NDLI: TaqMan real-time PCR assay for the detection and quantification of Sclerospora graminicola, the causal agent of pearl millet downy mildew

Content Provider	Springer Nature Link
Author	Babu, Bandamaravuri Kishore Sharma, Rajan
Copyright Year	2015
Abstract	Early detection and quantification of Sclerospora graminicola, causal agent of pearl millet downy mildew, from seed, plant and infested soil could help in initiating preventive measures to control the spread of the disease. Two sensitive assays, SgK PCR (conventional PCR) and SgTqK q-PCR (real-time PCR), were developed for the detection of S. graminicola. Both assays were targeted on the 28S region of the nuclear large subunit (nuLSU) of the rDNA cluster. In conventional PCR, the species-specific primers (SgK F/R) amplified a 436 bp product in S. graminicola. TaqMan real-time PCR specifically amplified an 86 bp fragment in diverse DNA samples of S. graminicola obtained from infected seed, root, leaves and infested soil. Both assays did not amplify or produce any fluorescent signal from DNA obtained from other test microbes or from healthy pearl millet leaves. The absolute quantity of target molecules (28S) in the DNA sample obtained from S. graminicola sporangia (6 × 108/ml) was estimated at 25 pg/μl and the copy number was calculated as 2.69 × 108 molecules/μl. The copy number of target molecules in a diploid nucleus (2n) of S. graminicola was estimated to be ~27 molecules/nucleus. This assay can be used as a rapid and efficient detection tool for the identification and diagnosis of S. graminicola, and will be helpful in ensuring safe exchange of S. graminicola-free pearl millet germplasm across borders.
Starting Page	149
Ending Page	158
Page Count	10
File Format	PDF
ISSN	09291873
Journal	European Journal of Plant Pathology
Volume Number	142
Issue Number	1
e-ISSN	15738469
Language	English
Publisher	Springer Netherlands
Publisher Date	2015-02-06
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	q-PCR Species-specific primers Downy mildew Obligate pathogen Sclerospora graminicola Plant Pathology Plant Sciences Ecology Agriculture Life Sciences
Content Type	Text
Resource Type	Article
Subject	Plant Science Horticulture Agronomy and Crop Science

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TaqMan Real-time PCR Assay For The Detection And Quantification Of Sclerospora Graminicola, The Causal Agent Of Pearl Millet Downy Mildew

1sequence of its-2 amplified from pearl millet downy mildew samples.

Studies on some aspects of pearl millet downy mildew pathogen (sclerospora graminicola)

Isolation and evaluation of bacterial endophytes against Sclerospora graminicola (Sacc.) Schroet, the causal of pearl millet downy mildew

Characterization of a hydroxyproline-rich glycoprotein in pearl millet and its differential expression in response to the downy mildew pathogen Sclerospora graminicola

Cellulysin induces downy mildew disease resistance in pearl millet driven through defense response

Characterisation Of Pathogenic And Molecular Diversity In Sclerospora Graminicola, The Causal Agent Of Pearl Millet Downy Mildew

Characterisation Of Pathogenic And Molecular Diversity In Sclerospora Graminicola, The Causal Agent Of Pearl Millet Downy Mildew

Variation In Pathogenicity Among Single-oospore Isolates Of Sclerospora Graminicola, The Causal Organism Of Downy Mildew In Pearl Millet

TaqMan real-time PCR assay for the detection and quantification of Sclerospora graminicola, the causal agent of pearl millet downy mildew

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TaqMan Real-time PCR Assay For The Detection And Quantification Of Sclerospora Graminicola, The Causal Agent Of Pearl Millet Downy Mildew

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TaqMan real-time PCR assay for the detection and quantification of Sclerospora graminicola, the causal agent of pearl millet downy mildew