Abstract:The present study was conducted to identify the alternate hosts of new leaf curl virus disease of sunflower. In the present study several crops and weed hosts were cross inoculated with leaf curl virus of sunflower under laboratory through insect vector whitefly (Bemisia tabaci), further all inoculated samples were retested (3-4 weeks after inoculation) by molecular based Polymerse chain reaction diagnosis for the presence of virus. The results revealed that the causal virus of the disease was successfully transmitted from sunflower to sunflower (Helianthus annuus), tomato (Solanum lycopersicum) and tobacco (Nicotiana tabacum L) and weed hosts such as Acanthospermum hispidum, Amaranthus viridis and Parthenium hysterophorus in a short incubation period (2-3 weeks after inoculation), while on other hosts Chilli (Capsicum annuum L) and Datura stramonium, infection occurs in delayed incubation period. Further molecular analysis thorough polymerase chain reaction (PCR) diagnostic technique using virus specific primers also confirmed the presence of coat protein (CP) of leaf curl begomovirus in virus inoculated hosts viz., chilli, sunflower, tomato, and tobacco and weed hosts such as Acanthospermum hispidum, Amaranthus viridis, Datura stramonium and Parthenium hysterophorus. Thus, findings substantiate that the above hosts are major sources of the virus inoculum and served as potential alternate hosts of the disease during the off season.