The Importance of Phellogen Cells and their Structural Characteristics in Susceptibility and Resistance to Excoriation in Immature and Mature Potato Tuber (Solanum tuberosum L.) Periderm

Potato tuber (Solanum tuberosum L.) periderm maturation is animportant physiological process that directly affects the susceptibilityand development of resistance to costly excoriation (skinning-typewounds) at harvest. The objectives of this research were toidentify the specific types of cells and the cellular changesassociated with susceptibility and resistance to tuber excoriationin immature and mature tubers respectively. Epifluorescent microscopicexamination of immature tuber periderm (phellem, phellogen andphelloderm cells) from several genetically diverse cultivarshas shown that the cellular damage resulting from excoriationoccurs within the phellogen (cork cambium), a meristematic layerof cells that gives rise to neighbouring phellem and phellodermcells. Tuber excoriation is the result of the fracture of radialphellogen cell walls linking the skin (phellem) to the phelloderm.As the tuber periderm matures, phellogen cells become inactiveand the radial walls of these cells become more resistant tofracture; resistance to excoriation develops. Ultrastructuralstudies of immature tuber periderm show that radial walls ofactive phellogen cells are thin and fragile. During peridermmaturation, both radial and tangential phellogen cell wallsthicken as they strengthen and become resistant to fracture,thereby providing resistance to excoriation. These results refuteprevious theories of the physiological changes responsible forthe onset of resistance to tuber skinning injury. The combinedresults establish a paradigm whereby the thickening and strengtheningof tuber phellogen cell walls upon periderm maturation are thedeterminant for resistance to tuber excoriation. Copyright 2001Annals of Botany Company Cambium, meristematic, periderm, phellem, phelloderm, phellogen, potato, skinning, Solanum tuberosum L., 0tuber     

Molecular and cytological aspects of native periderm maturation in potato tubers

Mature native periderm that exhibits resistance to excoriation (RE) is the primary defense for potato tubers against abiotic and biotic challenges. However, little is known about the physiology of periderm maturation and associated gene expressions. In this study, periderm maturation events and associated gene expressions were determined in tubers of two diverse potato genotypes (NDTX4271-5R (ND) and Russet Burbank (RB); 2008 and 2009 crops) at four harvest maturities ranging from immature (non-senesced vines and low RE) to mature (senesced vines and high RE). Approximately 104 d after planting, the fine balance of accumulation and loss of periderm phellem cell layers showed signs of subsiding, indicating cessation of cell division by the phellogen. Phellogen radial cell walls thickened as periderm matured throughout the harvests, increasing RE/skin-set. In both genotypes, the cell cycle gene cyclin-dependent kinase B (StCDKB) rapidly down-regulated after the second harvest coinciding with apparent cessation of cell division. Expression patterns of genes encoding epidermal growth factor binding protein (StEBP) and cyclin-dependent kinase regulatory subunit (StCKS1At) were less indicative of phellogen inactivation and periderm maturation. Genes encoding the structural cell wall proteins extensin (StExt1) for ND and extensin-like (StExtlk) for ND and RB remained up-regulated respectively by the second harvest, suggesting involvement with completion of phellem cell accumulation and on-set of periderm maturation. The expression of genes encoding pectin methyl esterase (StPME), StExt1 and a cell wall strengthening “tyrosine-and lysine-rich protein” (StTLRP) increased in phellogen cells from later harvests of ND tubers, but were down regulated in RB tubers; this suggests roles in phellem cell generation and completion of delayed cell wall development in non-meristematic phellogen cells of ND, a red skinned phenotype. StCDKB and StPrePME genes were rapidly down-regulated by the third harvest for both genotypes. Collectively, these results suggest that down-regulation of these genes coordinates with on-set of periderm maturation and skin-set progression.     

