Claúdia Luizon Dias Leme1 WOOD ANATOMY OF SEVEN SPECIES KNOWN AS “PAU-PARA-TUDO” IN BRAZIL

Different medicinal plant species can be sold under the same common name. Considering the importance of the correct identification, this study aims to separate, using wood anatomy, seven species popularly known as pau-para-tudo. The results show that Drimys brasiliensis is separated from the others by the presence of tracheids. Capsicodendron dinisii hhas scalariform perforation plates and oil cells associated with the axial parenchyma. Axial parenchyma paratracheal vasicentric and in marginal bands beyond the rays’ width, can separate Osteophoeum platyspermum from Simaba cedron. Handroanthus serratifolius has the unique presence of the axial unilateral paratracheal parenchyma and storied cell elements (parenchyma, fibers and vessel elements). Rauvolfia sellowii and Leptolobium dasycarpum can be separated by the number of square/upright marginal ray cells, greater in Rauvolfia sellowii. Thus, this work shows that wood anatomy is a valuable tool for species separation, helps with the identification and consequently is important for the quality control of plant product. ANATOMIA DA MADEIRA DE SETE ESPECIES CONHECIDAS COMO “PAUPARA-TUDO” NO BRASIL RESUMO: Diferentes espécies medicinais de plantas podem ser vendidas com o mesmo nome popular. Considerando a importância de uma identificação correta, este estudo tem por objetivo separar, utilizando a anatomia de madeira, sete espécies conhecidas como pau-para-tudo. Os resultados mostram que Drimys brasiliensis é separada das outras espécies pela presença de traqueídes. Capsicodendron dinisii apresenta placa de perfuração escalariforme e células oleíferas associadas ao parênquima axial. Parênquima axial paratraqueal vasicêntrico e em faixas marginais alem da largura dos raios, separa Osteophoeum platyspermum de Simaba cedron. Handroanthus serratifolius é a única das espécies que apresenta parênquima axial paratraqueal unilateral e elementos estratificados (parênquima, fibras e elementos de vaso). Rauvolfia sellowii e Leptolobium dasycarpum podem ser separada pelo número de células marginais quadradas e eretas, maiores em Rauvolfia sellowii. Assim, este trabalho mostra que a anatomia de madeira é uma ferramenta valiosa para a identificação das espécies e é importante consequentemente para o controle de qualidade do produto vegetal.

Among all these species, Drimys brasiliensis stands out as an effective anti parasitic agent, according to studies of bark and stem done by Corrêa et al. (2011) and Claudino et al. (2013).Barrero et al. (2000) obtained essential oil from wood of D. winteri.Capsicodendron dinisii (synonym of Cinnamodendron dinisii Scwacke) is indicated as analgesic (painkiller) to treat toothache, and also to control vomiting, paralysis, as a stimulant for the treatment of migraines and to help with diseases of the uterus and vagina (RODRIGUES; CARVALHO, 2008).Andrade (2013), proved the hemolytic effect of the essential oil from the leaves, stem and bark of this species.Anti-malarial activity was described for Simaba cedron by several authors, among them Moretti et al. (1994) and Oliveira et al. (2015) and a novel pentacyclic C(19) quassinoid, cedronolactone E (1), was isolated from the wood of Simaba cedron by Hitotsuyanagi et al. (2001).Its structure was elucidated by interpretation of spectroscopic data.Rauvolfia sellowii has been described as having alkaloids in its composition, both in young stem and bark (PRESIBELLA et al., 2003;SIENNA, 2008;BARATTO et al., 2010).This species is also used in the treatment of diabetes, cholesterol control, and is also considered a hypotensive and digestive tonic (PRESIBELLA et al., 2003;SOUZA;FELFILI, 2006).Handroanthus serratifolius, in turn, whose one its synonym is Tabebuia serratifolia (Vahl) G.Nicholson, presents in both bark and stem, anti-inflammatory, antimicrobial, anti-allergic, healing and anti-tumor properties (MORAIS et al., 2005;BITENCOURT et al., 2014) and was described as having anti tripanossomial and leishmanial activity by González-Coloma et al. (2012).
Whereas the authenticity of the plant is essential for their safe use in folk medicine and for the production of drugs, this study aimed to separate through wood anatomy, considering qualitative and quantitative parameters, seven species popularly known as "paupara-tudo" (wood-for-everything): Drimys brasiliensis, Capsicodendron dinisii, Osteophloeum platyspermum, Rauvolfia sellowii, Simaba cedron, Handroanthus serratifolius and Leptolobium dasycarpum.
Histological sections were prepared on a microtome.Transverse and longitudinal sections (tangential and radial) were obtained with thickness ranging between 20 and 30 micrometers (µm).The sections underwent clarification process, followed by double-staining procedure using astra blue and safranin or just safranin (KRAUS; ARDUIN, 1997).The stained and natural sections were submitted to a dehydration process in an ethanol series, immersed in n-butyl acetate, and then mounted on permanent slides with synthetic resin.
The anatomical description, which involved the quantitative and qualitative analysis, followed the IAWA list of microscope features (IAWA COMMITTEE, 1989).It was considered for the quantitative analysis the following features: vessel diameter and frequency; ray frequency, ray height and width (in micrometers and number of cell).Measurements were performed using all species samples except Drimys brasiliensis, which has tracheids and absent vessel elements.Quantitative results are showed by descriptive parameters averages, standard deviation and maximum-minimum values.An Infinity 1 camera coupled to an Olympus 3X31 optical microscope (The Image Pro-Express 6.0 analysis program) was used and statistical analysis was undertaken following Silva and Azevedo (2002).

