Identification of endangered or threatened Costa Rican tree species by wood anatomy and fluorescence activity

Róger Moya1*, Michael C. Wiemann2 & Carlos Olivares3 1. Instituto Tecnológico de Costa Rica, Escuela de Ingeniería Forestal, Apartado 159-7050, Cartago, Costa Rica; [email protected] 2. Forest Products Laboratory, USDA Forest Service, One Gifford Pinchot Drive Madison, Wisconsin 53726-2398, USA; [email protected] 3. Instituto Tecnológico de Costa Rica, Escuela de Ingeniería Forestal, Apartado 159-7050, Cartago, Costa Rica; [email protected] * Corresponding author

Received 10-I-2012. Corrected 20-X-2012. Accepted 12-XI-2012.

Abstract: Identificación de especies de árboles en peligro o amenazadas de Costa Rica basada en la anatomía de la madera y fluorescencia. A total of 45 native Costa Rican tree species are threatened or in danger of extinction, but the Convention on International Trade Endangered Species (CITES) includes only eight of these in its Appendices. However, the identification of other species based on their wood anatomy is limited. The present study objective was to describe and to compare wood anatomy and fluorescence activity in some endangered or threatened species of Costa Rica. A total of 45 (22 endangered and 23 threatened with extinction) wood samples of these species, from the xylaria of the Instituto Tecnológico de Costa Rica and the Forest Products Laboratory in Madison, Wisconsin, were examined. Surface fluorescence was positive in eight species, water extract fluorescence was positive in six species and ethanol extract fluorescence was positive in 24 species. Almost all species were diffuse porous except for occasional (Cedrela odorata, C. fissilis, Cordia gerascanthus) or regular (C. salvadorensis and C. tonduzii) semi-ring porosity. A dendritic vessel arrange- ment was found in Sideroxylon capari, and pores were solitary in Guaiacum sanctum and Vantanea barbourii. Vessel element length was shortest in Guaiacum sanctum and longest in Humiriastrum guianensis, Minquartia guianensis and Vantanea barbourii. Finally, anatomical information and fluorescence activity were utilized to construct an identification key of species, in which fluorescence is a feature used in identification. Rev. Biol. Trop. 61 (3): 1133-1156. Epub 2013 September 01.

Key words: fluorescence, Costa Rican woods, tropical woods, wood identification key.

For its size (51 000km2) Costa Rica is one government decrees to stop the cutting of 18 of the most biodiverse countries in the world. commercial species (Costa Rica 1997) protect- Of its 10 000 taxa, 1 300 are endemic and ed by international conventions such as CITES, more than 25% of these are considered rare and actions limiting harvesting in certain areas (Burger 1980). Nonetheless, intensive defor- of the country (Costa Rica 1996a, 1996b). A estation began at the start of the twentieth cen- number of important timber-producing spe- tury, and reached levels in excess of 50 000ha cies have been identified by some experts as per year in the 1970s (Alvarez 1986). Natural threatened species due to their rarity (Jimenez forest decreased to only 24.4% by 1987 (Sán- 1999). Identification of these timber-producing chez et al. 2001). species using floral and tree characteristics Tree timber species receive the highest has been widely described (Jimenez 1999). In degree of protection, and two actions are used contrast, identification of these species based to protect them: the promulgation of laws and on their wood anatomy is limited to only a few

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1133 species: examples are the species included in from the Forest Products Laboratory (FPL), CITES Appendices I and II. The CITES Iden- Wisconsin-USA collection (MADw and SJRw) tification Guide-Tropical Wood (CITES 2002) and the Instituto Tecnológico de Costa Rica has a general identification key with illustra- (ITCR) collection (TECw). Their sample num- tions of the transverse sections of each species. bers are detailed in tables 1 and 2. In some The book is intended for non-experts (Gas- cases only one sample and slide was available, son et al. 2010) and its use is limited. More so it was necessary to prepare additional per- recently, the International Association of Wood manent slides of these species. To do this, a Anatomists (IAWA) published several wood block (1cm3) was obtained from a wood sample descriptions of tree timbers included in CITES and was softened in hot water. Tangential, radi- (Gasson 2011, Gasson et al. 2011). al and transverse sections were cut (12-15μm Recently, wood anatomical identification thick). These sections were stained with safra- has been accompanied by other analyses that nin and dehydrated with a series of alcohol (5 together with special methods, can achieve minutes each in 50, 70 and 95%); finally, sec- precise identification that are useful in identi- tions were rinsed and mounted on microscope fication of endangered or threatened species. slides. Furthermore, a small piece was cut from Some of these techniques are relatively easy each wood block to prepare macerated wood to implement, for example, wood density, using Franklin’s method (Ruzin 1999). surface fluorescence, or fluorescence of water or ethanol extracts. (Miller & Wiemann 2006, Wood anatomical description: The Guzman et al. 2008, Wiemann & Ruffinatto IAWA list (IAWA 1989) was used as the basis 2012). Studies of anatomical features accom- for choosing identification characteristics, with panied with complex statistical procedures, some modifications to allow for increased such as the use of multivariate analysis, helps accuracy and subsequent species level separa- to identify CITES species (Gasson et al. 2010, tion. The quantitative anatomical features that MacLanchlan & Gasson 2010). were measured were: length and diameter of The objective of the present study was to the fibers, lumen diameter, cell wall thickness, describe and compare wood anatomy, fluores- vessel length, diameter and frequency of pores, cence, and density of endangered or threatened solitary pore frequency, diameter of interves- species of Costa Rica, and to use these charac- sel pits, and height and width of rays. Fiber teristics to build an identification key. The key dimensions and vessel lengths were measured will allow monitoring of the use and commer- on macerated wood. Permanent slides were cialization of these tree species (endangered or used for measurement of the other anatomi- threatened) in Costa Rica, and perhaps will be cal characters. Qualitative anatomical features also useful to other countries in the region. were also determined using the IAWA List as a guide (IAWA 1989). MATERIAL AND METHODS Additional information: Other important Wood samples and permanent slides: A information about endangered or threatened total 45 timber species growing in Costa Rica species used was: endemic category if the spe- were analyzed from March to December of cies grows only in Costa Rica, wood density, 2010, 22 of which are considered as endan- traditional uses of species, and fluorescence. gered species by the Costa Rican Government Wood density (weight/volume) was measured (Costa Rica 1996a and 1996b, Costa Rica in the air-dry condition. The four traditional 1997) (Table 1). Another 23 timber species uses considered were light or heavy construc- studied are considered to be in threat of extinc- tion, flooring, furniture, and handicrafts. Heart- tion in this country (Jimenez 1999) (Table 2). wood fluorescence was observed directly the Wood samples of all the species were obtained surface, in water extract, and in ethanol extract,

