The formation and distribution of ice within dormant and deacclimated peach flower buds

Abstract A freeze-fixation technique was used to examine the distribution of ice crystals and the pattern of freezing in peach flower buds. In dormant buds, ice crystals formed at localized sites within the bud axis and scales. Ice crystal formation disrupted tissues and mechanical injury from repetitive freezethaw cycles was apparent. There was evidence of ice formation in the floral organs of dormant buds exposed to ?25°C but none observed in buds exposed to either ?5 or ?10°C. The distribution of ice crystals was different in deacclimated buds. In addition to large ice crystals within the subtending bud axis and scales, evidence of large crystals within the developing floral organs was noted. These crystals were most prominent in the lower portions of the developing flower and peduncle, and caused a separation of the epidermal layer from adjacent cells. The distribution of ice crystals within both dormant and deacclimated peach flower buds corroborated the results of previous thermal analysis experiments.     

Properties of peach flower buds which facilitate supercooling

Water in dormant peach (Prunus persica [L.] Batsch. var. `Harbrite') flower buds deep supercooled. Both supercooling and the freezing of water within the bud axis and primordium as distinct components depended on the viability of the bud axis tissue. The viability of the primordium was not critical. Supercooling was prevented by wounding buds with a dissecting needle, indicating that bud structural features were important. Bud morphological features appeared to prevent the propagation of ice through the vascular tissue and into the primordium. In dormant buds, procambial cells had not yet differentiated into xylem vessel elements. Xylem continuity between the bud primordium and adjacent tissues did not appear to be established until buds had deacclimated. It was concluded that structural, morphological, and physiological features of the bud facilitated supercooling.     

Immunogold localization of pectins and glycoproteins in tissues of peach with reference to deep supercooling

Living xylem tissues and floral buds of several species of woody plants survive exposure to freezing temperatures by deep supercooling. A barrier to water loss and the growth of ice crystals into cells is considered necessary for deep supercooling to occur. Pectins, as a constituent of the cell wall, have been implicated in the formation of this barrier. The present study examined the distribution of pectin in xylem and floral bud tissues of peach (Prunus persica). Two monoclonal antibodies (JIM5 and JIM7) that recognize homogalacturonic sequences with varying degrees of esterification were utilized in conjunction with immunogold electron microscopy. Results indicate that highly esterified epitopes of pectin, recognized by JIM7, were the predominant types of pectin in peach and were uniformly distributed throughout the pit membrane and primary cell walls of xylem and floral bud tissues. In contrast, un-esterified epitopes of pectin, recognized by JIM5, were confined to the outer surface of the pit membrane in xylem tissues. In floral buds, these epitopes were localized in middle lamellae, along the outer margin of the cell wall lining empty intercellular spaces, and within filled intercellular spaces. JIM5 labeling was more pronounced in December samples than in July/August samples. Additionally, epitopes of an arabinogalactan protein, recognized by JIM14, were confined to the amorphous layer of the pit membrane. The role of pectins in freezing response is discussed in the context of present theory and it is suggested that pectins may influence both water movement and intrusive growth of ice crystals at freezing temperatures.     

Characterization of the Arrest in Anther Development Associated with Gibberellin Deficiency of the gib-1 Mutant of Tomato

The role of gibberellins in flower bud development was investigated by studying the gib-1 mutant of tomato, Lycopersicon esculentum. This gibberellin-deficient mutant initiates flower buds, but floral development is not completed unless the mutant is treated with gibberellin. Treatment with other plant growth regulators does not induce normal flower development. Development of gib-1 flower buds, as measured by progress toward anthesis, ceases at a bud length of 2.5 millimeters; however, increase in size of the bud continues. Buds between 2.5 and 3.7 millimeters are developmentally arrested but still are capable of developing normally after treatment with gibberellic acid. Anthers of these developmentally arrested buds contain pollen mother cells that are in the G1 phase of premeiotic interphase. Following treatment of developmentally arrested buds with gibberellic acid, premeiotic DNA synthesis and callose accumulation in pollen mother cells are evident by 48 hours posttreatment, and within 66 hours, prophase I of meiosis- and meiosis-related changes in tapetum development are observable.     

