Molecular detection of Tomato leaf curl New Delhi virus on Momordica charantia Medicinal Plants, 2(2) May 2010 117 Molecular detection and identification of Tomato leaf curl New Delhi virus isolate causing yellow mosaic disease in Bitter gourd (Momordica charantia), a medicinally important plant in India A.K. Tiwari 1 , P.K. Sharma 2 , M.S. Khan 3 , S.K. Snehi 3 , S.K. Raj 3 and G.P. Rao 1 1 Sugarcane Research Station, Kunraghat, Gorakhpur 273 008, U.P., India 2 Department of Microbiology, Ch. C.S. University Meerut, U.P., India 3 Division of Virology, NBRI, Rana Pratap Marg, Lucknow 226 001, U.P., India ABSTRACT Bitter gourd (Momordica charantia) is extensively cultivated in India, China and South East Asia for vegetable and traditional medicines. A severe yellow mosaic disease on bitter gourd (M. charantia) with a significant disease incidence was observed during the survey of different locations of Eastern U.P., India in the year 2007. The disease consisted of yellow mosaic accompanied with slight curling on leaves. The whitefly (Bemisia tabaci ) population was also observed in the vicinity. The characteristic disease symptoms and whitefly population indicated the possibility of begomovirus infection. PCR was carried out using the total DNA isolated from infected leaf samples and a pair of begomovirus specific primers which resulted in the expected size (~800 bp) amplicon in 3/6 samples indicating presence of begomovirus. For further identification of the begomovirus, the PCR amplicons were cloned and sequenced (GenBank accession no. EU439260). The sequence data analysis revealed highest 98-93% similarities with several isolates of Tomato leaf curl New Delhi virus (ToLCNDV) at both nucleotide and amino acid levels. The phylogenetic analysis also showed closest relationships of the isolate (EU439260) with various variant of ToLCNDV. Based on highest sequence similarities and closest relationships with ToLCNDV the virus isolated from bitter gourd was considered as an isolate of Tomato leaf curl New Delhi virus. [Medicinal Plants 2010; 2(2) : 117-123] . Keywords : Momordica charantia, leaf curl disease, PCR detection, sequence similarities, phylogenetic analysis, Tomato leaf curl New Delhi virus. INTRODUCTION Bitter gourd (Momordica charantia L.) of the family Cucurbitaceae, also known as bitter melon is extensively cultivated in north eastern region of Uttar Pradesh (U.P.) India (Yadav et al., 2004). It is regarded as one of the world’s major vegetable crops and has great economic importance. Bitter gourds are very low in calories but with dense precious nutrients. It is an excellent source of vitamins B 1 , B 2 , and B 3 , C, magnesium, folic acid, zinc, phosphorus, manganese, and has high dietary Corresponding author : A.K. Tiwari E-mail : ajay_biotech2005@rediffmail.com fiber. It is rich in iron, contains twice the beta-carotene of broccoli, twice the calcium of spinach, and twice the potassium of a banana. In India, the plant is used in a traditional medicine wherever it is found. It is often used to treat diabetes (Halberstein and Saunders, 1978; Mossa, 1985; Zhang, 1992) as it contains insulin-like compound known as polypeptide p which has been suggested as insulin replacement in diabetic patients. Leaves, fruits and roots are also used to treat fever (Halberstein and Saunders, 1978; Singh, 1986; Gir´on et al ., 1991). The bitter gourd is also been used in reproductive health as an abortifacient, birth control agent, or to treat painful menstruation and to facilitate childbirth (West et al ., 1981; Mossa, 1985). The antiviral activity of M. charantia is one of the important Research Article