Widyanto Dwi Nugroho

Gibberellin is required for the formation of tension wood and stem gravitropism in Acacia mangium seedlings

BACKGROUND AND AIMS Angiosperm trees generally form tension wood on the upper sides of leaning stems. The formation of tension wood is an important response to gravitational stimulus. Gibberellin appears to be involved in the differentiation of secondary xylem, but it remains unclear whether gibberellin plays a key role in the formation of tension wood and plant gravitropism. Therefore, a study was designed to investigate the effects of gibberellin and of inhibitors of the synthesis of gibberellin, namely paclobutrazole and uniconazole-P, on the formation of tension wood and negative stem gravitropism in Acacia mangium seedlings. METHODS Gibberellic acid (GA(3)), paclobutrazole and uniconazole-P were applied to seedlings via the soil in which they were growing. Distilled water was applied similarly as a control. Three days after such treatment, seedlings were tilted at an angle of 45° from the vertical, and samples of stems were collected for analysis 2 weeks, 2 months and 6 months after tilting. The effects of treatments on the stem recovery degree (Rº) were analysed as an index of the negative gravitropism of seedlings, together the width of the region of tension wood in the upper part of inclined stems. KEY RESULTS It was found that GA(3) stimulated the negative gravitropism of tilted seedling stems of A. mangium, while paclobutrazole and uniconazole-P inhibited recovery to vertical growth. Moreover, GA(3) stimulated the formation of tension wood in tilted A. mangium seedlings, while paclobutrazole and uniconazole-P strongly suppressed the formation of tension wood, as assessed 2 weeks after tilting. CONCLUSIONS The results suggest that gibberellin plays an important role at the initial stages of formation of tension wood and in stem gravitropism in A. mangium seedlings in response to a gravitational stimulus.

Radial variations in the anatomical characteristics and density of the wood of Acacia mangium of five different provenances in Indonesia

The anatomical characteristics and density of wood were examined in 23-year-old Acacia mangium trees that had been planted in Yogyakarta, Indonesia. The seeds had been collected from trees of five different provenances. The distance from the pith of the boundary between juvenile and mature wood was also examined to clarify the maturity of the wood. Lengths of wood fibers near the pith and the distance from the pith of the boundary between juvenile and mature wood differed significantly among provenances. By contrast, other anatomical characteristics of the wood such as fiber wall area, fiber wall thickness, fiber diameter, vessel lumen area, vessel diameter, vessel frequency and wood density did not differ significantly among provenances. Wood density was strongly correlated with the area of fiber walls. Our observations suggest that Sidei and Daintree might be more appropriate provenances among those examined for the Acacia mangium tree-breeding programs in Indonesia that are aimed at improving wood quality, because these provenances are associated with longer initial wood fibers and narrower juvenile areas than the other provenances studied.

Localized cooling of stems induces latewood formation and cambial dormancy during seasons of active cambium in conifers

BACKGROUND AND AIMS In temperate regions, trees undergo annual cycles of cambial growth, with periods of cambial activity and dormancy. Environmental factors might regulate the cambial growth, as well as the development of cambial derivatives. We investigated the effects of low temperature by localized cooling on cambial activity and latewood formation in two conifers, Chamaecyparis obtusa and Cryptomeria japonica. METHODS A plastic rubber tube that contained cooled water was wrapped around a 30-cm-wide portion of the main stem of Chamaecyparis obtusa and Cryptomeria japonica trees during seasons of active cambium. Small blocks were collected from both cooled and non-cooled control portions of the stems for sequential observations of cambial activity and for anatomical measurements of cell morphology by light microscopy and image analysis. KEY RESULTS The effect of localized cooling was first observed on differentiating tracheids. Tracheids narrow in diameter and with significantly decreased cambial activity were evident 5 weeks after the start of cooling in these stems. Eight weeks after the start of cooling, tracheids with clearly diminished diameters and thickened cell walls were observed in these stems. Thus, localized low temperature induced narrow diameters and obvious thickening of secondary cell walls of tracheids, which were identified as latewood tracheids. Two months after the cessation of cooling, a false annual ring was observed and cambium became active again and produced new tracheids. In Cryptomeria japonica, cambial activity ceased earlier in locally cooled portions of stems than in non-cooled stems, indicating that the cambium had entered dormancy sooner in the cooled stems. CONCLUSIONS Artificial cooling of stems induced latewood formation and cessation of cambial activity, indicating that cambium and its derivatives can respond directly to changes in temperature. A decrease in the temperature of the stem is a critical factor in the control of cambial activity and xylem differentiation in trees.