1134 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 6 1 1 1 1 7 2 6 5 3 1 7 2 1 1 1 2 1 3 7 9 12 Permanent slides observed 6 1 1 1 2 7 2 6 7 7 1 1 7 2 1 1 5 2 7 6 7 1| observed Wood samples Wood ) 3 0.72 0.74 0.88 0.78 0.79 0.54 0.56 0.75 0.60 0.59 0.85 0.70 0.91 0.82 0.78 0.79 0.80 0.78 0.57 0.51 0.55 1.06 (g/cm Wood density* Wood Fu Fu Fu LC LC HC HC HC HC Fu & H Fu & H Fu & H Fu & H HC & Fl HC & Fl HC & Fl Fu & LC Traditional Traditional Fu, Fl & H Fu, Fl & H Fu, Fl & H Fu, Fl & H HC, Fl & H used of wood No No No No No No No No No No No No No No Yes Yes Yes Yes Yes Yes Yes Yes Endemic TABLE 1 TABLE Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Gymnosperm Gymnosperm Gymnosperm Wood species Wood (Jacq.) Dugand Endangered timber species in Costa Rica (decreed by government) species in Costa Rica (decreed timber Endangered Species King Cordia gerascanthus L. Cordia Zamora & Poveda costaricense Zamora Sclerolobium chocoensis Prance & Poveda N. Zamora Caryodaphnopsis burgeri Couratari scottmorii Prance fissilis Vell. Cedrela Standl. salvadorensis Cedrela Swietenia humilis Zucc . Swietenia pendula Benth. Swietenia macrophylla Swietenia (Pittier) Record & Mell Dipteryx panamensis (Pittier) Hymenolobium mesoamericanum H.C. Lima Myroxylon balsamum (L.) Harms Myroxylon Paramachaerium gruberi Briz. & Kiitgaard curuense N. Zamora Platymiscium Podocarpus costaricensis de Laub. Platymiscium parviflorum Benth. Platymiscium polystachyum pinnatum var. Platymiscium pinnatum (Jacq.) Dugand Platymiscium Podocarpus guatemalensis Standl. Parl. Podocarpus macrostachys Guaiacum sanctum L. Botanical Family Botanical Boraginaceae Caesalpiniaceae Caryocaraceae Lauraceae Lecythidaceae Meliaceae Mimosaceae Papilionaceae Podocarpaceae Zygophyllaceae Source: Law decree No. 23700-MINAE, 25167-MINAE and 25663-MINAE. Source: Law decree content. at 12% of moisture density was determined Wood Legend * Fl=flooring, Fu=furniture and H=handicrafts. HC=Heavy construction, construction, LC=Light

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1135 8 1 6 3 1 6 2 9 1 1 1 8 1 2 2 9 3 1 2 1 2 5 9 Permanent slides observed 2 1 7 1 3 1 9 7 5 5 6 3 2 6 1 6 7 1 3 1 2 7 8 observed Wood samples Wood ) 3 0.37 0.79 1.01 0.59 0.63 0.89 0.44 0.44 0.78 0.69 0.91 0.53 0.66 0.72 0.64 0.48 0.98 0.75 0.68 0.78 0.78 1.01 0.95 (g/cm Wood Density* Wood Fu LC LC LC HC HC HC HC FU & H FU & H of wood LC & Fl LC & Fl LC & Fl LC & Fl HC & Fl HC & Fl HC & Fl HC & Fl LC, Fl & H HC, Fl & H HC, Fl & H HC, Fl & H HC, Fl & Ha Traditional used Traditional No No No No No No No No No No No No No No No No No No No Yes Yes Yes Yes Endemic TABLE 2 TABLE Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Angiosperm Gymnosperm Wood species Wood Timber species considered to be in threat of extinction in Costa Rica of extinction species considered to be in threat Timber (Donn. Sm.) Britton & Rose (Donn. Sm.) Britton (Donn. Sm.) Harms Species Donn. Sm. (Wild.) Ladd. (Wild.) (Triana) Ducke (Triana) Qualea paraensis Ducke Dussia macroprophyllata Dussia macroprophyllata Prumnopitys standleyi capari (A. DC.) Pittier Sideroxylon Dalbergia retusa Hemsl. retusa Dalbergia Minquartia guianensis Aubl. Cedrela tonduzii C.DC. Cedrela Cedrela odorata L. Cedrela Lecythis ampla Miers. Lecythis Oerst. pterocarpa Oreomunnea Couratari guianensis Aubl. Vantanea barbourii Standl. Vantanea Humiriastrum guianensis Cuatrec. costaricense Standl. & L.O. Williams versicolor Standl. & L.O. Tachigali Prioria copaifera Griseb. Pittier purpurea Poveda, N. Zamora & P.E. Sánchez & P.E. Copaifera camibar Poveda, N. Zamora Cynometra hemitomophylla Mora oleifera Dwyer Copaifera aromatica Tabebuia guayacan (Seem.) Hemsl. Tabebuia Astronium graveolens Jacq. Astronium Family Botanical Source: Jimenez 1999. Source: Jimenez content. at 12% of moisture density was determined Wood * Legend: Fl=flooring, Fu=furniture and H=handicrafts. HC=Heavy construction, construction, LC=Light Vochysiaceae Podocarpaceae Sapotaceae Papilionaceae Olacaceae Meliaceae Juglandaceae Lecythidaceae Humiriaceae Caryocaraceae Caesalpiniaceae Bignoniaceae Anacardiaceae

