Graduate Thesis Or Dissertation
 

Plagiogravitropic growth in rooted stem cuttings of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) related to indole-3-acetic acid and cytokinins

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  • Endogenous concentrations of indole-3-acetic acid (IAA) and cytokinins were compared to the tendency for plagiogravitropjc growth in rooted stein cuttings of Douglas-fir. A chromatographic method was developed for the purification and quantitation of trace levels of IAA. Two elutions with reverse-phase high performance liquid chromatography (HPLC) using a fluorescence detector resulted in a recovery of about 50%. The IAA peak was confirmed by gas chromatography/mass-spectrometry. Affinity chromatography using monoclonal antibodies raised against zeatjn rjbosjde and isopentenyladenosine, combined with HPLC and radioimmunoassay were used here for quantitatjon of cytokinins. Buds, apical needles and stems of branch terminals of eight year-old clones (clone 12-plagiogravitropic growth and clone 15-orthogravitropic growth), collections for quantitation of IAA and cytokinins were made in September, December, January, and February. The highest concentration of IAA in both clones was found in buds collected in December. Differences in IAA concentration between clones from samples collected in January and February were not significant. Significant differences were found in September and December. A high concentration of a cytokinin conjugate which was determined to be a hexose conjugate of zeatin riboside (ZX) but not zeatin ribose-O-glucoside, was found in the fall and winter buds. Some significant differences in cytokinin concentrations were found between these two clones. The interaction of ZX and ZR in buds on the four collection dates could indicate a role for ZX in the plagiogravitropic mechanism. Clonal differences in IAA concentrations between the morphological upper and lower halves of stem sections of rooted cuttings were not significant. The highest concentration of IAA was detected in the section of stem sampled immediately above the stem area exhibiting the most plagiogravitropjc curvature. The failure to find differences of IAA concentrations between morphological upper and lower halves of curvature did not support the related Cholodny-Went hypothesis that higher concentration of IAA was on the upper half. However, there is some evidence to suggest that lateral distribution of IAA could be occurring in rooted cuttings and that compression wood might be associated with this asymmetrical distribution of IAA. The curvature of rooted cuttings may also be directly affected by root growth potential. Higher concentration of endogenous IAA in December bud may be linked to high mortality of seedlings lifted and outplanted before December. The high levels of ZX in fall buds may be an inactive, storage form of biologically active zeatin riboside.
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