The Dead Sea is one of the most saline lakes on earth ( approximately 340 g/liter salinity) and is approximately 10 times saltier than the oceans. Eurotium herbariorum, a common fungal species, was isolated from its water. EhHOG gene, encoding a mitogen-activated protein kinase (MAPK) that plays an essential role in the osmoregulatory pathway in yeast and many other eukaryotes, was isolated from E. herbariorum. The deduced amino acid sequences of EhHOG indicated high similarity with homologous genes from Aspergillus nidulans, Saccharomyces cerevisiae, and Schizosaccharomyces pombe and contained a TGY motif for phosphorylation by MAPK kinase. When EhHOG was expressed in S. cerevisiae hog1Delta mutant, the growth and aberrant morphology of hog1Delta mutant was restored under high osmotic stress condition. Moreover, intracellular glycerol content in the transformant increased to a much higher level than that in the mutant during salt-stress situations. hog1Delta mutant complemented by EhHOG outperformed the wild type or had higher genetic fitness under high Li(+) and freezing-thawing conditions. The present study revealed the putative presence of a high-osmolarity glycerol response (HOG) pathway in E. herbariorum and the significance of EhHOG in osmotic regulation, heat stress, freeze stress, and oxidative stress. The Dead Sea is becoming increasingly more saline while the fungi living in it evolutionarily adapt to its high-saline environment, particularly with the extraordinarily high Li(+) concentration. The Dead Sea is potentially an excellent model for studies of evolution under extreme environments and is an important gene pool for future agricultural genetic engineering prospects.