1136 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 as described in the IAWA List (IAWA 1989). and Tachigali versicolor it was yellow, yellow- Surface fluorescence was determined in a dark- ish or weak yellow (Table 3). room from freshly prepared (planed or scraped) Water extract fluoresced in only nine spe- transverse and/or longitudinal surfaces exposed cies (Table 3). The fluorescence was yellow in to a low intensity, long wave ultraviolet light Copaifera species, H. mesoamericanum and P. (around 365nm). The specimens were recorded purpurea and green in Cordia gerascanthus, as either fluorescente (noting color and intensi- A. graveolens, M. balsamum and Platymis- ty) or not fluorescente. Only specimens which cium species (Table 3). C. gerascanthus and exhibited a definite yellow, green, orange or Platymiscium species did not show surface blue fluorescence were recorded as fluores- fluorescence, but they did show water extract cente. For water and ethanol extracts, color and fluorescence (Table 3). intensity of fluorescence were scored. Froth Ethanol extract fluorescence was found in tests were conducted and were scored as posi- 31 of the species. The fluorescence was green- tive (high intensity) if one minute after shaking ish blue or yellow, weak or light green, purple, vigorously, froth was present and covered the yellowish or bluish (Table 3). Several species entire surface of the solution, negative if all (Cedrela odorata, Cedrela fissilis, Cedrela ton- froth had disappeared, and variable if froth was duzii, Couratari guianensis, Couratari scott- still present around the edge of the test tube but morii, Dalbergia retusa, Dipteryx panamensis, did not extend over the entire surface of the Guaiacum sanctum, Oreomunnea pterocarpa, liquid column. , Prioria copaifera, Qualea paraensis, Swietenia humilis, Swietenia mac- RESULTS rophylla, Tabebuia guayacan and Vantanea barborii) had positive ethanol extract fluores- General aspects: 22% (ten species) were cence, but negative surface or water extract Papilionaceae, 18% (eight species) were Cae- fluorescence (Table 3). salpiniaceae, 13% (six species) were Meliace- ae, 9% (four species) were Podocarpaceae, 7% General features of endangered or (three species) were Lecythidaceae, 4% (two threatened species: Porosity: Diffuse porosity species) each were Caryocaraceae or Humiria- was common in endangered or threatened spe- ceae; other families represented the 27% (12 cies, being found in 39 species. Three species species) of endangered or threatened timber were both diffuse and semi-ring porous (C. species endemic to Costa Rica. Most of these species are used in heavy or light construction odorata, C. fissilis and C. gerascanthus) and or handicrafts; however, species in the Melia- two species were semi-ring porous (Cedrela ceae are utilized for furniture manufacturing. salvadorensis and C. tonduzii). Sideroxylon High wood density was a characteristic of most capari was unique with a radial or diagonal of the species, and the main use of the lumber pore pattern (Fig. 1a). Almost all of the spe- was construction (Table 1 and 2). cies had solitary pores and pore multiples. However, the percentage of solitary pores Fluorescence test: Surface fluorescence was higher than 85% in G. sanctum and V. was positive in 11 species. This represented barbourii, so these species can be classified as 24% of the total number of species. Fluores- pores exclusively solitary. Higher frequency of cence in , Copaifera multiple pores (>15 pores/mm2) was measured aromatica, Astronium graveolens, Mora ole- in three species: C. guianensis, P. purpurea ifera and Myroxylon balsamum was green, and T. guayacan. Pores frequency was the greenish, or weak green, whereas in Dussia highest in Guaiacum sanctum, and M. bal- macroprophyllata, Lecythis ampla, Hymenolo- samo. Lower pore frequencies (<2 pores/mm2) bium mesoamericanum, Peltogyne purpurea were observed in C. tonduzii, D. retusa, D.

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1137 30 30 30 32 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30-32 30-32 30-35 VRP* 30-31-32 30-31-32-33 ------+ + + + + + + + + + + Vestured 6.0 9.4 5.4 1.6 3.8 2.0 9.0 2.6 2.8 6.9 4.2 7.1 5.4 5.8 5.6 3.0 2.5 2.7 6.2 1.7 8.7 4.0 4.2 DIP 11.4 12.3 12.0 10.4 T T T T T T T A G G G G G G G G G G G G G G G G G-T G-T G-T Deposits S S S S S S S S S S S S S S S S S S S S S S S S S Sc Sc PP 66 89 58 230 DV 106 318 116 102 114 118 204 101 243 261 206 201 140 164 144 210 150 140 105 188 129 174 152 Vessels elements characteristics elements Vessels 93 LV 344 263 408 441 287 930 311 352 463 450 225 153 352 366 244 202 374 332 267 369 240 462 428 357 267 1018 PF 3.0 5.9 3.8 3.6 1.8 1.3 1.3 9.9 1.8 2.0 3.7 2.2 5.8 3.9 1.8 3.3 2.1 3.7 5.1 4.7 3.6 4.3 19.4 12.7 10.8 23.2 16.3 92 85 56 44 79 43 62 96 81 99 70 65 93 69 61 27 75 86 72 66 75 77 79 34 87 66 56 PSP TABLE 3 TABLE S S D D D D D D D D D D D D D D D D D D D D D D D-S D-S D-S arrangement Porosity and ------EEF green purple yellow variable greenish greenish yellowish yellowish yellowish light green light light green light weak green weak green weak green weak green weak green weak yellow greenish blue greenish blue greenish blue ------WEF weak green weak green weak yellow weak yellow weak yellow Heartwood fluorescence ------SF greenish greenish greenish greenish yellowish weak green weak yellow weak yellow greenish blue Results of fluorescence test and vessels characteristics of endangered and threat of extinction species in Costa Rica of extinction and threat of endangered test and vessels characteristics Results of fluorescence

Species Parkia pendula Paramachaerium gruberi Myroxylon balsamum Myroxylon pterocarpa Oreomunnea Mora oleifera Minquartia guianensis Lecythis ampla Lecythis Humiriastrum guianensis H. mesoamericanum Guaiacum sanctum Dussia macroprophyllata Dipteryx panamensis Dalbergia retusa Dalbergia Cynometra hemitomophylla Couratari scottmorii Couratari guianensis Cordia gerascanthus Cordia Copaifera camibar Copaifera aromatica Cedrela tonduzii Cedrela Cedrela salvadorensis Cedrela Cedrela fissilis Cedrela Cedrela odorata Cedrela Astronium graveolens Astronium Caryocar costaricense Caryodaphnopsis burgeri Anthodiscus chocoensis

