Effects of long-term storage on the stability of OpMNPV DNA contained in TM Biocontrol-1.
Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV) DNA was extracted from samples representing 10 lots of TM Biocontrol-1 stored at -10 degrees 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. pseudotsugata 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.
Reed
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Natural Resources Canada, Pacific Forestry Centre, Victoria, BC, Canada V8Z 1M5.
Otvos
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Reardon
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Ragenovich
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Williams
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D000818:Animals; D001681:Biological Assay; D015139:Blotting, Southern; D004279:DNA, Viral / Q000032:analysis*; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D010572:Pest Control, Biological / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D013048:Specimen Handling / Q000379:methods; D013997:Time Factors
D004279:DNA, Viral