Energy requirement for foliage construction depends on tree size inyoung Picea abies trees

 The relationship between stand biomass production, and tree age and size is generally a curve with a maximum. To understand why wood production decreases in the final stages of stand development, the influence of increasing tree size on foliage chemical composition and substrate requirement for foliage construction in terms of glucose [CC, g glucose (g dry mass)^{–  1}] was investigated in the evergreen conifer Picea abies (L.) Karst. Because it was already known that irradiance affects both foliage morphology and chemistry in this species, and it was expected that the foliage in large overstory trees would intercept on average more light than that in saplings in understory, irradiance was measured in the sampling locations and included in the statistical models. CC of needles increased with increasing total tree height (TH) and was independent of relative irradiance. A major reason for increasing CC with increasing TH was a greater proportion of carbon-rich lignin in the needles in large trees. However, lignin did not fully account for the observed changes in CC, and it was necessary to assume that certain other carbon-rich secondary metabolites such as terpenes also accumulate in the foliage of large trees. Enhanced requirements for needle mechanical strength as evidenced by greater lignin concentrations in large trees were attributed to increased water limitations with increasing tree height. Because water relations may also control the sink capacities for assimilate usage, apart from the mechanical requirements, they may provide an explanation for the accumulation of other energetically expensive compounds in the needles as well. Biomass partitioning within the shoot was another foliar parameter modified in response to increasing tree size. The proportion of shoot axes, which serve to provide needles with mechanical support and to supply them with water, decreased with increasing TH. This may limit water availability in the needles, and/or manifest a lower water requirement of the needles containing proportionally more supporting and storage substances, and consequently, less physiologically active compounds such as proteins. Probably the same factors which caused CC of the needles to depend on TH, were also responsible for greater CC of the shoot axes in larger trees. These results collectively suggest that increasingly more adverse water relations with increasing tree size may provide a mechanistic explanation for the decline in foliar biomass and its functional activity during stand ageing. Received: 9 April 1996 / Accepted: 14 January 1997     

Time constant for water transport in loblolly pine trees estimated from time series of evaporative demand and stem sapflow

 The use of stem sap flow data to estimate diurnal whole-tree transpiration and canopy stomatal conductance depends critically upon knowledge of the time lag between transpiration and water flux through the stem. In this study, the time constant for water movement in stems of 12-year-old Pinus taeda L. individuals was estimated from analysis of time series data of stem water flux and canopy transpiration computed from mean daytime canopy conductance, and diurnal vapor pressure deficit and solar radiation measurements. Water uptake through stems was measured using a constant-heat sapflow probe. Canopy transpiration was correlated to stem uptake using a resistance-capacitance equation that incorporates a time constant parameter. A least-squares auto-regression determined the parameters of the resistance-capacitance equation. The time constants for ten loblolly pine trees averaged 48.0 (SE = 2.0) min and the time lag for the diurnal frequency averaged 47.0 (SE = 2.0) min. A direct-cross correlation analysis between canopy transpiration and sap flow time series showed maximum correlation at an approximately 30 min lag. Residuals (model-predicted minus actual stem flow data) increased with increasing soil moisture depletion. While the time constants did not vary significantly within the range of tree sizes studied, hydraulic resistance and capacitance terms were individually dependent on stem cross-sectional area: capacitance increased and resistance decreased with stem volume. This result may indicate an inverse adjustment of resistance and capacitance to maintain a similar time constant over the range of tree sizes studied.     

Underground water use by Eucalyptus trees in an arid climate

 Expanding the area of tree plantations in the Negev desert of Israel requires prior quantification of the water resources in small watersheds. Combined hydrological and physiological measurements were used to study a “liman” (floodwater irrigated plot) in the Negev highlands, where Eucalyptus and other tree species are grown successfully. The amount of water flowing into the liman, surplus water flowing out of the liman, temporal soil moisture distribution, and water uptake by the trees were determined. Depending on rainfall intensity and distribution during the season, the liman received 2 – 3 times the total seasonal rainfall. Although the study was conducted during a year with a negligible amount of rainfall, the transpiration rate was closely correlated with potential transpiration throughout the year. The amount of water extracted from the soil was less than the time-integrated transpiration rate from the trees, suggesting that a water source other than soil water storage was available to the trees. We suggest that the trees extracted water from the rock fractures and/or utilized the lateral flows over the rock/soil interface.     

