Association of a Monopartite Begomovirus with Yellow Mosaic Disease of Pumpkin (Cucurbita maxima) in India.

Pumpkin (Cucurbita maxima) is an economically important vegetable crop of the family Cucurbitaceae. Yellow mosaic in C. maxima was observed in and around Lucknow, India, during the winter season of 1999-2000. High populations of whiteflies (Bemisia tabaci) were associated with diseased plants. Whiteflies were allowed to feed on diseased C. maxima plants and placed on healthy C. maxima seedlings. Symptoms of yellow mosaic disease were successfully reproduced in the seedlings. These results suggested a possible involvement of a geminivirus with the disease. Total DNA was extracted from infected and uninfected C. maxima seedlings by the CTAB method. DNA samples were used as templates in polymerase chain reaction (PCR) using degenerate primers A (5'-TAATATTACCKGWKGVCCSC-3') and B (5'-TGGACYTTRCAWGGBCCT TCACA-3'), which have been designed to amplify a region of about 500 to 600 nt from DNA-A of a majority of begomoviruses (1). The PCR products obtained were analyzed on a 1.5% agarose gel. The major amplicon obtained from the DNA of infected plants was about 550 bp as expected from these primers. However, no such amplicon was observed in DNA samples from uninfected plants. The amplicon obtained from PCR was cloned in a suitable cloning vector. Three separate clones were sequenced and the corrected sequence data (GenBank Accession No. AF381977) was compared with the corresponding sequences of other known geminiviruses. The highest identity (88%) was with a strain of monopartite Tomato leaf curl virus (TLCV) from southern India (GenBank Accession No. U38239). The first 34 nucleotides showed significant identity (91% and 97%, respectively) with Althea rosea enation mosaic virus (GenBank Accession No. AF014881) and TLCV strain D2 reported from Australia (GenBank Accession No. AF084006), both of which are monopartite begomoviruses. These results establish the provisional identity of the pathogen causing yellow mosaic disease on pumpkin in India as a closely related strain of the monopartite TLCV. The yellow leaf disease of muskmelon in Thailand and watermelon leaf curl disease in Pakistan also were recently reported to be caused by an Indian strain of TLCV (2,3). However, to our knowledge this is the first report of detection of a monopartite geminvirus causing yellow mosaic disease on pumpkin in India. References: (1) D. Deng et al. Ann. Appl. Biol. 125:327, 1994. (2) S. H. Khan et al. Plant Dis. 84: 102, 2000. (3) K. Sarmretwanich, Plant. Dis. 84:707, 2000.