Päivi L.H. Rinne

Chilling of Dormant Buds Hyperinduces FLOWERING LOCUS T and Recruits GA-Inducible 1,3-β-Glucanases to Reopen Signal Conduits and Release Dormancy in Populus

This work identifies 10 putative Populus orthologs of Arabidopsis genes that encode structurally different 1,3-β-glucanases and shows that they localize at and around plasmodesmata. These enzymes are differently regulated by daylength, temperature, GA3, and GA4, providing a mechanistic explanation of how cell communication is modulated during the dormancy cycling in synchrony with the seasons. In trees, production of intercellular signals and accessibility of signal conduits jointly govern dormancy cycling at the shoot apex. We identified 10 putative cell wall 1,3-β-glucanase genes (glucan hydrolase family 17 [GH17]) in Populus that could turn over 1,3-β-glucan (callose) at pores and plasmodesmata (PD) and investigated their regulation in relation to FT and CENL1 expression. The 10 genes encode orthologs of Arabidopsis thaliana BG_ppap, a PD-associated glycosylphosphatidylinositol (GPI) lipid-anchored protein, the Arabidopsis PD callose binding protein PDCB, and a birch (Betula pendula) putative lipid body (LB) protein. We found that these genes were differentially regulated by photoperiod, by chilling (5°C), and by feeding of gibberellins GA3 and GA4. GA3 feeding upregulated all LB-associated GH17s, whereas GA4 upregulated most GH17s with a GPI anchor and/or callose binding motif, but only GA4 induced true bud burst. Chilling upregulated a number of GA biosynthesis and signaling genes as well as FT, but not CENL1, while the reverse was true for both GA3 and GA4. Collectively, the results suggest a model for dormancy release in which chilling induces FT and both GPI lipid-anchored and GA3-inducible GH17s to reopen signaling conduits in the embryonic shoot. When temperatures rise, the reopened conduits enable movement of FT and CENL1 to their targets, where they drive bud burst, shoot elongation, and morphogenesis.

CENL1 Expression in the Rib Meristem Affects Stem Elongation and the Transition to Dormancy in Populus

We investigated the short day (SD)–induced transition to dormancy in wild-type hybrid poplar (Populus tremula × P. tremuloides) and its absence in transgenic poplar overexpressing heterologous PHYTOCHROME A (PHYA). CENTRORADIALIS-LIKE1 (CENL1), a poplar ortholog of Arabidopsis thaliana TERMINAL FLOWER1 (TFL1), was markedly downregulated in the wild-type apex coincident with SD-induced growth cessation. By contrast, poplar overexpressing a heterologous Avena sativa PHYA construct (P35S:AsPHYA), with PHYA accumulating in the rib meristem (RM) and adjacent tissues but not in the shoot apical meristem (SAM), upregulated CENL1 in the RM area coincident with an acceleration of stem elongation. In SD-exposed heterografts, both P35S:AsPHYA and wild-type scions ceased growth and formed buds, whereas only the wild type assumed dormancy and P35S:AsPHYA showed repetitive flushing. This shows that the transition is not dictated by leaf-produced signals but dependent on RM and SAM properties. In view of this, callose-enforced cell isolation in the SAM, associated with suspension of indeterminate growth during dormancy, may require downregulation of CENL1 in the RM. Accordingly, upregulation of CENL1/TFL1 might promote stem elongation in poplar as well as in Arabidopsis during bolting. Together, the results suggest that the RM is particularly sensitive to photoperiodic signals and that CENL1 in the RM influences transition to dormancy in hybrid poplar.

Dormancy cycling at the shoot apical meristem: transitioning between self-organization and self-arrest.

To survive winter deciduous perennials of the temperate zones cease growth and acquire a cold-acclimated state. Timing of these events is guided by sensory systems in the leaves that register critical alterations in photoperiod. Growth cessation on its own is not sufficient to develop adequate freezing tolerance, which requires entry of the shoot apical meristem (SAM) into dormancy. To fully appreciate perennial dormancy as a precondition for cold acclimation it is necessary to assess how it is brought about in a timely fashion, what the nature of it is, and how it is released. Short day (SD) exposure results in growth cessation, bud set, dormancy establishment at the SAM, and a moderate to high level of freezing tolerance. Subsequent chilling releases the SAM from dormancy and enhances freezing tolerance further. Recent investigations indicate that dormancy is a state of self-arrest that is brought about by an enzyme-based system which disrupts the intrinsic signal network of the SAM. Release from this state requires a complimentary enzyme-based system that is preformed during SD and mobilized by chilling. These findings are in agreement with the paradigm of dormancy cycling, which defines the seasonal alternations at the SAM as transitions between states of self-organization and self-arrest. The success of this survival strategy is based on the adequate scheduling of a complex array of events. The appreciation is growing that this involves signal cascades that are, mutatis mutandis, also recruited in floral evocation in many annuals, including Arabidopsis. A heuristic model is presented of dormancy cycling at the SAM, which depicts crucial molecular and cellular events that drive the cycle.

Plant lipid bodies and cell-cell signaling: a new role for an old organelle?

