Anatomical and Morphological Features of Scots Pine Heartwood Formation in Two Forest Types in the Middle Taiga Subzone

Currently, there is no consensus on how growing conditions affect the heartwood formation in Scots pine. Comparing the results obtained by different authors is difficult due to methodology differences and poor descriptions of the objects used. We selected two sample plots in (1) a blueberry pine forest on nutrient-rich and moist soil and (2) a lichen pine forest on nutrient-poor and dry soil and performed their detailed characterization. The sample plots were located 22 km apart in the middle taiga subzone (Karelia Republic, northwest Russia). In each sample plot, we selected five dominant trees (model trees), from which we took cores at different trunk heights (0.3, 1.5, 4.5, 7.5 and 10.5 m). The cores were treated with 2,6-dichlorophenolindophenol to identify the heartwood zone. Additionally, samples were taken to study the structural features of the transition zone between sapwood and heartwood. In both forest types, the number of heartwood rings depended on the cambium age, and the patterns of parenchyma cell death did not differ in the transition zone. These facts point to a predominantly internal regulation of the heartwood formation in Scots pine. The heartwood radius and its proportion on the cross-sections were significantly higher in the blueberry pine forest than in the lichen pine forest, despite the relative values of the annual ring width. Further research is needed to develop successful Scots pine heartwood width models under a wide range of conditions.

Evaluation of reference genes and characterization of the MYBs in xylem radial change of Chinese fir stem

The radial change (RC) of tree stem is the process of heartwood formation involved in complex molecular mechanism. Chinese fir (Cunninghamia lanceolata (Lamb.) Hook.), an evergreen species, is an important fast-growing timber tree in southern China. In this study, the top four stable genes (IDH, UBC2, RCA and H2B) were selected in RC tissues of 15 years old Chinese fir stem (RC15) and the genes (H2B, 18S, TIP41 and GAPDH) were selected in RC tissues of 30 years old Chinese fir stem (RC30). The stability of the reference genes is higher in RC30 than in RC15. Sixty-one MYB transcripts were obtained on the PacBio Sequel platform from woody tissues of one 30 years old Chinese fir stem. Based on the number of MYB DNA-binding domain and phylogenetic relationships, the ClMYB transcripts contained 21 transcripts of MYB-related proteins (1R-MYB), 39 transcripts of R2R3-MYB proteins (2R-MYB), one transcript of R1R2R3-MYB protein (3R-MYB) belonged to 18 function-annotated clades and two function-unknown clades. In RC woody tissues of 30 years old Chinese fir stem, ClMYB22 was the transcript with the greatest fold change detected by both RNA-seq and qRT-PCR. Reference genes selected in this study will be helpful for further verification of transcript abundance patterns during the heartwood formation of Chinese fir.

Liquid Chromatographic/Mass Spectrometric Study on the Role of Beech (Fagus sylvatica L.) Wood Polyphenols in Red Heartwood Formation

The present research focused on the analysis of European beech (Fagus sylvatica L.) wood polyphenols in respect to red heartwood formation, which is a significant color and technological defect of the species. For the first time, high-performance liquid chromatography/tandem mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) were applied for the investigation of the structure and the radial distribution of polyphenols. Altogether 125 compounds were characterized by their MSn spectra, of which 71 were tentatively identified, including procyanidins (dimers to pentamers), flavonoids (taxifolin, naringenin, isorhamnetin, (epi)afzelechin), (+)-catechin, (–)-epicatechin) and their glycoside conjugates, phenolic acids (gallic acid, vanillic acid, syringic acid) and their glycosides as well as gallic acid derivatives, many of which were identified for the first time in beech wood. It was found that the concentration of many compounds increased at the color boundary. In situ polyphenol synthesis and metabolism were clearly evidenced at the color boundary. Red heartwood contained only free aglycones (syringic acid, taxifolin, naringenin, isorhamnetin, naringenin, syringic acid). Contrary to earlier assumptions, the MALDI-TOF analysis did not indicate the presence of oxidized high-molecular-weight polymeric polyphenols in the red heartwood tissues. The role of individual compounds in the formation of the red heartwood chromophores are discussed.

