Wound-induced gene expression of putrescine N-methyltransferase in leaves of Nicotiana tabacum.

Putrescine N-methyltransferase (PMT) catalyzes the first committed step in the biosynthesis of pyrrolinium ring-containing alkaloids. Earlier studies have indicated that PMT gene expression is restricted to root tissue in Solanaceus plant species. During the analysis to further elucidate factors that govern the regulation of alkaloid synthesis, evidence was found for a novel expression pattern dictated by the 5'-flanking region of at least two members of the PMT-gene family. A 627-bp DNA fragment upstream of the NtPMT3 gene was fused to the beta-glucuronidase (GUS) reporter gene and used to produce stable transgenic lines of Nicotiana tabacum. Fluorometric and histochemical assays conducted on transgenic plants indicated high expression levels in root tissue and, in agreement with previous studies, no expression was detected in leaves. However, expression was observed in leaves when they were mechanically wounded. This expression was highly localized around the wound site and showed little evidence of long distance signaling, including lack of responsiveness to jasmonic acid. Expression was transient, with maximum levels immediately after wounding and diminishing after approximately 2-4 h. RT-PCR analysis of mRNA isolated from wild-type plants also indicated upregulation of PMT expression in leaves upon wounding as well as very low transcript levels in unwounded leaves. Low levels of PMT activity were detected in leaf tissue, which did not increase significantly upon wounding.