Nematodes are the dominant soil animals of the Antarctic Dry Valleys and are capable of surviving desiccation and freezing in an anhydrobiotic state. Genes induced by desiccation stress have been successfully enumerated in nematodes; however, little is known about gene regulation by Antarctic nematodes that can survive multiple types and incidences of environmental stress. In order to reveal the molecular response of anhydrobiotic survival, we investigated the genetic response of an Antarctic nematode, Plectus murrayi, which can survive desiccation and freezing. Using molecular approaches, we provide insight into the regulation of desiccation-induced transcripts during different stages of stress survival under conditions characteristic of the Antarctic Dry Valley environment. Our study revealed that exposure to slow desiccation and freezing plays an important role in the transcription of stress, metabolism and signal transduction-related genes and improves desiccation and freezing survival compared with nematodes exposed to fast desiccation and freezing. Temporal analyses of gene expression showed that pre-exposure to mild stress promotes survival of harsher stress. Our results further show that exposure to slow dehydration not only improves extreme desiccation survival but also promotes enhanced cold tolerance. We also provide evidence that slow dehydration can enhance freeze tolerance in an Antarctic nematode. Expression profiling of P. murrayi transcripts is an important step in understanding the genome-level response of this nematode to different environmental stressors.