Abstract: Suppression subtractive hybridization (SSH) was utilized for the isolation of cDNA fragments for Euonymus japonicus 'Zhuzi' differentially expressed genes, and forward suppression subtractive cDNA Library of cold-regulated gene was constructed. The seedlings of Euonymus japonicus 'Zhuzi' were treated with low temperature as tester and untreated seedlings as driver. Subtractive cDNA Library was differently screened through cDNA macroarray, six hundreds and four cDNA clones were identified as cold specifically induced or highly expressed. After sequencing of 84 cDNA clones and removing redundant cDNAs, 36 cold-regulated unique cDNA clones were obtained. The results showed that 12 cDNA clones are expected to be novel genes, because no sequence homology with any known sequences was found in GenBank databases. This indicated that it is possible to use SSH and cDNA macroarray in the isolation of cold-regulated genes.

Key words: Euonymus japonicus 'Zhuzi', Cold-induced gene, Suppression subtractive hybridization (SSH), cDNA macroarray