Periodicity of response to abscisic acid in lateral buds of willow (Salix viminalis L.)

Aseptically cultured lateral buds of Salix viminalis L. collected from field-grown trees exhibited a clear periodicity in their ability to respond to exogenous abscisic acid (ABA). Buds were kept unopened by ABA only when the plants were dormant or entering dormancy. Short days alone did not induce bud dormancy in potted plants but ABA treatment following exposure to an 8-h photoperiod prevented bud opening although ABA treatment of buds from long-day plants did not. Naturally dormant buds taken from shoots of field-grown trees and cultured in the presence of ABA opened following a chilling treatment. In no cases were the induction and breaking of dormancy and response to ABA correlated with endogenous ABA levels in the buds.Abbreviations ABA abscisic acid - GA₃ gibberellic acid - HPLC high-performance liquid chromatography - LD long day - MeABA methyl ABA - PAR photosynthetically active radiation - SD short day     

Two novel transcripts expressed in pea dormant axillary buds

To elucidate the molecular mechanism of apical dominance, the expression patterns of genes that are preferentially expressed in dormant axillary buds of pea (Pisum sativum L. cv. Alaska) seedlings were investigated. We isolated two cDNA clones, cPsAD1 and cPsAD2 whose corresponding genes were named PsAD1 and PsAD2, from a cDNA library of dormant axillary buds using the differential display method. The deduced amino acid sequence of PsAD1 contains 87 residues and is rich in glycine residues in the amino terminal region. A search of the protein databases failed to find any sequences similar to PsAD1 protein except for the glycine-rich region. Northern blot analyses showed that PsAD1 mRNA mainly accumulated in dormant axillary buds and that its amount rapidly decreased after decapitation of the terminal bud. In situ hybridization analyses indicated that PsAD1 mRNA was localized in the apical meristem, procambia, and leaf primordia in dormant axillary buds that were competent to grow out but whose growth was temporarily suspended. That is, the expression of the PsAD1 gene is closely associated with the dormancy of axillary buds. The deduced amino acid sequence of PsAD2 contains 98 amino acid residues and is not similar to those of previously characterized proteins. PsAD2 mRNA accumulated in dormant axillary buds, roots, mature leaflets and elongated stems, suggesting that PsAD2 is involved in not only the dormancy of axillary buds but also the non-growing state in various tissues.     

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.     

The Morphogenesis of Regeneration Buds in Hordeum bulbosum L.--A Perennial Grass

The tillering phase in Hordeum bulbosum L. is terminated whenthe newly-formed axillary buds no longer emerge as tillers,but differentiate into dormant regeneration buds. The patternof development of the axillary buds differs during the tilleringphase and the post-tillering phase. During the former, accumulationof leaf primordia corresponds to the age of the bud, i.e., leafnumber per bud increases basipetally along the shoot. Duringthe post-tillering phase, leaf number per bud decreases basipetallyfrom the base of the future bulb internode. This transitionis brought about by an acceleration in the rate of accumulationof leaf primordia which is more sustained in the buds situatedcloser to the base of the bulb internode. These positional differencesin the morphogenesis of the regeneration buds are reflectedin their physiological responses during the relaxation of dormancyand activation of the buds.     

Amplification of ABA biosynthesis and signaling through a positive feedback mechanism in seeds

