Optical properties of bud scales and protochlorophyll(ide) forms in leaf primordia of closed and opened buds

The transmission spectra of bud scales of 14 woody species and the 77 K fluorescence emission spectra of the innermost leaf primordia of closed and opened buds of 37 woody species were studied. Pigment concentrations were determined in some species. Bud scales had low transmittance between 400 and 680 nm with a local minimum around 680 nm. Transmittance increased steeply above 680 nm and was > 80% in the 700-800 nm spectral region. Significant protochlorophyllide (Pchlide) accumulation was observed in leaf primordia of tightly packed, closed buds with relatively thick, dark bud scales. In common ash (Fraxinus excelsior L.) and Hungarian ash (Fraxinus angustifolia Vahl.), the innermost leaf primordia of the closed buds contained protochlorophyll (Pchl) and Pchlide (abbreviated as Pchl(ide)), but no chlorophyll. We observed Pchl(ide) forms with emission maxima at 633, 643 and 655 nm in these leaves. Complete transformation of Pchlide(655) (protochlorophyllide form with maximum emission at 655 nm) into Chlide(692) (chlorophyllide form with maximum emission at 692 nm) occurred after irradiation for 10 s. The innermost leaf primordia of the buds of four species (flowering ash (Fraxinus ornus L.), horse chestnut (Aesculus hippocastanum L.), tree of heaven (Ailanthus altissima P. Mill.) and common walnut (Juglans regia L.)) contained Pchl(ide)(633), Pchl(ide)(643) and Pchlide(655) as well as an emission band at 688 nm corresponding to a chlorophyll form. The Pchlide(655) was fully photoactive in these species. The outermost leaf primordia of these four species and the innermost leaf primordia of 28 other species contained all of the above described Pchl(ide) forms in various ratios but in small amounts. In addition, Chl forms were present and the main bands in the fluorescence emission spectra were at 690 or 740 nm, or both. The results indicate that Pchl(ide) accumulation occurs in leaf primordia in near darkness inside the tightly closed buds, where the bud scales and the external leaf primordia function as optical filters.     

Terminal bud failure of black cottonwood (Populus trichocarpa) exposed to salt-laden winter storms

At coastal sites, trees are exposed to marine aerosols that may cause foliar necrosis and shoot dieback, which can result in deformed crowns and contorted stems. A six-year study of leaf primordia in terminal buds of black cottonwood trees (Populus trichocarpa Torr. & Gray) on Heimaey Island off the south coast of Iceland was undertaken to elucidate the physiological events associated with salt-deposition-related bud failure. Leaf and bud lengths, dry mass, water content and chloride concentrations were monitored and related to four phenological stages: (1) bud set; (2) dormancy induction; (3) dormancy release; and (4) bud break. The trees set buds in July and shed their leaves by late September. Leaf primordia generally stopped growing by September 10 +/- 22 days and attained midwinter water content in late September. Leaf growth commenced in the terminal buds by March 2 +/- 16 days, but mean dates of bud swelling and bud break were April 29 +/- 19 and May 10 +/- 12 days. In summer and until November, chloride concentrations in leaf primordia were low, but increasing. Chloride concentrations remained stable from December to February, even though the dormant trees were exposed to large amounts of marine aerosols. In February and March, three events occurred more or less simultaneously: (1) leaf extension growth commenced; (2) chloride concentration surged in the leaf primordia; and (3) the leaf primordia began to hydrate. Following dormancy release, growth and hydration of leaf primordia were negatively related to chloride concentration in the leaf primordia, with inhibition of leaf growth, tissue hydration and chloride acquisition occurring at a chloride concentration threshold estimated at 7.3 mg Cl- g(-1) tissue water. Necrosis of leaf primordia was observed above 14 mg Cl- g(-1) tissue water. Growth and hydration of leaves at bud break in mid-May was explained by a three-parameter logistic model of chloride concentration in leaf primordia at the end of March. By mid-May, 90% of all buds remained non-necrotic, but only 56% the terminal buds had broken. Salt alone explained the observed growth suppression of leaf primordia in the buds and the resultant failure of terminal buds to break by mid-May.     

