Xylogenesis: Coniferous Trees of Temperate Forests Are Listening to the Climate Tale during the Growing Season But Only Remember the Last Words!

The complex inner mechanisms that create typical conifer tree-ring structure (i.e. the transition from large, thin-walled earlywood cells to narrow, thick-walled latewood cells) were recently unraveled. However, what physiological or environmental factors drive xylogenesis key processes remain unclear. Here, we aim to quantify the influence of seasonal variations in climatic factors on the spectacular changes in the kinetics of wood cell differentiation and in the resulting tree-ring structure. Wood formation was monitored in three sites over 3 years for three coniferous species (Norway spruce [Picea abies], Scots pine [Pinus sylvestris], and silver fir [Abies alba]). Cell differentiation rates and durations were calculated and related to tracheid final dimensions and corresponding climatic conditions. On the one hand, we found that the kinetics of cell enlargement and the final size of the tracheids were not explained by the seasonal changes in climatic factors. On the other hand, decreasing temperatures strongly constrained cell wall deposition rates during latewood formation. However, the influence of temperature was permanently written into tree-ring structure only for the very last latewood cells, when the collapse of the rate of wall deposition was no longer counterbalanced by the increase of its duration. Our results show that the formation of the typical conifer tree-ring structure, in normal climatic conditions, is only marginally driven by climate, suggesting strong developmental control of xylogenesis. The late breakage of the compensatory mechanism at work in the wall deposition process appears as a clue to understand the capacity of the maximum latewood density to record past temperature conditions.

The effects of throughfall exclusion on xylogenesis of balsam fir.

A 20-40% reduction in soil moisture is projected for the boreal forest of Eastern Canada for the period 2070-99 relative to 1971-2000. In order to better predict the effects of a reduced water supply on the growth of balsam fir (Abies balsamea (L.) Mill.), a dominant tree species of the boreal forest, we simulated 2 consecutive years of summer droughts (starting in July) by means of throughfall exclusion. Four 100-m(2) plots were established in 2010 with polyethylene sheets maintained 1.3-2 m aboveground and redirecting the water outside the plots. Wood microcores were extracted weekly from mature trees from April to October 2011 to analyse the time dynamics of wood formation in that year. The number of tracheids formed during and before treatment and their anatomical characteristics were determined through microscopic analyses. The growth of lateral and terminal branches and the water potential of balsam fir seedlings were also monitored. Throughfall exclusion significantly reduced soil water content by 5.8% in 2010 and 10.5% in 2011. Xylogenesis was affected significantly by the treatment. Tracheids were 16.1% smaller in diameter and their cell wall was 14.1% thicker during both years. The treatment delayed by more than a week the start of the tracheid differentiation process in the second year with a concomitant decrease (26%) in the number of tracheids produced. The seedlings displayed a 32% reduction in growth and a 40% reduction in leaf water potential. Our results suggest that a future regime of increased frequency and intensity of droughts could have negative effects on the duration of xylogenesis and the production of xylem cells in balsam fir.

A rapid decrease in temperature induces latewood formation in artificially reactivated cambium of conifer stems.

BACKGROUND AND AIMS: Latewood formation in conifers occurs during the later part of the growing season, when the cell division activity of the cambium declines. Changes in temperature might be important for wood formation in trees. Therefore, the effects of a rapid decrease in temperature on cellular morphology of tracheids were investigated in localized heating-induced cambial reactivation in Cryptomeria japonica trees and in Abies firma seedlings. METHODS: Electric heating tape and heating ribbon were wrapped on the stems of C. japonica trees and A. firma seedlings. Heating was discontinued when 11 or 12 and eight or nine radial files of differentiating and differentiated tracheids had been produced in C. japonica and A. firma stems, respectively. Tracheid diameter, cell wall thickness, percentage of cell wall area and percentage of lumen area were determined by image analysis of transverse sections and scanning electron microscopy. KEY RESULTS: Localized heating induced earlier cambial reactivation and xylem differentiation in stems of C. japonica and A. firma as compared with non-heated stems. One week after cessation of heating, there were no obvious changes in the dimensions of the differentiating tracheids in the samples from adult C. japonica. In contrast, tracheids with a smaller diameter were observed in A. firma seedlings after 1 week of cessation of heating. Two or three weeks after cessation of heating, tracheids with reduced diameters and thickened cell walls were found. The results showed that the rapid decrease in temperature produced slender tracheids with obvious thickening of cell walls that resembled latewood cells. CONCLUSIONS: The results suggest that a localized decrease in temperature of stems induces changes in the diameter and cell wall thickness of differentiating tracheids, indicating that cambium and its derivatives can respond directly to changes in temperature.