1138 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 - - - - 30 30 30 30 30 30 30 30 30 30 30 30 30 VRP* 30-32-35 ------+ + + + + + + + + Vestured - - - - 1.3 3.6 2.8 1.6 2.5 5.5 6.1 6.1 6.6 2.3 6.4 9.1 7.7 DIP 11.0 - - - - T T A A G G G G G G G G G G Deposits - - - - S S S S S S S S S S S S S Sc PP ), LV=Length of vessels (µm), DV=Diameter ), LV=Length 2 - - - - 51 86 211 DV 118 172 137 189 170 166 177 167 172 168 149 Vessels elements characteristics elements Vessels - - - - LV 315 313 150 349 240 371 240 306 226 249 172 285 308 1160 - - - - PF 4.5 3.9 6.8 5.2 8.4 3.8 2.4 2.8 2.9 3.1 1.6 7.2 13.3 10.6 ) - - - - 95 69 31 70 64 39 62 41 77 75 59 79 81 29 PSP - - - - D D D D D D D D D D D D D D-DE-R arrangement Porosity and TABLE 3 ( Continued TABLE - - - - - EEF bluish yellow greenish greenish greenish greenish yellowish light green light light green light light green light weak green weak green weak green ------WEF light green light light green light light green light light green light Heartwood fluorescence ------SF yellow yellowish Results of fluorescence test and vessels characteristics of endangered and threat of extinction species in Costa Rica of extinction and threat of endangered test and vessels characteristics Results of fluorescence polystachyum Species Legend: SF=surface fluorescence, WEF=Water extract fluorescence, EEF: Ethanol extract fluorescence. D=diffuse-porous, S=semi-ring porous, DE=Vessels in dendritic S=semi-ring porous, DE=Vessels D=diffuse-porous, extract fluorescence, EEF: Ethanol fluorescence. WEF=Water Legend: SF=surface fluorescence, in radial or diagonal pattern, PSP=Percentage of solitary pores, PF=Pores frequency (pores/mm pattern, R=Vessels Vantanea barbourii Vantanea Tachigali versicolor Tachigali Tabebuia guayacan Tabebuia Swietenia macrophylla Swietenia Swietenia humilis Swietenia Sideroxylon capari Sideroxylon Podocarpus costaricensis Sclerolobium costaricense Sclerolobium var. pinnatum var. P. Podocarpus guatemalensis Prumnopitys standleyi Podocarpus macrostachys Qualea paraensis Platymiscium pinnatum Platymiscium Prioria copaifera Platymiscium parviflorum . Platymiscium (µm), PP=Perforation plates, S=simple perforations plate, Sc=scalariform perforations plate, Deposits: T=tyloses, G=gums, A=absent, DIP=diameter of intervascular pits (µm), of intervascular A=absent, DIP=diameter T=tyloses, G=gums, Deposits: perforations plate, Sc=scalariform plate, S=simple perforations (µm), PP=Perforation plates, pits Vessel-ray 31: borders; similar to intervessel pits in size and shape throughout the ray cell, pits with distinct Vessel-ray class: 30: according to IAWA pitting VRP=Vessels-ray gash-like) (scalariform, pits horizontal simple: pits with much reduced borders to apparently Vessel-ray 32: pits rounded or angular, simple: with much reduced borders to apparently absent, “+” feature 1989). “-“ anatomical rows (IAWA, to marginal pits restricted Vessel-ray pits of two distinct sizes or types in the same ray cell, 35: Vessel-ray 33: to vertical, present, “?” unknown feature. feature anatomical Platymiscium curuense Platymiscium Peltogyne

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1139 Fig. 1. (a) Cross-section of Sideroxylon capari showing vessels in radial pattern, (b) tangential sections of Sclerolobium costaricense showing uniseriate rays, (c) Vessel-rays pitting of C. burgeri, (d) tracheids and cross-field pitting in radial sections of Prumnopitys standleyi and (e) tracheids and cross-field pitting in radial sections of Podocarpus guatemalensis. Bar: 500µm in (a) and (b) and 200µm in (c), (d) and (e). macrophylla, H. mesoamericanum, L. ampla Tyloses, deposits or gum were found in the and Platymiscium curuense. vessels of almost all of the angiosperm species Vessel lengths for these species varied (Table 3). Gum was the most common sub- from 93 to 1 160mm (Table 3). Vessel length stance in vessel lumina. Nevertheless, neither was shortest in G. sanctum and longest in H. substance was observed in three of the angio- guianensis, M. guianensis and V. barbourii. sperm species: D. macroprophyllata, Sclerolo- Small diameter vessels (from 50-100mm) were bium costaricense and V. barbourii. found G. sanctum, M. guianensis, M. balsamo, All angiosperm species had alternate S. capari and T. versicolor. Large diameter ves- polygonal pits. Their diameter was minute sels were found only in O. pterocarpa. Vessel (<4µm) in 14 species and large (>=10µm) in diameters from 100-300mm were found in the five species. Pits in the other species were from other species. 4 to 10µm in diameter (Table 3). Simple perforation plates were found in Vestured pits were found in all species almost all species (Table 3). However, H. guia- of (Caesalpixiaceae, Mimosaceae, nensis (with 10-20 bars), M. guianensis (with Papilionaceae) and in V. barbourii (Table 3). <=10 bars) and V. barbourii (with <=20 bars) Vessel-ray pits with distinct borders, similar to had scalariform perforations. intervessel pits in size and shape throughout

