Journal of Plant Pathology (2003) 85, 315-316

C. Ratti, G.R.G. Clover, C. Rubies-Autonell, R. Resca, A. Pisi, V.A. Harju and C.M. Henry (2003)
A multiplex RT-PCR assay to distinguish and characterize sugarbeet soil borne viruses
Journal of Plant Pathology 85 (4), 315-316
X Meeting, Italian Society for Plant Pathology (poster)
Abstract: Six different primer sets were developed to identify Polymyxa betae, Beet soil borne virus (BSBV) and Beet virus Q (BVQ) and distinguish between A, B and P types of Beet necrotic yellow vein virus (BNYVV) using a multiplex reverse-transcription polymerase chain reaction (RT-PCR). RNA was extracted from 80 infected samples from Asia, Europe and North America. Virus infection was determined using single step RT-PCR and the BNYVV type using an established single strand conformation polymorphism (SSCP) detection method. Partial sequences of RNA 2 from 16 isolates of the BNYVV A and B type were determined and aligned and two sets of PCR primers were designed which yielded a single 323 base-pair RT-PCR fragment from samples containing the A type and a 157 base-pair product from samples containing the B type. Published sequences from P. betae, BSBV RNA 2, BVQ RNA 3 and BNYVV RNA 5 were also aligned and specific RT-PCR assay for P. betae (261 base-pair), BSBV (456 base-pair), BVQ (521 base-pair) and BNYVV RNA5 (593 base-pair) were designed. Fragment length differed sufficiently to allow all tests be run in a single PCR tube. Results obtained using the new multiplex RT-PCR assay were consistent with those from the established SSCP method and from single RT-PCR method for all 80 reference samples. Future work will identify primers to detect Beet soil borne mosaic virus (BSBMV) isolates as part of the same multiplex system.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Database assignments for author(s): Valerie A. Harju, Gerard R.G. Clover

Research topic(s) for pests/diseases/weeds:
identification/taxonomy
surveys/sampling/distribution

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Beet necrotic yellow vein virus	Beet/sugarbeet (Beta vulgaris)
	Polymyxa betae	Beet/sugarbeet (Beta vulgaris)
	Beet soil-borne virus	Beet/sugarbeet (Beta vulgaris)
	Beet virus Q	Beet/sugarbeet (Beta vulgaris)