Abscisic acid is an essential hormone for seed dormancy. Our previous study using the plant gene switch system, a chemically induced gene expression system, demonstrated that induction of 9‐cis‐epoxycarotenoid dioxygenase (NCED), a rate‐limiting ABA biosynthesis gene, was sufficient to suppress germination in imbibed Arabidopsis seeds. Here, we report development of an efficient experimental system that causes amplification of NCED expression during seed maturation. The system was created with a Triticum aestivum promoter containing ABA responsive elements (ABREs) and a Sorghum bicolor NCED to cause ABA‐stimulated ABA biosynthesis and signaling, through a positive feedback mechanism. The chimeric gene pABRE:NCED enhanced NCED and ABF (ABRE‐binding factor) expression in Arabidopsis Columbia‐0 seeds, which caused 9‐ to 73‐fold increases in ABA levels. The pABRE:NCED seeds exhibited unusually deep dormancy which lasted for more than 3 months. Interestingly, the amplified ABA pathways also caused enhanced expression of Arabidopsis NCED5, revealing the presence of positive feedback in the native system. These results demonstrated the robustness of positive feedback mechanisms and the significance of NCED expression, or single metabolic change, during seed maturation. The pABRE:NCED system provides an excellent experimental system producing dormant and non‐dormant seeds of the same maternal origin, which differ only in zygotic ABA. The pABRE:NCED seeds contain a GFP marker which enables seed sorting between transgenic and null segregants and are ideal for comparative analysis. In addition to its utility in basic research, the system can also be applied to prevention of pre‐harvest sprouting during crop production, and therefore contributes to translational biology.     

Trehalose 6‐phosphate is involved in triggering axillary bud outgrowth in garden pea (Pisum sativum L.)

Trehalose 6‐phosphate (Tre6P) is a signal of sucrose availability in plants, and has been implicated in the regulation of shoot branching by the abnormal branching phenotypes of Arabidopsis (Arabidopsis thaliana) and maize (Zea mays) mutants with altered Tre6P metabolism. Decapitation of garden pea (Pisum sativum) plants has been proposed to release the dormancy of axillary buds lower down the stem due to changes in sucrose supply, and we hypothesized that this response is mediated by Tre6P. Decapitation led to a rapid and sustained rise in Tre6P levels in axillary buds, coinciding with the onset of bud outgrowth. This response was suppressed by simultaneous defoliation that restricts the supply of sucrose to axillary buds in decapitated plants. Decapitation also led to a rise in amino acid levels in buds, but a fall in phosphoenolpyruvate and 2‐oxoglutarate. Supplying sucrose to stem node explants in vitro triggered a concentration‐dependent increase in the Tre6P content of the buds that was highly correlated with their rate of outgrowth. These data show that changes in bud Tre6P levels are correlated with initiation of bud outgrowth following decapitation, suggesting that Tre6P is involved in the release of bud dormancy by sucrose. Tre6P might also be linked to a reconfiguration of carbon and nitrogen metabolism to support the subsequent growth of the bud into a new shoot.     

Analysis of Cycles of Dormancy and Growth in Pea Axillary Buds Based on mRNA Accumulation Patterns of Cell Cycle-Related Genes

Axillary buds of pea (Pisum sativum L. cv. Alaska) do not growon intact plants. Dormant axillary buds can be stimulated togrow rapidly after decapitation. Here, we isolated cDNAs ofPCNA, cyclinB, cyclinD, and cdc2 from pea. The mRNA expressionlevels of these genes were very low in dormant axillary buds,whereas they remarkably increased after decapitation. Basedon the mRNA accumulation patterns of these genes, we found thatmost cells in dormant axillary buds are arrested at the G₁ phasein the cell cycle. There are four buds at the second node onpea seedlings. After decapitation, mRNAs became abundant inthe large and small buds and were kept during the following3 d. After 4 d, mRNAs were still present in the large bud, butnot in the small bud. However, after removal of the large bud,the mRNA levels started to increase again in the small bud.These mRNA accumulation patterns were the same as those afterthe first decapitation. These results suggested that most cellsin axillary buds at the second node are arrested at the G₁]phase again and have the capacity to undergo multiple cyclesof dormancy and growth. Moreover, in situ hybridization analysesdemonstrated that PCNA mRNA increased in all parts of the axillarybuds after decapitation. (Received October 31, 1997; Accepted December 11, 1997)     

