Qualitative Anatomical Characteristics and Fiber Quality of Tapped Styrax sumatrana Wood Grown in North Sumatra, Indonesia

: This study investigated the qualitative anatomical characteristics and ﬁ ber quality of tapped Styrax sumatrana wood to facilitate its further utilization. The transverse surface of the tapped S. sumatrana was light or greyish brown in the sapwood and reddish brown in the heart-wood. The resin canals of the tapped wood were formed along the growth rings at the boundaries between the heartwood and sapwood. Furthermore, microscopic analyses revealed an irregular out-line and rounded epithelial cells at the edges of the intercellular traumatic canal (TC). Approximately 8–16 epithelial cells surrounded the resin canals. The ﬁ bers in the tapped S. sumatrana were generally thin-walled, whereas those near the TC were thick-walled. Moreover, S. sumatrana were di ﬀ use-porous and exhibited intermediately distinct or indistinct growth ring boundaries. The vessels were mainly radial multiples and clusters of 3–5. In addition, they showed a diagonal-to-radial pa tt ern arrangement and a few tangential bands. Deposits were observed in some vessels in the tapped part but were absent in the untapped part. The ﬁ ber length of the tapped S. sumatrana wood was classi ﬁ ed as moderate to extensive and categorized as second-grade pulp quality. Finally, the cell walls were classi ﬁ ed as thin to moderate.


Introduction
Plants can produce exudates that usually come out when plants are injured.The exudates cover the wound and harden after being exposed to air [1,2].Resin, latex, and gum are the common exudates used.Resin is a type of exudate consisting of terpenoids or phenolics and is secreted by internal or surface glands of plant tissues [3,4].
One of the plant genera that produce resinous exudates is Styrax spp.The Styrax genus, which belongs to the family Styracaceae, comprises approximately 130 species [5].It has major habitats in the tropical, subtropical, and temperate regions of Asia, Europe, and America [6].Three Styrax species have been identified in Southeast Asia: Styrax benzoin, Styrax paralleloneurum synonymous with Styrax sumatrana, and Styrax tonkinesis.
The Styrax spp. is one of the most important plants in North Sumatra and has been utilized to produce a resin called benzoin.Benzoin has high economic value as a raw material for cosmetics, perfume binders, preservatives, and raw materials for pharmaceutical drugs, a natural antioxidant resource, and flavor enhancers in the food industry [7][8][9].The Styrax spp., apart from producing resin to be used for various uses, as mentioned, the wood also has the potential to be used for various purposes, such as light construction, furniture, and handy craft materials [9].
S. sumatrana can reach a height of 35 m and a diameter at breast height (DBH) of 40-60 cm.This wood is suitable for light construction, furniture manufacturing, handicrafts, pulp, and paper [6][7][8][9].S. sumatrana, known as the kemenyan, haminjon toba, or benzoin tree, is a commercial tree endemic to North Sumatra, Indonesia.The area of Styrax plantation in North Sumatra was 23,172 ha, with a total resin production of 8845 tons [10].
The S. sumatrana resin is harvested through mechanical treatment of the stem.A few holes with a depth of 3-5 cm are made in the stems using a sharp object and patted every four days for three months.The resin is subsequently harvested 3-4 months after treatment [11].The productive period of this tree as a resin producer is approximately 20 years, and Sumatra residents typically use it for firewood after its productive years.Following the productive phase, the resin tapping process leaves distinctive tapping scars on the wood of the Styrax tree.
Intercellular traumatic canals (TCs) may develop in response to various types of injuries, such as physical damage, infections, and insect attacks.Typically, these canals consist of epithelial cells producing natural resin and are organized in tangential bands or exist as a singular canal [12].Intercellular traumatic resin canals are found in various species, such as the Meliaceae family, due to mechanical stimuli [12], Tsuga sieboldii by piercing metal pins around the stem [13], and Picea engelmannii due to the attack of Dendroctonus rufipennis [14].In addition, Deng et al. [15] revealed that traumatic resin canals in Styrax are formed by parenchymal cells in the secondary xylem by schizolysigeny after injury.
Previous studies have demonstrated the anatomical characteristics of Styrax wood grown in Indonesia.Pasaribu et al. [16] reported that S. sumatrana generally shows distinctive anatomical characteristics with S. paralleloneurum.S. sumatrana and S. paralleloneurum showed multiseriate rays with three to six and three to four seriate rays, respectively.Moreover, S. paralleloneurum showed a large intercellular traumatic resin (TC), whereas S. sumatrana had a small TC in the growth ring.However, the fiber quality of both species was categorized as first-grade pulp quality.
S. sumatrana grown in North Sumatra is commonly wounded by resin tapping, which may affect its anatomical characteristics.So far, the anatomical characteristics of tapped S. sumatrana wood remain understudied.Therefore, the present study investigated the qualitative anatomical characteristics and fiber quality of tapped S. sumatrana according to the IAWA list for hardwood identification to provide valuable information on wood identification and quality to facilitate further utilization of the species.