1140 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 the ray cell, were common. They were found barbourii. Distinctly bordered pits were com- in almost all species. Other pits shapes were mon in both radial and tangential fiber walls. found in C. guianensis (Table 3). Vessel-ray The tracheids of species of the Podocarpaceae pits were restricted to marginal rows in C. cos- had, as expected, bordered pits in radial walls. taricense. Vessel-ray pits with much reduced Many of the angiosperm species had storied borders to apparently simple, with pits rounded fibers (Table 4). Crystals were only found in or angular and horizontal (scalariform, gash- the fibers of P. gruberi. like) to vertical (palisade) were found in A. graveolens and Caryodaphnopsis. burgeri (Fig. Ray parenchyma: Ray height over 1mm 1c), C. guianensis, and O. pterocarpa. Vessel- was observed in five species (C. costaricense, ray pits were restricted to marginal rows in V. C. gerascanthus, M. guianensis, P. purpurea barbourii. L. ampla had vessel-ray pits with and Q. paraensis), representing 11% of the spe- much reduced borders to apparently simple: the cies. Rays were exclusively uniseriate in eight pits were horizontal (scalariform, gash-like) to species (Fig. 1b): four Podocarpaceae (Fig. 1d vertical (palisade). and 1e) and four angiosperms (G. sanctum and Platymiscium species). Rays 1-3 cell in width Fibers: as expected, gymnosperm spe- were the most common, represented by 23 cies tracheids were longer than most of the species (51% of total species) (Table 4). Large angiosperm fibers, being almost 2mm length. rays (over 8 cells in width) were observed in However, the fiber length of V. barbourii was eight species (18% of total species) (Table 4). the longest of any species at 2.45mm. Fiber The rays of C. aromatica, C. gerascanthus and length varied from 1.50 to 1.95mm in C. P. purpurea were 4-10 seriate. Ray frequency costaricense, C. burgeri, C. gerascanthus, D. varied from 2 to 20 rays per mm (Table 4). The macroprophyllata, H. mesoamericanum, H. lowest frequencies were found in C. burgeri and guianensis, M. guianensis and M. balsamum. L. ampla and the highest ones (>14 rays/mm) The shortest fiber lengths were found in G. were in Cynometra hemitomophylla, G. sanctum sanctum, although C. odorata, C. salvador- and H. guianensis. Rays composed mostly of ensis, D. retusa, O. pterocarpa and S. capari procumbent cells were observed in 17 species also had short fibers (Table 4). Fiber lumen (38% of total species) and heterogeneous rays diameter was widest in D. macroprophyllata, were in 19 species (42% of total species). Nine O. pterocarpa, H. guianensis, P. macrostachys species had both homogeneous and heteroge- and V. barbourii, and narrowest in T. guayacan. neous rays (Table 4). Storied rays were found Narrow fibers (<15µm) were also found in C. in 14 species (Table 4). Two heights were found fissilis, C. guianensis, G. sanctum, Paramach- in D. macrophylla, P. purpurea and Swietenia aerium gruberi and S. costaricense. The lumen species. Crystals were observed in ray cells of diameter varied from 15 to 55µm in other 27 species (60% of total species); in 11 species species (Table 4). Fibers with very thick-walls (>7µm) were found in D. retusa, H. guianensis, (24%) crystals were found in the marginal ray M. guianensis, M. balsamum, S. capari and cells only (Table 4). Silica bodies were present V. barbourii. Thin-walled fibers were found in Anthodiscus chocoensis, Couratari species, in species of Cedrela and in C. guianensis, L. ampla, Q. paraensis and T. versicolor. Sheath P. copaifera, S. costaricense and S. humilis. cells were only observed in C. gerascanthus. Cell wall thickness of other endangered and threatened species varied from 3 to 7µm (Table Axial parenchyma: Apotraqueal paren- 4). Septate fibers were observed only in A. chyma was present in 21 species (48% of total graveolens, G. sanctum and Swietenia spe- species) (Table 5). Parenchyma diffuse and cies (Table 4). In the angiosperms, fibers with diffuse-in-aggregates was seen in D. retusa, simple to minutely bordered pits were most G. sanctum, M. guianensis and S. capari, common in H. guianensis, L. ampla and V. and it was the only parenchyma type in C.

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1141 ------S(+) S(+) S(+) S(+) R (C) U+P(-) U+ P(-) U+ P(-) U+ P(-) U+ P(-) U+ P(-) U+ P(-) U+ P(-) U+ P(+) ------+ + + + + + + +- +- RS Ht Ht Ht Ht Ht Ht Ht Ht Ht Ht Ht Ht Ht Ht RT Ho Ho Ho Ho Ho Ho Ho Ho-Ht Ho-Ht Ho-Ht Ho-Ht Ho-Ht Ho-Ht Ho-Ht Ho-Ht RF 3.2 7.8 6.1 2.0 6.4 6.7 3.7 4.8 3.5 5.8 6.2 6.8 6.8 3.6 5.7 6.6 7.0 6.6 5.9 2.8 9.5 11.6 12.4 14.9 16.6 19.7 10.0 10.4 14.4 Ray parenchyma 1 1 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 1-3 RW 3-10 4-10 4-10 4-10 1-3, 4-10 1-3, 4-10 1-3, 4-10 1-3, 4-10 1-3, U (>) 1-3, U (>) 1-3, U (>) 67 >1 >1 >1 RH 850 580 292 360 390 590 250 190 224 380 306 342 335 440 134 194 330 202 323 400 480 535 170 360 290>1 ------+ + + + + + -+ -+ FS ------+ + +? PFRT TABLE 4 TABLE BD BD GTF SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD ------+ SF -+ ------+ FC Fibers or tracheids 5.3 3.6 6.6 3.4 3.1 2.7 6.5 3.3 4.6 5.1 4.2 4.6 2.5 2.1 5.3 2.5 7.7 4.3 6.3 6.4 6.2 5.5 4.0 7.5 7.0 6.9 3.6 6.3 3.9 WCT LD 11.6 24.1 22.1 21.3 19.1 17.0 20.1 17.4 24.4 19.2 22.6 16.7 23.7 17.8 15.0 15.3 13.1 19.2 19.4 16.3 32.7 14.4 25.8 15.5 18.2 17.3 29.3 22.8 14.2 FL 1.7 1.2 1.8 1.2 1.2 1.2 1.5 0.9 1.1 1.6 1.4 1.3 0.9 1.4 0.9 1.3 0.6 1.1 1.0 1.7 1.3 1.6 1.3 1.7 1.7 0.8 0.5 1.8 0.9 Fiber or tracheid and ray parenchyma characteristics of species that are endangered or considered to be in threat of extinction in Costa Rica of extinction to be in threat or considered are endangered of species that characteristics and ray parenchyma Fiber or tracheid Spcies Caryocar costaricense Astronium graveolens Astronium Caryodaphnopsis burgeri Copaifera camibar Copaifera aromatica Cedrela tonduzii Cedrela Anthodiscus chocoensis Cedrela odorata Cedrela Parkia pendula Cordia gerascanthus Cordia Couratari scottmorii Cynometra hemitomophylla Cedrela salvadorensis Cedrela Cedrela fissilis Cedrela Peltogyne purpurea Peltogyne Couratari guianensis retusa Dalbergia Platymiscium curuense Platymiscium Dipteryx panamensis Dussia macroprophyllata Mora oleifera Humiriastrum guianensis ampla Lecythis Minquartia guianensis balsamum Myroxylon pterocarpa Oreomunnea Guaiacum sanctum H. mesoamericanum Paramachaerium gruberi

