E.urophylla×E.grandis seedlings were inoculated with 10 strains of eucalyptus endophytic fungi by irrigating the fungus suspension, and then subjected to low temperature stress 30 days later. The results showed that compared with the control the soluble sugar (SS) content increased by 41.11%, 11.11%, 32.78%, 38.89% and 6.67% in treatment 1,8,5,7,10, respectively. The soluble protein (SP) content was 14.90%, 0.72%, 10.94%, 3.37%, 24.40% and 0.36% higher in treatment 1,2,5,6,7,10 than that in the control. The proline content was 28.77%,22.11%,20.16%,11.25%,5.48% and 1.57% higher in treatment 5,7,1,2,4,9 than that in the control. The chlorophyll content was 9.52%,7.48%,7.48%,6.80%,5.44%,3.40%,3.40%,2.72%,2.04% and 2.04% higher in treatment 6,8,10,1,9,5,7,4,2,3 than that in the control. After being inoculated with strain 1 of the fungus, the cold-resistant capability of eucalyptus was much improved in terms of nutrition characteristics. However, the more physiological characteristics may be needed to determine whether the fungus can be applied in production.