Heartwood and Sapwood Variation and Development in Chenshan Red-Heart Chinese Fir (Cunninghamia lanceolata (Lamb.) Hook)

Chenshan red-heart Chinese fir is a provenance of Cunninghamia lanceolata, with high-value red heartwood, which is widely used in high-quality furniture and construction. Yet, there is still little information on heartwood development of this tree for high-value decorative timber, which is essential to improve one's plantation management strategy. Here, we investigated the horizontal and vertical variation of heartwood and sapwood and simulated heartwood formation process using stem analysis method. We selected 15 sample trees from five plots of 20 m × 30 m in Chenshan red-heart Chinese fir plantations (9, 15, 26, 29, and 34 years old, respectively). The results showed that Chenshan red-heart Chinese fir stems began to form heartwood when the xylem diameter reached 4 to 8 cm. The heartwood diameter and area, as well as the sapwood area, all increased in the different-aged Chenshan red-heart Chinese firs with increasing xylem diameter and decreased with increasing tree height. As tree height increased, the red heartwood formation rate declined at all ages. Relationship analysis showed that xylem diameter was the most important factor influencing heartwood formation. Red heartwood rate at breast height could be modeled by logistic models. We concluded that heartwood formation began at about 7 years old, and the formation rate increased until peaking at 60 percent at 40 years old. In conclusion, it will be imperative to prolong the Chenshan red-heart Chinese fir rotation period from the currently common 25 years to about 40 years to achieve the maximum sustainable yield of high-value decorative timber.

Variations in heartwood formation and wood density as a function of age and plant spacing in a fast-growing eucalyptus plantation

The heartwood formation process is little known in fast growing plantation woods. Therefore, the aims of this study were to determine how planting spacing and tree age affect the formation and proportion of heartwood and sapwood, as well as the density of eucalyptus wood. Trees from a eucalyptus clonal plantation (Eucalyptus grandis × E. urophylla hybrid) cultivated in three spacings (3 × 1, 3 × 2 and 3 × 3 m) were sampled at 14, 27, 32, 53, 64 and 76 months of age. Heartwood percentage was quantified with Dimethyl yellow indicator, while the wood density was determined by X-ray densitometry. The heartwood percentage, wood volume, heartwood density and wood density were correlated with the different growth rates. The heartwood formation process started between 32 and 53 months, regardless of spacing. The heartwood proportion doubled with increasing age in the widest spacing and increased about four times in 3 × 1 and 3 × 2 m spacing. The planting spacing influenced the growth rates of the trees, but did not affect the heartwood and sapwood percentage or density. The greatest increase in density values occurred between the first year of growth (14 months) and the beginning of heartwood formation (53 months).

Lignification and cell wall thickening of ray parenchyma cells in Scots pine sapwood

Scots pine exhibits variations in ray anatomy, which are poorly understood. Some ray parenchyma cells develop thick and lignified cell walls before heartwood formation. We hypothesized that some stands and trees show high numbers of lignified and thick-walled parenchyma cells early in the sapwood. Therefore, a microscopic analysis of Scots pine sapwood from four different stands in Northern Europe was performed on Safranin — Astra blue-stained tangential micro sections from outer and inner sapwood areas. Significant differences in lignification and cell wall thickening of ray parenchyma cells were observed in the outer sapwood between all of the stands for the trees analyzed. On a single tree level, the relative lignification and cell wall thickening of ray parenchyma cells ranged from 4.3% to 74.3% in the outer sapwood. In the inner sapwood, lignification and cell wall thickening of ray parenchyma cells were more frequent. In some trees, however, the difference in lignification and cell wall thickening between inner and outer sapwood was small since early lignification, and cell wall thickening was already more common in the outer sapwood. Ray composition and number of rays per area were not significantly different within the studied material. However, only one Scottish tree had a significantly higher number of ray parenchyma cells per ray. The differences discovered in lignification and cell wall thickening in ray parenchyma cells early in the sapwood of Scots pine are relevant for wood utilization in general and impregnation treatments with protection agents in particular.

