|Balaji, Boovaraghan - PURDUE UNIVERSITY|
Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: July 29, 2002
Publication Date: July 29, 2002
Citation: Balaji, B., Bucholtz, D.L., Anderson, J.M. 2002. Yellow dwarf virus quantification by real-time pcr during disease development in resistant and susceptible plants. Phytopathology. Technical Abstract: Reliable detection and quantification of yellow dwarf virus (YDV) is a critical component in managing these viral diseases in small grain cereal crops. The method of choice usually is the ELISA using antisera against the coat protein that are specific to different YDV strains. Currently, Real-time Quantitative PCR was applied to measure the accurate quantification of gene expression. We are applying this technique to detect and quantify YDV using primers specific for BYDV-PAV and CYDV-RPV coat protein genes. Given the higher sensitivity of RT-PCR and the advantage of using a real time PCR instrument, we have successfully utilized this method to detect BYDV and CYDV and examined disease development in a YDV-resistant wheatgrass and susceptible oat by quantifying the level of virus. Wheatgrass possesses a high level of resistance to YDV strains. A YDV-resistant wheat line (P29), and a susceptible parent, 8138 were also used for this study. All plants were infested with viruliferous aphids and plants collected at regular intervals for 12 days. RNA from each harvest and random hexamers was used for first strand cDNA synthesis that was used as the template for Real-time PCR. The procedure described is reproducible and sensitive and has the potential to be used as a rapid diagnostic tool for BYDV and CYDV and as a means for examining mechanisms of virus resistance.