Materials
The basic information of the sampled tree was derived from a previous study [9].A 15-year-old S. sumatrana tree tapped by the local community for resin harvesting was used in this study.The tree was located at an altitude of approximately 900 m above sea level in Banuaji IV village in the Adiankoting sub-district, North Tapanuli regency, North Sumatra, Indonesia (2°0′10.08″N, 99°4′14.52″E).A wood disc with 11.5 cm in diameter and 10 cm in thickness was obtained from the diameter breast height.The air-dry specific gravity of the wood sample was 0.58.

Macroscopic and Microscopic Observation
The transverse surfaces of the wood disc underwent abrasion with coarse sandpaper (plate number 1 from Deerfos Co., Ltd. in Incheon, Korea) via a sanding machine (TW/BD-46 model, running at 2070 rpm and 450 W from Rexon Industrial Corp., Ltd. in Taichung, Taiwan).Following this initial sanding, the wood discs were manually sanded with fine sandpaper (CC-600Cw from Daesung Abrasive Co., Ltd. in Seoul, Korea).The macroscopic structures of the transverse and longitudinal surfaces were observed under a stereo microscope (SMZ 745T, Nikon, Tokyo, Japan) Here, three wood blocks with dimensions of 20 (L) × 20 (R) × 20 (T) mm 3 were extracted from the tapped and untapped parts near the bark of the wood discs for microscopic observation.The optical microscopy in the present study was performed according to Savero et al. [17].The samples were soaked in a mixture of glycerin and water (50:50) and heated on a heating plate for 30-45 min.Cross, tangential, and radial sections with approximately 20 µm thickness were prepared using a sliding microtome (Nippon Optical Works Co., Ltd., Nagano, Japan).All slices were stained with 1% safranin and 1% light-green solution.Additionally, they were dehydrated using a graded series of alcohol (50, 70, 90, 95, and 99%) and xylene.Permanent slides were prepared using Canada balsam.The stained sections were observed using an optical microscope (Eclipse Si, Nikon, Tokyo, Japan) connected to i-Solution-lite version 26.1 software (IMT i-Solution Inc., Burnaby, BC, Canada).

Cell Maceration
Matchstick-sized samples were prepared for maceration.The wood samples were macerated in a boiling solution containing 60% glacial acetic acid and 30% hydrogen peroxide at 80 °C for 1-2 h [18].The macerated samples were rinsed under running tap water until neutralization.Next, fibers were separated using a pin while washing in distilled water.The fibers were stained using 1% safranin solution for approximately 3 h and then washed with distilled water.They were subsequently arranged on a microscope slide, dripped with glycerin, and mounted with a cover glass for observation under an optical microscope (Eclipse Si, Nikon, Tokyo, Japan) connected to i-Solution Lite software (IMT i-Solution Inc., Burnaby, BC, Canada).

Measuring Dimensions of Vessels and Fibers, as Well as Their Derivative Value
The fiber length and diameter, lumen diameter, and vessel length and diameter were each measured using 25 cells.The derivative values of the fiber, such as the Runkel ratio (RR), felting power (FP), mulhsteph ratio (MR), coefficient rigidity (CR), and flexibility ratio (FR), were analyzed to elucidate the quality of the wood fiber for pulp and paper production.The fiber quality was determined as previously described [19], as shown in Table 1.The pulp quality classification is shown in Table 2. Fibers were subsequently separated using a pin while washing in distilled water.
Table 1.Fiber derivative values and their formula equation.

