Bark storage proteins (BSPs) accumulate in the inner bark parenchyma of many woody plants during autumn and winter. We investigated the effect of a short-day (SD) photoperiod on the accumulation of the 32-kilodalton bark storage protein of poplar (Populus deltoides Bart. ex Marsh.) under controlled environmental and natural growing conditions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein gel blot analysis revealed that 10 days of SD exposure (8 hours of light) resulted in a 20% increase in the relative abundance of the 32-kilodalton bark storage protein of poplar. After 17 days of SD exposure, the 32-kilodalton bark storage protein accounted for nearly one-half of the soluble bark proteins. In natural field conditions, accumulation of the 32-kilodalton bark storage protein was observed to start by August 18 (daylength 14.1 hours). Immunoprecipitation of in vitro translation products with anti-BSP serum revealed that the SD protein accumulation was correlated with changes in the pool of translatable mRNA. A survey of poplar clones from different geographic origins revealed the presence of the 32-kilodalton BSP in the dormant bark of all the clones tested. These results demonstrate that a SD photoperiod induces, whether directly or indirectly, rapid changes in woody plant gene expression, leading to the accumulation of BSP.
Treatment differences were observed in the in vitro adventitious shoot regeneration response from internodal explants of three genotypes of Populus deltoides cultured on media supplemented with five concentrations each of the cytokinins 6-benzyladenine, 2-isopentyladenine, and zeatin. For each of the three genotypes, the greatest number of shoots were consistently regenerated on media containing the cytokinin zeatin. Tissue necrosis resulted when explants from any of the three genotypes were cultured on media supplemented with 6-benzyladenine. A zeatin concentration by genotype interaction was also observed. Genotypic differences in shoot regeneration were observed for 16 genotypes of Populus deltoides when cultured on medium supplemented with 0.5 mgL(-1) zeatin. Six genotypes were highly recalcitrant and failed to regenerate shoots. The percent of explants regenerating was greater than 50% for four genotypes.
In a partial diallel mating design among 20 blue and 20 Engelmann spruce parents, the interspecific crosses were successful only with Engelmann spruce as the female parent. No viable seed were obtained from the reciprocal cross among the 60 full-sib families attempted. Under the conditions of artificial pollination and a controlled germination environment, an average of 0.3% of the seed germinated on a total seed basis across all 20 Engelmann spruce females. Many abnormalities were observed among the germinating hybrid seed, suggesting hybrid inviability also contributes to the low crossability between these two species. Isozyme analysis was used to confirm the interspecific hybrids between blue and Engelmann spruce based on the unique genotypic compositions of the hybrids relative to the two species. No natural F1 hybrids between blue and Engelmann spruce were observed in this study based on isozyme analysis of mature individuals or their seedling progeny. Analyses included samples of open-pollinated seed from blue and Engelmann spruce females located in an area where both species are present in close proximity, often side-by-side, and where pollen shed and female strobilus receptivity in the two species are coincident. In addition, there was evidence of possible gametic selection or hybrid inviability among the full-sib progeny based on deviations of observed from expected segregation ratios for progeny isozyme genotypes. Deviations tended to favor the allele more common to both species rather than the allele unique or more common to only one species. Also, the elevationally allopatric blue and Engelmann spruce subpopulations were less divergent genetically than the sympatric subpopulations.
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