Cambial activity related to tree size in a mature silver-fir plantation.

BACKGROUND AND AIMS: Our knowledge about the influences of environmental factors on tree growth is principally based on the study of dominant trees. However, tree social status may influence intra-annual dynamics of growth, leading to differential responses to environmental conditions. The aim was to determine whether within-stand differences in stem diameters of trees belonging to different crown classes resulted from variations in the length of the growing period or in the rate of cell production. METHODS: Cambial activity was monitored weekly in 2006 for three crown classes in a 40-year-old silver-fir (Abies alba) plantation near Nancy (France). Timings, duration and rate of tracheid production were assessed from anatomical observations of the developing xylem. KEY RESULTS: Cambial activity started earlier, stopped later and lasted longer in dominant trees than in intermediate and suppressed ones. The onset of cambial activity was estimated to have taken 3 weeks to spread to 90 % of the trees in the stand, while the cessation needed 6 weeks. Cambial activity was more intense in dominant trees than in intermediate and suppressed ones. It was estimated that about 75 % of tree-ring width variability was attributable to the rate of cell production and only 25 % to its duration. Moreover, growth duration was correlated to tree height, while growth rate was better correlated to crown area. CONCLUSIONS: These results show that, in a closed conifer forest, stem diameter variations resulted principally from differences in the rate of xylem cell production rather than in its duration. Tree size interacts with environmental factors to control the timings, duration and rate of cambial activity through functional processes involving source-sink relationships principally, but also hormonal controls.

Linking carbon supply to root cell-wall chemistry and mechanics at high altitudes in Abies georgei.

BACKGROUND AND AIMS: The mobile carbon supply to different compartments of a tree is affected by climate, but its impact on cell-wall chemistry and mechanics remains unknown. To understand better the variability in root growth and biomechanics in mountain forests subjected to substrate mass movement, we investigated root chemical and mechanical properties of mature Abies georgei var. smithii (Smith fir) growing at different elevations on the Tibet-Qinghai Plateau. METHODS: Thin and fine roots (0·1-4·0 mm in diameter) were sampled at three different elevations (3480, 3900 and 4330 m, the last corresponding to the treeline). Tensile resistance of roots of different diameter classes was measured along with holocellulose and non-structural carbon (NSC) content. KEY RESULTS: The mean force necessary to break roots in tension decreased significantly with increasing altitude and was attributed to a decrease in holocellulose content. Holocellulose was significantly lower in roots at the treeline (29·5 ± 1·3 %) compared with those at 3480 m (39·1 ± 1·0 %). Roots also differed significantly in NSC, with 35·6 ± 4·1 mg g(-1) dry mass of mean total soluble sugars in roots at 3480 m and 18·8 ± 2·1 mg g(-1) dry mass in roots at the treeline. CONCLUSIONS: Root mechanical resistance, holocellulose and NSC content all decreased with increasing altitude. Holocellulose is made up principally of cellulose, the biosynthesis of which depends largely on NSC supply. Plants synthesize cellulose when conditions are optimal and NSC is not limiting. Thus, cellulose synthesis in the thin and fine roots measured in our study is probably not a priority in mature trees growing at very high altitudes, where climatic factors will be limiting for growth. Root NSC stocks at the treeline may be depleted through over-demand for carbon supply due to increased fine root production or winter root growth.