Extractives elucidation of Taiwania cryptomerioides sapwood

Taiwania (Taiwania cryptomerioides Hayata) has long been regarded as a living fossil from the Tertiary period of Mesozoic Era for its distinguished yellowish-red color with purplish-pink streaks presented in its heartwood. With this elegant appearance that matches the color “red” for good fortune in the Taiwanese culture, Taiwania is supposed to be a popular wood in Taiwan where it is a native species of. Extractives contribute to the properties of wood. It is a fascinating subject to investigate extractives biosynthesis in the process of heartwood formation. Up to date, there is no phytochemistry study of Taiwania sapwood. In this study, three new sesquiterpenoids, Taiwania A (1), Taiwania B (2), and Taiwania C (3), together with 75 known compounds in the Taiwania sapwood. The structures of extractives were determined by analysis of spectroscopic data and comparison with the literatures. This study supported secondary reaction lignans could be found in sapwood that confirmed our previous research on the Taiwania-type of heartwood formation.

Physiological changes during heartwood formation induced by plant growth regulators in Dalbergia odorifera (Leguminosae)

There is a need to better understand and promote formation of ‘Jiang Xiang’, the highly valuable heartwood of Dalbergia odorifera T. Chen. Plant growth regulators are the most widely used and generally most effective promoters of heartwood formation. However, to date no method has been proved effective for D. odorifera as little is known about biochemical and physiological change during heartwood development. Four potential heartwood induction substances viz. ethephon, methyl jasmonate, salicylic acid and abscisic acid solutions were injected into the trunk of D. odorifera to determine the effect on heartwood formation and related physiological responses. Non-structural carbohydrates, wood properties, essential oil and histochemical and structural observation were assessed in the post-treatment period. As also observed in the formation of natural heartwood, growth regulator-induced Jiang Xiang production was accompanied by sapwood dehydration, non-structural carbohydrates consumption and synthesis of heartwood specific substances. These substances were synthesized in the ray and axial parenchyma and than transported into the vessels through the pits. As the heartwood substances accumulated, basic density and essential oil content increased gradually, thereby Jiang Xiang was finally produced. In this process, physiological parameters of discoloured sapwood gradually evolved to resemble those of natural heartwood. Ethephon-induced Jiang Xiang was closest to natural heartwood and its density, essential oil content, composition and relative amounts met the standards for high-quality Jiang Xiang, while the other inducers had unsatisfactory induction effects. This study supports the hypothesis that ethephon could be applied to induce formation of Jiang Xiang in D. odorifera.

A transcriptomic view to wounding response in young Scots pine stems

We studied the stress response of five-year-old Scots pine xylem to mechanical wounding using RNA sequencing. In general, we observed a bimodal response in pine xylem after wounding. Transcripts associated with water deficit stress, defence, and cell wall modification were induced at the earliest time point of three hours; at the same time, growth-related processes were down-regulated. A second temporal wave was triggered either at the middle and/or at the late time points (one and four days). Secondary metabolism, such as stilbene and lignan biosynthesis started one day after wounding. Scots pine synthesises the stilbenes pinosylvin and its monomethyl ether both as constitutive and induced defence compounds. Stilbene biosynthesis is induced by wounding, pathogens and UV stress, but is also developmentally regulated when heartwood is formed. Comparison of wounding responses to heartwood formation shows that many induced processes (in addition to stilbene biosynthesis) are similar and relate to defence or desiccation stress, but often specific transcripts are up-regulated in the developmental and wounding induced contexts. Pine resin biosynthesis was not induced in response to wounding, at least not during the first four days.