The polyextremophile adopts a dual adaptive strategy to long-term salinity stress, simultaneously accumulating compatible solutes and K.

The bacterium is an extremely halophilic alkalithermophile that can thrive under conditions of high salinity (3.3-3.9 M Na), alkaline pH (9.5), and elevated temperature (53°C). To understand the molecular mechanisms of salt adaptation in , it is essential to investigate the protein, mRNA, and key metabolite levels on a molecular basis. Based on proteome profiling of under 3.1, 3.7, and 4.3 M Na conditions compared to 2.5 M Na condition, we discovered that a hybrid strategy, combining the "compatible solute" and "salt-in" mechanisms, was utilized for osmotic adjustment dur ing the long-term salinity adaptation of . The mRNA level of key proteins and the intracellular content of compatible solutes and K support this conclusion. Specifically, employs the glycine betaine ABC transporters (Opu and ProU families), Na/solute symporters (SSS family), and glutamate and proline synthesis pathways to adapt to high salinity. The intracellular content of compatible solutes, including glycine betaine, glutamate, and proline, increases with rising salinity levels in . Additionally, the upregulation of Na/ K/ H transporters facilitates the maintenance of intracellular K concentration, ensuring cellular ion homeostasis under varying salinities. Furthermore, exhibits cytoplasmic acidification in response to high Na concentrations. The median isoelectric points of the upregulated proteins decrease with increasing salinity. Amino acid metabolism, carbohydrate and energy metabolism, membrane transport, and bacterial chemotaxis activities contribute to the adaptability of under high salt stress. This study provides new data that support further elucidating the complex adaptation mechanisms of under multiple extremes.IMPORTANCEThis study represents the first report of simultaneous utilization of two salt adaptation mechanisms within the class in response to long-term salinity stress.