Hydraulic architecture correlates with bud organogenesis and primary shoot growth in beech (Fagus sylvatica)

In beech (Fagus sylvatica L.), the number of leaf primordia preformed in the buds determines the length and the type (long versus short) of annual growth units, and thus, branch growth and architecture. We analyzed the correlation between the number of leaf primordia and the hydraulic conductance of the vascular system connected to the buds. Terminal buds of short growth units and axillary buds of long growth units on lower branches of mature trees were examined. Buds with less than four and more than five leaf primordia formed short and long growth units, respectively. Irrespective of the type of growth unit the bud was formed on, the occurrence of a large number of leaf primordia was associated with high xylem hydraulic conductance. Xylem conductance was correlated to the area of the outermost annual ring. These results suggest that organogenesis and primary growth in buds correlates with secondary growth of the growth units and thus with their hydraulic architecture. Possible causal relationships between the variables are discussed.     

Preformation in vegetative buds of Prunus persica: factors influencing number of leaf primordia in overwintering buds

We investigated the influence of bud position, cultivar, tree age, tree carbohydrate status, sampling date, drought and light exposure on the number of leaf primordia formed in dormant vegetative peach buds (Prunus persica (L.) Batsch) relative to the number of primordia formed after bud break (neoformed). During winter dormancy, vegetative peach buds from California and Italy were dissected and the number of leaf primordia recorded. Between leaf drop and bud break, the number of leaf primordia doubled from about five to about 10. Parent shoot length, number of nodes on the parent shoot, cross-sectional area of the parent shoot, bud position along the parent shoot and bud cross-sectional area were correlated with the number of leaf primordia. Previous season light exposure, drought and tree carbohydrate status did not affect the number of leaf primordia present. The number of leaf primordia differed significantly among peach varieties and tree ages at leaf drop, but not at bud break. Our results indicate that neoformation accounted for all shoot growth beyond about 10 nodes. The predominance of neoformed shoot growth in peach allows this species great plasticity in its response to current-season conditions.     

Freezing behaviors in leaf buds of cold-hardy conifers visualized by NMR microscopy

1H-Nuclear magnetic resonance (NMR) microscopy was used to study freezing behavior in wintering leaf buds of Momi fir (Abies firma Sieb. et Zucc.) and Japanese red pine (Pinus densiflora Sieb. et Zucc.). The images acquired predominantly reflected the density of mobile (i.e., non-ice) protons mainly from unfrozen water. By comparing images taken at various subfreezing temperatures, we determined which tissues produced the high and low temperature exotherms detected by differential thermal analyses. Typical extra-organ freezing was successfully imaged in leaf buds of A. firma. The bud scales readily froze at -7 degrees C, but shoot primordia remained supercooled to -14 degrees C in December buds and to -21 degrees C in March buds. The size of supercooled shoot primordia was reduced with decreasing temperature, indicating a gradual decrease in water content of the shoot primordia. In contrast, the signal from shoot primordia of P. densiflora disappeared between -7 and -14 degrees C, corresponding to the high temperature exotherm at -8 degrees C, indicating extracellular freezing of the shoot primordia. The xylem and bark tissues readily froze at -7 degrees C in A. firma and between -7 and -14 degrees C in P. densiflora. We conclude that NMR microscopy can noninvasively provide more spatially specific information about freezing behavior in leaf buds than traditional methods such as differential thermal analysis. In particular, it allows the organized and harmonized freezing behaviors in complex organs to be visualized directly thereby revealing the diversity of mechanisms involved in freezing behaviors.     

Masting in Fagus crenata and its influence on the nitrogen content and dry mass of winter buds

In Fagus, full-mast seeding years are invariably followed by at least one non-mast year. Both flower and leaf primordia develop during the summer within the same winter buds. Flower bud initiation occurs when the N content of developing seeds is increasing rapidly. We hypothesized that competition for nitrogen (N) between developing seeds and buds limits flower primordium formation in mast years and, hence, limits seed production in years following mast years. We tested this hypothesis in three Fagus crenata Blume forests at elevations of 550, 900 and 1500 m. Bud N concentration (N con), amount of N per bud (N bud) and dry mass per bud (DM) were compared between a mast year (2005) and the following non-mast year (2006), and between winter buds containing both leaf and flower primoridia (BF), which were formed during the non-mast year, and winter buds containing leaf primordia only (BL), which were formed in both mast and non-mast years. In addition, leaf numbers per shoot corresponding to the analyzed buds were counted, and the effect of masting on litter production was analyzed by quantifying the amounts of litter that fell in the years 2004 to 2007. The dry mass and N content of BF formed in 2006 by trees at both 550 and 1500 m were 2.1-3.4-fold higher than the corresponding amounts in BL, although the numbers of leaves per current-year shoot in 2007 that developed from the two bud types in the same individuals did not differ significantly. These results indicate that more N and carbohydrate are expended in producing BF than in producing BL. The amount of litter from reproductive organs produced in the mast year was similar to the amount of leaf litter at 900 and 1500 m, but three times as much at 550 m. Leaf numbers per shoot were significantly lower at all elevations in the mast year than in the non-mast years (and the amount of leaf litter at 550 and 1500 m tended to be lower in the mast year than in the non-mast years. In conclusion, preferential allocation of resources to seeds in the mast year reduced the availability of resources for flower primordium formation, and this may have accounted for the poor seed production in the following non-mast year.     

