Co-ordinated gene expression during phases of dormancy release in raspberry (Rubus idaeus L.) buds

Bud break in raspberry (Rubus idaeus L.) is often poor and uneven, with many of the subapical buds remaining in a dormant state. In order to determine the dormancy status of raspberry buds, an empirical measure of bud burst in a growth-permissive environment following exposure to chilling (4 degrees C cold storage) was developed. For cv. Glen Ample, percentage bud burst in intact canes and isolated nodes was recorded after 14 d. Isolated nodes (a measure of endodormancy) achieved 100% bud burst after approximately 1500 h chilling whereas buds on intact plants (combined endo- and paradormancy) required an additional 1000 h chilling. A microarray approach was used to follow changes in gene expression that occurred during dormancy transition. The probes for the microarrays were obtained from endodormant and paradormant raspberry bud cDNA libraries. The expression profiles of 5300 clones from these libraries were subjected to principal component analysis to determine the most significant expression patterns. Sequence analysis of these clones, in many cases, enabled their functional categorization and the development of hypotheses concerning the mechanisms of bud dormancy release. Thus a set of novel candidates for key dormancy-related genes from raspberry buds have been identified. Bud dormancy is fundamental to the study of plant developmental processes and, in addition, its regulation is of significant economic importance to fruit and horticultural industries.     

The dynamic nature of bud dormancy in trees: environmental control and molecular mechanisms

In tree species native to temperate and boreal regions, the activity-dormancy cycle is an important adaptive trait both for survival and growth. We discuss recent research on mechanisms controlling the overlapping developmental processes that define the activity-dormancy cycle, including cessation of apical growth, bud development, induction, maintenance and release of dormancy, and bud burst. The cycle involves an extensive reconfiguration of metabolism. Environmental control of the activity-dormancy cycle is based on perception of photoperiodic and temperature signals, reflecting adaptation to prevailing climatic conditions. Several molecular actors for control of growth cessation have been identified, with the CO/FT regulatory network and circadian clock having important coordinating roles in control of growth and dormancy. Other candidate regulators of bud set, dormancy and bud burst have been identified, such as dormancy-associated MADS-box factors, but their exact roles remain to be discovered. Epigenetic mechanisms also appear to factor in control of the activity-dormancy cycle. Despite evidence for gibberellins as negative regulators in growth cessation, and ABA and ethylene in bud formation, understanding of the roles that plant growth regulators play in controlling the activity-dormancy cycle is still very fragmentary. Finally, some of the challenges for further research in bud dormancy are discussed.     

Dormancy and spring development of lateral buds in mulberry (Morus alba)

Patterns of spring development of lateral buds of mulberry (Morus alba L. cv. Shin-ichinose) coppice shoots on 11-year-old low-pruned stumps varied in response to girdling, pruning and arching. The erect controls showed a weak acrotonic (apex-favoring) growth habit, in which the majority of the buds, including the basal ones, sprouted and elongated in mid- and late April, and hence there was a prolonged imposition of dominance on the upper laterals in mid- and late May. In contrast, early spring girdling or pruning enhanced the activity of the upper buds of the proximal (lower) halves of the girdled stems or of the pruned stems, resulting in considerable dominance of the laterals from such buds in late April. Arching markedly inhibited buds on the under side of the arched stems, leading to poor shoots. By late April, the buds on the adaxial (upper) side readily grew into new vertical shoots, which dominated over the lateral ones. When studied by a multiple-node-cutting test, increased length of segments of post-dormant mulberry stems was accompanied by decreased bud activity of the segments and by decreased breaking ability of the lower buds within the segments, suggesting the importance of roots in the weak acrotonic habit of the erect stem in spring. By contrast, the acropetal influences of the attached stems can in part affect dominance relationships, perhaps mediated through competition for factors translocated from the roots. Continuous basal applications of abscisic acid inhibited bud break and shoot growth of the postdormant stem segments, but these inhibitory effects could be reversed by applied gibberellic acid A₃ (GA₃). Two phases of lateral bud dormancy in erect mulberry coppice shoots were identified. The first was characterized by a smaller breaking capacity in the upper buds than in the lower ones and hence by a basitonic (base-favoring) gradient in bud growth potential. The second phase corresponded to a restoration of these capabilities in the upper buds and to a change towards a linear gradient in bud growth potential, with disappearance of the dormant condition, in February and March. This gradient change during dormancy release may represent the physiological basis for the weak acrotonic habit of erect mulberry stems in spring.     