Spatial and temporal changes in multiple hormone groups during lateral bud release shortly following apex decapitation of chickpea (Cicer arietinum) seedlings

Although the co-ordination of promotive root-sourced cytokinin (CK) and inhibitory shoot apex-sourced auxin (IAA) is central to all current models on lateral bud dormancy release, control by those hormones alone has appeared inadequate in many studies. Thus it was hypothesized that the IAA : CK model is the central control but that it must be considered within the relevant timeframe leading to lateral bud release and against a backdrop of interactions with other hormone groups. Therefore, IAA and a wide survey of cytokinins (CKs), were examined along with abscisic acid (ABA) and polyamines (PAs) in released buds, tissue surrounding buds and xylem sap at 1 and 4 h after apex removal, when lateral buds of chickpea are known to break dormancy. Three potential lateral bud growth inhibitors, IAA, ABA and cis -zeatin 9-riboside (ZR), declined sharply in the released buds and xylem following decapitation. This is in contrast to potential dormancy breaking CKs like trans -ZR and trans -zeantin 9-riboside 5'phosphate (ZRMP), which represented the strongest correlative changes by increasing 3.5-fold in xylem sap and 22-fold in buds. PAs had not changed significantly in buds or other tissues after 4 h, so they were not directly involved in the breaking of bud dormancy. Results from the xylem and surrounding tissues indicated that bud CK increases resulted from a combination synthesis in the bud and selective loading of CK nucleotides into the xylem from the root.     

The relationship between xylem differentiation and dormancy evolution in apricot flower buds (Prunus armeniaca L.): the influence of environmental conditions in two Mediterranean areas

Environmental and climatic conditions affect the flower bud growth, flowering and yield performance of fruit species. Temperature and water availability appear to be important factors for temperate fruit trees in mild climates. The aim of this research was to study the involvement of temperature regime on biological processes, such as xylem differentiation and dormancy evolution, in apricot flower buds (Prunus armeniaca L.). Over 3 consecutive years, biological and anatomical investigations were carried out in Tuscany (Italy) and Murcia (Spain) on two cultivars characterized by a different chilling requirement (CR): Currot, a traditional Spanish cultivar with a very low CR, and Stark Early Orange (SEO), a North American cultivar with a very high CR. Currot had a regular bearing, and was characterized by a synchronism between endodormancy release and xylem differentiation. On the other hand, SEO showed an irregular flower bud growth leading to flowering and fruit-set failure, even with a high level of chilling accumulation. No relationship was found in SEO between xylem development and flower bud growth reactivation. The de-synchronism between these processes could determine the poor adaptability to different environmental areas of SEO cultivar, and its very high CR is not the main hypothesised cause of the altered flower bud development.     

Cold hardiness in various organs and tissues of Rhododendron species and the supercooling ability of flower buds as the most susceptible organ

The freezing resistance of various organs and tissues was determined in 24 Rhododendron species (mainly Subgenus Tsutsutsi) having different ecological distributions. The order of hardiness for organ or tissue is as follows: leaf bud > wood ≧ bark > flower bud, and the flower bud is characterized as the most cold-susceptible organ. The relationship of killing temperature (KT) to northern distribution was the most significant in leaf buds compared to other organs and tissues. KTs of leaf buds for the most hardy species were ?45 °C (or below) and those for the most tender species were about ?23 °C, while KTs of flower buds were about ?28 °C for the former and ?16 °C for the latter. Although KTs of flower buds native to southwestern Japan were well correlated with the exothermic temperature distribution (ETD) of florets, those in the more northern species were generally lower than ETDs. The supercooling ability of flower buds appears to be sufficient to avoid the freezing stress since the extreme minimum temperature (EMT) at the northern limit of natural distribution for each tree species examined was not lower than the KT and ETD of the flower buds.     

The Formation and Distribution of Ice within Forsythia Flower Buds

Differential thermal analysis detected two freezing events when dormant forsythia (Forsythia viridissima Lindl.) flower buds were cooled. The first occurred just below 0°C, and was coincident with the freezing of adjacent woody tissues. The second exotherm appeared as a spike between −10 and −25°C and was correlated with the lethal low temperature. Although this pattern of freezing was similar to that observed in other woody species, differences were noted. Both direct observations of frozen buds and examination of buds freeze-fixed at −5°C demonstrated that ice formed within the developing flowers at temperatures above the second exotherm and lethal temperature. Ice crystals had formed within the peduncle and in the lower portions of the developing flower. Ice also formed within the scales. In forsythia buds, the developing floral organ did not freeze as a unit as noted in other species. Instead the low temperature exotherm appeared to correspond to the lethal freezing of supercooled water within the anthers and portions of the pistil.     