The Derivative Values of Fiber Equation
Runkel ratio (RR) 2

Macroscopic Characteristics in the Transverse and Longitudinal Surfaces
Figure 1 shows the transverse, tangential, and radial surfaces of an S. sumatrana wood disc wounded by tapping.S. sumatrana wood showed light or greyish-brown sapwood and reddish-brown heartwood on the transverse surface.The heartwood proportion was higher than that of sapwood (3:1).Pasaribu et al. [16] reported that untapped S. sumatarana and S. paralleloneurum were light brown or slightly grayish-brownish-yellow and that there was no color difference between heartwood and sapwood in either species.Similarly, Damayanti et al. [21] reported that S. benzoin was light brown or slightly grayishbrownish-yellow, whereas the heartwood and sapwood showed no color difference.S. sumatrana wood showed clear TCs, forming a complete ring and a resin pocket consisting of whitish substances in the earlywood (Figure 1A,C).The radial section showed clear dark lines representing a TC in the longitudinal direction (Figure 1D).In other hardwood species, Dünisch and Baas [12] mentioned that Meliaceae species had two kinds of channels between cells: single or small groups of channels (called local intercellular canals), and the other type formed thin bands of channels along the outer edge of the wood disc.The length of the TC was extended from millimeters to a couple of meters and, in some cases, could be extended down the entire axis of the tree.

Cross-Sections
Optical micrographs of cross-sections of the tapped and untapped parts of S. sumatrana are presented in Figure 2. The wood discs showed an intermediate between distinct and indistinct growth ring boundaries, with diffuse-porous vessels observed (Figure 2A,B).The vessels were mainly in radial multiples and clusters of three to five and partly solitary.They also showed an angular outline (Figure 2C,D).In addition, S. sumatrana tended to have a diagonally to radially arranged pattern and a few tangential vessel bands.Gum or deposits were observed in some vessels in the tapped part (Figure 2B), whereas they were absent in the untapped part of the wood discs (Figure 2A).The number of vessels and diameter of vessel lumina were 7-11 per mm 2 and 96-190 (131 ± 28) µm, respectively.
TCs were arranged in tangential bands resembling growth rings (Figure 2B,D,G).In addition, the TC showed an irregular outline and rounded parenchymal cells (epithelial cells) at the edges of the canal.Approximately 8-16 epithelial cells surrounded these resin canals (Figure 2F), and TCs fulfilled with resin are also shown in Figure 2G.The fibers in the tapped and untapped parts of S. sumatrana were generally thin-walled, whereas those near the TC were thick-walled (Figure 2E,F).The apotracheal axial parenchyma was diffused and diffused-in-aggregate, whereas the paratracheal axial parenchyma was vasicentric and banded, including reticulate, narrow, and marginal bands (Figure 2C-E).
Tapped S. sumatrana generally showed anatomical characteristics comparable to Styrax wood [16,21].These studies identified mechanical treatment during resin tapping as the cause of TC in Styrax wood and other hardwoods.Pasaribu et al. [16] reported that TCs were formed at the growth ring boundary of untapped S. sumatrana and S. paralleloneurum.However, the TC that formed in S. paralleloneurum was larger than that in S. sumatrana.Damayanti et al. [21] and Pasaribu et al. [16] attributed these differences in TC size to genetic differences or natural wounding during wood harvesting.Dünisch and Baas [12] observed that wounded Meliaceae species typically developed intercellular canals, which were commonly found either individually or in small clusters, often appearing as thin tangential bands.They also mentioned that there were thick-walled fibers and increased production of reddish to dark red-colored accessory compounds near the intercellular canals.Rounded parenchyma cells were frequent at the edges of intercellular canals and may extend into the secretion.