Ecotype-dependent control of growth,dormancy and freezing tolerance under seasonal changes in <Emphasis Type="Italic">Betula pendula</Emphasis> Roth

Woody plants in the temperate and boreal zone undergo annual cycle of growth and dormancy under seasonal changes. Growth cessation and dormancy induction in autumn are prerequisites for the development of substantial cold hardiness in winter. During evolution, woody plants have developed different ecotypes that are closely adapted to the local climatic conditions. In this study, we employed distinct photoperiodic ecotypes of silver birch (Betula pendula Roth) to elucidate differences in these adaptive responses under seasonal changes. In all ecotypes, short day photoperiod (SD) initiated growth cessation and dormancy development, and induced cold acclimation. Subsequent low temperature (LT) exposure significantly enhanced freezing tolerance but removed bud dormancy. Our results suggested that dormancy and freezing tolerance might partially overlap under SD, but these two processes were regulated by different mechanisms and pathways under LT. Endogenous abscisic acid (ABA) levels were also altered under seasonal changes; the ABA level was low during the growing season, then increased in autumn, and decreased in winter. Compared with the southern ecotype, the northern ecotype was more responsive to seasonal changes, resulting in earlier growth cessation, cold acclimation and dormancy development in autumn, higher freezing tolerance and faster dormancy release in winter, and earlier bud flush and growth initiation in spring. In addition, although there was no significant ecotypic difference in ABA level during growing season, the rates and degrees of ABA alterations were different between the ecotypes in autumn and winter, and could be related to ecotypic differences in dormancy and freezing tolerance.     

Release of apical dominance in potato tuber is accompanied by programmed cell death in the apical bud meristem

Potato (Solanum tuberosum) tuber, a swollen underground stem, is used as a model system for the study of dormancy release and sprouting. Natural dormancy release, at room temperature, is initiated by tuber apical bud meristem (TAB-meristem) sprouting characterized by apical dominance (AD). Dormancy is shortened by treatments such as bromoethane (BE), which mimics the phenotype of dormancy release in cold storage by inducing early sprouting of several buds simultaneously. We studied the mechanisms governing TAB-meristem dominance release. TAB-meristem decapitation resulted in the development of increasing numbers of axillary buds with time in storage, suggesting the need for autonomous dormancy release of each bud prior to control by the apical bud. Hallmarks of programmed cell death (PCD) were identified in the TAB-meristems during normal growth, and these were more extensive when AD was lost following either extended cold storage or BE treatment. Hallmarks included DNA fragmentation, induced gene expression of vacuolar processing enzyme1 (VPE1), and elevated VPE activity. VPE1 protein was semipurified from BE-treated apical buds, and its endogenous activity was fully inhibited by a cysteinyl aspartate-specific protease-1-specific inhibitor N-Acetyl-Tyr-Val-Ala-Asp-CHO (Ac-YVAD-CHO). Transmission electron microscopy further revealed PCD-related structural alterations in the TAB-meristem of BE-treated tubers: a knob-like body in the vacuole, development of cytoplasmic vesicles, and budding-like nuclear segmentations. Treatment of tubers with BE and then VPE inhibitor induced faster growth and recovered AD in detached and nondetached apical buds, respectively. We hypothesize that PCD occurrence is associated with the weakening of tuber AD, allowing early sprouting of mature lateral buds.     

Epigenetic repressor-like genes are differentially regulated during grapevine (Vitis vinifera L.) development

Grapevine sexual reproduction involves a seasonal separation between inflorescence primordia (flowering induction) and flower development. We hypothesized that a repression mechanism implicating epigenetic changes could play a role in the seasonal separation of these two developmental processes in grapevine. Therefore, the expression of five grapevine genes with homology to the Arabidopsis epigenetic repressor genes FERTILIZATION INDEPENDENT ENDOSPERM (FIE), EMBRYONIC FLOWER 2 (EMF2), CURLY LEAF (CLF), MULTICOPY SUPPRESSOR OF IRA 1 (MSI1) and SWINGER (SWN) was analyzed during the development of buds and vegetative and reproductive organs. During bud development, the putative grapevine epigenetic repressor genes VvCLF, VvEMF2, VvMSI1, VvSWN and VvFIE are mainly expressed in latent buds at the flowering induction period, but also detected during bud burst and inflorescence/flower development. The overlapping expression patterns of grapevine PcG-like genes in buds suggest that chromatin remodeling mechanisms could be operating during grapevine bud development for controlling processes such as seasonal flowering, dormancy and bud burst. Furthermore, the expression of grapevine PcG-like genes was also detected in fruits and vegetative organs, suggesting that epigenetic changes could be at the basis of the regulation of various proliferation–differentiation cell transitions that occur during grapevine development.     

Phases of dormancy in Yam tubers (Dioscorea rotundata)