DEVELOPMENTAL ANATOMY OF THE HORNED OAK GALL INDUCED BY CALLIRHYTIS CORNIGERA ON QUERCUS PALUSTRIS (PIN OAK)

The horned oak gall forms on twigs of Quercus palustris Muench. and is initiated when the wasp Callirhytis corrigera O.S. oviposits into the periderm or cortex of the twigs. Injury to phellogen as a consequence of oviposition results in dedifferentiation of phelloderm and underlying phloem tissue to form a wound-response phellogen. The wound-response phellogen encircles and compartmentalizes the wasp ovum and is continuous with the normal stem phellogen. This tissue arrangement forms the framework for the ensuing stages of gall development. Development of the insect larva coincides with the formation of a gall-matrix phellogen. This additional meristem is derived from locally hyperplastic stem phellogen where adjacent to the wound-response phellogen. The essentially parenchymatous, roughly spherical body of the gall, the gall matrix, is formed chiefly from derivatives of the gall-matrix and wound-response phellogens. In the immediate area of the wasp larva, the wound-response phellogen produces cells toward the larva that differentiate to form a vascularized, externally sclerified, horn-shaped chamber containing in its base the developing insect larva. The involvement of the vascular cambium is apparent during intermediate gall development stages. Vascular strands, produced transverse to the stem axis, penetrate the gall-matrix tissue and terminate near the base of the horn-like larval chamber. Vessel elements in the region of the gall that are not included in this tissue alteration often become heavily tylosed. Disruption of the vascular tissue through the galled area appears to be a primary cause of twig death.     

Histological analysis of the maturation of native and wound periderm in potato (Solanum tuberosum L.) Tuber

Maturation of potato (Solanum tuberosum L.) tuber native and wound periderm and development of resistance to periderm abrasion were investigated utilizing cytological and histochemical techniques. Both native and wound periderm consist of three different tissues: phellem, phellogen and phelloderm. It was previously determined that the phellogen walls of immature native periderm are thin and prone to fracture during harvest, leading to periderm abrasion (excoriation). Phellogen walls thicken and become less susceptible to fracture upon maturation of the periderm, leading to resistance to excoriation. We now demonstrate that phellogen cells of immature wound periderm also have thin radial walls and that wound periderm abrasion is due to fracture of these walls. Maturation of the wound periderm is also associated with an increase in the thickness of the phellogen radial walls. Histological analysis with ruthenium red and hydroxylamine-FeCI2, which stain unesterified and highly methyl-esterified pectins, respectively, indicates that the phellogen cell walls of native and wound periderm differ significantly regardless of the stage of maturity. Results obtained by staining with ruthenium red and hydroxylamine-FeCI2 imply that phellogen cell walls of immature native periderm contain methyl-esterified pectin, but are lacking in unesterified (acidic) pectins. Maturation of native periderm is accompanied by an apparent increase in unesterified pectins in the walls of phellogen cells, which may allow for the strengthening of phellogen cell walls via calcium pectate formation. Histological staining of the phellogen walls of wound periderm, on the other hand, implies that these walls are deficient in pectins. Moreover, maturation of wound periderm is not accompanied by an increase in unesterified pectins in these walls. Since peroxidase is known to catalyse the cross-linking of cell wall polymers, we stained native and wound periderm for the presence of peroxidase utilizing guaiacol as a substrate. Peroxidase staining was strong in the phellogen walls of both immature and mature native periderm and we could not detect any differences in staining between them. Peroxidase staining was weak in the phellogen walls of immature wound periderm and was not detectably different in mature wound periderm. Peroxidase data imply that there are distinct differences between native and wound periderm, though our data do not indicate that changes in peroxidase activity are involved in the development of resistance to periderm abrasion that occurs upon maturation of the periderm. However, we cannot rule out the involvement in this process of peroxidase isozymes that have low affinity for the substrates utilized here.     

