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.

Plants differ in tissue localization of nitrate reduction and assimilation. Some species reduce nitrate primarily in the leaves, whereas other species localize nitrate reduction and assimilation in the roots. We determined how nitrate assimilation is partitioned among leaves, stems and roots of poplar (Populus tremula L. × P. alba L.) by comparing tissue differences in in vivo nitrate reductase activity (NRA), nitrate reductase abundance and tissue nitrate concentration. Compared with stems or roots, NRA was greater in leaves, and the highest leaf NRA was found in young leaves. Leaf and root NRA increased with increasing nitrate supply, whereas stem NRA remained constant. Leaf NRA was at least 10-fold greater than root NRA at all external nitrate concentrations. Nitrate reductase abundance increased in all tissues with increasing nitrate availability, and nitrate reductase abundance was at least 10-fold greater in leaves than in stems or roots at all nitrate availabilities. Tissue nitrate concentration increased with increasing nitrate supply and was greater in roots than in stems and leaves. Photoperiod influenced NRA, with leaf NRA declining in nitrate-fertilized plants with short daily photoperiods (8-h). We conclude that different tissues of poplar vary in nitrate assimilation with little nitrate assimilation occurring in roots and the most nitrate assimilation taking place in leaves.

Background: Quantitative PCR (qPCR) is a widely used technique for gene expression analysis. A common normalization method for accurate qPCR data analysis involves stable reference genes to determine relative gene expression. Despite extensive research in the forest tree species Populus, there is not a resource for reference genes that meet the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) standards for qPCR techniques and analysis. Since Populus is a woody perennial species, studies of seasonal changes in gene expression are important towards advancing knowledge of this important developmental and physiological trait. The objective of this study was to evaluate reference gene expression stability in various tissues and growth conditions in two important Populus genotypes (P. trichocarpa “Nisqually 1” and P. tremula x P. alba 717 1-B4) following MIQE guidelines. Results: We evaluated gene expression stability in shoot tips, young leaves, mature leaves and bark tissues from P. trichocarpa and P. tremula. x P. alba grown under long-day (LD), short-day (SD) or SD plus low-temperatures conditions. Gene expression data were analyzed for stable reference genes among 18S rRNA, ACT2, CDC2, CYC063, TIP4-like, UBQ7, PT1 and ANT using two software packages, geNorm^PLUS and BestKeeper. GeNorm^PLUS ranked TIP4-like and PT1 among the most stable genes in most genotype/tissue combinations while BestKeeper ranked CDC2 and ACT2 among the most stable genes. Conclusions: This is the first comprehensive evaluation of reference genes in two important Populus genotypes and the only study in Populus that meets MIQE standards. Both analysis programs identified stable reference genes in both genotypes and all tissues grown under different photoperiods. This set of reference genes was found to be suitable for either genotype considered here and may potentially be suitable for other Populus species and genotypes. These results provide a valuable resource for the Populus research community.

CRISPR‐Cas based genome editing technologies hold great potential for genetic research and bioengineering in trees. CRISPR‐Cas9 was previously demonstrated to be a highly efficient genome editing system in poplar (Zhou et al., 2015). However, these applications only used the nuclease activity of Cas9 for targeted mutagenesis. By contrast, Cas9‐derived base editors can install specific base changes in a plant genome.