Photosynthetic leaf area modulates tiller bud outgrowth in sorghum

Shoot branches or tillers develop from axillary buds. The dormancy versus outgrowth fates of buds depends on genetic, environmental and hormonal signals. Defoliation inhibits bud outgrowth indicating the role of leaf‐derived metabolic factors such as sucrose in bud outgrowth. In this study, the sensitivity of bud outgrowth to selective defoliation was investigated. At 6 d after planting (6 DAP), the first two leaves of sorghum were fully expanded and the third was partially emerged. Therefore, the leaves were selectively defoliated at 6 DAP and the length of the bud in the first leaf axil was measured at 8 DAP. Bud outgrowth was inhibited by defoliation of only 2 cm from the tip of the second leaf blade. The expression of dormancy and sucrose‐starvation marker genes was up‐regulated and cell cycle and sucrose‐inducible genes was down‐regulated during the first 24 h post‐defoliation of the second leaf. At 48 h, the expression of these genes was similar to controls as the defoliated plant recovers. Our results demonstrate that small changes in photosynthetic leaf area affect the propensity of tiller buds for outgrowth. Therefore, variation in leaf area and photosynthetic activity should be included when integrating sucrose into models of shoot branching.     

AtPER1 enhances primary seed dormancy and reduces seed germination by suppressing the ABA catabolism and GA biosynthesis in Arabidopsis seeds

Seed is vital to the conservation of germplasm and plant biodiversity. Seed dormancy is an adaptive trait in numerous seed‐plant species, enabling plants to survive under stressful conditions. Seed dormancy is mainly controlled by abscisic acid (ABA) and gibberellin (GA) and can be classified as primary and secondary seed dormancy. The primary seed dormancy is induced by maternal ABA. Here we found that AtPER1, a seed‐specific peroxiredoxin, is involved in enhancing primary seed dormancy. Two loss‐of‐function atper1 mutants, atper1‐1 and atper1‐2, displayed suppressed primary seed dormancy accompanied with reduced ABA and increased GA contents in seeds. Furthermore, atper1 mutant seeds were insensitive to abiotic stresses during seed germination. The expression of several ABA catabolism genes (CYP707A1, CYP707A2, and CYP707A3) and GA biosynthesis genes (GA20ox1, GA20ox3, and KAO3) in atper1 mutant seeds was increased compared to wild‐type seeds. The suppressed primary seed dormancy of atper1‐1 was completely reduced by deletion of CYP707A genes. Furthermore, loss‐of‐function of AtPER1 cannot enhance the seed germination ratio of aba2‐1 or ga1‐t, suggesting that AtPER1‐enhanced primary seed dormancy is dependent on ABA and GA. Additionally, the level of reactive oxygen species (ROS) in atper1 mutant seeds was significantly higher than that in wild‐type seeds. Taken together, our results demonstrate that AtPER1 eliminates ROS to suppress ABA catabolism and GA biosynthesis, and thus improves the primary seed dormancy and make the seeds less sensitive to adverse environmental conditions.     

A morphological study of the development of the second inflorescences in strawberry (Fragaria×ananassa Duch.)

To clarify the timing of the differentiation of the first and second inflorescences in strawberry (Fragaria × ananassa Duch.), morphological changes on shoot apices during short day and low night temperature treatments were observed by scanning electron microscopy (SEM) and optical microscopy. Axillary buds just below the first inflorescence (axillary bud 1) became visible when sepal primordia of the primary flower were differentiated. By this time, other axillary buds had already developed. Axillary bud 1 developed four leaf primordia, and then a differentiated inflorescence at its summit. The phase transition of shoot apices from the vegetative to the reproductive phase may therefore trigger the differentiation of axillary bud 1 which is destined to develop into extension crowns.     

Anatomical and Histochemical Changes of Norway Spruce Buds Induced by Simulated Acid Rain

The study was focused on changes of anatomical and histochemical parameters of buds of 4-year-old Norway spruce (Picea abies L. Karst) trees subjected to simulated acid rain (SAR). Solutions of pH 2.9 and 3.9 were applied by spraying on shoot and/or by watering for two years. No macroscopic changes of buds or needles were observed in connection with SAR application and the only induced change was 2-week earlier onset of bud break in all treated variants compared to the control. Two-year treatment caused decrease in number of leaf primordia and increase in number of living bud scales in treated dormant buds while these parameters remained unchanged in the control buds. Treatments with solution of pH 2.9 caused decrease of flatness of bud apical meristem during the vegetative season. Increased activity of non-specific esterase in treated buds occurred during dormancy and bud break and the enhanced accumulation of phenolic compounds was detected at the beginning of shoot growth. Changes in histochemical parameters of bud tissues were induced mainly by spraying of shoots and can thus be qualified as primary damage.     