Differential growth and recovery rates following defoliation in related deciduous and evergreen trees

 Deciduous larches, Larix spp., and evergreen pines, Pinus spp., are sympatric Pinaceae conifers. Adjacent monocultures of 10-year-old Larix decidua Mill. and Pinus resinosa Ait. were subjected to single-season artificial defoliation by clipping from 0% to 99% of each needle. Survival, above-ground productivity, and architecture were measured for 36 months. P. resinosa and L. decidua exhibited differential relationships with defoliation intensity and recovery time. Two months after treatment, defoliation reduced larch height growth but had no effect on radial growth. By contrast, P. resinosa stem radial growth was reduced immediately, but height growth was not decreased until the following year. Pine leader growth and above-ground biomass following 66% defoliation never recovered to control values or 33% defoliated pines. Conversely, defoliated larch quickly recovered from an initial growth loss to eliminate all treatment effects on biomass. The plasticity in architectural response found in larch, but not pine, might partially account for defoliation tolerance. Both P. resinosa and L. decidua exhibited non-linear responses to defoliation. These patterns may be caused partially by the uneven distribution of nutrients within needles, rather than a simple function of leaf area lost to defoliators. Concentrations of 13 nutrients in P. resinosa were highest either in the mid- (Ca, Mg, S, Zn, B, Mn, Fe, Al and Na) or basal- (N, P, K, and Cu) section. The relatively low nutrient content in needle tips may contribute to similar biomass productivity between trees defoliated 33% and controls. Removal of needle mid-sections significantly reduced whole-plant productivity. In contrast, L. decidua nutrients are concentrated in the distal sections. Nutrient concentrations were generally highest in larch. Our results agree with an emergent prediction of the carbon/nutrient balance theory that defoliation more severely reduces growth of evergreen than deciduous species. These results are discussed within the physiological, ecological and evolutionary context of allocation theory, with implications for natural resource management and plant-insect interaction theory. Received: 6 April 1995 / Accepted: 29 August 1995     

A cytoskeletal basis for wood formation in angiosperm trees: the involvement of microfilaments

The cortical microfilament (MF) component of the cytoskeleton within axial elements of the secondary vascular system of the angiosperm tree, Aesculus hippocastanum L. (horse-chestnut) was studied using transmission electron microscopy of ultrathin sections and indirect immunofluorescence microscopy of actin in thick sections. As seen by electron microscopy, MF bundles have a net axial orientation within fusiform cambial cells and their secondary vascular derivatives (i.e. in the axial xylem and phloem parenchyma, xylem fibres, vessel and sieve elements, and companion cells). Immunofluorescence studies, however, reveal that this axial orientation can be more accurately described as a helix of extremely high pitch; it is a persistent feature of all axial secondary vascular elements during their development. Helical MF arrays are the only arrangement seen in secondary phloem cells. However, in addition to helices, other MF arrays are seen in secondary xylem cells. For example, fibres possess ellipses of MFs associated with simple-pit formation, and vessel elements possess circular arrays of MFs that associate with the developing inter-vessel bordered pits, ray–vessel contact pits, and with the perforation plate. Linear MF arrays are seen co-oriented with the developing tertiary wall-thickenings in vessel elements. The possible roles of MFs during the cytodifferentiation of secondary vascular cells is discussed, and compared with that of microtubules. Received: 7 June 1999 / Accepted: 23 December 1999     

Above- and below-ground characteristics associated with wind toppling in a young Populus plantation

 Damage from a dormant-season windstorm in a 3-year-old Populus research trial differed among four clones and three spacings and between monoclonal and polyclonal plots. Clonal differences in susceptibility to toppling (or leaning) were associated with both above- and below-ground characteristics. Susceptible clones had less taper in the lower stem and more weight in branches on the upper stem. The most susceptible clone also had the most above-ground biomass per unit of cross-sectional root area. The other susceptible clone had the least root system development in the windward quadrants. Wind toppling was least at the closest spacing. Apparently, mutual support was more important than individual tree characteristics from which the most damage would be expected at the closest spacing. Differences between paired trees of the same clone and spacing which did or did not topple were primarily associated with distribution of root systems by compass quadrant or depth. At the closest spacing where crown sway would have been minimized, trees which did not topple had greater cross-sectional root area in the windward direction than trees which did topple. At the widest spacing where crown sway would have been greatest, windfirm trees had greater cross-sectional root area than non-windfirm trees in both the windward and leeward directions. Toppling was reduced in polyclonal plots; this reduction may have been the result of more rapid stand differentiation in the polyclonal plots or reduction in the “domino effect” by inclusion of more windfirm clones in the mixture. Received: 23 October 1995 / Accepted: 22 February 1996     

