Seasonal nitrogen remobilization and the role of auxin transport in poplar trees

Seasonal nitrogen (N) cycling in Populus, involves bark storage proteins (BSPs) that accumulate in bark phloem parenchyma in the autumn and decline when shoot growth resumes in the spring. Little is known about the contribution of BSPs to growth or the signals regulating N remobilization from BSPs. Knockdown of BSP accumulation via RNAi and N sink manipulations were used to understand how BSP storage influences shoot growth. Reduced accumulation of BSPs delayed bud break and reduced shoot growth following dormancy. Further, 13N tracer studies also showed that BSP accumulation is an important factor in N partitioning from senescing leaves to bark. Thus, BSP accumulation has a role in N remobilization during N partitioning both from senescing leaves to bark and from bark to expanding shoots once growth commences following dormancy. The bark transcriptome during BSP catabolism and N remobilization was enriched in genes associated with auxin transport and signaling, and manipulation of the source of auxin or auxin transport revealed a role for auxin in regulating BSP catabolism and N remobilization. Therefore, N remobilization appears to be regulated by auxin produced in expanding buds and shoots that is transported to bark where it regulates protease gene expression and BSP catabolism.

Cold Acclimation and Alterations in Dehydrin-like and Bark Storage Proteins in the Leaves of Sibling Deciduous and Evergreen Peach

Seasonal changes in cold tolerance and proteins were studied in the leaves of sibling deciduous and evergreen peach [Prunus persica (L.) Batsch]. Freezing tolerance [defined as the subzero temperature at which 50% injury occurred (LT50)] was assessed using electrolyte leakage. Proteins were separated by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. Electroblots were probed with anti-dehydrin and anti-19-kD peach bark storage protein (BSP) antibodies. Leaf LT50 decreased successively from -5.8 °C on 18 Aug. to -10.3 °C in the evergreen genotype and from -7.0 °C to -15.0 °C in the deciduous genotype by 14 Oct. Protein profiles and immunoblots indicated the accumulation of a 60- and 30-kD protein during cold acclimation in the leaves of deciduous trees; however, levels of these proteins did not change significantly in the evergreen trees. Immunoblots indicate that the 60-kD protein is a dehydrin-like protein. Gel-electrophoresis and immunoblots also indicated that the 19-kD BSP progressively disappeared from summer through fall in leaves of deciduous peach but accumulated to a high level in bark tissues. A similar inverse relationship was not evident in evergreen peach.

Changes in Ultrastructure of Bark Cells and Slippage of Bark in Poplar Plants when Grown under Short-day Photoperiods

The development of bud dormancy in poplar plants is initiated by short-day photoperiods (SD). During the development of bud dormancy, there was a gradual increase in the force required to peel off the bark from the stems. We measured the force required for bark peeling and investigated the cellular changes associated with this phenomenon. Stem samples were collected from plants which had been grown under SD for different period of time up to 10 weeks. At each sampling date, the forces required to peel off the bark were measured by a tensiometer. At the same time, samples were fixed to examine ultrastructural changes by transmission electron microscopy. We have observed that there was a significant increase in the force (in Newtons) required to peel off bark from poplar stems when the development of dormancy was initiated by SD treatment. Many ultrastructural changes were observed, including the accumulation of bark storage proteins, the break down of the central vacuole to form many small vacuoles, thickened cell walls, etc. Efforts have been made to relate ultrastructural alterations to changes in the force required for bark peeling.

Bark storage Protein in Peach, Plum, and Cherry Trees

The main form of nitrogen reserves during overwintering are amino acids and proteins. Specific proteins called bark storage proteins (BSP) have been characterized in many tree species. To identify BSPs in `O'Henry' peach, `Angeleno' plum, and `Early Burlat' cherry trees, samples of bark were collected from January through December 1993 from trees growing under field conditions in Santiago, Chile. SDS-PAGE analyses were used to characterize the seasonal variation on the protein pattern on the bark of those Prunus species. A 60 kDa BSP was identified in the bark of all three species, which corresponds to the main protein present in the bark during the winter. This protein may play an important role as a nitrogen reserve in these fruit trees.

COLD ACCLIMATION IN THE LEAVES OF SIBLING DECIDUOUS AND EVERGREEN PEACH: ALTERATIONS IN DEHYDRIN AND BARK STORAGE PROTEINS

Seasonal pattern of cold tolerance and proteins were studied in the leaves of sibling deciduous and evergreen peach (Prunus persica). In contrast to deciduous peach that undergoes endodormancy in fall, evergreen peach does not (leaves are retained and shoot tips elongate under favorable conditions) (Arora et al., Plant Physiol. 99:1562-1568). Cold tolerance (LT50) was assessed using electrolyte leakage method. Proteins were separated by SDS-PAGE. Electroblots were probed with anti-dehydrin (Dr. T. Close) and anti-19 kD, peach bark storage protein (BSP) antibodies. LT50 of leaves successively increased from about -7C (18 Aug.) to -15C and -11.5C (23 Oct.) in deciduous and evergreen genotypes, respectively. The most apparent change in the protein profiles was the accumulation of a 60-kD protein during cold acclimation in the leaves of deciduous trees; however, it did not change significantly in evergreen peach. Immunoblots indicate that 60-kD protein is a dehydrin protein. PAGE and immunoblots indicated that 19-kD BSP disappeared progressively during summer through fall in the leaves of deciduous peach, but accumulated to large amounts in bark tissues. Similar inverse relationship for its accumulation in leaf vs. bark tissue was not evident in evergreen peach. Results indicate that BSP expression may be regulated by altered source/sink relationship.