Multiplication of a granulosis virus isolated from the potato tuber moth in a new established cell line of Phthorimaea operculella

Abstract

A newly established cell line was obtained from the culture of embryonic cells of the potato tuber moth Phthorimaea operculella in low temperature conditions (19 degrees C) using modified Grace's medium supplemented with 10% fetal bovine serum. The population doubling time was about 80 h when cells were cultivated at 19 degrees C and 38 h at 27 degrees C. The cell line had a relatively homogeneous population consisting of various sized spherical cells. The cells were cultivated for more than 25 passages. Their polypeptidic profile was different from profiles of other P. operculella cell lines we previously described and from other lepidopteran cells. The new cell line was designated ORS-Pop-95. The complete replication of the potato tuber moth granulosis virus (PTM GV) was obtained in vitro by both viral infection and DNA transfection. PTM GV multiplied at a significant level during several passages of the cell line that was maintained at 19 degrees C. As long as the cells were maintained at 19 degrees C, virus multiplication could also be obtained at the same rate at 27 degrees C. To compare PTM GV multiplied both in vivo and in vitro, we used morphological identification, serological, DNA probe diagnosis and endonuclease digest profile analysis and confirmed the identity of the virus.

The Potato Tuberworm: A Literature Review of Its Biology, Ecology, and Control

“…Microbial Control Insect diseases caused by bacteria, viruses, and nematodes have been developed as microbial pesticides to control insect pests not only in the field but also in the storage (Reed 1969(Reed , 1971Reed and Springett 1971;Hunter 1973;Briese and Mende 1983;Alcázar et al 1992;BenSalah and Aalbu 1992;Das et al 1992;Kroschel et al 1996b;Lery et al 1997;Roux and Baumgartner 1998;Setiawati et al 1999). However, microbial control of P. operculella is not yet developed for massive commercial use although some authors have indicated the potential use of those pathogens in the future (Lacey and Arthurs 2005;Arthurs et al 2008).…”

Baculovirus host-range

“…Studies into these processes are facilitated by the availability of cultured cells that support the entire life cycle of the virus. For baculoviruses robust replication in cell culture has been achieved for NPVs infecting Lepidoptera (39,54,116,121) and to a limited extent for GVs (40,77,98,133). (30,32).…”

Untitled

“…Six established lepidopteran cell lines were used in this study. Two cell lines obtained from embryonic cells of P. operculella, ORS-Pop-93 (Po93) (Léry et al, 1995) and ORS-Pop-95 (Po95) (Léry et al, 1997) were cultured in modified Grace's medium (Léry and Fédière, 1990) containing 10% fetal bovine serum (FBS). Po93 was maintained at 27 • C, and was routinely subcultured every 5 to 7 days when 1 to 2 × 10 6 cells were seeded in each 25 cm 2 flask.…”