Cultivable halotolerant ice-nucleating bacteria and fungi in coastal precipitation

Ice-nucleating particles (INPs) represent a rare subset of aerosol particles that initiate cloud droplet freezing at temperatures above the homogenous freezing point of water (-38 degrees C). Considering that the ocean covers 71 % of the Earth's surface and represents a large potential source of INPs, it is imperative that the identities, properties and relative emissions of ocean INPs become better understood. However, the specific underlying drivers of marine INP emissions remain largely unknown due to limited observations and the challenges associated with isolating rare INPs. By generating isolated nascent sea spray aerosol (SSA) over a range of biological conditions, mesocosm studies have shown that marine microbes can contribute to INPs. Here, we identify 14 (30 %) cultivable halotolerant ice-nucleating microbes and fungi among 47 total isolates recovered from precipitation and aerosol samples collected in coastal air in southern California. Ice-nucleating (IN) isolates collected in coastal air were nucleated ice from extremely warm to moderate freezing temperatures (-2.3 to -18 degrees C). While some Gammaproteobacteria and fungi are known to nucleate ice at temperatures as high as -2 degrees C, Brevibacterium sp. is the first Actinobacteria found to be capable of ice nucleation at a relatively high freezing temperature (-2.3 degrees C). Air mass trajectory analysis demonstrates that marine aerosol sources were dominant during all sampling periods, and phylogenetic analysis indicates that at least 2 of the 14 IN isolates are closely related to marine taxa. Moreover, results from cell-washing experiments demonstrate that most IN isolates maintained freezing activity in the absence of nutrients and cell growth media. This study supports previous studies that implicated microbes as a potential source of marine INPs, and it additionally demonstrates links between precipitation, marine aerosol and IN microbes.