Great progress has recently been made in cryobiology. One field, however, has been neglected: the temporal sequence of the effects of photoperiod and temperature, and their relative importance in cold hardening. This is relevant to the question of importance of diapause in cold-hardiness. Denlinger (1991) outlined the categories of such relations and stressed a great need for further detailed research. A survey of studies done over the past decade revealed many gaps in the evidence and the ambiguous nature of the data on the photoperiodic regulation of cold-hardiness. We hope that this review will stimulate further research in this field. Among several directions where research is most needed we have stressed (1) simultaneous recording of changes in survival and dynamics of suspected cryoprotectants (stressed also by Danks, 1996), (2) checking the regulation of different phases of cold hardening, and (3) discrimination between direct and indirect (mediated via neuroendocrine system) effects of environmental cues on cold hardening.
The aim of this study was to investigate the relationship between antioxidant enzymes and reactive oxygen species production in diapausing larvae of the European corn borer, Ostrinia nubilalis (Lepidoptera: Pyralidae) kept at 5°C, -3°C and -16°C for two weeks. The amount of hydrogen peroxide (H2O2), activity of antioxidant enzymes, copper zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutases (MnSOD) and catalase (CAT) in whole body homogenates, as well as the electron paramagnetic resonance (EPR) spectroscopy of this insect's whole body were analysed. A higher level of melanin radical and lower CuZnSOD and CAT activities were found in larvae kept at -3°C than at 5°C and -16°C. At the same temperature (-3°C) an elevated H2O2 concentration was recorded. A possible regulatory role of H2O2 at -3°C, which is the temperature that triggers freezing tolerance, is suggested.