Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi

Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi

11 páginas, ilustraciones y tablas estadísticas.

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Main Authors: Pathak, Ekta, El-Borai, F. E., Campos-Herrera, R., Johnson, E. G., Stuart, Robin J., Graham, J. H., Duncan, L. W.
Format: artículo biblioteca
Language:English
Published: Elsevier 2012
Subjects:Arthrobotrys, Catenaria, Entomopathogenic nematodes, Nematophagous fungi, Parasitism, Quantitative real-time PCR,
Online Access:http://hdl.handle.net/10261/60314
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spelling dig-ica-es-10261-603142020-05-19T11:12:44Z Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi Pathak, Ekta El-Borai, F. E. Campos-Herrera, R. Johnson, E. G. Stuart, Robin J. Graham, J. H. Duncan, L. W. Arthrobotrys Catenaria Entomopathogenic nematodes Nematophagous fungi Parasitism Quantitative real-time PCR 11 páginas, ilustraciones y tablas estadísticas. Entomopathogenic nematodes (EPNs) are important pathogens of soilborne insects and are sometimes developed commercially to manage insect pests. Numerous nematophagous fungal species (NF) prey on nematodes and are thought to be important in regulating natural or introduced EPN populations. However, nematophagy by these fungi in nature cannot be inferred using existing methods to estimate their abundance in soil because many of these fungi are saprophytes, resorting to parasitism primarily when certain nutrients are limiting. Therefore, we developed an assay to quantify NF DNA in samples of nematodes. Species-specific primers and TaqMan probes were designed from the ITS rDNA regions of Arthrobotrys dactyloides, Arthrobotrys oligospora, Arthrobotrys musiformis, Gamsylella gephyropagum and Catenaria sp. When tested against 23 non-target fungi, the TaqMan real-time PCR assay provided sensitive and target-specific quantification over a linear range. The amount of A. dactyloides or Catenaria sp. DNA in 20 infected nematodes, measured by real-time PCR, differed between fungal species (P ¼ 0.001), but not between experiments (P > 0.05). However, estimates of relative NF parasitism using a bioassay with 20 nematodes infected by either species, differed greatly (P < 0.001) depending on whether the fungi were alone or combined in the samples used in the assay. Tests done to simulate detection of NF DNA in environmental samples showed that, for all species, background genomic DNA and/or soil contaminants reduced the quantity of DNA detected. Nested PCR was ineffective for increasing the detection of NF in environmental samples. Indeed, real-time PCR detected higher amounts of NF DNA than did nested PCR. The spatial patterns of NF parasitism in a citrus orchard were derived using real-time PCR and samples of nematodes extracted from soil. The parasitism by Catenaria sp. was positively related to the abundance of both heterorhabditid and steinernematid EPNs. The possible significance of the associations is ambiguous because NF attack a broad range of nematode taxa whereas EPNs are a small minority of the total nematode population in a soil sample. These studies demonstrate the potential of real-time PCR to study the role of NF parasitism in soil food webs. No. 691, 2010e2011 cycle Peer reviewed 2012-11-15T11:54:02Z 2012-11-15T11:54:02Z 2012 artículo http://purl.org/coar/resource_type/c_6501 Fungal Biology 116: 563-573(2012) 1878-6146 http://hdl.handle.net/10261/60314 10.1016/j.funbio.2012.02.005 en http://dx.doi.org/10.1016/j.funbio.2012.02.005 none Elsevier
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 Arthrobotrys
Catenaria
Entomopathogenic nematodes
Nematophagous fungi
Parasitism
Quantitative real-time PCR
Arthrobotrys
Catenaria
Entomopathogenic nematodes
Nematophagous fungi
Parasitism
Quantitative real-time PCR
spellingShingle Arthrobotrys
Catenaria
Entomopathogenic nematodes
Nematophagous fungi
Parasitism
Quantitative real-time PCR
Arthrobotrys
Catenaria
Entomopathogenic nematodes
Nematophagous fungi
Parasitism
Quantitative real-time PCR
Pathak, Ekta
El-Borai, F. E.
Campos-Herrera, R.
Johnson, E. G.
Stuart, Robin J.
Graham, J. H.
Duncan, L. W.
Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi
description 11 páginas, ilustraciones y tablas estadísticas.
format artículo
topic_facet Arthrobotrys
Catenaria
Entomopathogenic nematodes
Nematophagous fungi
Parasitism
Quantitative real-time PCR
author Pathak, Ekta
El-Borai, F. E.
Campos-Herrera, R.
Johnson, E. G.
Stuart, Robin J.
Graham, J. H.
Duncan, L. W.
author_facet Pathak, Ekta
El-Borai, F. E.
Campos-Herrera, R.
Johnson, E. G.
Stuart, Robin J.
Graham, J. H.
Duncan, L. W.
author_sort Pathak, Ekta
title Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi
title_short Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi
title_full Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi
title_fullStr Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi
title_full_unstemmed Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi
title_sort use of real-time pcr to discriminate parasitic and saprophagous behaviour by nematophagous fungi
publisher Elsevier
publishDate 2012
url http://hdl.handle.net/10261/60314
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