology
Species specific primers
Steinernema
TaqMan probe
Entomopathogenic nematode
Heterorhabditis
Quantitative real-time PCR
Soil ecology
Species specific primers
Steinernema
TaqMan probe
spellingShingle Entomopathogenic nematode
Heterorhabditis
Quantitative real-time PCR
Soil ecology
Species specific primers
Steinernema
TaqMan probe
Entomopathogenic nematode
Heterorhabditis
Quantitative real-time PCR
Soil ecology
Species specific primers
Steinernema
TaqMan probe
Campos-Herrera, R.
Johnson, E. G.
El-Borai, F. E.
Stuart, Robin J.
Graham, J. H.
Duncan, L. W.
Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays
description 14 páginas, ilustraciones y tablas estadísticas
format artículo
topic_facet Entomopathogenic nematode
Heterorhabditis
Quantitative real-time PCR
Soil ecology
Species specific primers
Steinernema
TaqMan probe
author Campos-Herrera, R.
Johnson, E. G.
El-Borai, F. E.
Stuart, Robin J.
Graham, J. H.
Duncan, L. W.
author_facet Campos-Herrera, R.
Johnson, E. G.
El-Borai, F. E.
Stuart, Robin J.
Graham, J. H.
Duncan, L. W.
author_sort Campos-Herrera, R.
title Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays
title_short Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays
title_full Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays
title_fullStr Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays
title_full_unstemmed Long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time PCR assays
title_sort long-term stability of entomopathogenic nematode spatial patterns in soil as measured by sentinel insects and real-time pcr assays
publisher Association of Applied Biologists
publishDate 2011
url http://hdl.handle.net/10261/60177
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