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ARS Home » Midwest Area » Columbia, Missouri » Biological Control of Insects Research » Research » Publications at this Location » Publication #66497


item Grasela, James
item McIntosh, Arthur

Submitted to: In Vitro Cellular And Developmental Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/3/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary: The objective of this study was to determine the ability of a baculovirus of the velvetbean caterpillar, Anticarsia gemmatalis, to infect seven established insect cell lines. This has important implications in the development of an optimal insect cell line for the commercial production and use of this virus as a biological insecticide. Tobacco budworm cells produced the highest virus level five days after infection with the virus. The New World bollworm and the velvetbean caterpillar cell lines also showed large virus levels five days after infection. In the cabbage looper cell line the virus concentration was detected only at low levels. However, the virus was unable to replicate in cotton boll weevil cells. A nonradioactive DNA probe confirmed the above different virus levels in the seven cell lines. The results of this study are important since it further defines the range in which this baculovirus is able to infect other insect cells.

Technical Abstract: A clone of Anticarsia gemmatalis multiple nuclear polyhedrosis virus AgMNPV, derived from a wildtype isolate (Hondrina, Brazil) and designated AgMNPV-CL4-3A1, was used to determine the host range of this virus in six established lepidopteran cell lines: Anticarsia gemmatalis (BCIRL-AG-AM1), Helicoverpa zea (BCIRL-HZ-AM1), Heliothis virescens (BCIRL-HV-AM1), Helicoverpa armigera (BCIRL-HA-AM1), Trichoplusia ni (TN-CL1), Bombyx mori (BMN), and a coleopteran cell line Anthonomus grandis (BRL-AG-1). Based on temporal studies of TCID-50 values, BCIRL-HV-AM1 cells gave the highest extracellular virus (ECV) titer (9.69 x 10**6 TCID-50/ml) at 120 h postinoculation followed by BCIRL-HA-AM1 cells (7.59 x 10**5 TCID-50/ml) at 96 h postinoculation and BCIRL-AG-AM1 cells (2.78 x 10**5 TCID-50/ml) at 120 h postinoculation. In addition, a low ECV titer of 1.98 x 10**3 TCID-50/ml was detected from TN-CL1 cells 96 hr postinoculation. Using a random primed digoxigenin-dUTP labeled DNA probe of the clone, dot blot hybridization studies confirmed the above findings on extracellular virus production in cell lines. Dot-blot hybridization also detected the presence of viral DNA in BMN and BCIRL-HZ-AM1 cells which were otherwise refractile to virus as determined by TCID-50. BRL-AG-1 cells were nonpermissive to AgMNPV-CL4-3A1 based on both TCID-50 and hybridization results. AgMNPV-CL4-3A1 and the wildtype AgMNPV had similar restriction profiles that were different from AcMNPV.