Xylem development in prunus flower buds and the relationship to deep supercooling

Xylem development in eight Prunus species was examined and the relationship to deep supercooling assessed. Dormant buds of six species, P. armeniaca, P. avium, P. cerasus, P. persica, P. salicina, and P. sargentii deep supercooled. Xylem vessel elements were not observed within the dormant floral primordia of these species. Instead, discrete bundles containing procambial cells were observed. Vascular differentiation resumed and xylem continuity was established during the time that the capacity to deep supercool was lost. In P. serotina and P. virginiana, two species which do not supercool, xylem vessels ran the length of the inflorescence and presumably provided a conduit for the spread of ice into the bud. The results support the hypothesis that the lack of xylem continuity is an important feature of buds which deep supercool.     

Fruit Regulates Bud Sprouting and Vegetative Growth in Field-Grown Loquat Trees (Eriobotrya japonica Lindl.): Nutritional and Hormonal Changes

The effects of fruit on bud sprouting and vegetative growth were compared on fruiting and defruited loquat trees from fruit set onward. Carbohydrate and nitrogen content in leaves and bark tissues and hormone concentrations were studied during the fruit development and vegetative growth periods. On defruited trees, a significant proportion of buds sprouted in winter, whereas buds from fruiting trees sprouted only in the spring when fruit reached its final size. Furthermore, when panicles were completely removed in autumn, the buds also sprouted. In addition, fruit directly affected vegetative growth by reducing shoot length. An effect of sink removal (flower or fruit) promoting bud sprouting, regardless of the season, was then demonstrated. Neither soluble sugar concentration nor nitrogen fraction concentration in leaves or bark tissues was related to bud sprouting, but a certain nutritional imbalance was observed during the most active period of fruit development. Moreover, fruit sink activity significantly modified hormone content by increasing indole-3-acetic acid (IAA) and reducing zeatin concentrations, resulting in a higher IAA/zeatin ratio parallel to the lower bud sprouting intensity. Therefore, these changes caused by fruit removal are all related to vegetative growth, but there is no evidence that they are responsible for bud burst.     

Population dynamics of western flower thrips (Thysanoptera: Thripidae) in nectarine orchards in British Columbia

Development of a control strategy for thrips attacking nectarine trees depends on an understanding of their phenology, distribution, and life history as related to characteristics of nectarine orchards. To this end, we compared the overwintering behavior, distribution, and abundance of western flower thrips, Frankliniella occidentalis (Pergande), among 11 nectarine orchards located in the dry central interior of British Columbia, Canada, during 1993 and 1994. Western flower thrips emerged from areas not previously used for agriculture (wild areas) and from within orchards before trees were out of dormancy. Flight of thrips within and around orchards peaked during early bud development, with a second major peak several weeks later after husk fall as the next generation emerged. Orchards protected from wild areas by other orchards had the lowest densities of thrips in buds. Density estimates of western flower thrips on trees were not affected by location of trees within orchards or buds within trees, but most thrips were found in the most developed buds on a tree at any one time. Thrips were not found within buds until petal was first visible on the buds. Larval feeding on buds at early petal fall resulted in serious surface russetting of fruit.     

Endogenous IAA levels and development of coffee flower buds from dormancy to anthesis

Dormant coffee (Coffea arabica L.) flower buds require water stress to stimulate regrowth. A xylem specific water-soluble dye, azosulfamide, was used to quantify water uptake of buds after their release from dormancy by water stress. In non-stressed flower buds, the rate of water uptake was generally slower and variable compared to stressed flower buds, where the rate of uptake tripled from 1 to 3 days after rewatering and preceded the doubling of fresh and dry weight of buds. Free, ester and amide IAA levels of developing flower buds were measured by gas chromatography-mass spectrometry-selective ion monitoring using an isotope dilution technique with [¹³C₆]IAA as an internal standard. Throughout development, the majority of IAA was present as amide IAA. The proportions of amide and free IAA increased one day after plants were released from water stress, and preceded the doubling of fresh and dry weight. Free and conjugated IAA content per bud remained stable during the period of rapid flower growth until one day before anthesis.Abbreviations FW fresh weight - IAA indole 3-acetic acid - HPLC high performance liquid chromatography - GC-MS-SIM gas chromatography-mass spectrometry selected ion monitoring - NAA naphthalene acetic acid - IBA indole butyric acid     

The effect of summer shading on flower bud morphogenesis in apricot (Prunus armeniaca L.)

