A cDNA clone corresponding to a transcript that was induced by auxin in tomato seedling roots encodes a 223 amino acid protein belonging to the proteinase inhibitor II family of tomato and potato. The TR8 open reading frame has an N-terminal region with characteristics of a signal peptide and three copies of a 64 amino acid segment that is also present in multiple copies in other PI-II family members. Comparison to other serine proteinase inhibitors indicates that all three domains are trypsin-specific. Transcripts homologous to TR8 increased in abundance within 24 h of auxin treatment and continued to increase through 72 h, at which point the induction was approximately ten-fold. Auxin induction was observed in roots and hypocotyls, but not in cotyledons or epicotyls. Induction may therefore be correlated with lateral and adventitious root initiation, which occurs over a similar time frame in the same tissues. RFLPs detected with the TR8 probe were mapped to the long arm of chromosome 11.