The transcriptome data of Bursaphelenchus xylophilus (B. xylophilus) at –5 ℃ was analyzed, and the low temperature induced differentially expressed gene Bx-daf-2 was screened out. The co-expressed genes of low- temperature-stressed transcriptome and Bx-daf-2 RNAi-treated transcriptome were screened, and it was found that Bx-tre-1 was regulated by Bx-daf-2. The function of Bx-tre-1 in B. xylophilus under low temperature stress was researched further. The results showed that Bx-tre-1 involved in the response of low temperature stress in B. xylophilus and its expression down-regulated, which caused the activity of Bx-TRE enzyme to decrease, and then the trehalose content increased. Under low temperature stress, the Bx-TRE enzyme activity of Bx-tre-1 RNAi group decreased, trehalose content and the survival rate increased, indicating that the increase of trehalose content could contribute to the improvement of the survival rate of B. xylophilus under low temperature stress. The above results indicated that Bx-daf-2 and Bx-tre-1 coordinately regulate the increase of trehalose content, thus promote the resistance of B. xylophilus against low temperature.