Purification of Tomato leaf curl Bangalore virus and production of polyclonal antibodies

Tomato leaf curl Bangalore virus (ToLCBV) was successfully transmitted using viruliferous whiteflies (Bemisia tabaci) to several tomato varieties and hybrids, viz. Arka vikas, Pusa ruby, Rashmi, Rakshitha and Swaraksha. Hybrid Rashmi took 100% uniform infection, which was selected for further propagation of the virus. ToLCBV was also transmitted to Nicotiana benthamiana; the rate of infection was 30-40% in spite of two inoculations with 10-15 viruliferous B. tabaci/plant. The virus was purified and polyclonal antisera were produced in rabbits by injecting purified native virus and recombinant coat protein (rCP) of ToLCBV. The antisera of ToLCBV and ToLCBV rCP had a titre of 1:8000 and 1:400 respectively, when tested with infected leaf extract of tomato. Both sources of antisera successfully detected begomovirus infections in reservoir host plants, viz. Parthenium hysterophorus, Acanthospermum hispidum, Ageratum conyzoides, and fieldcollected tomato samples and cross-reacted with hibiscus leaf curl in DAC-ELSIA