Multiplex PCR assay for the simultaneous detection and differentiation of Olpidium bornovanus, O. brassicae, and O. virulentus

Herrera-Vásquez, J. A.; Cebrián, María del Carmen; Alfaro-Fernández, A.; Córdoba-Sellés, M. C.; Jordá, C.

doi:10.1016/j.mycres.2009.01.007

Cited by 29 publications

(24 citation statements)

References 23 publications

(31 reference statements)

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“…Differentiation of these two species is only possible at a molecular level. Application of a multiplex PCR based test, using specific primers (Herrera-Vásquez et al 2009) for each of the two Olpidium species, clearly identified the fungus captured by B. pekinensis in the olive orchard soil as O. brassicae.…”

Section: Discussionmentioning

confidence: 99%

“…A primer specific for O. virulentus (OLPvirF) and a primer specific for O. brassicae (OLPbraF) were used together with one common reverse primer (OLPR) (Herrera-Vásquez et al 2009) in a multiplex PCR assay. DNA of O. brassicae and O. virulentus used as positive controls in PCR assays were kindly supplied by Herrera-Vasquez.…”

Section: Olpidium Spp Recovery and Molecular Identificationmentioning

confidence: 99%

“…In this work we applied a PCR-based test developed by Herrera-Vásquez et al (2009) that allows the discrimination of O. brassicae from O. virulentus through the size of the generated amplicons. Olpidium species present in the olive orchard soil were characterized and used to test their capacity to transmit OMMV, a variant designated as OMMV L11 and a construct OMMV/OMMV L11.…”

Section: Introductionmentioning

confidence: 99%

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Evidence of olive mild mosaic virus transmission by Olpidium brassicae

et al. 2011

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Transmission of three strains of OMMV by an Olpidium sp. was evaluated and compared. The three strains were 1) an OMMV wild type (WT) recovered from olive trees, 2) an OMMV variant (L11) obtained after 15 serial passages of single local lesions induced in Chenopodium murale plants, and 3) a construct OMMV/OMMVL11 in which the coat protein (CP) gene replaced that of the wild type. A single-sporangial culture derived from Chinese cabbage (Brassica pekinensis) used as a bait plant grown in soil of an olive orchard, was identified as Olpidium brassicae based on the size and sequence of the generated amplicon in PCR specific tests. Each of the three virus strains was soil transmitted to cabbage roots in the absence of the fungus at similar rates of 30 to 40%. Separate plant inoculation by O. brassicae zoospores incubated with each viral strain resulted in enhanced transmission of OMMV, reaching 86% of infection whereas that of the other two strains remained practically unaffected at ca. 34%. Binding assays showed that the amount of virus bound to zoospores, estimated spectrophotometrically, was 7% in the case of OMMV, and practically nil in the case of the other two viral strains. Substitution of the coat protein (CP) gene of OMMV by that of the OMMV L11 strain, drastically reduced viral transmissibility in the presence of zoospores to the level of that observed in their absence. Our data shows that OMMV soil transmission is greatly enhanced by O. brassicae zoospores and that the viral CP plays a significant role in this process, most likely by facilitating virus binding and later entrance into the host plant roots.

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Section: Discussionmentioning

confidence: 99%

Section: Olpidium Spp Recovery and Molecular Identificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Evidence of olive mild mosaic virus transmission by Olpidium brassicae

et al. 2011

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“…A primer specific for O. virulentus (OLPvirF) and a primer specific for O. brassicae (OLPbraF) were used together with one common reverse primer (OLPR) in a multiplex PCR assay (Herrera-Vasquez et al 2009). Amplified products were separated by electrophoresis in 1 % agarose gel in 1× TBE buffer (0.1 M Tris, 0.09 M boric acid, 1 mM EDTA, pH 8.4) at 80 V and stained with 0.2 μg ml −1 ethidium bromide.…”

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confidence: 99%

Olive mild mosaic virus transmission by Olpidium virulentus

et al. 2015

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The ability of Olpidium virulentus to vector Olive latent virus 1 (OLV-1), Olive mild mosaic virus (OMMV) and Tobacco necrosis virus D (TNV-D) was evaluated. Transmission assays involved zoospore acquisition of each virus, inoculation onto cabbage plant roots followed by viral detection. Assays revealed that these viruses are transmitted in the absence of the fungus, but the transmission rates of OMMV are much higher when OMMV is incubated with O. virulentus zoospores prior to inoculation, while the transmission rates of each OLV-1 and TNV-D do not change when they are incubated with the fungus. Our data shows that O. virulentus is an efficient vector of OMMV, greatly enhancing virus transmissibility, but not that of OLV-1 nor TNV-D. Prior studies have shown that another fungus, O. brassicae, also enhances OMMV transmission. To our knowledge, this is the first report of a plant virus, OMMV, being efficiently transmitted by two different species of fungi.

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“…No obstante, el virus nunca se transmitió a plantas sanas regadas con el percolado obtenido del riego de plantas únicamente infectadas con PepMV, lo que pone de manifiesto que el virus no se transmite con el agua de riego sin la presencia del vector, contrariamente a los resultados obtenidos en otros estudios que determinan la distribución del virus en sistemas hidropónicos (Fakhro et al, 2005;Schwarz et al, 2007). Asimismo, Herrera-Vásquez et al, 2009b) en el agua de drenaje obtenida tras el riego de plantas infectadas con dicho agente, mientras que PepMV estaba presente únicamente en las muestras de agua de drenaje procedente de plantas infectadas con el virus y el hongo vector.…”

Section: Discussionunclassified

Epidemiología y variabilidad patogénica del virus del mosaico del pepino dulce (Pepino mosaic virus). Nuevas enfermedades asociadas a su presencia (torrao o cribado)

Fernandez¹,

Olvido²

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Multiplex PCR assay for the simultaneous detection and differentiation of Olpidium bornovanus, O. brassicae, and O. virulentus

Cited by 29 publications

References 23 publications

Evidence of olive mild mosaic virus transmission by Olpidium brassicae

Evidence of olive mild mosaic virus transmission by Olpidium brassicae

Olive mild mosaic virus transmission by Olpidium virulentus

Epidemiología y variabilidad patogénica del virus del mosaico del pepino dulce (Pepino mosaic virus). Nuevas enfermedades asociadas a su presencia (torrao o cribado)

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