Climate change and the regulation of wood formation in trees by temperature

Key messageA better understanding of the influence of environmental conditions on wood formation should help to improve the radial growth of trees and to prepare for climate change.AbstractThe cambial activity of trees is associated with seasonal cycles of activity and dormancy in temperate zones. The timing of cambial reactivation in early spring and dormancy in autumn plays an important role in determination of the cambial growth and the environmental adaptivity of temperate trees. This review focuses on the temperature regulation of the timing of cambial reactivation and xylem differentiation and highlights recent advances of bud growth in relation to cambial activity of temperate trees. In addition, we discuss relationships between the timing of cambial reactivation, start of xylem differentiation and changes in levels of storage materials to identify the source of the energy required for cell division and differentiation. We also present a summary of current understanding of the effects of rapid increases and decreases in temperature on cambial activity, by localized heating and cooling, respectively. Increases in temperature from late winter to early spring influence the physiological processes that are involved in the initiation of cambial reactivation and xylem differentiation both in localized heated stems and under natural conditions. Localized cooling has a direct effect on cell expansion, the thickening of walls of differentiating tracheids, and the rate of division of cambial cells. A rapid decrease in temperature of the stem might be the critical factor in the control of latewood formation and the cessation of cambial activity. Therefore, temperature is the main driver of cambial activity in temperate trees and trees are able to feel changes in temperature through the stem. The climate change might affect wood formation in trees.

Xylogenesis in Trees: From Cambial Cell Division to Cell Death

Abstract Wood is produced by the cambium of stems of trees. The cambial activity of trees exhibits seasonal cycles of activity and dormancy in temperate and cool zones. The timing of cambial reactivation in the early spring plays an important role in the determination of the quantity of wood and the environmental adaptivity of trees. Numerous studies have demonstrated that the cambial reactivation is controlled by the temperature of stems. As soon as cambial cells lose the ability to divide, they start to differentiate into secondary xylem cells. Finally, cell death occurs in secondary xylem cells, but its timing is different among the types and positions of cells. During formation of the secondary wall in tracheids or wood fibers, the cellulose microfibrils change their orientation progressively to form thick cell walls with a highly organized structure. In addition, secondary xylem cells form modifications of the cell wall, such as helical thickening and pits, by the localized deposition of cellulose microfibrils. There are considerable evidences that the dynamics of cortical microtubules are closely related to the orientation and localization of newly deposited cellulose microfibrils in the differentiating secondary xylem cells, indicating that microtubules control the structure of cell wall, thereby wood quality.

Gibberellin mediates the development of gelatinous fibres in the tension wood of inclined Acacia mangium seedlings.

BACKGROUND AND AIMS Gibberellin stimulates negative gravitropism and the formation of tension wood in tilted Acacia mangium seedlings, while inhibitors of gibberellin synthesis strongly inhibit the return to vertical growth and suppress the formation of tension wood. To characterize the role of gibberellin in tension wood formation and gravitropism, this study investigated the role of gibberellin in the development of gelatinous fibres and in the changes in anatomical characteristics of woody elements in Acacia mangium seedlings exposed to a gravitational stimulus. METHODS Gibberellin, paclobutrazol and uniconazole-P were applied to the soil in which seedlings were growing, using distilled water as the control. Three days after the start of treatment, seedlings were inclined at 45 ° to the vertical and samples were harvested 2 months later. The effects of the treatments on wood fibres, vessel elements and ray parenchyma cells were analysed in tension wood in the upper part of inclined stems and in the opposite wood on the lower side of inclined stems. KEY RESULTS Application of paclobutrazol or uniconazole-P inhibited the increase in the thickness of gelatinous layers and prevented the elongation of gelatinous fibres in the tension wood of inclined stems. By contrast, gibberellin stimulated the elongation of these fibres. Application of gibberellin and inhibitors of gibberellin biosynthesis had only minor effects on the anatomical characteristics of vessel and ray parenchyma cells. CONCLUSIONS The results suggest that gibberellin is important for the development of gelatinous fibres in the tension wood of A. mangium seedlings and therefore in gravitropism.