Detection of the endophyte Discula umbrinella in buds and twigs of Fagus sylvatica

Endophytic fungi have been isolated from buds and twigs of beech trees collected at four different sites in Switzerland. Discula umbrinella. was recovered at high frequencies from the bud scales and the twig pieces contiguous to the buds, but was virtually absent from the rolled up leaves enclosed by the scales. In addition to infection by air-borne inoculum, thalli of D. umbrinella may grow from the twigs into the leaf tissues.     

Effect of GA(4 + 7) on the initiation and development of the inflorescence bud of evergreen azalea

Apical buds of evergreen azalea (Rhododendron sp.) were treated with GA(4 + 7) at different stages of development. Treatment of vegetative buds stimulated shoot growth, slightly delayed both flower initiation and development, but increased the number of flower primordia. Treatment at the time of floral transition induced bud abortion at an early stage of the reproductive development. Treatment of inflorescence buds which contained at least one complete flower substituted for chilling in overcoming dormancy and prevented inflorescence bud abortion.     

Nitrogen sources for current-year shoot growth in 50-year-old sessile oak trees: an in situ (15)N labeling approach

We used long-term in situ (15)N labeling of the soil to investigate the contribution of the two main nitrogen (N) sources (N uptake versus N reserves) to sun shoot growth from bud burst to full leaf expansion in 50-year-old sessile oaks. Recovery of (15)N by growing compartments (leaves, twigs and buds) and presence of (15)N in phloem sap were checked weekly. During the first 2 weeks following bud burst, remobilized N contributed ~90% of total N in growing leaves and twigs. Nitrogen uptake from the soil started concomitantly with N remobilization but contributed only slightly to bud burst. However, the fraction of total N due to N uptake increased markedly once bud burst had occurred, reaching 27% in fully expanded leaves and 18% in developed twigs. In phloem sap, the (15)N label appeared a few days after the beginning of labeling and increased until the end of bud burst, and then decreased at full leaf expansion in June. Of all the shoot compartments, leaves attracted most of the absorbed N, which accounted for 68% of new N in shoots, whereas twigs and new buds accounted for only 28 and 3%, respectively. New N allocated to leaves increased from unfolding to full expansion as total N concentration in the leaves decreased. Our results underline the crucial role played by stored N in rapid leaf growth and in the sustained growth of oak trees. Any factors that reduce N storage in autumn may therefore impair spring shoot growth.     

Characterization and identification of leaf morphology of Populus deltoides Bartr.clones

Leaves are of fundamental importance to plants,representing their facility to generate power and are the sensing units of plants towards the environment.An attempt was made to characterize and compare the variations of leaf morphology of various Populus deltoides Bartr.clones by studying the winter buds and other leaf parameters of fully developed leaves.To achieve these objectives,forty-three exotic and indigenous clones of P.deltoides Bartr.were evaluated for different parameters.On the basis of various morphological characteristics the results reveal that each clone has a distinct color pattern of leaves.Different colors observed in these clones varied from light green through green to dark green.Two distinct lengths of the leaf apex were found,i.e.,short and long; as well both acuminate and acute apex types were found.Erratic distribution of serration of leaves was also found.In this study,the morphological traits of leaves provided discriminatory grounds for separating various populations of P.deltoides Bartr.clones.Winter bud studies indicate that different clones vary considerably with regard to shape,color,shape of leaf scars and exudation.     