1142 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 ------S(+) R (C) S(+)v U+ (-) U+ P(-) U+ P(-) ------+ + + + +- +- RS Ht Ht Ht Ht Ht RT Ho Ho Ho Ho Ho Ho Ho Ho Ho Ho Ho-Ht RF 8.9 6.0 9.6 4.2 5.6 7.7 5.9 4.7 4.1 7.0 8.1 9.5 8.2 11.0 11.1 12.1 Ray parenchyma 1 1 1 1 1 1 1 1 1-3 1-3 1-3 1-3 1-3 1-3 RW 1-3, 4-10 1-3, 4-10 71 55 90 RH 111 170 160 160 550 250 190 224 227 650 330 147 260, >1 ------+ + + + + -+ -+ -+ -+ FS ) ------+ PFRT T T T T BD BD GTF SBD SBD SBD SBD SBD SBD SBD SBD SBD SBD TABLE 4 ( Continued TABLE ------+ SF - + ------FC Fibers or tracheids 3.6 4.4 4.9 6.3 3.1 4.6 2.9 5.7 4.4 2.7 2.9 3.6 3.8 8.9 3.1 6.8 WCT 6.2 LD 21.5 18.4 19.9 31.5 35.4 26.5 21.8 31.4 18.3 14.6 18.8 21.0 17.0 17.7 33.1 FL 1.2 1.0 1.1 2.4 3.2 2.9 1.0 2.5 1.1 1.1 1.0 0.8 0.9 0.8 0.9 1.6 Fiber or tracheid and ray parenchyma characteristics of species that are endangered or considered to be in threat of extinction in Costa Rica of extinction to be in threat or considered are endangered of species that characteristics and ray parenchyma Fiber or tracheid polystachyum Spcies Platymiscium parviflorum Platymiscium Platymiscium pinnatum Platymiscium var. pinnatum var. P. Podocarpus costaricensis Podocarpus guatemalensis Podocarpus macrostachys Prioria copaifera Prumnopitys standleyi Qualea paraensis Sclerolobium costaricense Sclerolobium Swietenia humilis Swietenia macrophylla Swietenia guayacan Tabebuia Sideroxylon capari Sideroxylon Tachigali versicolor Tachigali barbourii Vantanea Fiber: FL=Fiber or tracheids length (mm), LD=lumen or tracheids diameter (µm), WCT=wall cell thick (µm), FC=Fibers crystals, SF=Septate fibers, GTF=ground tissue fiber, WCT=wall cell thick (µm), FC=Fibers crystals, SF=Septate fibers, GTF=ground tissue fiber, (µm), Fiber: FL=Fiber or tracheids length (mm), LD=lumen diameter Ray FS=Fibers stored. tangential, and radial both in common pits PFRT=Fiber T=tracheids, pit, bordered distinctly BD=Fibers with pits, bordered minutely to SBD=simple Ho=homogenous, RS=ray Ht=heterogeneous, type, RT=ray (ray/mm), RF=ray frequency uniseriate, (µm), U(>)=mainly width cell (µm), RW=ray RH=ray height parenchyma: present. feature absent, “+”anatomical feature “-“ anatomical bodies presence, S=Silica cells, P=procumbent U=crystals in upright cells, stored, R(C)=crystals presence in ray,

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1143 Radial canals Radial canals Radial Traumatic canals Traumatic Vasicentric tracheids Vasicentric Crystals in enlarged cells Crystals in enlarged Others anatomical features Others anatomical Axial & ray canals & sheath cells Axial & ray canals Axial & ray canals & sheath cells Axial & ray canals - - - + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (+) + (-+) + (-+) + (-+) + (-+) + (-+) + (-+) AP (C) AP + (+), S(+) 2 2 2 >8 3-8 3-8 3-8 3-8 3-8 3-8 3-8 3-8 3-8 3-8 3-8 5-8 5-8 3-5 3-5 3-8 5-8 3-4 Cell 2, 3-4 2 & 3-4 2 & 3-4 2 & 3-4 2 & 3-4 3-8 & >8 3-8 & >8 ------+ ves + ves + ves + ves + wae + wae + wae + wae Stored +- wae - - - - - Nar Nar Nar Nar Mar Mar Mar Mar Mar Mar Mar Mar Mar Mar Wid Wid Wid Nar-Mar Nar-Ret-Mar Nar-Ret-Mar Nar-Ret-Scal Wid-Ret-Mar Wid-Nar-Ret-Scal TABLE 5 TABLE Banded Parenchyma Axial parenchyma - - - - Vc Vc Sca Sca Sca Sca Con Sca-Vc Vc-Con Sca & Vc Sca & Vc Sca & Vc Sca-Vc-Loz Sca, Vc & Con Sca, Loz-alif & Con Loz-alif Loz-alif & Con Loz-alif Loz-alif, Con, Vc Vc, Loz-alif & Con Vc, Loz-alif Sca, Vc, Con & Uni Sca, Loz-alif & Con Uni Loz-alif Loz-alif, win-alif & Uni Loz-alif, Sca, Loz-alif & wing-alif Sca, Loz-alif Parenchyma arrangement Parenchyma Vc, Loz-alif, Win-alif & Uni Win-alif Vc, Loz-alif, Vc, Loz-alif, win-alif & Con Vc, Loz-alif, ------Diff Diff Diff Diff Diff Diff Diff Diff Diff Type Diff-Agg Diff-Agg Diff-Agg Diff-Agg ------+ + + + + + + + + + + + + AP Fiber, ray parenchyma and axial parenchyma characteristics of endangered and threatened species in Costa Rica and threatened of endangered characteristics parenchyma and axial ray parenchyma Fiber, Cedrela odorata Cedrela Astronium graveolens Astronium Caryodaphnopsis burgeri fissilis Cedrela Caryocar costaricense Anthodiscus chocoensis salvadorensis Cedrela Cordia gerascanthus Cordia Couratari guianensis Cedrela tonduzii Cedrela Copaifera aromatica Copaifera camibar Couratari scott-morii Cynometra hemitomophylla Dalbergia retusa Dalbergia Dipteryx panamensis Guaiacum sanctum Dussia macroprophyllata H. mesoamericanum Humiriastrum guianensis Myroxylon balsamum Myroxylon pterocarpa Oreomunnea Paramachaerium gruberi Parkia pendula Mora oleifera Lecythis ampla. Lecythis Minquartia guianensis Peltogyne purpurea Peltogyne curuense Platymiscium

