Chimeric constructs containing the promoter and upstream sequences of Ha hsp17.6 G1, a small heat shock protein gene, reproduced in transgenic tobacco (Nicotiana tabacum) its unique seed-specific expression patterns previously reported in sunflower. These constructs did not respond to heat shock, but were expressed without exogenous stress during late zygotic embryogenesis coincident with seed desiccation. Site-directed mutagenesis of its distal and imperfect heat shock element strongly impaired in vitro heat shock transcription factor binding and transgene expression in seeds. Deletion analyses of upstream sequences indicated the contribution of additional cis-acting elements with either positive or negative effects on transgene expression. These results show differences in the transcriptional activation through the heat shock element of small heat shock protein gene promoters in seeds compared with the heat shock response. In addition, they suggest that heat shock transcription factors and other distinct trans-acting factors cooperate in the regulation of Ha hsp17.6 G1 during seed desiccation.This work was funded by the Spanish Comisión Interministerial de Ciencia y Tecnología (grant no. BIO96–0474) and by Junta de Andalucía (grant no. CVI148).Peer reviewed