Based on the latex transcriptome date, a cDNA sequence of rapid alkalinization factor (RALF) family gene was identified by RT–PCR. The cDNA is 767 bp in size with a 411 bp open reading frame and encodes 136 amino acids. BLASTX searches of GenBank showed that the deduced amino acid sequences of RALF gene was similar to AtRALF34，thus was named as HbRALF34. Expression profiles of HbRALF34 in different tissues of Hevea brasiliensis clone Reyan 7–33–97 and in response to ethylene (ET), jasmonic acid (JA), wounding, hydrogen peroxide (H2O2), high salt, low temperature and drought were assayed via real–time quantitative PCR (RT–qPCR). The results suggested that HbRALF34 was ubiquitously expressed in the leaves, flowers, barks and latex of rubber tree, and its highest expression was in latex. Compared with healthy rubber tree, the expression level of HbRALF34 was increased in latex and bark of tapping panel dryness rubber tree. Moreover, the expression of HbRALF34 was induced by ET, JA, H2O2 and wounding treatments, but was inhibited by high salt stress, low temperature and drought treatments. All these results suggest that HbRALF34 is likely involved in signal transduction, regulating rubber biosynthesis and latex flow, as well as responding to stress in Hevea brasiliensis.