Induction and expression of mutations in mammalian cells in the absence of DNA synthesis and cell division

The induction of mutations by the alkylating agent ethyl methanesulfonate (EMS) was determined with Chinese hamster ovary cells maintained in serum-free medium to arrest DNA synthesis and cell division. The arrested cultures were treated with EMS and maintained in serum-free medium for various time intervals post-treatment before serum containing medium was added to initiate DNA synthesis and cell division. The concentration-dependent increase in 6-thioguanine-resistant mutants in the arrested cultures was similar to that found with exponentially dividing cultures when serum was added to the arrested cultures immediately after the EMS treatment; the time course of phenotypic expression was also similar with both cultures. In addition, maintenance of the arrested cultures in serum-free medium for up to 18 days post-treatment resulted in no change in the mutant frequency. This suggests that the mutagenic damage is not removed in these arrested cultures. Furthermore, maintenance of the arrested state for increasing time intervals before serum addition results in decreases in the time necessary for maximum phenotypic expression. Cultures maintained in serum-free medium for 16 days after mutation treatment show complete expression of the mutations with no need for subculture. This last result suggests that the mutagenic damage induced by EMS in Chinese hamster ovary cells is not removed and that this damage results in both the induction and expression of mutation in the absence of DNA replication.     

Selection for pesticide resistance in Aphytis

Field-collected populations of Aphytis holoxanthus DeBach (Hymenoptera: Aphelinidae), a parasite of Chrysomphalus aonidum (L.) (Homoptera: Diaspididae), were tested for malathion and for azinphosmethyl tolerance, and selected for increased resistance. Initial tolerance for malathion was not significantly different between populations, and could not be significantly increased by 30 selection cycles. Two field populations showed relatively high levels of azinphosmethyl tolerance, which were further increased by selection to 24–48 times that of a susceptible laboratory population.The differences in response to the two organophosphorus insecticides are discussed in relation to differences in their modes of application, detoxification by insects and the genetic bottleneck experienced by an imported species.