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Title: Assessing the movement of Cucurbit yellow stunting disorder virus in susceptible and tolerant cucumber germplasms using serological and nucleic acid based methods

Author
item EID, S - AMERICAN UNIV BEIRUT
item ATAMIAN, H - AMERICAN UNIV BEIRUT
item ABOU-JAWDAH, Y - AMERICAN UNIV BEIRUT
item Havey, Michael

Submitted to: Journal of Phytopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/29/2007
Publication Date: 8/1/2008
Citation: Eid, S., Atamian, H.S., Abou-Jawdah, Y., Havey, M.J. 2008. Assessing the movement of Cucurbit yellow stunting disorder virus in susceptible and tolerant cucumber germplasms using serological and nucleic acid based methods. Journal of Phytopathology. 156:438-445.

Interpretive Summary: Cucurbit yellow stunting disorder virus (CYSDV) is an emerging virus causing significant yield losses in cucurbits. Simple and reliable detection and quantification methods are important for disease management. In a susceptible cucumber plants, CYSDV was detected 5 days post-inoculation (DPI) by two techniques, RT-PCR or by tissue blot immunoassay (TBIA). At 8-9 DPI CYSDV was detected by dot blot immunoassay (DBIA) or ELISA. For CYSDV quantification, real-time RT-PCR was the most sensitive method. DBIA was more sensitive than ELISA and equally sensitive to nucleic acid hybridization. Time course studies at 3, 5, 8 and 14 DPI, using TBIA, revealed that tolerance to CYSDV in three cucumber germplasms was not correlated with restricted or delayed virus movement. These detection and quantification methods for CYSDV will be of interest to cucumber breeders and pathologists in the public and private sectors as an aid for the incorporation of tolerance to CYSDV into elite cucumbers.

Technical Abstract: Cucurbit yellow stunting disorder virus (CYSDV) is an emerging virus causing significant yield losses in cucurbits. Simple but reliable detection and quantification methods constitute an important support to disease management. In a susceptible germplasm CYSDV was detected 5 days post-inoculation (DPI) by RT-PCR or by tissue blot immunoassay (TBIA), and 8-9 DPI by dot blot immunoassay (DBIA) or ELISA. For CYSDV quantification, real-time RT-PCR was the most sensitive method and gave the best linear range of detection, over four orders of magnitude as compared to about two orders of magnitude with DBIA and nucleic acid hybridization. DBIA was more sensitive than ELISA and equally sensitive to nucleic acid hybridization with a non-radioactively labeled cDNA probe. Time course studies at 3, 5, 8 and 14 DPI, using TBIA, revealed that tolerance to CYSDV in three tolerant cucumber germplasms was not correlated with restricted or delayed virus movement.