Skip to main content
ARS Home » Northeast Area » Frederick, Maryland » Foreign Disease-Weed Science Research » Research » Publications at this Location » Publication #147103
Title: DEVELOPMENT OF A REAL-TIME PCR ASSAY TO DETECT RALSTONIA SOLANACEARUM RACE 3, BIOVAR 2 (BV2) IN GERANIUM PLANTS

Author
item NIKOLAEVA, E. - RUSSIAN RES. INST. PHYTO.
item MATVEEEVA, E. - RUSSIAN RES. INST. PHYTO.
item Schaad, Norman

Submitted to: American Phytopathological Society Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: 3/15/2003
Publication Date: 6/1/2003
Citation: Nikolaeva, E., Matveeva, E., Schaad, N.W. 2003. Development of a real-time PCR assay to detect ralstonia solanacearum race 3, biovar 2 (BV2) in geranium plants. Phytopathology. 93:S65

Interpretive Summary:

Technical Abstract: Ralstonia solanacearum (Rs) race 3, bv2 infects potato and geranium under cool temperatures. The organism is highly regulated in the US and EU. We have developed and tested the sensitivity of a real-time BIO-PCR assay for specific detection of Rs race 3, bv2 in geranium plants. Healthy stems (100 or 1,000) spiked with a single, infected petiole, were soaked in water for 2 h and the liquid used for direct PCR and for BIO-PCR. For BIO-PCR enrichment, 100ul samples were plated onto mSMSA agar for 24-48 h and the plate washings used for PCR. PCR was done in a Smart Cycler using two different sets of primers and probe: 1) Rs race 3, bv2-specific RSC-F, RSC-R primers and RSC-P probe (Ozakman and Schaad, unpublished); and, 2) Rs species-specific RS-I-F, RS-I-R primers and RS-P probe (Weller et al., 2000). None of the samples were positive by a direct PCR assay. With BIO-PCR, both sets of primers and probe detected 1 infected petiole in 100 healthy stems, but only RSC-F, RSC-R primers and RSC-P probe were able to detect 1 infected petiole in 1000 healthy stems (Ct 36.1).