Gene Induction of Stilbene Biosynthesis in Scots Pine in Response to Ozone Treatment, Wounding, and Fungal Infection

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Abstract
TheS-adenosyl-l-methionine:pinosylvin-O-methyltransferase (PMT)2 gene was sequenced from Scots pine (Pinus sylvestris). The open reading frame is arranged in two exons spaced by one 102-bp intron. Promoter regulatory elements such as two “CAAT” boxes and one “TATA” box were identified. Several cis-regulatory elements were recognized: stress-responsive elements (Myb-responsive elements) as well as G, H, and GC boxes. Moreover, elicitor-responsive elements (W boxes) and a sequence resembling the simian virus 40 enhancer core were found. In phloem and needles of control trees, the transcripts of stilbene synthase (STS) and PMT were hardly detectable. Increased ozone fumigation up to 0.3 μL L−1 enhanced the transcript level of STSand PMT in needles but not in healthy phloem. Wounding, e.g. mock inoculation, of stem-phloem was characterized by a transient increase in STS and PMT transcripts, which was more pronounced in the case of fungal inoculation. Combination of fungal-challenge or mock treatment with ozone resulted in a positive interaction at 0.3 μL L−1. Scots pine stilbene formation appeared to be induced via STS andPMT gene expression upon ozone and fungal stress as well as wounding. The broad stress-responsiveness is in agreement with the range of various cis-acting elements detected in the STSand PMT promoters.