Radial and Tangential Sections
Optical micrographs of the radial and tangential sections of tapped S. sumatrana are shown in Figures 3 and 4, respectively.Procumbent body ray and square marginal cells were observed in the radial section (Figure 3A).Epithelial cells were observed at the edges of the TC, whereas a rejuvenated ray was observed within the TC (Figure 3B).The vessels contained scalariform perforation plates (Figure 3C).The width of the rays was uniseriate to multiseriate (two to four seriates) (Figure 4A), and the fibers were non-septated and septated with distinctly bordered pits (Figures 4A,B) in the tangential section.The intervessel pits were alternating (Figure 4C), and the vessel ray pits had distinct borders in size and shape, similar to the intervessel pits (Figure 4D).Chambered prismatic oxalate crystals were observed in the axial (Figure 5A) and ray (Figure 5B) parenchyma.Silica grains were also observed in the ray cells (Figure 5C).Dickison and Phend [22]

Vessel and Fiber Characteristics
Optical micrographs of the cell components of tapped S. sumatrana are shown in Figure 6.The vessel elements were tubes with tails at both ends (Figure 6A), with a few vessels showing parallel and opposite arrangements of the perforation plates on the tips of their elements (Figures 4, 5 and 6A).The length of the vessels ranged from 436 to 965 µm, and the average was 721 ± 171 µm, showing feature 53 in the IAWA list for hardwood identification [23].The vessel length of S. sumatrana in the present study was shorter than that of Styrax wood in previous studies [16,21,24].Pasaribu et al. [16] reported a S. sumatrana vessel length of 396-1449 µm and an average of 1031 ± 178 µm.In addition, the vessel length of S. paralleloneurum was 671-1374 µm, and the average was 1026 ± 187 µm.In addition, the vessel length of S. paralleloneurum was 671-1374 µm, and the average was 1026 ± 187 µm.Damayanti et al. [21] reported S. benzoin and S. paralleloneurum vessel element lengths of 1135 ± 168 µm and 1055 ± 166 µm, respectively.Machado et al. [24] reported that the vessel lengths of Styrax latifolium, Styrax martii, Styrax leprosus, and Styrax camporum from Brazil were 970 ± 197 µm, 873 ± 197 µm, 853 ± 157 µm, and 738 ± 121 µm, respectively.
The dimensions and derivative values of the tapped S. sumatrana wood fibers are listed in Table 3.The fiber length was 1390.8 µm, which is categorized into moderate to extensive based on the Hardwood List of the IAWA Committee [23].The fiber length of S. sumatrana was smaller than that of Styrax wood reported in previous studies.Pasaribu et al. [18] reported that the fiber length and diameter of untapped S. sumatrana ranged from 1525 to 2290 µm, and the average was 1860 ± 163 µm.Damayanti et al. [21] reported an S. benzoin fiber length of 1930 ± 184 µm and an S. paralleloneurum fiber length of 1870 ± 139 µm.Machado et al. [24] reported that the fiber lengths of Styrax latifolium, Styrax martii, and Styrax leprosus were 1904 ± 317 µm, 1798 ± 262 µm, and 1811 ± 294 µm, respectively.The fiber diameter was 29.23 µm, whereas the wall thickness and fiber lumina diameter were 5.62 µm and 17.99 µm, respectively.The fiber wall of S. sumatrana in the present study was classified as thin to moderate based on the Hardwood List of the IAWA Committee [23].Damayanti et al. [21] mentioned that the fiber wall thickness and lumen diameter of tapped S. benzoin were 2.3 ± 0.4 and 32.5 ± 3.1, respectively.The fiber walls of untapped S. sumatrana and tapped S. benzoin in previous studies were distinctively thinner than those of tapped S. sumatrana in the present study.Pasaribu et al. [16] reported that the fiber and lumina diameter of untapped S. sumatrana ranged from 25 to 48 µm and from 20 to 43 µm, respectively, and the average was 35 ± 3 µm in fiber diameter and 31± 3 µm in lumina diameter.In addition, the authors also mentioned that the fiber wall thickness of S. sumatrana ranged from 1 to 3 µm with an average of 2.3 ± 0.4 µm.RR is the double-cell-wall thickness ratio to the lumen diameter.A lower RR value is more favorable for producing high-quality fiber pulp, which is essential for achieving complete flatness and adequate fiber bonding in pulp sheets [25].In the present study, the average RR of S. sumatrana wood was 0.64, and the RR ranged from 0.42 to 1.00, indicating a third-grade pulp quality.The RR in the present study demonstrated a lower pulp grade than that of Styrax wood reported in previous studies.Pasaribu et al. [18] reported a 0.15 RR in S. sumatrana and S. paralleloneurus, demonstrating the potential production of firstgrade pulp.Damayanti et al. [21] reported that the RR of both tapped S. benzoin and untapped S. paralleloneurus was 0.14, which was another indicator of first-grade pulp.
The FP refers to the ratio of fiber length to diameter.A higher FP indicates a greater inter-fiber bonding and tear strength, which positively correlates with the tensile strength of paper sheets [25].The FP of S. sumatrana wood was 46.38 and ranged from 34.73 to 57.40, showing third-grade pulp quality.The FPs of S. sumatrana and S. paralleloneurus were 52.42 and 47.70, respectively, in a previous study, indicating third-grade pulp quality [16].The FP of tapped S. benzoin and untapped S. paralleloneurus revealed third-grade pulp quality at 52.09 and 53.12, respectively, in another study [21].
The MR refers to the ratio of the surface area of the fiber wall to the total surface area of the fiber.A smaller MR value leads to greater density in pulp sheets with high mechanical strength [25].The MR of tapped S. sumatrana wood ranged from 50.73% to 74.98% and averaged 62.13% in this study, showing third-grade pulp quality.This MR demonstrated a lower pulp quality grade than that of Styrax wood reported in previous studies.Pasaribu et al. [18] reported an S. sumatrana and S. paralleloneurus MR of 24.35 and 23.72, respectively, demonstrating first-grade pulp quality.Similarly, the MR of tapped S. benzoin and untapped S. paralleloneurus showed first-grade pulp quality at 23.26 and 22.94, respectively [21].
The coefficient of rigidity (CR) is the ratio of the cell wall thickness to the fiber diameter, and a higher CR of a fiber reduces tensile strength [25].The CR of tapped S. sumatrana ranged from 0.15 to 0.25, with an average value of 0.19 in the present study, showing thirdgrade pulp quality.However, this quality class was considerably lower than that of other Styrax woods reported in previous studies.The CR of untapped S. sumatrana, tapped S. benzoin, and untapped S. paralleloneurus was approximately 0.06, showing first-grade pulp quality [16,21].
The FR is the ratio of the lumen to fiber diameter.Here, the FR of tapped S. sumatrana ranged from 0.50 to 0.70, with an average of 0.61.However, it was lower than that of other Styrax wood reported in previous studies.The FR of untapped S. sumatrana, tapped S. benzoin, and untapped S. paralleloneurus was approximately 0.88, showing first-grade pulp quality [16,21].
The total score of tapped S. sumatrana wood fibers in the present study demonstrated a second-grade pulp quality.In contrast, the total scores of untapped S. sumatrana, tapped S. benzoin, and untapped S. paralleloneurus showed first-grade pulp quality in previous studies [16,21].