Trophic control of bud break in peach (Prunus persica) trees: a possible role of hexoses

Vegetative buds of peach (Prunus persica L. Batsch.) trees act as strong sinks and their bud break capacity can be profoundly affected by carbohydrate availability during the rest period (November-February). Analysis of xylem sap revealed seasonal changes in concentrations of sorbitol and hexoses (glucose and fructose). Sorbitol concentrations decreased and hexose concentrations increased with increasing bud break capacity. Sucrose concentration in xylem sap increased significantly but remained low. To clarify their respective roles in the early events of bud break, carbohydrate concentrations and uptake rates, and activities of NAD-dependent sorbitol dehydrogenase (SDH), sorbitol oxidase (SOX) and cell wall invertase (CWI) were determined in meristematic tissues, cushion tissues and stem segments. Only CWI activity increased in meristematic tissues shortly before bud break. In buds displaying high bud break capacity (during January and February), concentrations of sorbitol and sucrose in meristematic tissues were almost unchanged, paralleling their low rates of uptake and utilization by meristematic tissues, and indicating that sorbitol and sucrose play a negligible role in the bud break process. Hexose concentrations in meristematic tissues and glucose imported by meristematic tissues correlated positively with bud break capacity, suggesting that hexoses are involved in the early events of bud break. These findings were confirmed by data for buds that were unable to break because they had been collected from trees deprived of cold. We therefore conclude that hexoses are of greater importance than sorbitol or sucrose in the early events of bud break in peach trees.     

Photoperiodic control of seasonal development and dormancy in tropical stem-succulent trees

Tropical stem-succulent trees store large quantities of water in their trunks yet remain leafless during the early and mid dry season. In contrast to most other tropical trees, bud break of vegetative buds is not induced in fully hydrated stem succulents between the winter solstice and the spring equinox by leaf abscission, abnormal rain showers or irrigation. Vegetative buds of leafless trees are therefore in a state of endo-dormancy similar to that of temperate perennial plants during early winter. Highly synchronous bud break regularly occurs soon after the spring equinox, often weeks before the first rainfalls of the wet season. These observations suggested that endo-dormancy and bud break might be induced by declining and increasing photoperiods after the autumn and spring equinoxes, respectively. In phenological field observations, we confirmed highly synchronous bud break after the spring equinox in many trees of five stem-succulent species in the northern and southern hemispheres. Shoot growth of potted saplings of Plumeria rubra L. was arrested by a decline in day length below 12 h after the autumn equinox, but continued in saplings maintained in a 13-h photoperiod. Conversely, exposure to a 13-h photoperiod induced bud break of dormant apical buds in saplings and cuttings in January, whereas plants maintained in the natural day length of < 11.7 h remained dormant. Photoperiodic control of endo-dormancy of vegetative buds in stem succulents is thus supported by field observations and experimental variation of the photoperiod. At low latitudes, where annual variation of day length is less than 1 h, bud dormancy is induced and broken by variations in photoperiod of less than 30 min.     

Effects of near-lethal heat stress on bud break, heat-shock proteins and ubiquitin in dormant poplar (Populus nigra Charkowiensis x P. nigra incrassata)

We assessed the effects of near-lethal heat stress on bud break, heat-shock proteins (HSPs) and ubiquitin in hybrid poplar (Populus nigra (L.) Charkowiensis x P. nigra (L.) incrassata). Shoots, with 10-15 buds each, were collected from September to March and exposed to temperatures between 20 and 60 degrees C for 2 h. Shoots were then placed in a greenhouse at 18-22 degrees C with supplemental light and cumulative bud break was recorded over a 4-week period. Samples of bud tissues were collected during and up to 96 h after heat treatment for protein analysis. De novo synthesis of proteins was monitored by exposing excised buds to [(35)S]-methionine for 3 h before, during, or after heat treatment. Heat treatments of 40-45 degrees C resulted in both a release from endodormancy and a decrease in thermal units needed for bud break during ecodormancy. The response to near-lethal heat stress was complex and was affected by intrinsic thermal sensitivity. Heat treatments were least effective during August and became progressively more effective as endodormancy progressed. In the later stages of ecodormancy, a heat treatment of 45 degrees C either inhibited bud break or killed the buds. Although temperatures of 42.5 to 45 degrees C inhibited incorporation of [(35)S]-methionine into proteins for at least 48 h, several HSPs were synthesized in response to temperatures of 40-45 degrees C. Immunoblots indicated that one of the heat-induced proteins was immunologically related to HSP70. Increases in free and conjugated forms of ubiquitin were also observed in response to heat treatment. Production of HSPs and ubiquitin, however, was not consistently associated with the heat treatments that induced the highest percentage of bud break. The roles of heat-induced protein degradation, HSPs, and ubiquitin in overcoming dormancy by near-lethal heat stress are discussed.     

