Author
POLSTON, JANE - UNIV. FLORIDA | |
Hladky, Laura | |
AKAD, FOUAD - UNIV. FLORIDA | |
Wintermantel, William - Bill |
Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 5/5/2008 Publication Date: 8/1/2008 Citation: Polston, J.E., Hladky, L.L., Akad, F., Wintermantel, W.M. 2008. First Report of Cucurbit Yellow Stunting Disorder Virus in Cucurbits in Florida. Plant Disease. 92:1251 Interpretive Summary: In August and September of 2007, watermelon plants (Cucumis lanatus L.) in commercial fields in Manatee and Hillsborough Counties in west central Florida began exhibiting a range of symptoms including stunting, deformation, interveinal chlorosis, leaf mottling and spotting. Leaves of plants had large populations of silverleaf whitefly (Bemisia tabaci biotype B). Leaf samples were collected from ten watermelon plants showing different combinations of symptoms. Total RNA was extracted and subjected to reverse transcription (RT)-PCR for the presence of criniviruses using primers specific to regions of the genes encoding the coat protein and HSP70h genes of CYSDV, and for the presence of begomoviruses using two pairs of primers designed to amplify portions of both the begomovirus A and B components. RT-PCR amplified the expected fragment of the coat protein and HSP70h genes from two of the symptomatic plants and from CYSDV-infected control plants, whereas no fragment was obtained from RNA extracted from leaves of healthy controls. The coat protein amplicon was sequenced from one of the Manatee Co. isolates and the sequenced portion was found to be 100% identical to isolates from Texas, California, Jordan and France, and shared 99% identity with a well characterized CYSDV isolate from Spain, but only 91% with an isolate from Iran. The begomovirus primer pair pBL1v2040 and PCRc154 produced a 678 bp amplicon with 94% sequence identity to isolates of CuLCrV from California and Arizona. Since the initial appearance of CYSDV numerous additional fields have also been confirmed infected in the Manatee and Hillsborough County areas using both molecular and serological methods. The emergence of CYSDV in Florida follows the recent identification of CYSDV in California and Arizona, USA and Sonora, Mexico in 2006. The close relationship between the appearance of CYSDV in Florida as well as CuLCrV suggests the possibility of movement on transplants. The rapid and extensive movement of CYSDV throughout cucurbit production regions in the United States warrants concern for all cucurbit production in the southern U.S. Technical Abstract: In August and September of 2007, watermelon plants (Cucumis lanatus L.) in commercial fields in Manatee and Hillsborough Counties in west central Florida began exhibiting a range of symptoms including stunting, deformation, interveinal chlorosis, leaf mottling and spotting. Leaves of plants had large populations of silverleaf whitefly (Bemisia tabaci biotype B). Leaf samples were collected from ten watermelon plants showing different combinations of symptoms. Total RNA was extracted using RNeasy mini kit (Qiagen, Valencia, CA) and subjected to reverse transcription (RT)-PCR for the presence of criniviruses using primers specific to regions of the genes encoding the coat protein and HSP70h genes of CYSDV, and for the presence of begomoviruses using two pairs of primers, AC1048 and AV494, a degenerate primer pair designed to amplify a region of the coat protein gene on the begomovirus A component, and PBL1v2040 and PCRc154, a degenerate primer pair designed to amplify a region of the hypervariable region of the begomovirus B component. RT-PCR amplified the expected 394 bp fragment of the coat protein gene from two of the symptomatic plants and from CYSDV-infected control plants, whereas no fragment was obtained from RNA extracted from leaves of healthy controls. Similarly, the 175 bp HSP70h fragment was amplified from the same symptomatic melons and from CYSDV infected control plants, but no fragment was obtained from RNA extracted from leaves of healthy controls. The coat protein amplicon was sequenced from one of the Manatee Co. isolates (GenBank Accession No. Pending) and the 344 nt sequenced portion of the amplicon was found to be 100% identical to isolates from Texas, California, Jordan and France, and shared 99% identity with a well characterized CYSDV isolate from Spain, but only 91% with an isolate from Iran. The begomovirus primer pair pBL1v2040 and PCRc154 produced a 678 bp amplicon that is consistent with the presence of a bipartite begomovirus in all ten samples. Sequence analysis of four of the amplicons revealed that all 678-bp amplicons had 94% sequence identity to isolates of CuLCrV from California and Arizona. Since the initial appearance of CYSDV numerous additional fields have also been confirmed infected in the Manatee and Hillsborough County areas using both molecular and serological methods. The emergence of CYSDV in Florida follows the recent identification of CYSDV in California and Arizona, USA and Sonora, Mexico in 2006. The close relationship between the appearance of CYSDV in Florida as well as CuLCrV suggests the possibility of movement on transplants. The rapid and extensive movement of CYSDV throughout cucurbit production regions in the United States warrants concern for all cucurbit production in the southern U.S. |