Isolation and Characterization of Baculoviruses from Three Major Lepidopteran Pests in the Semi-Arid Tropics of India

Abstract

Baculoviruses were isolated from three major lepidopteran pests, Helicoverpa armigera, Spodoptera litura and Amsacta albistriga in the semi-arid tropics during natural epizootic conditions at ICRISAT fields, Patancheru, Andhra Pradesh, India. Biological, morphological and biochemical analysis identified these isolates as Nucleopolyhedroviruses (NPVs). Scanning electron microscopy of the occlusion bodies (OBs) purified from diseased larvae revealed polyhedral particles of size approximately 0.5-2.5μm [Helicoverpa armigera Nucleopolyhedrovirus (HearNPV)], 0.9-2.92μm [Spodoptera litura Nucleopolyhedrovirus (SpltNPV)] and 1.0-2.0μm [Amsacta albistriga Nucleopolyhedrovirus (AmalNPV)] in diameter. Transmission electron microscopy of thin sections of OBs of the three isolates revealed up to 5-8 multiple bacilliform shaped particles packaged within a single viral envelope. The dimensions of these particles were 277.7×41.6nm for HearNPV, 285.7×34.2nm for SpltNPV and 228.5×22.8nm for AmalNPV. Each of HearNPV and AmalNPV contained up to 6 nucleocapsids and SpltNPV contained up to 7 nucleocapsids per envelope. The estimated molecular weights of the purified OB (polyhedrin) protein of the three NPVs were 31.29-31.67kDa. Virus yield (OBs/larva) was 5.18±0.45×10(9) for HearNPV, 5.73±0.17×10(9) for SpltNPV and 7.90±0.54×10(9) for AmalNPV. The LC50 values of various NPVs against 2nd and 3rd instar larvae indicated 2.30×10(4) and 1.5×10(5) OBs/ml for HearNPV, 3.5×10(4) and 2.4×10(5) OBs/ml for SpltNPV and 5.6×10(4) and 3.96×10(5) OBs/ml for AmalNPV. The lethal time required to cause 50% mortality (LT50) for these three species were also defined. This study has shown that the NPVs infecting three major lepidopteran pests in India are multiple NPVs, and they have good potential to use as biocontrol agents against these important pests.