Canadian Forest Service Publications

Effects of long-term storage on the stability of OpMNPV DNA contained in TM Biocontrol-1. 2003. Reed, C.; Otvos, I.S.; Reardon, R.; Ragenovich, I.; Williams, H.L. Journal of Invertebrate Pathology 84: 104-113.

Year: 2003

Issued by: Pacific Forestry Centre

Catalog ID: 23600

Language: English

Availability: Order paper copy (free), PDF (download)

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Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV) DNA was extracted from samples representing 10 lots of TM Biocontrol-1 stored at –10°C for 5–15 years and digested with the restriction enzymes BglII, PstI, and SalI. DNA from the OpMNPV virus strain (MEM-75-STANDARD) used to produce the TM Biocontrol-1 lots was also extracted and digested. No restriction fragment length polymorphisms were observed in any of the samples and there was no evidence of DNA degradation. This indicates that long-term cold storage of TM Biocontrol-1 had no adverse effect on the quality of the OpMNPV DNA. In addition to the expected >23 kb OpMNPV DNA, extracts from lots 4a, 5b, and 6 contained 10 additional nucleic acid segments, ranging in size from 0.9 to 4.2 kb. The electrophoretic profile of these segments was characteristic of O. seudotsugata cypovirus (OpCPV). RNase A/DNase I treatment showed that the nucleic acid contaminants were composed of RNA, suggesting that lots 4a, 5b, and 6 contained OpCPV as well as OpMNPV. Bioassay results have shown that there is a decrease in efficacy of stored TM biocontrol-1, but this did not appear to be directly correlated with the length of time in storage.