Wounding coordinately induces cell wall protein, cell cycle and pectin methyl esterase genes involved in tuber closing layer and wound periderm development

Little is known about the coordinate induction of genes that may be involved in agriculturally important wound-healing events. In this study, wound-healing events were determined together with wound-induced expression profiles of selected cell cycle, cell wall protein, and pectin methyl esterase genes using two diverse potato genotypes and two harvests (NDTX4271-5R and Russet Burbank tubers; 2008 and 2009 harvests). By 5 d after wounding, the closing layer and a nascent phellogen had formed. Phellogen cell divisions generated phellem layers until cessation of cell division at 28 d after wounding for both genotypes and harvests. Cell cycle genes encoding epidermal growth factor binding protein (StEBP), cyclin-dependent kinase B (StCDKB) and cyclin-dependent kinase regulatory subunit (StCKS1At) were induced by 1 d after wounding; these expressions coordinated with related phellogen formation and the induction and cessation of phellem cell formation. Genes encoding the structural cell wall proteins extensin (StExt1) and extensin-like (StExtlk) were dramatically up-regulated by 1-5 d after wounding, suggesting involvement with closing layer and later phellem cell layer formation. Wounding up-regulated pectin methyl esterase genes (StPME and StPrePME); StPME expression increased during closing layer and phellem cell formation, whereas maximum expression of StPrePME occurred at 5-14 d after wounding, implicating involvement in later modifications for closing layer and phellem cell formation. The coordinate induction and expression profile of StTLRP, a gene encoding a cell wall strengthening "tyrosine-and lysine-rich protein," suggested a role in the formation of the closing layer followed by phellem cell generation and maturation. Collectively, the genes monitored were wound-inducible and their expression profiles markedly coordinated with closing layer formation and the index for phellogen layer meristematic activity during wound periderm development; results were more influenced by harvest than genotype. Importantly, StTLRP was the only gene examined that may be involved in phellogen cell wall thickening after cessation of phellogen cell division.     

Developmental stages and fine structure of surface callus formed after debarking of living lime trees (Tilia sp.)

Wounding of trees by debarking during the vegetative period sometimes results in the formation of callus tissue which develops over the entire wound surface or on parts of it. This light and transmission electron microscopy study of living lime trees found that the formation of such a surface callus is subdivided into three stages. During the first stage, numerous cell divisions take place in regions where differentiating xylem remains at the wound surface after debarking. This young callus tissue consists of isodiametric parenchymatous cells. Cambium cells, sometimes also remaining at the wound surface, collapse and do not contribute to callus formation. During the second stage, cells in the callus undergo differentiation by forming a wound periderm with phellem, phellogen and phelloderm. In the third stage, a cambial zone develops between the wound periderm and the xylem tissue laid down prior to wounding. This process is initiated by anticlinal and periclinal divisions of a few callus cells only. Later this process extends tangentially to form a continuous belt of wound cambium. Subsequently, this cambium produces both wound xylem and wound phloem and thus contributes to further thickening.     

Polyamine levels in buds and twigs of Tilia cordata from dormancy onset to bud break

The fluctuations of free and bound polyamines (PAs) were studied in vegetative buds and underlying twigs of linden (Tilia cordata L.) from August to May, to assess the connection between PA levels and seasonal cycles of growth and dormancy. Outer and inner bud scales and shoot tips (short shoot tips with leaf initials in contiguous short internodes) were analyzed separately, as were phloem with cortex and xylem with pith tissue from twigs. Seasonal variations in PA levels were present in buds and twigs during the research period. The most abundant PA in buds and twigs in free and bound forms was spermidine followed by putrescine. PA amounts were low in buds and twigs in autumn. In twig tissues, free PAs were predominant whereas in bud scales, bound PAs accumulated over free PAs in autumn, first in inner scales and later in outer scales as well. PA levels did not increase dramatically during the onset of dormancy in autumn but lower temperatures and probable cold hardening correlated positively with bound PAs in bud scales. In shoot tips with leaf initials, and contiguous short internodes, free putrescine and spermidine levels rose simultaneously with bud burst and new growth, while bound PAs diminished quite radically from temporary bud scales and from growing shoot tips.     