Wood description
Drimys brasiliensis (Figure 1A-B minutely bordered pits.d)Axial parenchyma unilateral paratracheal and aliform with short and long confluences, usually forming bands.e).Rays predominantly biseriate, composed exclusively of procumbent cells.f).Rays and axial elements storied (Table 1 and 2).Leptolobium dasycarpum (Figure 3 F-H): a)Growth ring boundaries indistinct or absent.b)Vessels diffuse; solitary and multiple of 2-5 cells, simple perforations plate, alternate intervessel vestured pits and vesselray pitting with distinct borders similar to intervessel pits.c)Fibers thick walled with simple to minutely bordered pits.d)Axial parenchyma apotracheal diffuse, paratracheal aliform and aliform confluent.e)Rays predominantly three-seriate, body ray cells procumbent, mostly with one row of square/upright marginal cells.f)Prismatic crystals in radial and axial parenchyma cells.
The measured anatomical parameters (quantitative data) are on Table 1: Vessel diameter (µm), Vessel frequency (mm 2 ), Ray width (µm), Ray width in number of cells, Ray height in micrometers (µm), Ray height in number of cells, and Ray frequency (linear millimeter).Most of the parameters showed significant difference at 5% of probability.

DISCUSSION
Wood anatomy reveals, most of the time, anatomical features associated with specific groups of plants, since several taxa may have combinations of typical microscopic anatomical features (BAAS et al., 2000).
D. brasiliensis (Winteraceae), characterized by uniseriate and multiseriate rays, where the multiseriate rays are larger in height and width than the uniseriate ones (Figure 1 A-B), is a vesselless wood.The presence of tracheids and absence of vessel elements easily separated D. brasiliensis from the others species.
C. dinisii, with vessels predominantly solitary (Figure 1A) and axial parenchyma displayed similarly to R. sellowii and H. serratifolius.However, it is possible to separate C. dinisii from the others by the presence of the scalariform perforation plate (Figure 1G), intervessel pits opposite and the oil cells in the axial parenchyma (Figure 1 D-F).Scalariform perforation plates were also described by Majcher (2013) for C. dinisii.According to Wilson (1965) and Carlquist (1988), C. dinisii belongs to a basal family, Canellaceae, which, considering the trends of secondary xylem, presents primitive vessels elements with scalariform perforation plate.Quantitative data, such as vessel diameter, width and frequency of rays, were also helpful to separate these species, since C. dinisii has smaller vessels, narrower and more frequent rays than the others (Table 1 and 2).O. platyspermum is characterized by paratracheal vasicentric axial parenchyma in narrow bands or lines (Figure 2A) which marks the growth ring boundaries as discussed by Lisboa et al. (1984Lisboa et al. ( , 1987)).These features, besides the ray width (Table 1) can separate it from S.  cedron, which shows a banded axial parenchyma.Only simple perforations were observed for this species.
The two species, O. platyspermum and S. cedron, are separated from the others for not having diffuse apotracheal parenchymal.Scalariform perforation plates and laticifers or tanniniferous tubes, although described for O. platyspermum by Insidewood (2004), were not observed in the samples.R. sellowi has indistinct or absent growth ring boundaries, vessels with diffuse distribution, solitary and multiple vessels, simple perforation plate, paratracheal parenchyma, rays with 4 or more cells wide, body ray with procumbent cells and 1-4 marginal rows of square/ upright cells (Figure 2 C-G), as described by Alves andAngyalossy-Alfonso (2000, 2002).R. sellowii and L. dasycarpum can be separated considering the number of square/upright marginal cells, since R. sellowii (Figure 2 D-G), has a larger number of cells, also, the uniseriate rays can be as wide as the multiseriate rays, differently from L. dasycarpum (Figure 3G).For S. cedron, on the other hand, exclusively procumbent cells on the rays was observed, and uniseriate and multiseriates rays (Figure 3B) have distinct size as well as in D. brasiliensis (Figure 1 A-B).