BACKGROUND AND AIMS: The control of dormancy in yam (Disocorea spp.) tubers is poorly understood and attempts to shorten the long dormant period (i.e. cause tubers to sprout or germinate much earlier) have been unsuccessful. The aim of this study was to identify and define the phases of dormancy in Dioscorea rotundata tubers, and to produce a framework within which dormancy can be more effectively studied. METHODS: Plants of 'TDr 131' derived from tissue culture were grown in a glasshouse simulating temperature and photoperiod at Ibadan (7 degrees N), Nigeria to produce tubers. Tubers were sampled on four occasions: 30 d before shoot senescence (149 days after planting, DAP), at shoot senescence (179 DAP), and twice during storage at a constant 25 degrees C (269 and 326 DAP). The development of the apical shoot bud was described from tissue sections. In addition, the responsiveness of shoot apical bud development to plant growth regulators (gibberellic acid, 2-chloroethanol and thiourea) applied to excised tuber sections was also examined 6 and 12 d after treatment. KEY RESULTS AND CONCLUSIONS: Three phases of tuber dormancy are proposed: Phase I, from tuber initiation to the appearance of the tuber germinating meristem; Phase II, from the tuber germinating meristem to initiation of foliar primordium; and Phase III, from foliar primordium to appearance of the shoot bud on the surface of the tuber. Phase I is the longest phase (approx. 220 d in 'TDr 131'), is not affected by PGRs and is proposed to be an endo-dormant phase. Phases II and III are shorter (<70 d in total), are influenced by PGRs and environmental conditions, and are therefore endo-/eco-dormant phases. To manipulate dormancy to allow off-season planting and more than one generation per year requires that the duration of Phase I is shortened.     

Lateral Bud Dormancy in the Blackcurrant Ribes nigrum (L.)

Previous investigations have shown that the inhibition of lateralbuds in blackcurrants soon after their formation is due to apicaldominance and this is followed by leaf inhibition. The presentstudies have revealed that bud scales are the main cause ofinhibition even before leaf fall. The inhibitor level assayedby the wheat coleoptile test shows large seasonal changes; itis maximal in early autumn and minimal in spring. Inhibitor levels are reduced by low-temperature treatment inparallel with the promotion of dormancy breaking; temperatureswell below freezing, applied for very short times, almost certainlyact through the killing of bud scales in the same way as doesrapid high-temperature treatment. Other treatments causing budbreak in the presence of high inhibitor levels are: continuousillumination, ethylene vapour, and application of gibberellicacid. It is suggested that bud inhibition in the blackcurrant is dueto the formation of an inhibitor produced by the leaves andaccumulated in the bud scales. The inhibitor (probably abscisicacid) may then interact with internal gibberellin levels, summerand winter dormancy differing only quantitatively.     

THE RELATION OF AGE AND BUD BREAK TO THE DETERMINATION OF PHYLLOTAXY IN CATALPA SPECIOSA

Catalpa speciosa is interesting in that the phyllotaxy of the lateral shoots can be either decussate or whorled. The pattern appears to be correlated with the number of buds which emerge. The arrangement is not related to dormancy; nor does the stage of development of the bud at the time of bud break influence the phyllotaxy. The control of bud break appears to be related to the position of the bud on the stem. Possible mechanisms controlling the lateral shoot pattern are discussed.     

Abscisic acid catabolism enhances dormancy release of grapevine buds

The molecular mechanism regulating dormancy release in grapevine buds is as yet unclear. It was formerly proposed that dormancy is maintained by abscisic acid (ABA)‐mediated repression of bud–meristem activity and that removal of this repression triggers dormancy release. It was also proposed that such removal of repression may be achieved via natural or artificial up‐regulation of VvA8H‐CYP707A4, which encodes ABA 8′‐hydroxylase, and is the most highly expressed paralog in grapevine buds. The current study further examines these assumptions, and its experiments reveal that (a) hypoxia and ethylene, stimuli of bud dormancy release, enhance expression of VvA8H‐CYP707A4 within grape buds, (b) the VvA8H‐CYP707A4 protein accumulates during the natural transition to the dormancy release stage, and (c) transgenic vines overexpressing VvA8H‐CYP707A4 exhibit increased ABA catabolism and significant enhancement of bud break in controlled and natural environments and longer basal summer laterals. The results suggest that VvA8H‐CYP707A4 functions as an ABA degrading enzyme, and are consistent with a model in which the VvA8H‐CYP707A4 level in the bud is up‐regulated by natural and artificial bud break stimuli, which leads to increased ABA degradation capacity, removal of endogenous ABA‐mediated repression, and enhanced regrowth. Interestingly, it also hints at sharing of regulatory steps between latent and lateral bud outgrowth.     

Freezing exposure releases bud dormancy in Betula pubescens and B. pendula

Bud dormancy in woody plants is released by long-term exposure to non-freezing chilling temperatures, whereas freezing temperatures have been considered to have little or no effect. However, the present results demonstrate that short-term exposure to freezing can release bud dormancy in Betula pubescens (Ehrh.) and B. pendula (Roth). Short-term freezing during the dormancy induction phase improved the release of bud dormancy only if an adequate level of dormancy had been reached. In fully dormant or chilled plants both the percentage and the speed of bud-burst increased, the more so the lower the temperature. Our results rule out the possibility that endogenous abscisic acid could be directly involved in the physiological control of bud dormancy release. The fast, easily applicable method presented here for bud dormancy release could further investigations into the biochemical and biophysical background to the process. The mechanisms of bud dormancy release and its relationship to cold acclimation are discussed in the light of these results, as also are the implications of the findings for modelling of bud dormancy.