Hans Beeckman

Tyloses and phenolic deposits in xylem vessels impede water transport in low-lignin transgenic poplars: a study by cryo-fluorescence microscopy

Of 14 transgenic poplar genotypes (Populus tremula × Populus alba) with antisense 4-coumarate:coenzyme A ligase that were grown in the field for 2 years, five that had substantial lignin reductions also had greatly reduced xylem-specific conductivity compared with that of control trees and those transgenic events with small reductions in lignin. For the two events with the lowest xylem lignin contents (greater than 40% reduction), we used light microscopy methods and acid fuchsin dye ascent studies to clarify what caused their reduced transport efficiency. A novel protocol involving dye stabilization and cryo-fluorescence microscopy enabled us to visualize the dye at the cellular level and to identify water-conducting pathways in the xylem. Cryo-fixed branch segments were planed in the frozen state on a sliding cryo-microtome and observed with an epifluorescence microscope equipped with a cryo-stage. We could then distinguish clearly between phenolic-occluded vessels, conductive (stain-filled) vessels, and nonconductive (water- or gas-filled) vessels. Low-lignin trees contained areas of nonconductive, brown xylem with patches of collapsed cells and patches of noncollapsed cells filled with phenolics. In contrast, phenolics and nonconductive vessels were rarely observed in normal colored wood of the low-lignin events. The results of cryo-fluorescence light microscopy were supported by observations with a confocal microscope after freeze drying of cryo-planed samples. Moreover, after extraction of the phenolics, confocal microscopy revealed that many of the vessels in the nonconductive xylem were blocked with tyloses. We conclude that reduced transport efficiency of the transgenic low-lignin xylem was largely caused by blockages from tyloses and phenolic deposits within vessels rather than by xylem collapse.

Influence of drought on tree rings and tracheid features of Pinus nigra and Pinus sylvestris in a mesic Mediterranean forest

We investigated the influence of climate on the ring width and xylem anatomy of two co-occurring pines (Pinus nigra Arn. and P. sylvestris L.) in the mountains of east-central Spain in order to test their utility for dendroclimatic reconstructions. We developed chronologies of ring width, mean lumen diameter and mean cell-wall thickness (in the earlywood, latewood, and the total annual ring) and the number of cells between 1960 and 2006. Drought, expressed as the standardized precipitation-evapotranspiration index (SPEI), was the main climatic driver of tree radial growth, although trees were also sensitive to temperature (negative effect in previous autumn and current summer) and precipitation (with a general positive effect). P. sylvestris response was stronger to climate of the current year, whereas the effect of previous-year climate was more important for P. nigra. Warm and dry summers reduced ring width, tracheid lumen, and wall thickness in both species, whereas warm winter-spring temperatures had the opposite effect, primarily for P. sylvestris. Previous-year or early-season conditions mainly affected earlywood features, whereas latewood was more responsive to summer climate. Overall, climate appeared to be a stronger limiting factor for P. sylvestris. During periods of drought, cell-wall thickness was reduced while lumen width increased in the latewood of P. sylvestris. This could compromise its hydraulic safety against drought-induced cavitation as our site was close to the southern and dry edge of the species distribution area. Our results suggest that anatomical variables record different and stronger climate information than ring width variables, especially in P. sylvestris. Reconstruction models for SPEI at the 3-month scale were developed for July–August and September–October using principal components regression. The best models included anatomical and width variables of both pine species suggesting that tracheid chronologies can be useful for drought reconstructions especially at mesic sites or with species that encode a mixed drought and temperature-precipitation signal.

Seeing Central African forests through their largest trees

Large tropical trees and a few dominant species were recently identified as the main structuring elements of tropical forests. However, such result did not translate yet into quantitative approaches which are essential to understand, predict and monitor forest functions and composition over large, often poorly accessible territories. Here we show that the above-ground biomass (AGB) of the whole forest can be predicted from a few large trees and that the relationship is proved strikingly stable in 175 1-ha plots investigated across 8 sites spanning Central Africa. We designed a generic model predicting AGB with an error of 14% when based on only 5% of the stems, which points to universality in forest structural properties. For the first time in Africa, we identified some dominant species that disproportionally contribute to forest AGB with 1.5% of recorded species accounting for over 50% of the stock of AGB. Consequently, focusing on large trees and dominant species provides precise information on the whole forest stand. This offers new perspectives for understanding the functioning of tropical forests and opens new doors for the development of innovative monitoring strategies.

