Development of real-time PCR (TaqMan®) assays for the detection and quantification of Botrytis cinerea in planta

Suárez, Mariana; Walsh, Kathryn; Boonham, Neil; O'Neill, T. M.; Pearson, Simon; Barker, Ian K.

doi:10.1016/j.plaphy.2005.07.003

Cited by 118 publications

(111 citation statements)

References 40 publications

(58 reference statements)

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“…F. graminearum was identified, from Leslie and Summerell (2006). DNA from 75 F. graminearum strains, morphologically identified, was extracted using a CTAB method (Suarez et al, 2005), modified in this work using seven days fungal mycelium (100−200 mg) and adding 1 μl of proteinase K (20 mg/ml) to CTAB-grinding buffer shortly before the use. F. graminearum DNA was analyzed with specific primers Fg16F/Fg16R under the conditions described by Nicholson et al (1998), to determine the identity of all the strains.…”

mentioning

confidence: 99%

Difference in Chemotype Composition of Fusarium graminearum Populations Isolated from Durum Wheat in Adjacent Areas Separated by the Apennines in Northern-Central Italy

Purahong¹,

Tonti²,

Salomoni³

et al. 2011

The Plant Pathology Journal

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Chemotype composition of Fusarium graminearum strains, isolated from durum wheat kernels from naturally FHB infected fields in Northern and Central Italy, was investigated by multiplex PCR. The different climatic and environmental conditions of the two examined areas separated by the Apennines affected the composition of chemotypes. 15Ac-DON chemotype was predominant in both the sub areas. Nivalneol chemotype was more frequent in the warmer sub area.

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mentioning

confidence: 99%

Difference in Chemotype Composition of Fusarium graminearum Populations Isolated from Durum Wheat in Adjacent Areas Separated by the Apennines in Northern-Central Italy

Purahong¹,

Tonti²,

Salomoni³

et al. 2011

The Plant Pathology Journal

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“…A number of nucleic-acid-based methods for the detection of B. cinerea in plants are considered highly specific, although they have not been screened for their specificity against nontarget Botrytis species (Brouwer et al 2003;Gachon and Saindrenan 2004;Mehli et al 2005;CadleDavidson 2008;Celik et al 2009;Sanzani et al 2012). In addition, none of the methods described that have considered nontarget Botrytis species discriminated B. cinerea from the phylogenetically related species B. pelargonii or B. fabae, which cause leaf-base necrosis in Pelargonium and chocolate spot in broad bean, respectively (Røed 1949;Suarez et al 2005;Spotts et al 2008;Tomlinson et al 2010). As B. fabae does not attack pelargonium plants (data not shown) and B. pelargonii has never been reported to attack pelargoniums since the description of Røed (1949), we believe that it would be unlikely to detect these species in pelargonium samples.…”

Section: Discussionmentioning

confidence: 99%

“…Accurate quantification of a pathogen in diseased plant tissues is an important means for the evaluation of the degree of host susceptibility and it provides essential information for breeding programs for plant resistance. Several PCR and real-time PCR assays have been developed for detection and quantification of B. cinerea in many types of samples (Rigotti et al 2002;Suarez et al 2005;Cadle-Davidson 2008;Diguta et al, 2010;Tomlinson et al 2010;Sanzani et al 2012). …”

Section: Introductionmentioning

confidence: 99%

A real-time PCR assay for detection and quantification of Botrytis cinerea in Pelargonium x hortorum plants, and its use for evaluation of plant resistance

