【Background】Tetrastichus brontispae (Ferriere), is a parasitoid that progressively improved the prevention of damage caused by the Brontispa longissima (Gestro). Its cold tolerance plays an important role in its population establishment and control efficiency. Thus, exploring the tolerance genes of this pest induced by cold temperatures could provide important clues for the study of cold tolerance mechanism of this parasitoid.【Method】In this study, cDNA libraries was constructed using forward and reverse suppression subtractive hybridization (SSH). Here, cDNA from cold-acclimated insects were used as testers against the cDNA of noneacclimated insects used as drivers and vice versa, cold-acclimated insects as drivers with none-acclimated insects as testers. Identification of positive differential fragments by white-blue plague selection and those confirmed clones were randomly selected for sequencing analysis, of which the results were aligned with GenBank database via BLASTX programme. 【Result】The specific cDNA subtractive library of T.brontispae under low temperature was successfully constructed. The data showed that 40 and 4 high quality ESTs defined from the forward and reverse suppression subtractive libraries were selected of which the length for the inserted sequences varied between 200 bp and 700 bp. The 44 high quality ESTs of subject were highly homogenous with that of other known species and the main function of these ESTs involved in metabolism, cell structure, signal transduction, amino and protein synthesis. We speculated that there were at least 5 different candidate genes that might be related to the cold tolerance of T.brontispae, including HSP proteins, trehalose-phosphate synthase, glutamine synthetase, ATP synthase subunit beta, and NADH dehydrogenase (ubiquinone).【Conclusion and significance】This study is important for future research on the expression of cold hardness of T.Brontispae, and lays some theoretical basis on the molecular mechanism of cold hardness of T.brontispae.