The effect of prolonged flooding on the bark of mangrove trees

Growth and physiological response of woody plants to flooding have been analyzed in detail; however, relatively few studies have been oriented towards the effects of water immersion on cambial activity and wood and bark anatomy of trees that are growing in prolonged flooding conditions. These studies are important to understand the possible effects of predicted sea level rising in mangroves as a consequence of global warming. We studied five species growing in a mangrove forest, sampling three to six trees of each species, in sites that have the longest flooding period. Differences in bark appearance and phloem structure between the submerged stem portion and the portion of the stem above the water surface exist in all species. Although aerenchyma formation and stem hypertrophy are the most common events related to flooding, each type of tissue responded differently. Annona glabra L., Laguncularia racemosa (L.) Gaertn f. and Hibiscus tiliaceus L. developed rythidome. Avicennia germinans (L.) Stearn developed rythidome only in the submerged stem portion. Phyllanthus elsiae Urb., developed one periderm in both stem portions. Species that developed rythidome also developed aerenchyma between periderms and in the phellem. H. tiliaceus and P. elsiae, showed the highest values for anatomical phloem and periderm characters below water surface, while an inverse tendency was observed in A. glabra and L. racemosa, suggesting that prolonged flooding modifies vascular cambium and phellogen differently. Results indicate that sea level rising would affect distribution of the species according to their specific flooding tolerance.     

Buds buried in bark: the reason why <Emphasis Type="Italic">Quercus suber</Emphasis> (cork oak) is an excellent post-fire epicormic resprouter

Key message

Cork oak has buds protected by the full thickness of its substantial phellem, thus explaining why it is the only European tree that can epicormically resprout after higher intensity fire.

Abstract

Epicormic resprouting has various ecological advantages over basal resprouting. However, after higher intensity fires epicormic resprouting is rare as it is difficult for trees and shrubs to keep both their buds and vascular cambia alive. Quercus suber (cork oak) is the only European tree that can resprout epicormically after higher intensity fires. Q. suber develops very thick bark and it has been assumed, without anatomical evidence, that the bark protects the epicormic buds. We investigated if developmental anatomy could explain why Q. suber is an excellent post-fire epicormic resprouter. We examined buds from mature Q. suber trees, macroscopically using a stereo microscope and microscopically using semi-thin microtome sections. Q. suber produced buds in the foliage leaf axils and the bud scale axils. With the commencement of extensive phellem (cork) production the base of the epicormic buds remained at, or just below, the level of the phellogen and thus cork began to bury the buds, although a narrow tube connected each bud to the bark surface. Q. suber epicormic buds became deeply buried in the phellem and would be protected from heat by the full phellem thickness. With its rapid and substantial development of phellem Q. suber had well-protected epicormic buds even in relatively small diameter stems. These results provide the anatomical evidence to show why Q. suber is a noted epicormic resprouter after crown fire.

     

Frosthärtegradienten entlang der Knospenfolge auf Eschentrieben

Zusammenfassung Die Manna-Esche,Fraxinus ornus, weist eine deutliche akrotone Knospenförderung auf. Die Terminalknospe ist stets die größte Knospe, zwei kleinere Beiknospen, die normalerweise nicht austreiben, sitzen ihr unmittelbar an. Die übrigen Seitenknospen sind entsprechend der dekussierten Blattstellung paarweise auf jedem Knoten inseriert und nehmen in ihrer Größe von oben nach unten ab. An der jeweiliger Triebbasis befinden sich außerdem noch winzige Ersatzknospen.Dieser Knospenfolge entspricht ein Gradient in der Kälteresistenz. Die Terminalknospe ist immer die kälteempfindlichste Knospe, die Ersatzknospen sind immer am resistentesten. Im zeitigen Frühjahr, wenn die Pflanze ihre Aktivität aufzunehmen beginnt, geht die winterliche Kälteresistenz in jenen Knospen am schnellsten verloren, die am frühesten zum Austrieb kommen. Nicht austreibende Knospen, wie beispielsweise die subterminalen Beiknospen und die basalen Ersatzknospen, verlieren ebenfalls etwas von ihrer Resistenz, bleiben aber während der ganzen Vegetationsperiode der Pflanze deutlich widerstandsfähiger als jene Knospen, die sich entwickeln.