The effect of bagging branches on levels of endophytic fungal infection in Japanese beech leaves

The species composition of the endophytic mycobiota in leaves of Japanese beech trees (Fagus crenata) and the sources for leaf infections were studied in a forest reserve situated in central eastern Honshu, Japan. To clarify the mechanism of infection of leaves, half of the branches were covered with polyethylene bags and species composition and levels of endophytic fungal infection were then compared with those of unbagged controls. Isolations were carried out from the leaves, petioles, and current‐year twigs of both, bagged and unbagged branches. Additionally, species composition was detected in overwintered terminal buds of beech trees and in the leaves of potted seedlings that had been placed in the field in different seasons. The species assemblage of the unbagged leaves, petioles, and current‐year twigs was dominated by Mycosphaerella buna, Ascochyta fagi, Periconiella sp., and Tritirachium sp. Other frequently recovered species were Xylaria sp., Phomopsis sp., and Tubakia dryina. Mycosphaerella buna and A. fagi were never isolated from leaves on bagged branches. A. fagi was, however, detected on both bagged and unbagged petioles and current‐year twigs at comparatively low isolation frequencies. The detection of Periconiella sp. on all occasions in both bagged and unbagged leaves was a characteristic feature that differs from those of the other three dominant endophytic fungi. The fungus was also detected without significant differences in bagged and unbagged petioles and current‐year twigs on most sampling dates. Furthermore, Periconiella sp. was isolated from immature twigs inside the bud scales. Tritirachium sp. was frequently detected in unbagged leaves and petioles and in both bagged and unbagged current‐year twigs, and rarely in bagged leaves and petioles, but was never recovered from terminal buds. The results of the potted seedling experiments revealed that all four dominant species had airborne inocula. The infection of leaves by M. buna occurs exclusively by airborne propagules, i.e. ascospores in spring and conidia in autumn. In Periconiella sp. hyphal growth of the fungus from immature twigs inside the buds into the leaf tissues was suggested in addition to infection by airborne inocula. Tritirachium sp. hyphae were suggested to grow from previous‐ to current‐year twigs. Ascochyta fagi was present in the outermost scales of overwintered terminal buds, but no systemic growth of the fungus into the petioles and current‐year twigs was observed. Our technique of covering the branches before new leaves unfolded was effective in preventing infection by airborne inocula of endophytic fungi.     

路易斯安娜鸢尾组培和苗期生长规律初步研究

以路易斯安娜鸢尾顶芽为外植体,利用不同激素配比诱导不定芽,定期测量组培苗株高、蓬径、主茎长和叶片数,观察组培苗的染色体数、叶片结构和开花性状等。结果表明:路易斯安娜鸢尾顶芽诱导率最高的培养基为MS+6-BA 2.0 mg.L-1+NAA 0.5 mg.L-1,产生有效生产苗最多的培养基为MS+6-BA 0.5 mg.L-1+NAA 0.2 mg.L-1,最佳生根培养基为1/2MS+NAA 0.5 mg.L-1+活性碳0.5 g.L-1。2年生组培苗平均株高达到71.6 cm,平均蓬径达到60.9 cm,叶片维持在6~12片,冬季仍有绿叶。组培苗染色体个数、叶片通气组织、开花性状等均与母株一致,表明路易斯安娜鸢尾组培苗可保持其母株的观赏性状和水生习性,可通过组培快速繁殖大规模生产优质种苗。     

Shoot development,chlorophyll, gas exchange and carbohydrates in lychee seedlings (Litchi chinensis)

Shoot growth, chlorophyll concentrations, gas exchange and starch concentrations were studied in lychee (Litchi chinensis Sonn.) seedlings of cultivar "Wai Chee" grown in a heated greenhouse at Nambour in subtropical Australia (27 degrees S). We also examined the effects of shoot defoliation and root pruning on leaf expansion. Shoot growth showed a rhythmic cycle under constant greenhouse conditions, with a mean duration of flushing of 20 days and an interval of 10 days over three cycles. Shoots and leaves expanded in a sigmoidal pattern to about 80 mm and 500 cm(2), respectively, for each flush. Starch concentrations of the lower stem and roots decreased as the young red leaves expanded, and increased as the fully expanded leaves turned dark green. Chlorophyll concentrations and net CO(2) assimilation rate were highest in the fully expanded dark green leaves. Removing 50% of the area of each fully expanded leaf had little effect on the expansion of younger leaves, but total biomass of defoliated plants was only 60% of that of controls. In contrast, removing half the roots just before bud swelling reduced final leaf area by 80%. We conclude that the young shoot has relatively low rates of photoassimilation until the leaves are fully expanded and dark green, and depends on assimilates from elsewhere in the plant. During leaf expansion, translocation of assimilates to the shoot occurred at the expense of the roots.     