Autotrophic and heterotrophic respiration in needle fir and Quercus-dominated stands in a cool-temperate forest, central Korea.

To investigate annual variation in soil respiration (R (S)) and its components [autotrophic (R (A)) and heterotrophic (R (H))] in relation to seasonal changes in soil temperature (ST) and soil water content (SWC) in an Abies holophylla stand (stand A) and a Quercus-dominated stand (stand Q), we set up trenched plots and measured R (S), ST and SWC for 2 years. The mean annual rate of R (S) was 436 mg CO(2) m(-2) h(-1), ranging from 76 to 1,170 mg CO(2) m(-2) h(-1), in stand A and 376 mg CO(2) m(-2) h(-1), ranging from 82 to 1,133 mg CO(2) m(-2) h(-1), in stand Q. A significant relationship between R (S) and its components and ST was observed over the 2 years in both stands, whereas a significant correlation between R (A) and SWC was detected only in stand Q. On average over the 2 years, R (A) accounted for approximately 34% (range 17-67%) and 31% (15-82%) of the variation in R (S) in stands A and Q, respectively. Our results suggested that vegetation type did not significantly affect the annual mean contributions of R (A) or R (H), but did affect the pattern of seasonal change in the contribution of R (A) to R (S).

Mitochondrial bioenergetics linked to the manifestation of programmed cell death during somatic embryogenesis of Abies alba.

The present work reports changes in bioenergetic parameters and mitochondrial activities during the manifestation of two events of programmed cell death (PCD), linked to Abies alba somatic embryogenesis. PCD, evidenced by in situ nuclear DNA fragmentation (TUNEL assay), DNA laddering and cytochrome c release, was decreased in maturing embryogenic tissue with respect to the proliferation stage. In addition, the major cellular energetic metabolites (ATP, NAD(P)H and glucose-6-phosphate) were highered during maturation. The main mitochondrial activities changed during two developmental stages. Mitochondria, isolated from maturing, with respect to proliferating cell masses, showed an increased activity of the alternative oxidase, external NADH dehydrogenase and fatty-acid mediated uncoupling. Conversely, a significant decrease of the mitochondrial K (ATP)(+) channel activity was observed. These results suggest a correlation between mitochondrial activities and the manifestation of PCD during the development of somatic embryos. In particular, it is suggested that the K (ATP)(+) channel activity could induce an entry of K(+) into the matrix, followed by swelling and a release of cytochrome c during proliferation, whereas the alternative pathways, acting as anti-apoptotic factors, may partially counteract PCD events occurring during maturation of somatic embryos.

Cambial activity and intra-annual xylem formation in roots and stems of Abies balsamea and Picea mariana.

BACKGROUND AND AIMS: Studies on xylogenesis focus essentially on the stem, whereas there is basically no information about the intra-annual growth of other parts of the tree. As roots strongly influence carbon allocation and tree development, knowledge of the dynamics of xylem production and maturation in roots at a short time scale is required for a better understanding of the phenomenon of tree growth. This study compared cambial activity and xylem formation in stem and roots in two conifers of the boreal forest in Canada. METHODS: Wood microcores were collected weekly in stem and roots of ten Abies balsamea and ten Picea mariana during the 2004-2006 growing seasons. Cross-sections were cut using a rotary microtome, stained with cresyl violet acetate and observed under visible and polarized light. The number of cells in the cambial zone and in differentiation, plus the number of mature cells, was counted along the developing xylem. KEY RESULTS: Xylem formation lasted from the end of May to the end of September, with no difference between stem and roots in 2004-2005. On the contrary, in 2006 a 1-week earlier beginning of cell differentiation was observed in the stem, with cell wall thickening and lignification in roots ending up to 22 d later than in the stem. Cell production in the stem was concentrated early in the season, in June, while most cell divisions in roots occurred 1 month later. CONCLUSIONS: The intra-annual dynamics of growth observed in stem and roots could be related to the different amount of cells produced by the cambium and the patterns of air and soil temperature occurring in spring.