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A gradient in cold-resistance of ash bud sequences

Summary Terminal buds of ash (Fraxinus ornus L.) twigs are more developed than the two additional subterminal buds and the axillary buds inserted lower on the twig. The lateral buds gradually decrease in size from the tip to the base of the twig.A gradient of cold-resistance was established in the bud sequence of ash twigs. The terminal bud is the most frost-sensitive, and the inactive substitutional buds are the most resistant. As soon as the trees become active at the beginning of spring, the high degree of cold-resistance achieved during winter is reduced first in those buds which are the earliest to burst. The subterminal and the most basal buds, which, as a rule, do not open, partially lose their cold-resistance in spring and summer, however they remain markedly more resistant during this time than growing buds.In conclusion: InFraxinus ornus the more a bud is developed, the less it becomes cold-resistant and the sooner it loses the resistance it had.

     

Age-dependent bark photosynthesis of aspen twigs

The photosynthetic performance of trembling aspen (Populus tremula L.) twigs and leaves was studied in relation to selected structural features of aspen bark. PFD transmittance of intact periderm was reduced by about 90% in current-year twigs through peridermal thickening. However, because of drastic changes within the bark microstructure, PFD transmittance increased in 1-year-old twig segments up to 26% of the incident PFD. On a unit surface area basis, the chlorophyll content of young twigs (425 mg Chl m^-2) almost reached that of leaves (460 mg Chl m^-2). The chlorophyll content of aspen bark chlorenchyma was clearly age-dependent, even increasing in current-year twigs with advancing internodal age. The low bark chlorophyll a/b ratios (about 2.6 compared with 3.9 in leaves) indicate that bark chloroplasts are shade-adapted. Positive net photosynthesis was not found in aspen twigs, but apparent respiration was distinctly reduced in the light due to light-driven carbon refixation (bark photosynthesis) within the chlorenchymal tissues. Under constant microclimatic conditions, dark respiration rates were strongly correlated with stem-internal CO₂ refixation. In accordance with increasing dark respiration rates, the efficiency of this carbon recycling was generally greater in the metabolically more active, younger twig segments than in older segments; carbon refixation rates reached up to 80% of dark respiration values. At least in young twigs and branches and thus in the light-exposed outer parts of tree crowns, respiratory CO₂ losses by the tree skeleton could efficiently be reduced. Refixation of carbon dioxide may be of great importance for carbon budgets in the environmentally controlled or pathogen-induced leafless states of deciduous aspen trees.     

FLOODWATER TEMPERATURE AND STEM LENTICEL HYPERTROPHY IN MANGIFERA INDICA (ANACARDIACEAE)

One-year-old seedling trees of Mangifera indica L. cv. Peach were exposed to floodwater temperatures of 15, 22.5, or 30 C for at least 13 days. Immediately prior to flooding, and at daily intervals thereafter, trees were visually examined for evidence of lenticel hypertrophy. Although lenticel hypertrophy was first apparent after 5 days of submergence at 30 C, and after 6 days at 22.5 C, the mean number of days of flooding until lenticel hypertrophy was first observed was 6.6 and 8.1 for the 30 and 22.5 C treatments, respectively. Even after 28 days, lenticel hypertrophy did not occur on plants flooded at 15 C. Initial stages of lenticel hypertrophy were characterized by development of intercellular spaces in the phellem and production of additional phellem tissue by increased phellogen activity. Later stages of hypertrophy were characterized by development of intercellular spaces in the phellem and cortex. Tree survival was not affected by floodwater temperature or lenticel hypertrophy.     

Wound healing and induced resistance in potato tubers

It was demonstrated that biogenic elicitors, arachidonic acid and chitosan, locally and systemically stimulated wound healing in potato tuber tissues by increasing the number of wound periderm layers, accelerating the development of cork cambium (phellogen), and inducing proteinase inhibitors. The signal molecules, jasmonic and salicylic acids, had different effects on the development of wound periderm: jasmonic acid locally and systemically stimulated potato wound healing and elevated the level of proteinase inhibitors, whereas salicylic acid did not have any effect on wound healing and even blocked the formation of proteinase inhibitors.     