NODE COUNTING IN AXILLARY BUDS OF NICOTIANA TABACUM CV. WISCONSIN 38, A DAY-NEUTRAL PLANT

A mature, quiescent, primary axillary bud on the main axis of a flowering Nicotiana tabacum cv. Wisconsin 38 plant, when released from apical dominance and before forming its terminal flower, produced a number of nodes which was dependent upon its position on the main axis. Each bud produced about one more node than the next bud above it. The total number of nodes produced by an axillary bud was about 6 to 8 greater than the number of nodes present above this bud on the main axis. At anthesis of the terminal flower on the main axis, mature, quiescent, primary axillary buds had initiated 7 to 9 leaf primordia while secondary axillary buds, sometimes present in addition to the primary ones, had initiated 4 to 5 leaf primordia. When permitted to grow out independently, primary and secondary axillary buds located at the same node on the main axis produced the same number of nodes before forming their terminal flowers. In contrast, immature primary axillary buds which had produced only 5 leaf primordia and which were released from apical dominance prior to the formation of flowers on the main axis produced only as many nodes as would be produced above them on the main axis by the terminal meristem, i.e., “extra” nodes were not produced. Therefore, it is the physiological status of the plant and not the number of nodes on the bud at the time of release from apical dominance that influenced the node-counting process of a bud. When two axillary buds were permitted to develop on the same main axis, each produced the same number of nodes as single axillary buds developing at these nodes. Thus, the counting process in an axillary bud of tobacco is independent of other buds. Axillary buds on main axes of plants that had been placed horizontally produced the same number of nodes as identically-positioned axillary buds on vertical plants, indicating that gravity does not play a major role in the counting, by an axillary bud, of the nodes on the main axis.     

Development of shoot inHydrangea macrophylla I. Terminal and axillary buds

The structure of shoots, in particular of winter buds, ofHydrangea macrophylla was examined. The non-flower-bearing shoot is usually composed of a lower and an upper part, between which a boundary is discernible by means of a distinctly short internode. This internode is the lowermost of the upper part, and it is usually shorter than the internodes immediately above and below, although the internodes tend to shorten successively from the proximal to the distal part of the shoot. Variations exist in the following characters among the terminal bud, the axillary bud on the lower part of the shoot and the axillary bud on the upper part: (1) length of bud; (2) character of the outermost pair of leaf primordia; (3) degree of development of secondary buds in the winter bud; and (4) the number of leaf primordia. Usually, the terminal bud contains several pairs of foliage leaf primordia with a primordial inflorescence at the terminal of the bud, but the axiallary bud contains only the primordia of foliage leaves in addition to a pair of bud scales.     

Ontogeny of proventitious epicormic buds in Quercus petraea. I. In the 5 years following initiation

 The persistence of large epicormic shoots is one of the main factors that reduces timber quality and value in Quercus petraea. The early phases of epicormic shoot formation, i.e. the initiation of the epicormic buds, their survival and their proliferation over the years, are not clearly understood. In the present work, we studied the initiation of the axillary buds giving rise to epicormic buds and shoots, and followed their behaviour during the first 5 years using both scanning electron microscopy and light microscopy. Two types of proventitious epicormic buds have been identified. The first type has small axillary buds associated with the rings of bud-scale scars which are found at the base and tip of each growth unit. These buds are made of a terminal meristem surrounded only by scales; no leaf primordium is detected. During the second and third years of epicormic life, meristematic areas appear in the scale axil. Progressively, the meristematic areas organize into secondary bud primordia composed solely of the terminal meristem surrounded by scales. The second type of epicormic bud has secondary buds produced by a large axillary bud when this large bud either developed into a shoot or partially abscised. The epicormic potential in Q. petraea is characterized by a balance between the epicormic buds in apparent rest, enclosing meristematic areas and secondary bud primordia, and their mortality over the years. Received: 22 January 1998 / Accepted: 8 May 1998