The genetic resolution of juvenile canopy structure and function in a three-generation pedigree of Populus

 A genetic approach to the understanding of tree architecture is to cross trees of contrasting features and to study their segregating F₂ progenies. For this purpose, members of a 3-generation pedigree, combining Populus trichocarpa, P. deltoides, and their F₁ and F₂ offspring, were grown side by side in a clonally replicated plantation. At 2 and 3 years of growth, tree architecture was analyzed at the stem, branch, and leaf levels. In all generations, proleptic branches were more numerous, longer, and had more and larger leaves than sylleptics initiated in the same year. The analysis of variance revealed significant genotypic effects on growth, branch and leaf biometrics in the F₂ family, with broad-sense heritabilities (H²) ranging from 0.50 to 0.80 for most traits. For branch and leaf traits, the H² values were found to vary among branch types and crown positions. In year 2, the degree of genetic control was stronger for sylleptics than proleptics and for upper than lower crown positions. These patterns were followed in year 3, except that H² values were more a function of position within crown, as a consequence of increased competition among trees. The genetic correlations between branch/leaf morphology and stem growth were also a function of branch type and crown position. Generally, traits on proleptics or at upper positions were more tightly correlated with height growth, whereas those on sylleptics or at lower positions, with basal area growth. By year 3, proleptic traits showed increased genetic correlations with both height and radial growth. The implications of these results for the construction of ideotypes are discussed. Received: 1 December 1995     

Chemical composition of apoplastic transport barriers in relation to radial hydraulic conductivity of corn roots (Zea mays L.)

The hydraulic conductivity of roots (Lp_r) of 6- to 8-d-old maize seedlings has been related to the chemical composition of apoplastic transport barriers in the endodermis and hypodermis (exodermis), and to the hydraulic conductivity of root cortical cells. Roots were cultivated in two different ways. When grown in aeroponic culture, they developed an exodermis (Casparian band in the hypodermal layer), which was missing in roots from hydroponics. The development of Casparian bands and suberin lamellae was observed by staining with berberin-aniline-blue and Sudan-III. The compositions of suberin and lignin were analyzed quantitatively and qualitatively after depolymerization (BF₃/methanol-transesterification, thioacidolysis) using gas chromatography/mass spectrometry. Root Lp_r was measured using the root pressure probe, and the hydraulic conductivity of cortical cells (Lp) using the cell pressure probe. Roots from the two cultivation methods differed significantly in (i) the Lp_r evaluated from hydrostatic relaxations (factor of 1.5), and (ii) the amounts of lignin and aliphatic suberin in the hypodermal layer of the apical root zone. Aliphatic suberin is thought to be the major reason for the hydrophobic properties of apoplastic barriers and for their relatively low permeability to water. No differences were found in the amounts of suberin in the hypodermal layers of basal root zones and in the endodermal layer. In order to verify that changes in root Lp_r were not caused by changes in hydraulic conductivity at the membrane level, cell Lp was measured as well. No differences were found in the Lp values of cells from roots cultivated by the two different methods. It was concluded that changes in the hydraulic conductivity of the apoplastic rather than of the cell-to-cell path were causing the observed changes in root Lp_r. Received: 17 March 1999 / Accepted: 22 June 1999     

