Development of real-time RT-PCR assays for the detection of Cucumber vein yellowing virus (CVYV) and Cucurbit yellow stunting disorder virus (CYSDV) in the whitefly vector Bemisia tabaci

Gil-Salas, Francisco M.; Morris, J.; Colyer, Alison; Budge, Giles E.; Boonham, Neil; Cuadrado, I. M.; Janssen, Dirk

doi:10.1016/j.jviromet.2007.05.032

Cited by 40 publications

(38 citation statements)

References 39 publications

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“…The thermal cycling conditions were composed of an initial denaturalization step at 95 °C for 3 min, followed by 45 cycles at 95 °C for 10 s, 58 °C for 20 s. Melting curve analysis and gel electrophoresis were performed to confirm specificity of the product. Relative transcriptomic expressions of the selected genes were statistically analyzed using methods from Gil-Salas et al (2007). This method calculates the Mean Normalized Expression using the average of Cq values and slope-intercept point from each regression analysis, including the calculation of variance and errors.…”

Section: Strain Culture Media and Growth Conditionmentioning

confidence: 99%

“…Data were normalized using both housekeeping genes, in independent statistical analysis, and Mean Normalized Expression was calculated according to Gil-Salas et al (2007). In the analysis, actin and β-Tubulin genes were selected as the most stable candidate gene for normalization (Mamarabadi et al 2008;Dekkers et al 2012).…”

Section: Comparison Of Gene and Protein Accumulation Pattern For Ungementioning

confidence: 99%

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Proteomic profiling of Botrytis cinerea conidial germination

et al. 2014

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Botrytis cinerea is one of the most relevant plant pathogenic fungi. The first step during its infection process is the germination of the conidia. Here, we report on the first proteome analysis during the germination of B. cinerea conidia, where 204 spots showed significant differences in their accumulation between ungerminated and germinated conidia by two-dimensional polyacrylamide gel electrophoresis and qPCR. The identified proteins were grouped by gene ontology revealing that the infective tools are mainly preformed inside the ungerminated conidia allowing a quick fungal development at the early stages of conidial germination. From 118 identified spots, several virulence factors have been identified while proteins, such as mannitol-1-phosphate dehydrogenase, 6,7-dimethyl-8-ribityllumazine synthase or uracil phosphoribosyltransferase, have been disclosed as a new potential virulence factors in botrytis whose role in pathogenicity needs to be studied to gain new insights about the role of these proteins as therapeutic targets and virulence factors.

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Section: Strain Culture Media and Growth Conditionmentioning

confidence: 99%

Section: Comparison Of Gene and Protein Accumulation Pattern For Ungementioning

confidence: 99%

Proteomic profiling of Botrytis cinerea conidial germination

et al. 2014

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“…RT-qPCR assays have been developed for CYSDV and CCYV but not for LIYV or BPYV. These assays are either based on the TaqMan Ò or SYBR Green chemistry [1,26,67]. RT-qPCR has been used for sensitive virus detection and quantitation of viral titer.…”

Section: Diagnosismentioning

confidence: 99%

“…RT-qPCR has been used for sensitive virus detection and quantitation of viral titer. Gil-Salas et al [26] developed an assay for CYSDV and Cucumber vein yellowing virus (CVYV) detection. Subsequently, the same group quantified CYSDV titer in co-infection with CVYV [27].…”

Section: Diagnosismentioning

confidence: 99%

Whitefly-transmitted criniviruses of cucurbits: current status and future prospects

Abrahamian

Abou‐Jawdah

2013

VirusDis.

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In the past decade, crinviruses have gained interest due to their rapid widespread and destructive nature for cucurbit cultivation. Several members of the genus Crinivirus are considered emerging viruses. Currently, four criniviruses: Beet pseudo-yellows virus, Cucurbit chlorotic yellows virus, Cucurbit yellow stunting disorder virus and Lettuce infectious yellows virus have been reported to infect field-or greenhouse-grown cucurbits. Apart from their cucurbit hosts, criniviruses infect other cash crops and weeds. Criniviruses are exclusively transmitted by whiteflies. The virion titer and the vector genus or species complex are predominant factors affecting virus transmission. These criniviruses maintain genetic stability with limited intra-species variability. They share similar core genome structure and replication strategies with some variations in the non-core proteins and downstream replication processes. Management of the diseases induced by criniviruses relies on integrated disease management strategies and on resistant varieties, when available. This review will cover their epidemiology, molecular biology, detection and management.

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“…Real-time PCR was performed using dual labelled probes designed to confirm that all the samples were B. tabaci (Gil-Salas et al 2007) and to discriminate between B and Q biotypes (the assay targets a single nucleotide polymorphism (SNP) in the mitochondrial cytochrome oxidase I (mtCOI) gene) (Jones et al 2008). Two forward primers, a reverse primer and a FAM-labelled probe were designed to detect TYLCV and TYLCSV in viruliferous whitefly ( Table 1).…”

mentioning

confidence: 99%

First record of the Q Biotype of the sweetpotato whitefly, Bemisia tabaci, intercepted in the UK

et al. 2012

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For the UK, Bemisia tabaci poses a threat primarily to protected vegetable crops due to the transmission of several plant-pathogenic viruses. There are at least 24 different biotypes of B. tabaci that cannot be differentiated through morphological traits. The B (Middle East-Asia Minor 1 species) and Q (Mediterranean species) biotypes are widely considered to be the most important and, as such, the ability to rapidly and precisely biotype B. tabaci interceptions is vital when developing effective control strategies. Intercepted adult/pupal B. tabaci received from the UK Plant Health and Seeds Inspectorate (PHSI) during 2002-2003 (n060) and 2010-2011 (n042) were both biotyped and tested for the presence of Tomato yellow leaf curl virus (TYLCV) and Tomato yellow leaf curl Sardinia virus (TYLCSV) using a real-time PCR assay based on TaqMan® chemistry. The positive results indicated that during 2002-2003 the Q biotype comprised 68.3 % of the interceptions whilst in 2010-2011 it comprised 66.7 % of the B. tabaci samples intercepted. Only three of the B biotypes collected during 2002-2003 were positive for TYLCSV, two originating from Israel and the other of unknown origin. The implications in regards to pest management of the insect are discussed.

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Development of real-time RT-PCR assays for the detection of Cucumber vein yellowing virus (CVYV) and Cucurbit yellow stunting disorder virus (CYSDV) in the whitefly vector Bemisia tabaci

Cited by 40 publications

References 39 publications

Proteomic profiling of Botrytis cinerea conidial germination

Proteomic profiling of Botrytis cinerea conidial germination

Whitefly-transmitted criniviruses of cucurbits: current status and future prospects

First record of the Q Biotype of the sweetpotato whitefly, Bemisia tabaci, intercepted in the UK

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