Expanded Supercooling Capacity With No Cryoprotectant Accumulation Underlies Cold Tolerance of the European Grapevine Moth.

The European grapevine moth, Lobesia botrana (Denis et Schiffermuller), is a serious invasive pest that causes significant losses to the flowers and fruits of grapes in most of the world. This multivoltine pest passes the winter as the third-generation diapausing pupa. The current study was designed to compare nondiapausing (first and second generations) and diapausing pupae (third generation) and to investigate the relationship among cold tolerance, the supercooling point (SCP), and diapause development of the third-generation diapausing pupae. The lethal temperatures (LTs) for the three generations were determined using 24-h exposure at subzero temperatures. The mean SCP of the pupae was estimated at approx. -22.6°C, the lowest level of which (-23.7°C) was recorded in the well-developed diapausing pupae in February. The highest level of cold tolerance was also recorded in February. There were no significant differences among the temperatures required to kill 30, 50, and 90% of the pupae. The temperatures significantly decreased from October onward and reached the lowest levels in February during which the lowest SCP and the highest cold tolerance were observed in the diapausing pupae. No significant differences were found in the cryoprotectant levels, among the diapausing and nondiapausing pupae, and the diapause development. The highest activity of cAMP-dependent protein kinase (AMPK) was recorded in the late diapause in February. The findings suggested a relationship among SCP depression, cold tolerance enhancement, and diapause development. A bimodal cold-tolerance strategy (freeze-intolerant and freeze-tolerant) was found to be a feature of the pupae.