Influence of temperature on the dynamics of ATP,ADP and non-adenylic triphosphate nucleotides in vegetative and floral peach buds during dormancy

The nucleotides test of endodormancy, which is based on the capacity of tissues to synthesize ATP and non-adenylic triphosphate nucleotides (NTP), cannot be used for floral buds, and it is of questionable use for vegetative buds. In an attempt to find an alternative test, we examined whether the dormancy state of vegetative and floral buds of trees exposed to different temperature conditions during the rest period is directly related to their ATP, ADP and NTP concentrations and ATP/ADP ratio. Once the buds had entered endo- or paradormancy, the nucleotide concentrations and the ATP/ADP ratio were low in the vegetative primordia and very low in the floral primordia. Only after the action of chilling, when the buds were considered to have completed the endodormancy and paradormancy phases, did the nucleotide concentrations increase, accompanied by a steep rise in ATP/ADP ratio. We conclude that the ATP/ADP ratio could be used to characterize the bud dormancy state by comparison with critical values of 1.5 for vegetative primordia and 1.0 for floral primordia.     

Variation in the chemical composition of green leaves and leaf litters from three deciduous tree species growing on different soil types

The chemical composition of green leaves and leaf litters of sweet chestnut (Castanea sativa), oak (Quercus robur) and beech (Fagus sylvatica) were determined for 26 sites grouped into high fertility (HF) and low fertility (LF) soils according to base saturation and N-mineralization potentials. Measurements were made of total carbon, acid detergent fibre (ADF), Klason lignin, holo-cellulose, sugar constituents of hemicellulose and phenylpropanoid derivatives of lignin, and nutrient concentrations (N, Ca, P, Mg, K and Mn). Leaf and litter constituents varied within and between species according to soil groups, but beech showed contrasting responses to oak and chestnut. Beech leaves had lower ADF, lignin and cellulose on HF soils than LF soils, whereas oak and chestnut leaves had higher ADF, lignin and cellulose on HF than the LF soils. Conversely, the same constituents in beech leaf litter were higher on HF soils than LF soils, but lower in oak and chestnut leaf litter on HF soils than LF soils. The phenylpropanoid derivatives of lignin and sugar constituents of hemicellulose also showed similar variations in relation to soil groups with contrasting patterns for in leaves and litters. Re-absorption of N from leaves before litter fall was negatively correlated with soil N mineralization potential for beech (highest on LF soils) but showed an unexpected, positive relationship for oak and chestnut (highest on HF soils). These intra-specific differences of leaf and litter chemistry in relation to soil fertility status are unprecedented and largely unexplained. The observed patterns reflect phenotypic responses to soil type that result in continuum of litter quality, within and between tree species, that have been shown in related studies to significantly influence litter decomposition rates.     

Seasonal variation in the photosynthetic capacity of a willow (Salix cv. Aquatica gigantea) canopy. 2 Comparison of the structure and function of chloroplasts at different levels in the canopy

Development of leaf chloroplast ultrastructure at five levels in a willow (Salix cv. Aquatica gigantea) canopy was followed during one growing season in the field. Changes in chloroplast ultrastructure were compared with the rate of CO(2) uptake of the same leaves. The highest rates of CO(2) uptake were recorded in young leaves exposed to full available sunlight. In these leaves, the area of the grana stacks was less than 20% of the total chloroplast area and the degree of thylakoid stacking was less than 1.5. The chloroplasts of these leaves contained large amounts of starch and small amounts of plastoglobuli. As the canopy grew and the leaves in the lower parts of the canopy became shaded, the structure of the chloroplast thylakoids gradually changed. In leaves at the two lowest levels of the canopy, the degree of stacking at the end of the growing season was close to 2 and correspondingly the rate of CO(2) uptake was low. The areas of grana stacks and plastoglobuli in these chloroplasts increased and were about 30 and 10% of the chloroplast area, respectively, by the end of the growing season. The increase in the degree of thylakoid stacking was caused by increased biosynthesis of grana lamellae, which in general were thinner than the lamellae of young leaves. The length of the stroma lamellae did not change with leaf age. Morphometric measurements showed that the structure of the chloroplasts in leaves 160 cm above ground was dynamic and responsive to environmental conditions so that photosynthetic capacity remained high for 7 weeks despite an increase in leaf shading.