Effects of the Indole-3-Acetic Acid (IAA) Transport Inhibitors N-1-Naphthylphthalamic Acid and Morphactin on Endogenous IAA Dynamics in Relation to Compression Wood Formation in 1-Year-Old Pinus sylvestris (L.) Shoots

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Abstract
Both N-1-naphthylphthalamic acid (NPA) and methyl-2-chloro-9-hydroxyfluorene-9-carboxylic acid (CF) inhibit the polar transport of indole-3-acetic acid (IAA) and, therefore, are attractive tools for investigating IAA9s role in the regulation of plant growth. Ringing an intact conifer shoot with lanolin containing NPA or CF induces the formation of compression wood above the ring. This induction has been attributed to a postulated accumulation of IAA above the application site of the IAA transport inhibitor, but the validity of this postulation has never been confirmed. Using gas chromatography-selected ion monitoring-mass spectroscopy with [13C6]IAA as an internal standard, we measured the levels of endogenous free and conjugated IAA in 1-year-old Pinus sylvestris (L.) shoots ringed with NPA or CF. The level of free IAA was dramatically decreased below the ring, indicating that the polar transport of endogenous IAA was inhibited by the treatment. However, the free IAA level above the ring, where compression wood was formed, was also slightly lower than in control shoots. The lack of IAA accumulation above the site of the IAA transport inhibitor could not be explained by an increase in IAA conjugation. Furthermore, the turnover of [2–14C]IAA, measured using high-performance liquid chromatography with on-line radioactivity monitoring, was the same in NPA-treated and control shoots. The decrease in IAA level above a NPA or CF ring is attributed to these substances being transported acropetally and interfering with polar IAA transport along the shoot. It is concluded that compression wood formation above a NPA or CF ring is not associated with an overall increase in cambial region IAA level or increased IAA turnover. Instead, we suggest that acropetally transported NPA and CF induce compression wood formation by interacting with the NPA receptor in differentiating tracheids, thereby locally increasing IAA in these cells.