Development of Real-Time PCR for the Rapid Detection of Episomal <i>Banana streak virus</i> (BSV)

Delanoy, M.; Salmon, Michel; Kummert, J.; Frison, Emile; Lepoivre, Philippe

doi:10.1094/pdis.2003.87.1.33

Cited by 27 publications

(24 citation statements)

References 20 publications

(19 reference statements)

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“…Real-time PCR methods have been described able to distinguish eBSV sequences from those of episomal BSV (Delanoy et al, 2003), and with real-time PCR now becoming more routine in Africa, this technique offers promise to distinguish eDBVs from sequences originating from viral particles. A further method reported as being able to distinguish viral particles from EPRVs is rolling circle amplification (RCA) (Rector et al, 2004), but our experience with yam indicates that it cannot be used for rapid indexing purposes as it amplifies plant sequences at low frequency.…”

Section: Diagnostics To Distinguish Edbvs From Dbvsmentioning

confidence: 99%

The prevalence of badnaviruses in West African yams (Dioscorea cayenensis-rotundata) and evidence of endogenous pararetrovirus sequences in their genomes

et al. 2014

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A comparison of serological and nucleic acid techniques was used to investigate whether the PCRpositives were sequences amplified from badnavirus particles or putative endogenous badnavirus sequences in the yam genome. Protein A sandwich-enzyme-linked immunosorbent assay (PAS-ELISA) with badnavirus polyclonal antisera detected cross-reacting viral particles in only 60% (92 of 153) of the CIRAD collection samples analyzed, in contrast to the aforementioned 81% by PCR.Immunosorbent electron microscopy (ISEM) of virus preparations of a select set of 16 samples, representing different combinations of positive and negative PCR and PAS-ELISA results, identified bacilliform particles in 11 of these samples. Three PCR-positive yam samples from Burkina Faso (cv. Pilimpikou) were identified in which no viral particles were detected by either PAS-ELISA or ISEM.Southern hybridisation results using a yam badnavirus RT-RNaseH sequence (Gn155Dr) as probe, supported a lack of badnavirus particles in the cv. Pilimpikou and identified their equivalent sequences to be of plant genome origin. Probe Gn155Dr, however, hybridised to viral particles and plant genomic DNA in three D. rotundata samples from Guinea. These results represent the first data demonstrating the presence of integrated sequences of badnaviruses in yam. The implications of this for virus-indexing, and breeding and multiplication of seed yams are discussed. IntroductionYam (Dioscorea species) is the fourth most important food tuber crop in the world after potato, sweet potato, and cassava (FAO, 2012). In West Africa, it is the second most important food crop after cassava by value and production (FAO, 2012;Scarcelli et al., 2006). It plays an essential role in food security and income generation for smallholders, particularly in West Africa which produces about 95% of the world's total yam production (Asiedu and Sartie, 2010; IITA, 2012;Mignouna et al., 2008 Several surveys on yam viruses suggest that badnaviruses are the most prevalent globally (Bousalem et al., 2009; Eni et al., 2008a, b;Galzi et al., 2013;Kenyon et al., 2008). Badnavirus particles were first reported in yam in association with a flexuous virus, causing internal brown spot disease in D. alata and D. cayenensis in the Caribbean (Harrison and Roberts, 1973;Mantell and Haque, 1978). Two decades later yam badnaviruses were characterised by their nucleic acid and serological properties; particles isolated from D. alata and D. bulbifera were partially characterised and named informally as Dioscorea alata bacilliform virus (DaBV) and Dioscorea bulbifera bacilliform virus (DbBV) Phillips et al., 1999). These viruses were reported to induce leaf distortions and veinal chlorosis , although others found that often infected plants show no marked symptoms (Kenyon et al., 2008;Seal and Muller, 2007). Sensitive virus diagnostic tests are required to enable the identification of virus-free seed yams, and will underpin current efforts in West Africa to generate and multiply disease-free yam planting material (I...