Conventional tree height–diameter relationships significantly overestimate aboveground carbon stocks in the Central Congo Basin

Policies to reduce emissions from deforestation and forest degradation largely depend on accurate estimates of tropical forest carbon stocks. Here we present the first field-based carbon stock data for the Central Congo Basin in Yangambi, Democratic Republic of Congo. We find an average aboveground carbon stock of 162 ± 20  Mg  C  ha(-1) for intact old-growth forest, which is significantly lower than stocks recorded in the outer regions of the Congo Basin. The best available tree height-diameter relationships derived for Central Africa do not render accurate canopy height estimates for our study area. Aboveground carbon stocks would be overestimated by 24% if these inaccurate relationships were used. The studied forests have a lower stature compared with forests in the outer regions of the basin, which confirms remotely sensed patterns. Additionally, we find an average soil carbon stock of 111 ± 24  Mg  C  ha(-1), slightly influenced by the current land-use change.

Successive Cambia: A Developmental Oddity or an Adaptive Structure?

Background Secondary growth by successive cambia is a rare phenomenon in woody plant species. Only few plant species, within different phylogenetic clades, have secondary growth by more than one vascular cambium. Often, these successive cambia are organised concentrically. In the mangrove genus Avicennia however, the successive cambia seem to have a more complex organisation. This study aimed (i) at understanding the development of successive cambia by giving a three-dimensional description of the hydraulic architecture of Avicennia and (ii) at unveiling the possible adaptive nature of growth by successive cambia through a study of the ecological distribution of plant species with concentric internal phloem. Results Avicennia had a complex network of non-cylindrical wood patches, the complexity of which increased with more stressful ecological conditions. As internal phloem has been suggested to play a role in water storage and embolism repair, the spatial organisation of Avicennia wood could provide advantages in the ecologically stressful conditions species of this mangrove genus are growing in. Furthermore, we could observe that 84.9% of the woody shrub and tree species with concentric internal phloem occurred in either dry or saline environments strengthening the hypothesis that successive cambia provide the necessary advantages for survival in harsh environmental conditions. Conclusions Successive cambia are an ecologically important characteristic, which seems strongly related with water-limited environments.

Tree ring analysis of Brachystegia spiciformis and Isoberlinia tomentosa: Evaluation of the enso-signal in the miombo woodland of eastern Africa

The value of growth rings as proxy data for climate reconstruction was studied in two miombo woodland species in eastern Africa. Growth rings, marked by terminal parenchyma, were visually detectable on carefully prepared stem discs of Isoberlinia tomentosa and Brachystegia spiciformis, dominant species of the miombo woodland in north-western Tanzania. However, the presence of multiple growth ring anomalies rendered cross-dating of the growth ring series between trees difficult. Cross-dating succeeded for eight out of thirteen samples for Isoberlinia tomentosa, but was unsuccessful for Brachystegia spiciformis. A mean series of 38 years was calculated for Isoberlinia tomentosa only. Monthly precipitation, monthly maximum air temperature and monthly SOI-value (Southern Oscillation Index) correlated significantly with tree ring widths of the mean series. These correlations are strong indicators of the annual character of the growth rings. They also suggest that Isoberlinia tomentosa provides an appropriate paleoclimatic record for dendroclimatic reconstruction.

Dendrochronology in the dry tropics: the Ethiopian case

Dendrochronology is developing outside temperate and boreal regions. Over the past decade substantial progress has been made in Mediterranean and wet tropical regions. However, research in dry tropical regions, notably those of sub-Saharan Africa, has remained fragmentary. Here, we try to identify the unique challenges and opportunities of dendrochronology in the dry tropics. First, we briefly review the status of dendrochronology outside temperate and boreal regions with an emphasis on sub-Saharan Africa. Subsequently, we focus upon one of those areas where dendrochronology in the dry tropics is at the forefront of scientific advance: Ethiopia. A detailed review of tree ring studies in the lowlands and highlands highlights the complexity of ring formation and made us identify four major types of growth ring expression: anatomically not distinct rings, multiple rings per year, annual rings and multiple missing rings. This complex tree growth behaviour is associated with large-scale variations in precipitation regime (unimodal to multimodal) and relatively small-scale variations in tree sensitivity to water availability. Literature results are used to develop a scheme that can be used to predict differences in growth ring formation along gradients in these two factors. Because of the exceptional growth sensitivity of and the importance of local site conditions (topography, biological factors, etc.) for most trees sampled, those growing at the limits of their ecological amplitude are prone to possess multiple rings per year or multiple missing rings. In such circumstances, site selection should not always take place at the limits of the ecological amplitude of a species, but may sometimes have to be diverted to more mesic environments. Successful studies are now appearing, such as those reporting correlations between tree ring chronologies and Blue Nile river flows.