Sampling conditions for reliable routine detection by enzyme-linked immunosorbent assay of three ilarviruses in fruit trees

Enzyme-linked immunosorbent assay (ELISA) was used to test plum trees for prune dwarf (PDV), Prunus necrotic ringspot (NRSV) and apple mosaic (ApMV) viruses, cherry trees for PDV and NRSV, and apple trees for ApMV. Optimum conditions were determined for sampling in large-scale surveys for these viruses. All three viruses were detected throughout the growing season in individual samples of young leaves, or twigs with newly formed buds. However, when single infected leaves were combined with different numbers of healthy leaves, tests were most successful for all three viruses early in the growing season. PDV was detected in 1/40 (infected/total leaves) cherry leaves in April and May and 1/40 plum leaves until July, whereas NRSV was detected in 1/20 cherry leaves until July and 1/20 plum leaves until May. ApMV was detected in 1/20 apple or plum leaves until June but was detected less readily in mature leaves after June than either NRSV or PDV. There was no evidence of uneven distribution of virus infection in the trees. The viruses were detected in leaf samples kept for 8 wk at 3°C but freezing was less reliable for storage especially with ApMV. ApMV was detected in tests on plants held for several weeks at 25°C, and PDV and NRSV in plants held at 30°C.     

FRUITING AND TWIG DIAMETER IN CITRUS

On orange and grapefruit trees, fruiting was associated with an increase in diameter of the twig. On twigs with lateral fruit, this increase was restricted to that part of the twig below the point of attachment of the fruit. Possibly, fruit development induces orientation of vascular tissues and, hence, translocation toward the fruit.     

Overwintering form of the causal agent of shot hole disease in Khorasan Razavi, Iran

Shot hole disease of stone fruit trees caused by some plant pathogenic fungi is a major constraint to stone fruit production worldwide where the trees are grown. Identification of the causal agents of the disease and their overwintering forms in stone fruit trees of Khorasan Razavi was necessary for disease management programs. Buds, twigs, fallen leaves and fruits were collected from the infected peach, apricot, nectarine and almond trees in winter 2007. The samples were superficially disinfested in 1% sodium hypochlorite for 2-3 min and then in 70% ethanol for 45 sec. Two to three fragments of 4x4 mm from each tissue were separately cultured on 2% water agar and potato dextrose agar (PDA), and purified on PDA. Just a pathogenic fungal species, Wilsonomyces corpophilus was isolated from the infected buds and twigs. No microorganism was isolated from the fallen leaves and fruits collected from underneath of the infested stone fruit trees. Pathogenicity of the fungus was examined on detached shoots of current year of four varieties of stone fruit trees. Fungal discs were placed under the bark of the bud base. Control shoots were similarly treated with sterile PDA discs. Inoculated shoots were placed in a humid growth chamber at 25 degrees C. Fungal hyphae appeared at 30 days post inoculation. Control shoots were asymptomatic. Pathogenicity intensities or lesion lengths were significantly different among the four varieties tested. A completely randomised design with five replicates was employed to measure the number of spores in infested buds and twigs of each variety of stone fruit tree. The samples were sliced and placed into a glass tube of centrifuge containing 3 ml of sterile distilled water. They were mixed on a vortex mixer for 30-40 min and centrifuged at 3000 rpm for 5 min. Pelleted material from each sample was suspended in 500 microl of sterile distilled water and the spores were counted using a hemocytometre. Results revealed that the fungus overwinters as hyphae and conidia in the infected buds, and as hyphae and globular chlamydospores in twig lesions.     