The aim of this investigation was to assess whether imposed summer shading treatments in apricot (Prunus armeniaca L.) can affect the main phenological phases related to the floral morphogenesis (floral differentiation, xylogenesis), flower bud growth and quality in terms of bud capacity to set fruit. Experimental trials were carried out on fully-grown trees of ‘San Castrese’ and ‘Stark Early Orange’ cultivars characterized by different biological and agronomical traits to which shadings were imposed in July and August. Histological analysis was carried out from summer onwards in order to determine the evolution of floral bud differentiation, and the acropetal progression of primary xylem differentiation along the flower bud axis. Periodical recordings to evaluate the bud drop, blooming time, flowering and fruit set rates were performed also. These shade treatments determined a temporary shutdown of floral differentiation, slowed xylem progression up to the resumption of flower bud growth and a reduced entity of flowering and fruit set. These events were particularly marked in ‘San Castrese’ cultivar, which is well known for its adaptability to different climatic conditions. These findings suggest that adequate light penetration within the canopy during the summer season could be the determining factor when defining the qualitative traits of flower buds and their regular growth, and ultimately to obtain good and constant crops.     

Ontogeny of the proventitious epicormic buds in Quercus petraea.

In the present work, we described the fate of proventitious epicormic buds on the trunks of 40-year-old Quercus petraea trees and in parallel the vascular trace they produced in the wood. Our results show that small and large individual epicormic buds can survive as buds for 40 years and that both are composed of a terminal meristem and scales. Meristematic areas are detected in the scale axils of small buds; in addition to these meristems the large buds also have secondary bud primordia. The small buds are connected to the pith of the main stem by a unique trace, whereas the large buds are connected by one or multiple traces. A single trace might imply that the whole bud is still alive and multiple traces might indicate that the terminal meristem has died. In the latter case, each trace is connected to a secondary bud of the large bud. The buds found in a cluster are composed of a terminal meristem and scales with axillary meristems in the scale axils. A cluster is connected to the pith of a stem either by a unique trace when it seems to be the result of partial abscission of an epicormic shoot or multiple traces when it might have originated from an epicormic bud in which the terminal meristem has died. Whatever the type of the bud, the vascular trace in the bark is composed of a cambium, secondary xylem and parenchyma cells and the trace present in the wood had parenchyma cells with vestiges of secondary xylem. Each year, the vascular trace should be produced in the bark by the cambium of the tree but not by the bud itself. On 40-year-old Q. petraea, we observed a proliferation of epicormic buds and in parallel a multiplication of the number of vascular traces in the trunk, but the knots caused by the traces of epicormic buds in the wood, either as individuals or in clusters, are minor since their colours are only slightly darker than those of woody rays and they are less than 2 mm in diameter. The knots will appear when epicormic buds develop into shoots. Received: 30 March 1999 / Accepted: 09 June 1999     

Direct effects of retinoic acid on the development of the tail bud in chick embryos

Retinoic acid (RA) has been reported to induce vascular lesions and haematoma formation in the vicinity of the tail bud during the critical period for inducing abnormalities of tail bud development in hamsters (Wiley, '83; Tibbles and Wiley, '88), mice (Tibbles and Wiley, '88) and chicken embryos (Jelinek and Kistler, '81). Experiments were conducted to determine whether or not these vascular lesions were the primary cause of the malformations which they accompanied. Chick embryos were exposed for varying lengths of time to several dosages of RA. Primitive streaks or tail buds from treated embryos were then excised prior to vascularization and transplanted to the coelomic walls of untreated host embryos. The grafts were harvested at 3 or 6 days after grafting and processed for histological examination. Observations of serial sections of controls showed that the primitive streak and early (stage 13-14) tail bud were able to form neural tubes and a variety of other structures including ganglia, nerve fibres, and kidney tubules. Treatment of donor embryos with RA prior to grafting, however, affected the frequency and characteristics of the neural tubes and other tissues developing in the grafts. The effects of RA on development were correlated with both the dosage and length of exposure to the teratogen prior to grafting. Since the grafts were made before the appearance of blood vessels in the tail buds, we have concluded that the effects of RA on the development of tail bud tissues, and especially the secondary neural tube, are direct and are not mediated solely through the disruptive effects of vascular lesions seen in intact embryos.     

Passion fruit flowers: Kunitz trypsin inhibitors and cystatin differentially accumulate in developing buds and floral tissues

In order to better understand the physiological functions of protease inhibitors (PIs) the PI activity in buds and flower organs of passion fruit (Passiflora edulis Sims) was investigated. Trypsin and papain inhibitory activities were analyzed in soluble protein extracts from buds at different developmental stages and floral tissues in anthesis. These analyses identified high levels of inhibitory activity against both types of enzymes at all bud stages. Intriguingly, the inhibitory activity against both proteases differed remarkably in some floral tissues. While all organs tested were very effective against trypsin, only sepal and petal tissues exhibited strong inhibitory activity against papain. The sexual reproductive tissues (ovary, stigma-style and stamen) showed either significantly lower activity against papain or practically none. Gelatin–SDS–PAGE assay established that various trypsin inhibitors (TIs) homogenously accumulated in developing buds, although some were differentially present in floral organs. The N-terminal sequence analysis of purified inhibitors from stamen demonstrated they had homology to the Kunitz family of serine PIs. Western-blot analysis established presence of a ∼60 kDa cystatin, whose levels progressively increased during bud development. A positive correlation between this protein and strong papain inhibitory activity was observed in buds and floral tissues, except for the stigma-style. Differences in temporal and spatial accumulation of both types of PIs in passion fruit flowers are thus discussed in light of their potential roles in defense and development.     