et al. 2015

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A reliable EvaGreen-based qPCR assay was developed for the relative quantification of fungal and plant DNAs in the Botrytis cinerea-Pelargonium× hortorum pathosystem. Primers, designed on RPB2 (DNA-dependent RNA polymerase subunit II) gene sequences directed the amplification of a specific 149 bp product for B. cinerea. No cross-reactivity was observed in DNA extracted from several nontarget fungi and bacterial, and in pelargonium plants, with the exception of the closely related species Botryotinia pelargonii and Botrytis fabae. This assay allowed for quantification of as little as 1.55 pg fungal DNA and 9.95 pg plant DNA, and detection of the pathogen in both symptomatic and asymptomatic pelargonium plants. The qPCR protocol is suitable for studying cultivar resistance and induced resistance in pelargonium plants, which requires accurate quantification of the pathogen in diseased host tissue. This assay allowed us to establish that the Pelargonium×hortorum cvs. 'XL Boomerang' and 'Eroica 2000' showed high foliar resistance to B. cinerea with respect to the susceptible cv. 'Fernando', and that spray applications of 0.3 mM acibenzolar-S methyl markedly protect pelargonium plants against gray mold. The protection for plants treated with chitosan (0.2 %) did not reach significance.

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“…The volume of DNA was quantified using a nano drop ND-1000 Spectrophotometer (Applied Biosystems). A quantitative polymerase chain reaction (qPCR) was performed with the extracted DNA using scar primers of B. cinerea as designed by Suarez et al, (2005) following an established protocol (e.g. [2,38,39] for B. cinerea.…”

Section: Determination Of Biomass Of Systemic B Cinereamentioning

confidence: 99%

“…A quantitative polymerase chain reaction (qPCR) was performed with the extracted DNA using scar primers of B. cinerea as designed by Suarez et al, (2005) following an established protocol (e.g. [2,38,39] for B. cinerea. A master mix containing 12.5µl qPCR master mix (Qiagen UK), 1µl of each forward and reverse scar primers (Invitrogen), and 5µl of water was prepared and 20µl was aliquoted into each well.…”

Section: Determination Of Biomass Of Systemic B Cinereamentioning

confidence: 99%

Bi-Directional Interaction between Systemic Botrytis Cinerea, and Aphid Myzus Persicae on Lettuce Plant

Sm¹,

Mu²,

Lawan³

et al. 2017

BJSTR

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Bi-directional interaction between economically important fungal pathogen Botrytis cinerea and aphid Myzus persicea causes serious economic losses to the crop plants resulting in loss of both time and money spend. In bi-direction interaction the fungal pathogen and insect herbivores, can both interact directly with the plant and interact indirectly with each other as they struggle to compete for the resources of the plant. We investigated the bi-directional interaction between the necrotrophic and economically important fungal pathogen Botrytis cinerea Pers Fr (Helotiales Sclerotiniaceae) and the aphid Myzus persicae (Hemiptera Aphididae) on plant host, lettuce Lactuca sativa (Asteraceae: Compositae). Botrytis cinerea on host plant causes a brownish discoloration of the leaf petiole accompanied by the rotting of the leaves, however, presence of aphids on lettuce plants causes' economic damage directly through injury and indirectly through virus transmission, resulting in wilting and head contamination.In this study, it was found that negative interaction between B. cinerea and Myzus persicae was established. The presence of fungal pathogen B. cinerea and Myzus persicae stressed the host lettuce plant, resulting in significant reduction in the rate of photosynthesis and chlorophyll fluorescence, and reduction in the dry shoot and root weight of the lettuce plant. The study established that aphid population growth rate and the number of B. cinerea lesions decreased when both were present on the same host plant.

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Development of real-time PCR (TaqMan®) assays for the detection and quantification of Botrytis cinerea in planta

Cited by 118 publications

References 40 publications

Difference in Chemotype Composition of Fusarium graminearum Populations Isolated from Durum Wheat in Adjacent Areas Separated by the Apennines in Northern-Central Italy

Difference in Chemotype Composition of Fusarium graminearum Populations Isolated from Durum Wheat in Adjacent Areas Separated by the Apennines in Northern-Central Italy

A real-time PCR assay for detection and quantification of Botrytis cinerea in Pelargonium x hortorum plants, and its use for evaluation of plant resistance

Bi-Directional Interaction between Systemic Botrytis Cinerea, and Aphid Myzus Persicae on Lettuce Plant

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