Cavitation of intercellular spaces is critical to establishment of hydraulic properties of compression wood of Chamaecyparis obtusa seedlings

BACKGROUND AND AIMS When the orientation of the stems of conifers departs from the vertical as a result of environmental influences, conifers form compression wood that results in restoration of verticality. It is well known that intercellular spaces are formed between tracheids in compression wood, but the function of these spaces remains to be clarified. In the present study, we evaluated the impact of these spaces in artificially induced compression wood in Chamaecyparis obtusa seedlings. METHODS We monitored the presence or absence of liquid in the intercellular spaces of differentiating xylem by cryo-scanning electron microscopy. In addition, we analysed the relationship between intercellular spaces and the hydraulic properties of the compression wood. KEY RESULTS Initially, we detected small intercellular spaces with liquid in regions in which the profiles of tracheids were not rounded in transverse surfaces, indicating that the intercellular spaces had originally contained no gases. In the regions where tracheids had formed secondary walls, we found that some intercellular spaces had lost their liquid. Cavitation of intercellular spaces would affect hydraulic conductivity as a consequence of the induction of cavitation in neighbouring tracheids. CONCLUSIONS Our observations suggest that cavitation of intercellular spaces is the critical event that affects not only the functions of intercellular spaces but also the hydraulic properties of compression wood.

Three-Dimensional Imaging of Cambium and Secondary Xylem Cells by Confocal Laser Scanning Microscopy

Wood is one of the most important renewable natural resources. The quantity and quality of wood are determined by the activity of cambium, xylem differentiation, and cell death. This chapter provides protocols for visualizing the structure of cambium and secondary xylem cells, dynamic changes in cortical microtubules (MTs) and cell organelles, such as nuclei and vacuoles, and the detection of fluorescence spectra of autofluorescence of cell walls and nuclear DNA fragmentation by confocal laser scanning microscopy (CLSM) with fluorescent dye. CLSM is a powerful tool for the construction of three-dimensional (3D) images of the process of wood formation in trees.

Effect of epicuticular wax crystals on the localization of artificially deposited sub-micron carbon-based aerosols on needles of Cryptomeria japonica

Elucidation of the mechanism of adsorption of particles suspended in the gas-phase (aerosol) to the outer surfaces of leaves provides useful information for understanding the mechanisms of the effect of aerosol particles on the growth and physiological functions of trees. In the present study, we examined the localization of artificially deposited sub-micron-sized carbon-based particles on the surfaces of needles of Cryptomeria japonica, a typical Japanese coniferous tree species, by field-emission scanning electron microscopy. The clusters (aggregates) of carbon-based particles were deposited on the needle surface regions where epicuticular wax crystals were sparsely distributed. By contrast, no clusters of the particles were found on the needle surface regions with dense distribution of epicuticular wax crystals. Number of clusters of carbon-based particles per unit area showed statistically significant differences between regions with sparse epicuticular wax crystals and those with dense epicuticular wax crystals. These results suggest that epicuticular wax crystals affect distribution of carbon-based particles on needles. Therefore, densely distributed epicuticular wax crystals might prevent the deposition of sub-micron-sized carbon-based particles on the surfaces of needles of Cryptomeria japonica to retain the function of stomata.

Stem gravitropism and tension wood formation in Acacia mangium seedlings inclined at various angles

Background and Aims In response to a gravitational stimulus, angiosperm trees generally form tension wood on the upper sides of leaning stems in order to reorientate the stems in the vertical direction. It is unclear whether the angle of inclination from the vertical affects tension wood formation. This study was designed to investigate negative gravitropism, tension wood formation and growth eccentricity in Acacia mangium seedlings inclined at different angles. Methods Uniform seedlings of A. mangium were artificially inclined at 30°, 45°, 60° and 90° from the vertical and harvested, with non-inclined controls, 3 months later. We analysed the effects of the angle of inclination on the stem recovery angle, the anatomical features of tension wood and radial growth. Key Results Smaller inclination angles were associated with earlier stem recovery while stems subjected to greater inclination returned to the vertical direction after a longer delay. However, in terms of the speed of negative gravitopism towards the vertical, stems subjected to greater inclination moved more rapidly toward the vertical. There was no significant difference in terms of growth eccentricity among seedlings inclined at different angles. The 30°-inclined seedlings formed the narrowest region of tension wood but there were no significant differences among seedlings inclined at 45°, 60° and 90°. The 90°-inclined seedlings formed thicker gelatinous layers than those in 30°-, 45°- and 60°-inclined seedlings. Conclusion Our results suggest that the angle of inclination of the stem influences negative gravitropism, the width of the tension wood region and the thickness of gelatinous layers. Larger amounts of gelatinous fibres and thicker gelatinous layers might generate the higher tensile stress required for the higher speed of stem-recovery movement in A. mangium seedlings.