A marking technique for live fish eggs and larvae

 A new marking technique for live fish eggs and larvae was proposed to elucidate the larval biology and adult breeding ecology of wild fish. In the laboratory, females of a freshwater goby Rhinogobius sp. OR were abdominally injected with one of three coloring agents—brilliant blue FCF, rose Bengal, or β-carotene—before their oviposition. The rose Bengal proved lethal to adult fish. The other two dyes had little effect on adult mortality. With these two treatments, there were negative effects on neither fecundity nor egg mortality, resulting in normally developed larvae. The brilliant blue FCF stained eggs and larvae greenish blue whereas the staining effect of β-carotene was unclear. The timing of injection was important in effective staining of eggs and reducing the risk of miscarriage. In conclusion, the brilliant blue FCF was the more useful marker. We discuss what this method can show us about the ecology of wild fish and how this method can be applied to field study. Received: March 6, 2002 / Revised: July 11, 2002 / Accepted: August 14, 2002 RID="*" ID="*" Present address: Center for Marine Environmental Studies, Ehime University, 3 Bunkyo-cho, Matsuyama 790-8577, Japan (e-mail: nokuda@sci.ehime-u.ac.jp) Acknowledgments I am grateful to K. Karino, M. Kohda, and A. Moriyama for giving us valuable advice and to M. Inoue and H. Miyatake for their field assistance. This study was financially supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists. Correspondence to:Noboru Okuda     

Correspondence of environmental tolerances with leaf and branch attributes for six co-occurring species of broadleaf evergreen trees in northern California

 For the angiosperm dominants of northern California’s mixed evergreen forests, this study compares the display of photosynthetic tissue within leaves and along branches, and examines the correspondence between these morphological attributes and the known environmental tolerances of these species. Measurements were made on both sun and shade saplings of six species: Arbutus m e n z i e s i i (Ericaceae), C h r y s o l e p i s c h r y s o p h y l l a (Fagaceae), L i t h o c a r p u s d e n s i f l o r u s (Fagaceae), Quercus c h r y s o l e p i s (Fagaceae), Quercus w i s l i z e n i i (Fagaceae), and Umbellularia c a l i f o r n i c a (Lauraceae). All species had sclerophyllous leaves with thick epidermal walls, but species differed in leaf specific weight, thickness of mesophyll tissues and in the presence of a hypodermis, crystals, secretory idioblasts, epicuticular deposits, and trichomes. The leaves of Arbutus were 2 – 5 times larger than those of C h r y s o l e p i s, L i t h o c a r p u s and Umbellularia and 4 – 10 times larger than those of both Quercus species. Together with differences in branch architecture, these leaf traits divide the species into groups corresponding to environmental tolerances. Shade-tolerant C h r y s o l e p i s, L i t h o c a r p u s, and Umbellularia had longer leaf lifespans and less palisade tissue, leaf area, and crown mass per volume than the intermediate to intolerant Arbutus and Quercus. Having smaller leaves, Quercus branches had more branch mass per leaf area and per palisade volume than other species, whereas Arbutus had less than other species. These differences in display of photosynthetic tissue should contribute to greater growth for Quercus relative to the other species under high light and limited water, for Arbutus under high light and water availability, and for C h r y s o l e p i s, L i t h o c a r p u s, and Umbellularia under limiting light levels. Accepted: 22 March 1996     

Variability of some phenolic acids in phloem of 1-year-old shoots of Scots pine trees growing with Heterobasidion annosum (Fr.) Bref.

 HPLC chromatographic analyses of some phenolic acids in phloem of 1-year-old shoots sampled from 32 trees of eight Polish provenances of Scots pine (Pinus sylvestris L.) growing under conditions of annosum root [Heterobasidion annosum (Fr.) Bref.] are discussed. Considerable quantitative and qualitative differentiation was found among individual trees. The variability of trees was estimated with regard to the level of phenolic acids and correlations were established in order to assess the character of their joint occurrence in shoot phloem. In view of pathogen presence, the content of phenolic acids varies between individuals depending upon the genotype of pine, the stage of development of the disease and upon the effect of tree growth conditions. Received: 23 April 1997 / Accepted: 12 September 1997     

A novel glycoprotein obtained from Chlorella vulgaris strain CK22 shows antimetastatic immunopotentiation