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Section: Diagnostics To Distinguish Edbvs From Dbvsmentioning

confidence: 99%

The prevalence of badnaviruses in West African yams (Dioscorea cayenensis-rotundata) and evidence of endogenous pararetrovirus sequences in their genomes

et al. 2014

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“…Using crude preparation of tissues, apple stem grooving virus, potato virus Y and banana streak virus were detected by RT-PCR or real-time PCR (Marinho et al, 1998;Chandelier et al, 2001;Delanoy et al, 2003). Besides, simple and rapid methods for nucleic acid liberation towards suitable and sensitive detection of some viruses and viroids by PCR have been described by Thomson and Dietzgen (1995) and Nakahara et al (1999), respectively.…”

Section: Discussionmentioning

confidence: 99%

“…The DNA extraction efficiency of this crude preparation method was further assessed by semi-quantitative evaluation of phytoplasma DNA using real-time PCR. The application of crude preparation of infected plants has been previously tested for virus detection by PCR amplification (Marinho et al, 1998;Marbot et al, 2002;Delanoy et al, 2003); furthermore a simple plant tissue preparation without homogenization suitable for detection of viruses by PCR was described by Thomson and Dietzgen (1995).…”

Section: Introductionmentioning

confidence: 99%

A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of ‘Candidatus Phytoplasma mali’

Aldaghi

Massart

Dutrecq

et al. 2009

Journal of Virological Methods

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a b s t r a c tDifferent PCR protocols have been established for detection of European fruit trees phytoplasmas; however the majority of the procedures for extracting phytoplasma DNA are complex, time consuming, and expensive, with a risk of contamination or loss of target DNA. In present study, a crude extract preparation method previously used to detect other plant pathogens was adapted to samples from apple trees infected by 'Candidatus Phytoplasma mali'. End-point and real-time PCR detection of 'Ca. P. mali' were used to compare this extraction procedure with an established method for efficient extraction of purified DNA. The crude extract proved fully adequate for phytoplasma detection in samples from 86 in vitro and 35 in vivo apple shoots or plants and 10 periwinkle plants. High inter-and intra-run reproducibility was obtained for phytoplasma detection with different TaqMan MGB-or SYBR Green-based real-time PCR protocols applied to the crude extracts. Real-time PCR applied to serially diluted crude and purified extracts revealed the same phytoplasma detection limit (dilution up to 10 5 ). All results confirm the suitability of this simple, quick, efficient extraction technique for accurate detection of 'Ca. P. mali' in different types of apple and periwinkle samples.

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“…Realtime quantitative PCR is a method that provides accurate and reproducible quantification of gene copies, does not require post-PCR sample handling and results in much faster and higher throughput assays (Heid et al, 1996). Recently, real-time quantitative RT-PCR was successfully applied to detecting plant pathogens (Eun et al, 2000;Roberts et al, 2000;Cullen et al, 2001;Korimbocus et al, 2002;Balaji et al, 2003;Delanoy et al, 2003;Nicolaisen, 2003;Schena et al, 2004;Pico et al, 2005), but there have been no reports on identifying RBSDV with real-time quantitative RT-PCR.…”

Section: Introductionmentioning

confidence: 99%

Improved method for assaying maize plant resistance to maize rough dwarf disease by artificial inoculation and real-time RT-PCR

et al. 2006

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The study of maize rough dwarf disease (MRDD) and breeding for resistance requires inoculation of maize plants by means of planthoppers. The plant age, insect density and inoculation duration are main factors in the success of maize rough dwarf disease inoculation. These parameters were tested using a susceptible maize inbred line Ye478. Using one or two-leaf plants, 15 planthoppers per plant and a five day inoculation duration, the line Ye478 was the most susceptible with 100% diseased plants; F112132 was moderately susceptible with 60% diseased plants and 90110 and F022411 were resistant without any disease. The results were consistent with those from six years of field studies. Using enzyme-linked immunosorbent assay (ELISA) and real-time quantitative RT-PCR, rice black-streaked dwarf virus was detected in severely diseased plants. The plants were rated from 0 to 3 according to their symptoms at the time of flowering. Plants scoring 0, 1 and 2 could not be distinguished by ELISA, only by real-time quantitative RT-PCR. All of the plants with a score of 3 were positive by ELISA and real-time quantitative RT-PCR. The significant differences in the average viral contents in plants with different symptom ratings could be distinguished by using real-time RT-PCR.

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Development of Real-Time PCR for the Rapid Detection of Episomal Banana streak virus (BSV)

Cited by 27 publications

References 20 publications

The prevalence of badnaviruses in West African yams (Dioscorea cayenensis-rotundata) and evidence of endogenous pararetrovirus sequences in their genomes

The prevalence of badnaviruses in West African yams (Dioscorea cayenensis-rotundata) and evidence of endogenous pararetrovirus sequences in their genomes

A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of ‘Candidatus Phytoplasma mali’

Improved method for assaying maize plant resistance to maize rough dwarf disease by artificial inoculation and real-time RT-PCR

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