Plant lipid droplets are found in seeds and in post-embryonic tissues. Lipid droplets in seeds have been intensively studied, but those in post-embryonic tissues are less well characterised. Although known by a variety of names, here we will refer to all of them as lipid bodies (LBs). LBs are unique spherical organelles which bud off from the endoplasmic reticulum, and are composed of a single phospholipid (PL) layer enclosing a core of triacylglycerides. The PL monolayer is coated with oleosin, a structural protein that stabilizes the LB, restricts its size, and prevents fusion with adjacent LBs. Oleosin is uniquely present at LBs and is regarded as a LB marker. Although initially viewed as simple stores for energy and carbon, the emerging view is that LBs also function in cytoplasmic signalling, with the minor LB proteins caleosin and steroleosin in a prominent role. Apart from seeds, a variety of vegetative and floral structures contain LBs. Recently, it was found that numerous LBs emerge in the shoot apex of perennial plants during seasonal growth arrest and bud formation. They appear to function in dormancy release by reconstituting cell-cell signalling paths in the apex. As apices and orthodox seeds proceed through comparable cycles of dormancy and dehydration, the question arises to what degree LBs in apices share functions with those in seeds. We here review what is known about LBs, particularly in seeds, and speculate about possible unique functions of LBs in post-embryonic tissues in general and in apices in particular.

Shoot meristems of deciduous woody perennials: self-organization and morphogenetic transitions

Shoot apical meristems of deciduous woody perennials share gross structural features with other angiosperms, but are unique in the seasonal regulation of vegetative and floral meristems. Supporting longevity, flowering is postponed to the adult phase, and restricted to some axillary meristems. In cold climates, photoperiodic timing mechanisms and chilling are recruited to schedule end-of-season growth arrest, dormancy cycling and flowering. We review recently uncovered generic meristem properties, perennial meristem fate, and the role of CENL1, FT1 and FT2 in bud formation and flowering. We also highlight novel findings, suggesting that dormancy release is mediated by mobile lipid bodies that deliver enzymes to plasmodesmata to recover symplasmic communication and meristem function.

The embryonic shoot: a lifeline through winter

The tiny vascular axis of the embryo emerges post-embryonically as an elaborate and critical infrastructure, pervading the entire plant system. Its expansive nature is especially impressive in trees, where growth and development continue for extended periods. While the shoot apical meristem (SAM) orchestrates primary morphogenesis, the vascular system is mapped out in its wake in the provascular cylinder, situated just below the emerging leaf primordia and surrounding the rib meristem. Formation of leaf primordia and provascular tissues is incompatible with the harsh conditions of winter. Deciduous trees of boreal and temperate climates therefore enter a survival mode at the end of the season. However, to be competitive, they need to maximize their growth period while avoiding cellular frost damage. Trees achieve this by monitoring photoperiod, and by timely implementation of a survival strategy that schedules downstream events, including growth cessation, terminal bud formation, dormancy assumption, acquisition of freezing tolerance, and shedding of leaves. Of central importance are buds, which contain an embryonic shoot that allows shoot development and elongation in spring. The genetic and molecular processes that drive the cycle in synchrony with the seasons are largely elusive. Here, we review what is known about the signals and signal conduits that are involved, the processes that are initiated, and the developmental transitions that ensue in a terminal bud. We propose that addressing dormancy as a property of the SAM and the bud as a unique shoot type will facilitate our understanding of winter dormancy.

Axillary buds are dwarfed shoots that tightly regulate GA pathway and GA-inducible 1,3-β-glucanase genes during branching in hybrid aspen

Highlight Axillary buds uniquely regulate gibberellin (GA) pathway genes, enabling them to stay inhibited but simultaneously poised for growth. Decapitation promotes expression of GA-inducible 1,3-β-glucanase genes that function to reinvigorate symplasmic connections to the stem.

Perennial Life Style of Populus : Dormancy Cycling and Overwintering

Deciduous trees in boreal and temperate areas are strictly conditioned by the environment, especially by photoperiod and temperature. However, it is in particular the successful submission to these conditions that has rewarded them with long-life spans. A crucial strategy to ensure growth over many seasons is to timely assume dormancy and a level of hardiness that permits survival through winter. A consensus is emerging that dormancy, although traditionally regarded as a systemic feature, is a property of the shoot apical meristem (SAM). This chapter discusses our current understanding of the regulatory mechanisms that drive the annual cycles of dormancy and acclimation.

Long and short photoperiod buds in hybrid aspen share structural development and expression patterns of marker genes

Highlight Short photoperiod and apical dominance trigger a shared developmental bud programme at terminal and axillary positions, while the capacity to establish photoperiod-induced dormancy is lost in maturing para-dormant axillary buds.

Mapping symplasmic fields at the shoot apical meristem using iontophoresis and membrane potential measurements.

Microinjections of fluorescent dyes have revealed that the shoot apical meristem (SAM) is dynamically partitioned into symplasmic fields (SFs), implying that plasmodesmata (Pd) are held shut at specific locations in the proliferating cellular matrix. The SFs are integrated into a coherent morphogenetic unit by exchange of morphogens and transcription factors via gating Pd between adjacent SFs, and by ligand-receptor interactions that operate across the extracellular space. We describe a method for the real-time mapping of SF in the SAM by iontophoresis and membrane potential measurements.