Fluctuations of cambial activity in relation to precipitation result in annual rings and intra-annual growth zones of xylem and phloem in teak (Tectona grandis) in Ivory Coast

BACKGROUND AND AIMS Teak forms xylem rings that potentially carry records of carbon sequestration and climate in the tropics. These records are only useful when the structural variations of tree rings and their periodicity of formation are known. METHODS The seasonality of ring formation in mature teak trees was examined via correlative analysis of cambial activity, xylem and phloem formation, and climate throughout 1·5 years. Xylem and phloem differentiation were visualized by light microscopy and scanning electron microscopy. KEY RESULTS A 3 month dry season resulted in semi-deciduousness, cambial dormancy and formation of annual xylem growth rings (AXGRs). Intra-annual xylem and phloem growth was characterized by variable intensity. Morphometric features of cambium such as cambium thickness and differentiating xylem layers were positively correlated. Cambium thickness was strongly correlated with monthly rainfall (R(2) = 0·7535). In all sampled trees, xylem growth zones (XGZs) were formed within the AXGRs during the seasonal development of new foliage. When trees achieved full leaf, the xylem in the new XGZs appeared completely differentiated and functional for water transport. Two phloem growth rings were formed in one growing season. CONCLUSIONS The seasonal formation pattern and microstructure of teak xylem suggest that AXGRs and XGZs can be used as proxies for analyses of the tree history and climate at annual and intra-annual resolution.

Cambial Growth Season of Brevi-Deciduous Brachystegia spiciformis Trees from South Central Africa Restricted to Less than Four Months

We investigate cambial growth periodicity in Brachystegia spiciformis, a dominant tree species in the seasonally dry miombo woodland of southern Africa. To better understand how the brevi-deciduous (experiencing a short, drought-induced leaf fall period) leaf phenology of this species can be linked to a distinct period of cambial activity, we applied a bi-weekly pinning to six trees in western Zambia over the course of one year. Our results show that the onset and end of cambial growth was synchronous between trees, but was not concurrent with the onset and end of the rainy season. The relatively short (three to four months maximum) cambial growth season corresponded to the core of the rainy season, when 75% of the annual precipitation fell, and to the period when the trees were at full photosynthetic capacity. Tree-ring studies of this species have found a significant relationship between annual tree growth and precipitation, but we did not observe such a correlation at intra-annual resolution in this study. Furthermore, a substantial rainfall event occurring after the end of the cambial growth season did not induce xylem initiation or false ring formation. Low sample replication should be taken into account when interpreting the results of this study, but our findings can be used to refine the carbon allocation component of process-based terrestrial ecosystem models and can thus contribute to a more detailed estimation of the role of the miombo woodland in the terrestrial carbon cycle. Furthermore, we provide a physiological foundation for the use of Brachystegia spiciformis tree-ring records in paleoclimate research.

A survey of the systematic wood anatomy of the Rubiaceae

Recent insight in the phylogeny of the Rubiaceae, mainly based on macromolecular data, agrees better with wood anatomical diversity patterns than previous subdivisions of the family. The two main types of secondary xylem that occur in Rubiaceae show general consistency in their distribution within clades. Wood anatomical characters, especially the fibre type and axial parenchyma distribution, have indeed good taxonomic value in the family. Nevertheless, the application of wood anatomical data in Rubiaceae is more useful in confirming or negating already proposed relationships rather than postulating new affinities for problematic taxa. The wood characterised by fibre-tracheids (type I) is most common, while type II with septate libriform fibres is restricted to some tribes in all three subfamilies. Mineral inclusions in wood also provide valuable information with respect to systematic relationships.