In this study, yeasts were isolated from Macadamia ternifolia honey by traditional culture methods, identified by morphological observation and molecular biological technology, and its growth characteristics were studied. The results showed that 6 strains with ester flavor were isolated from 3 M. ternifolia honey samples, numbered as JG01-8, JG01-15, JG01-H4, JG02-9, JG02-11 and JG03-1, respectively. After identification, the strain JG01-8 was identified as Hannaella luteola, the strain JG01-H4 was identified as Sporobolomyces carnicolor, and the strains JG01-15、JG02-9、JG02-11 and JG03-1 were identified as Zygosaccharomyces rouxii, which were all non-Saccharomyces yeasts. The tolerance of strain JG01-H4 was better than others, which could tolerate ethanol volume fraction 9%, pH 2.0, NaCl content 25% and glucose content 60%, and it was potential strain for fermentation in high permeability environment.