Isolation of mitochondria from embryogenic cultures of Picea abies (L.) Karst. and Abies cephalonica Loud.: characterization of a K+(ATP) channel.

A valuable method to isolate and purify mitochondria from embryonal masses of two coniferous species (Picea abies [L.] Karst. and Abies cephalonica Loud.) is described. Crude mitochondria from both species were shown to be intact, oxygen consuming (with malate plus glutammate, succinate and NADH as substrates) and well coupled (respiratory control ratio ca. 4). The oxidation of the substrates was only partially KCN-insensitive (alternative oxidase) in some cases. However, these fractions were contaminated by membranes (e.g. plasmalemma, tonoplast, Golgi and endoplasmic reticulum). After purification by a discontinuous Percoll gradient (18, 23, 40%, v/v), three mitochondrial populations were separated. The 0/18 interface fraction was composed mainly of broken and uncoupled mitochondria, while the other two (18/23 and 23/40 interface fractions) contained intact and coupled mitochondria, but only 23/40 interface fraction revealed to be better purified starting from both coniferous embryonal masses. In the latter purified fraction, the presence of a cyclosporin A-sensitive K (ATP) (+) channel was demonstrated. These findings were discussed in the light of the potential use of these mitochondrial fractions in bioenergetic studies, or in the involvement of these organelles to stress response in conifers.

The positional distribution of cell death of ray parenchyma in a conifer, Abies sachalinensis.

We monitored the distribution of death of secondary xylem cells in a conifer, Abies sachalinensis. The cell death of tracheids, which are tracheary elements, occurred successively and was related to the distance from cambium. Thus, it resembled programmed cell death. By contrast, the death of long-lived ray parenchyma cells had the following features: (1) ray parenchyma cells remained alive for several years or more; (2) in many cases, no successive cell death occurred even within a given radial cell line of a ray; and (3) the timing of cell death differed among upper and lower radial cell lines and other lines of cells within a ray. These results indicate that the death of long-lived ray parenchyma cells involves a different process from the death of tracheids. The initiation of secondary wall formation and the lignification of ray parenchyma cells in the current year's annual ring were delayed in the upper and lower radial cell lines of a ray. In addition, the density of distribution and orientation of cortical microtubules in such cells were different from those in cells in other radial lines. Ray parenchyma cells in the previous year's annual ring within the upper and lower radial cell lines of a ray contained many starch grains. Our results indicate that positional information is an important factor in the control of the pattern of differentiation and, thus, of the functions of ray parenchyma cells that are derived from the same cambial ray cells.

Differentiation of terminal latewood tracheids in silver fir trees during autumn.

BACKGROUND AND AIMS: The differentiation of terminal latewood tracheids of silver fir (Abies alba) trees grown in Slovenia was investigated in autumn/winter 2001/2002. METHODS: The experimental trees were divided into three groups: one with narrow annual rings, width less than 1 mm; one with annual ring widths between 1 and 4 mm; and one group with broad rings larger than 4 mm. The differentiation of terminal latewood tracheids was investigated by light-, electron- and UV-microscopy in tissues sampled in October and November 2001 and March 2002. KEY RESULTS: In the middle of October, cambial divisions did not occur any more in any of the trees. In trees with narrow annual rings, cell wall deposition as well as lignification were completed in terminal latewood tracheids at this date, whereas in trees with annual ring widths of more than 1 mm these processes still continued. Electron microscopy as well as UV microscopy revealed an unlignified inner S(2) layer and the absence of S(3) and warty layers. With increasing distance from the cambium, wall formation and lignification gradually appeared to be completed. Samples of all trees taken in the middle of November only contained differentiated terminal latewood tracheids. At the structural and lignin topochemical level, November and March samples showed completed differentiation of walls of terminal latewood tracheids. CONCLUSIONS: In trees with broader annual rings, the final steps of differentiation of the youngest latewood tracheids near the cambium still continued during autumn, but were finished prior to winter. It was concluded from structural observations that duration of cambial activity is longer in trees with broad annual rings than in trees with narrow rings.