For H. serratifolius, it was observed that this species is easily separated from the others species by the presence of the unilateral paratracheal parenchyma (Figure 3D) and the stratification of the cell elements (Figure 3E).Most parameters analyzed match the descriptions of Alves andAngyalossy-Alfonso (2000, 2002), Trevizor (2011) and Albuquerque (2012), as the presence of growth ring boundaries, vessels with diffuse distribution, solitary and multiple vessels, simple perforation, axial parenchyma apotracheal and paratracheal with short to long confluences, usually forming bands.Although Albuquerque (2012) describes the species as with scarce paratracheal aliforme, homocelular rays with 2-3 procumbent cells wide and presence of storied elements.Reticulate, foraminate, and/or other types of multiple perforation plates, as described by Insidewood (2004), were not observed.
L. dasycarpum is easily separated from the others by the presence of vestured pits.The presence of prismatic crystals is also an important feature.Crystals were observed in the ray cells of R. sellowii and C. dinisii while for L. daysidarpum they were present also in axial parenchyma series (Figure 3C).Insidewood (2004) described prismatic crystals in ray cells of Cinnamodendron axillare (Canelaceae).
Although the qualitative anatomical analysis show several features previously highlighted, by Carlquist (1988) for D. brasiliensis, by Majcher (2013) for C. dinisii, by Lisboa et al. (1984Lisboa et al. ( , 1987) ) for O. platyspermum, by Detienne and Jacquet. (1983), Manieri (1958), Alves andAlgyalossy-Alfonso (2000, 2002) and Miller (2007) for R. sellowii, and, by Alves and Algyalossy-Alfonso (2000, 2002,) Léon (2009), Trevizor (2011) and Albuquerque (2012) for H. serratifolius, our results review some of the features, add description of species not yet investigated and give more emphasis to the quantitative features previously poorly explored.Thus, the results show that D. brasiliensis is separated from the others by the presence of tracheids.C. dinisii has scalariform perforations and oil cells associated with the axial parenchyma.The axial parenchyma paratracheal vasicentric and in marginal bands in O. platyspermum, beyond the rays width, can separate it from S. Cedron.H. serratofolius has the unique presence of the axial unilateral paratracheal parenchyma and storied cell elements.R. sellowii and L. dasycarpum can be separated from each other by the number of square/upright marginal ray cells, greater in R. sellowii.

CONCLUSIONS
When the seven species were compared, the uniqueness of some features is clear and shows how wood anatomy is a valuable tool to separate these species of distinct families.Drimys brasiliensis is separated from the others by the presence of tracheids.Capsicodendron dinisii has scalariform perforations plates and oil cells associated with the axial parenchyma.Axial parenchyma paratracheal vasicentric and in marginal bands beyond the rays' width, can separate Osteophoeum platyspermum fromSimaba cedron.Handroanthus serratifolius has the unique presence of the axial unilateral paratracheal parenchyma and storied cell elements (parenchyma, fibers and vessel elements).Rauvolfia sellowii and Leptolobium dasycarpum can be separated by the number of square/ upright marginal ray cells, greater in Rauvolfia sellowii.

TABLE 1
Anatomical quantitative parameters of the wood species.Vessel diameter (µm), VF -Vessel frequency (mm2), RW -Ray width (µm), RWC -Ray width in number of cells, RH -Ray height in micrometers (µm), RHC -Ray height in number of cells, and FR -Ray frequency (linear millimeter).The results are shown as: mean ± standard deviation, maximum value -minimum value.The Tukey Test at a 5% probability level was applied.The averages followed by the same letter do not differ statistically between themselves.

TABLE 2
Anatomical qualitative parameters for the separation of the species.