Conclusions
The qualitative anatomical characteristics of tapped wood in the present study showed general characteristics of Styrax woods.TC formation was observed along the growth rings in the tapped part of the wood disc.The TC showed an irregular outline with 8-16 epithelial cells at the edges of the canal.Tapped S. sumatrana generally exhibited thin-walled fibers, whereas thick-walled fibers were observed near the TC.Furthermore, deposits were observed in some vessels in the tapped part but not in the untapped part.
Chambered prismatic oxalate crystals were observed in both axial and ray parenchyma, whereas silica grains were observed in ray cells.The fiber length and cell wall of tapped S. sumatrana were classified as moderate to extensive and thin to moderate, respectively.Moreover, the derivative value of the tapped S. sumatrana wood fibers indicated second-grade pulp quality.
In conclusion, the formation of TCs aligned tangentially was the most distinct characteristic of tapped S. sumatrana wood.The fiber quality of the tapped wood was categorized as second grade, which is acceptable for industry applications.

Figure 1 .
Figure 1.Transverse (A-C) and radial surfaces (D) of a tapped Styrax sumatrana wood disc.The resin pocket can be seen in (C).The dark lines (arrows in (A,C,D)) in the transverse and radial surfaces indicate traumatic intercellular canal bands.Scale bars: 2 mm (A,C,D) and 30 mm (B).

Table 3 .
Dimension and derivative value of tapped S. sumatrana wood fiber.