ARS | Publication request: A qPCR assay for detection and quantification of Verticillium dahliae in spinach seed.

Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: May 2, 2010
Publication Date: June 1, 2010
Citation: Rauscher, G.M., Mou, B., Hayes, R.J., Koike, S.T., Maruthachalam, K., Subbarao, K.V., Klosterman, S.J. 2010. A qPCR assay for detection and quantification of Verticillium dahliae in spinach seed.. Phytopathology. 100:S107.

Technical Abstract: The fungus Verticillium dahliae is the causal agent of Verticillium wilt of lettuce and other specialty crops in the Salinas Valley of California. Spinach, another major specialty crop in California, is not affected by Verticillium wilt in commercial production. However, spinach seed infected with V. dahliae and planted in the Salinas Valley increases inoculum density and introduces exotic strains that may contribute to Verticillium wilt epidemics. The goal of this work is to develop a real time quantitative PCR (qPCR) assay for the detection and quantification of V. dahliae in spinach seed. The assay is based on the use of SYBR Green methodology with previously published primer sequences specific for the beta-tubulin gene of V. dahliae. Parallel plating and qPCR assays revealed that the qPCR assay can be used for reliable detection of V. dahliae in seed infected at the 10 percent level. Because the qPCR assay enables rapid and reliable detection of V. dahliae, the assay has implications as a useful tool to limit the spread of the pathogen