1144 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 Traumatic canals Traumatic Axial canals diffuse Axial canals Others anatomical features Others anatomical Helical thickening in vessels thickening Helical Vessels of 2 distinct diameter of 2 distinct Vessels - - - - - + (+) + (+) + (+) + (+) + (+) + (+) + (+-) + (-+) + (-+) + (-+) + (-+) AP (C) AP 2 2 2 < 4 < 4 < 4 3-4 5-8 5-8 3-8 3-8 Cell 2 & 3-4 2 & 3-4 2 & 3-4 5-8 & 8> < 4 and 5-15 ------+ ves + ves + ves + ves + wae Stored ) ------Nar Nar Nar Mar Mar Mar Wid-Nar Banded Parenchyma TABLE 5 ( Continued TABLE Axial parenchyma - - - - - Vc Sca Sca Sca-Vc Sca-Uni Loz-alif & Con Loz-alif & Con Loz-alif & Con Loz-alif Vc, Loz-alif & Uni Vc, Loz-alif Vc, Loz-alif & Con Vc, Loz-alif Parenchyma arrangement Parenchyma Vc, Loz-alif, win-alif & Con Vc, Loz-alif, ------Sca Sca Diff Diff Diff Diff Diff Diff Type Diff-Agg ------+ + + + + + + + + AP Fiber, ray parenchyma and axial parenchyma characteristics of endangered and threatened species in Costa Rica and threatened of endangered characteristics parenchyma and axial ray parenchyma Fiber, Vantanea barbourii Vantanea Loz-w=lozenge-winged, Loz-alif=lozenge-aliform: Vc=vasicentric, Con=confluent, in aggregates, Sca=scanty, Diff-Agg=diffuse Diff=diffuse. AP=apotraqueal parenchyma, AP(C)=crystals present in axial parenchyma band, S=Silica bodies present, Scal=Scalarifom band, ret=reticulate, Wid=wide Nar=narrow band, Mar=marginal, uni=unilateral, storied. elements storied, ae=axial present), ves=vessel elements (chambered Sideroxylon capari Sideroxylon Swietenia humilis Swietenia macrophylla Swietenia guayacan Tabebuia Tachigali versicolor Tachigali Prioria copaifera Prumnopitys standleyi Qualea paraensis costaricense Sclerolobium Platymiscium parviflorum. Platymiscium pinnatum Platymiscium polystachyum pinnatum var. P. Podocarpus costaricensis Podocarpus guatemalensis Podocarpus macrostachys

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1145 costaricense. Almost all the species of angio- Meliaceae species: Two genera of Melia- sperms had paratracheal parenchyma (Table 5). ceae were analyzed: four species of Cedrela Scanty paratracheal parenchyma was present (C. odorata, C. salvadorensis, C. fissilis and in sixteen species, and vasicentric parenchyma C. tonduzii), and two species of Swietenia (S. was found in A. graveolens, C. odorata, C. macrophylla and S. humilis). The wood anato- fissilis, C. tonduzii, C. gerascanthus and T. my of these species, especially C. odorata, C. versicolor. Three or more different paratracheal fissilis and S. macrophylla, has been described types were observed in 17 species (38% of total by several authors (Panshin 1933, White & species). Aliform and confluent parenchyma Gasson 2008). Four species of Meliaceae (C. was very common in several species. Banded salvadorensis, C. fissilis, S. macrophylla and S. parenchyma was observed in 30 species (68 humilis) are considered as endangered and their of total species; 73% of angiosperm species). cutting has been prohibited in the natural forest Parenchyma marginal or in wide bands (more in Costa Rica (Costa Rica 1996a). Felling of than 3 cells wide) were the most common the other species, C. odorata and C. tonduzii, is banded parenchyma (Table 5). Axial paren- permitted. Therefore, it is important to have a chyma was reticulate in Couratari species, C. method to separate the species. The species of hemitomophylla, L. ampla and O. pterocarpa. Swietenia are easily separated from species of Axial parenchyma was storied in 13 species Cedrela by anatomical features such as storied (Table 5). Fusiform cells were found in Platy- rays (Fig. 2g, h, i) and diffuse porosity in Swi- miscium species, Paramachaerium grugeri, etenia but not in Cedrela (Fig. 2a-2i). M. balsamo, P. copaifera, T. guayacan and T. Platymiscium species: some differences versicolor. Crystals were present in chambered were found among species of Platymiscium axial parenchyma in almost all gymnosperm include presence of traumatic canals in Platy- species (Table 5). They were found enlarged miscium parviflorum (Fig. 3a) but not in the in P. gruberi. A. chocoensis, A. graveolens, other species (Fig. 3b). Platymiscium pinnatum C. gerascanthus and Podocarpus species, T. var. polystachyum had irregularly storied rays guayacan did not have any crystals. Silica bod- (Fig. 4c) whereas in the other species storing ies were present in Couratari species. was well-defined (Fig. 4a, 4b and 4d). The parenchyma was paratracheal in P. pinnatum Other anatomical features: some spe- var. polystachyum but not in other Platymis- cies had other distinctive anatomical features, cium species. It was scanty, unilateral para- which are detailed in table 5. They could tracheal, winged-aliform in P. pinnatum var be used to facilitate wood identification. For polystachyum (Fig. 3b), but lozenge-aliform example, radial or axial canals are found in A. in the other species. We also found marginal chocoensis, C. costaricense, C. aromatica and parenchyma in P. pinnatum var polystachyum P. copaifera. Traumatic canals were observed (Fig. 3b). in C. camibar, H. guianensis and Platymis- cium pinnatum. Pores with two distinct diam- Couratari species: The two Couratari eters are found in D. retusa and Q. paraensis. species can be distinguished by several dif- C. gerascanthus was a unique species with ferences. C. scottmorii has narrow reticulate sheath cell in the rays. Finally, helical thicken- parenchyma bands that are two cells wide (Fig. ings were observed in some vessels elements 5b), whereas reticulate bands in C. guianensis of S. costaricense. are up to four cells wide (Fig. 5a). Rays are 1-3 seriate in C. scottmorii (Fig. 5d), but up Specific species: Major differences to 5-seriate in C. guianensis (Fig. 5c). The among species groups are included in the next frequency of silica bodies is different among few paragraphs. species, with, the highest frequency observed