 A glycoprotein extract (CVS), derived from the unicellular green alga Chlorella vulgaris, strain CK22, exhibited a pronounced antitumor effect against both spontaneous and experimentally induced metastasis in mice. Inhibition of tumor metastasis was enhanced when intratumor administration of CVS was followed by s.c. injection of CVS. Anti-metastatic immunopotentiation was observed in euthymic mice, but not in athymic nude mice. The antitumor activity of CVS was reflected in antigen-specific, T-cell-mediated immunity. Both CD4 and CD8 T cells contributed to the antimetastatic effects, as shown by in vivo depletion experiments with anti-T-cell subset antibodies. Furthermore, CVS caused the recruitment of T cells to the regional lymph nodes and their proliferation in these organs. The CD4-positive population, following CVS injection at the time of tumor rechallenge, displayed a pronounced increase in the proportion of T cells that were CD18 bright, CD44 bright, CD25⁺, CD54⁺, CD69⁺ or CD71⁺ in the lymph nodes. Thus, CVS induces T cell activation in peripheral lymph nodes in tumor-bearing mice. We conclude that CVS augments antimetastatic immunity through T cell activation in lymphoid organs and enhances recruitment of these cells to the tumor sites. Presurgical treatment with CVS might prevent metastasis or tumor progression. Received: 5 June 1997 / Accepted: 12 September 1997     

A berberine-aniline blue fluorescent staining procedure for suberin,lignin, and callose in plant tissue

Summary A fluorescent staining procedure to detect suberin, lignin and callose in plants has been developed. This procedure greatly improves on previous methods for visualizing Casparian bands in root exodermal and endodermal cells, and performs equally well on a variety of other plant tissues. Berberine was selected as the most suitable replacement forChelidonium majus root extract after comparing the staining properties of the extract with those of four of its constituent alkaloids. Aniline blue counterstaining efficiently quenched unwanted background fluorescence and nonspecific berberine staining, while providing a fluorochrome for callose. When used with multichambered holders which allow simultaneous processing of freehand sections, this efficient staining procedure facilitates morphological studies involving large numbers of samples.Abbreviations ISCC-NBS Inter-Society Color Council-National Bureau of Standards - UV ultraviolet light     

Natural and experimentally altered hydraulic architecture of branch junctions in Acer saccharum Marsh. and Quercus velutina Lam. trees

 The functional xylem anatomy and the hydraulic conductivity of intact and treated branch junctions of the diffuse-porous sugar maple (Acer saccharum Marsh.) were compared to those of the ring-porous black oak (Quercus velutina Lam.). Maple shoots possessed greater growth intensity than those of oak. The extensive growth of the maple trees resulted in about a two-fold increase in xylem production in the maple branches. Branches were altered by removing a patch of bark from the base of a branch (near a junction) leaving a bridge of bark on the upper or lower side of the branch. The experimentally treated branch junctions revealed that, in oak, most (up to 92%) of the water flows in the lower side of a branch, where most of the large vessels occurred. In maple, most of the conductive tissue was observed to form in the upper side of the branches, which was equally or more conductive than the lower side. A treatment of longitudinal, parallel scratches in the bark-bridge, which reduced earlywood vessel width, substantially decreased conductivity (to only 15%) in oak, but had no effect on conductivity in maple. In maple, such wounding stimulated more wood formation and increased conductivity. In both trees, a narrow bridge at the junction induced more wood formation and higher conductivity in the branch. The mechanisms controlling wood formation and water flow in branch junctions of ring- and diffuse-porous trees are discussed. Received: 14 February 1996 / Accepted: 27 May 1996     

Variable expression of tumor necrosis factor α in human malignant melanoma localized by in situ hybridization for mRNA

 Tumor necrosis factor α (TNFα) is a cytokine, produced by lymphocytes and monocytes, with cytotoxic activity against some but not all tumor cell lines. Resistance to the cytolytic effects of TNFα has been reported in cell lines with autocrine TNFα production. The purpose of this study was to investigate whether human primary malignant melanoma and tumor infiltrating lymphocytes produce TNFα in vivo. Optimal conditions for in situ hybridization for TNFα mRNA in paraffin-embedded tissue were established. Analysis of 13 primary malignant melanomas and 3 metastatic lesions with different degrees of immunohistochemical TNFα positivity demonstrated that, in some tumors, both melanoma cells and leukocytes contained TNFα mRNA and protein. These findings demonstrate variable production of TNFα in primary and metastatic melanoma in vivo.The previously described resistance to TNFα cytolytic activity may, therefore, be clinically important. Received: 18 December 1996 / Accepted: 12 June 1997