Casparian bands occur in the periderm of Pelargonium hortorum stem and root

Background and Aims

Casparian bands are characteristic of the endodermis and exodermis of roots, but also occur infrequently in other plant organs, for example stems and leaves. To date, these structures have not been detected in phellem cells of a periderm. The aim of this study was to determine whether Casparian bands occur in phellem cells using tests that are known to detect Casparian bands in cells that also contain suberin lamellae. Both natural periderm and wound-induced structures were examined in shoots and roots.

Methods

Using Pelargonium hortorum as a candidate species, the following tests were conducted: (1) staining with berberine and counterstaining with aniline blue, (2) mounting sections in concentrated sulphuric acid and (3) investigating the permeability of the walls with berberine as an apoplastic, fluorescent tracer.

Key Results

(1) Berberine–aniline blue staining revealed a modification in the radial and transverse walls of mature phellem cells in both stems and roots. Three days after wounding through to the cortex of stems, the boundary zone cells (pre-existing, living cells nearest the wound) had developed vividly stained primary walls. By 17 d, staining of mature phellem cells of wound-induced periderm was similar to that of natural periderm. (2) Mature native phellem cells of stems resisted acid digestion. (3) Berberine was excluded from the anticlinal (radial and transverse) walls of mature phellem cells in stems and roots, and from the wound-induced boundary zone.

Conclusions

Casparian bands are present in mature phellem cells in both stems and roots of P. hortorum. It is proposed that Casparian bands act to retard water loss and pathogen entry through the primary cell walls of the phellem cells, thus contributing to the main functions of the periderm.     

HETEROPHYLLY IN POPULUS GRANDIDENTATA (SALICACEAE) WITH EMPHASIS ON RESIN GLANDS AND EXTRAFLORAL NECTARIES

Big-tooth aspen displays heterophylly on twigs of saplings and on upper, vigorously growing twigs of larger trees. Early leaves, those present in overwintering buds which expand in spring, differ from late leaves, those that are mostly initiated and developed during the late spring and summer of the same year. The two sets of leaves differ in size, shape, number of teeth, number of marginal resin glands, and number and size of basilaminar extrafloral nectaries. Secretory structures are generally more prominent and active on late leaves. Marginal resin glands, resin-secreting stipules, and basilaminar nectaries of both early and late leaves are similar anatomically, having a secretory epidermis of cytoplasmically dense palisade-like cells separated from vascular bundle endings by several layers of isodiametric parenchyma. These secretory structures, and the dense mat of trichomes which cover the leaf until it fully expands, may help to protect the young leaves from insect damage. The basilaminar nectaries seem especially effective because they attract ants that probably discourage visits by other insects.     

Optimisation of spatial allocation patterns in lianas compared to trees used for support

There are only limited possibilities to study the competition between trees and lianas in the top canopy of tropical rain forests. Furthermore, the important question how the leaf traits are related to twig support is rarely studied, especially regarding growing space partitioning between the self-supporting and the climbing growth form. Our study used the hot-air balloon within the “Operation Canopee” in the Masoala National Park, Madagascar, to test the differences in spatial allocation patterns of leaves and twigs in lianas and tree parts used for support. The sampling design emphasised to obtain a common assembly of twigs and leaves from both, trees and lianas. The results from the top canopy were compared to the data from the understorey regarding biomass and nutrients in leaves and distal twigs. In the understorey the reduction in structural investment was much stronger in lianas than in trees. The results showed that lianas reduced carbon investment per volume, but increased leaf nitrogen concentration and leaf area ratio (LAR), the latter driven by a reduction in leaf mass per area (LMA). In the top canopy, lianas contributed about one third of the leaf area density of 3 m² m⁻³. For distal twigs, no relationship was found between twig biomass per volume and leaf area density for trees, but lianas balanced both structural parameters closely. The climbers benefit from the external support provided by the trees and optimise the area of assimilation tissue at low per volume investment for mechanical stability. Several traits such as low LMA and high leaf nitrogen concentrations together with higher LAR and optimised spatial investment advantage the climbing growth form and enable a fast acquisition of growing space. The results emphasize the necessity to consider spatial and structural features of growing space acquisition when dealing with plant competition. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.