Abstract
Simple Summary The alpine insect fauna of Aotearoa/New Zealand is unusual in that many species survive freezing solid at any time of the year. The physiological mechanisms that operate are not well-understood, but we do know the challenges imposed on cells when liquid water expands to form a solid (ice). Insects that are freeze tolerant start freezing at relatively warm temperatures with the help of ice nucleating agents that foster the slow formation of ice crystals outside cells. Here we consider the potential role of microbes living in the guts of alpine Aotearoa/New Zealand insects to facilitate this widespread ability to tolerate freezing. Many bacteria and fungi produce ice-nucleating agents that mediate formation of ice crystals at temperatures just below 0 degrees C. Some encourage ice formation on foliage with subsequent tissue damage, allowing microbial access to plant nutrients and causing crop damage. Other roles for ice-nucleating agents may include more positive outcomes for insects by facilitating controlled freezing at relatively warm temperatures. Insects that are freeze-tolerant start freezing at high sub-zero temperatures and produce small ice crystals. They do this using ice-nucleating agents that facilitate intercellular ice growth and prevent formation of large crystals where they can damage tissues. In Aotearoa/New Zealand the majority of cold adapted invertebrates studied survive freezing at any time of year, with ice formation beginning in the rich microbiome of the gut. Some freeze-tolerant insects are known to host symbiotic bacteria and/or fungi that produce ice-nucleating agents and we speculate that gut microbes of many New Zealand insects may provide ice-nucleating active compounds that moderate freezing. We consider too the possibility that evolutionary disparate freeze-tolerant insect species share gut microbes that are a source of ice-nucleating agents and so we describe potential transmission pathways of shared gut fauna. Despite more than 30 years of research into the freeze-tolerant mechanisms of Southern Hemisphere insects, the role of exogenous ice-nucleating agents has been neglected. Key traits of three New Zealand freeze-tolerant lineages are considered in light of the supercooling point (temperature of ice crystal formation) of microbial ice-nucleating particles, the initiation site of freezing, and the implications for invertebrate parasites. We outline approaches that could be used to investigate potential sources of ice-nucleating agents in freeze-tolerant insects and the tools employed to study insect microbiomes.