舞钢玉兰芽种类与成枝成花规律的研究

报道了舞钢玉兰芽的种类、分枝习性与成枝生长规律,拟花蕾、着生位置、解剖结构及其分化发育成花规律。从中发现：（1）当年生枝上有休眠芽、叶芽（侧叶芽和顶叶芽）、拟花蕾3种;（2）拟花蕾有缩台枝、芽鳞状托叶、雏枝、雏芽及雏蕾组成,因其外形似“花蕾“,称为“拟花蕾“;（3）缩台枝是枝与花着生的中间过滤枝变阶段,是由母枝顶端节间缩短、增粗的枝段和当年由雏枝生长的1次极短新枝所组成;（4）4-5月及7-8月前后两批形成的拟花蕾,均经过未分化发育期、花被分化发育期、雄蕊群分化发育期及离心皮雌蕊群分化发育期,各期均依次递后交错进行,但不逆转,也不能截然分开,直到翌春花分化发育全部结束,开花后才能结实;（5）芽鳞状托叶是托叶的变态,最外层薄革质,外面密被短柔毛,始落期6月中下旬,其余纸质--膜质,外面密被或疏被毛柔毛,翌春开花时脱落完毕;（6）雏蕾有雏梗、雏花及包被雏花的佛焰苞状托叶组成;（7）分枝习性与成枝生长规律与预生分枝及预生一同生分枝呈单阶无歧、单阶1歧生长规律,稀有单阶2歧生长规律。     

The expression of tgas118, encoding a defensin in Lycopersicon esculentum, is regulated by gibberellin.

A flower specific cDNA, tgas118, has been isolated after differential screening of a gib-1 anther cDNA library of Lycopersicon esculentum. The corresponding mRNA was present in all tissues analysed. Northern blot analysis revealed that in wild-type tomato the gene was predominantly expressed throughout flower development, while in the gibberellin (GA)-deficient mutant of tomato (gib-1) the abundance declined. Treatment of the mutant with GA resulted in an accumulation of the tgas118 mRNA within hours in leaf and bud tissues. In the leaf, GA1, GA3 and GA9 were effective in enhancing the expression while GA4 was not. In addition to GA, wounding and dehydration also increased the accumulation of tgas118 mRNA in leaf tissue. In situ hybridization showed that application of 50 ng GA3 bud(-1) induced a similar spatial expression of the tgas118 mRNA in gib-1 buds 24 h post treatment to that found in wild-type flower buds. The deduced TGAS118 protein displays up to 77% similarity with defensins and as its expression is up-regulated by stimuli such as wounding it is proposed that it may play a role in protection against pathogens.     

Expression of two gibberellin-regulated cDNAs during early flower development in tomato (Solanum lycopersicon). Effect of grafting and paclobutrazol

To study the role of translocation of gibberellin (GA) intermediates or bioactive GAs from other plant parts to buds during early flower development in tomato ( Solanum lycopersicon ), the effect of grafting and paclobutrazol (PAC) treatment on the expression of tgas100 and tgas118 , two GA-regulated mRNAs, was analysed. Both mRNAs accumulated in a dose-dependent fashion. Application of 0.5 ng GA₃ per bud to developmentally arrested flower buds of a GA-deficient mutant of tomato ( gib-1 ) induced tgas100 expression, while the tgas118 abundance increased. For obtaining normal flower development through anthesis in the mutant, a single GA₃ treatment was required of at least 5 ng GA₃ per bud. In wild-type flower buds, PAC decreased the abundance of tgas100 and tgas118 mRNAs either when PAC was sprayed on whole plants or directly applied to buds. When only the wild-type buds were treated with PAC, the expression profiles characteristic for untreated buds were not restored by translocation of endogenous GAs. Grafting of gib-1 scions onto wild-type donor plants did not result in normal flower development or expression profiles like in wild-type buds. We conclude that the role of GA transport in early flower development of tomato is negligible and that the GAs required for development have to be synthesized in the flower bud itself.