1146 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 Fig. 2. Transverse sections of (A) Swietenia macrophylla, (B) Swietenia humilis, (C) Cedrela odorata, (D) Cedrela fissilis, (E) Cedrela salvadorensis, (F) Cedrela tonduzii, and tangential sections of (G) Swietenia macrophylla, (H) Cedrela odorata and (I) Cedrela salvadorensis. Bar: 500µm in (A) - (F) and 200µm in (G) - (I).

in C. guianensis. The vessels-ray pits in C. Podocarpaceae species: We looked scottmorii have much reduced borders and the at two genera: three species of Podocarpus pits are rounded or angular, unlike those of C. (P. costaricensis, P. guatemalensis and P. mac- guianensis which are horizontal (scalariform, rostachys), and one species of Prumnopitys (P. gash-like) to vertical (palisade). standleyi). Axial parenchyma is present in P. macrostachys and P. costaricensis (Fig. 7a-7b) Copaifera species: The two Copaifera and is scanty in P. guatemalensis and Prum- species differed mainly by paratracheal paren- nopitys standleyi (Fig. 7c-7d). The highest chyma and ray dimensions. Paratracheal paren- proportion of axial parenchyma was observed chyma is more abundant in C. aromatica than in P. macrostachys (Fig. 7a), it was rare to in C. camibar. It is vasicentric 2-3 cells in moderately abundant in P. costaricensis (Fig. width in Copaifetra aromatica (Fig. 6a), but 7b) and scanty in P. guatemalensis (Fig. 7d). scanty paratracheal or vasicentric 1-2 cells in Another important difference between Podo- C. camibar (Fig. 6b). The rays are 1- 3 cells carpus and Prumnopitys is ray height. Rays wide and high in C. camibar (Fig. 6d), but were highest in P. macrostachys (5 to 10 cells) were commonly 4-10 seriate and low in C. aro- (Fig. 8a), but they were only 2-4 cells high in P. matica (Fig. 6c). There is no difference in the costaricensis (Fig. 8b). Ray frequency is high- resin canals of the species. est in P. guatemalensis (Fig. 7c). The rays of

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1147 Fig. 3. Transverse sections of (A) Platymiscium parviflorum showing traumatic canals, (B) Platymiscium pinnatum var polystachyum, (C) Platymiscium pinnatum and (D) Platymiscium curuense. Bar: 200 µm.

Fig. 4. Tangential sections of (A) Platymiscium parviflorum, (B) Platymiscium pinnatum, (C) Platymiscium pinnatum var polystachyum and (D) Platymiscium curuense. Bar: 200µm.

1148 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 Fig. 5. Cross and tangential sections of Couratari species. (A) cross-section of Couratari guianensis, (B) cross-section of Couratari scott-morii, (C) tangential section showing ray width in Couratari guianensis and (D) tangential section showing ray width in Couratari scott-morii. Bar: 200µm.

Fig. 6. Cross and tangential sections of Copaifera species. (A) cross-section of Copaifera aromatica, (B) cross-section of Copaifera camibar, (C) tangential section showing ray width in Copaifera aromatica and (D) tangential section showing ray width in Copaifera camibar. Bar: 500µm in (A) and (B), 200µm in (C) and (D).

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1149 Fig. 7. Transverse sections of (A) Podocarpus macrostachys, (B) Podocarpus costaricensis, (C) Podocarpus guatemalensis and (D) Prumnopitys standleyi. Bar: 200µm.

Fig. 8. Tangential sections of (A) Podocarpus macrostachys, (B) Podocarpus costaricensis, (C) Podocarpus guatemalensis and (D) Prumnopitys standleyi. Bar: 200µm.

1150 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 Key for identification

The following identification key was developed to distinguish among the 45 timber species considered to be endangered or threatened in Costa Rica. This key use microscopic wood features as well and fluorescence and wood density.

1. Gymnosperm ...... 3 2. Angiosperm ...... 6 3a. Parenchyma present and rays 2-4 cells in height ...... 4 3b. Parenchyma abundant sparse and rays 2-10 cells in height ...... 5 4a. Parenchyma abundant ...... Podocarpus macrostachys 4b. Parenchyma scanty ...... Podocarpus guatemalensis 5a. Parenchyma sparse ...... Podocarpus costaricensis 5b. Axial Parenchyma absent ...... Prumnopitys standleyi 6a. Negative fluorescence ...... 7 6b. Positive fluorescence ...... 14 7a. Pores solitarys and in groups ...... 8 7b. Pores solitary or pores in dendritic or diagonal and/or radial pattern ...... 13 8a. Scalariform perforations; ray height over 1 mm, axial parenchyma diffuse and diffuse-in-aggregates, paratracheal scanty; vasicentric tracheids ...... Minquartia guianensis 8b. Simple perforations ...... 9 9a. Semi-ring porous, parenchyma diffuse, paratracheal scanty, vasicentric and in marginal or in seemingly marginal bands ...... Cedrela salvadorensis 9b. Diffuse porous ...... 10 10a. Rays not storied ...... 11 10b. Ray storied, axial parenchyma apotraqueal diffuse, scanty , lozenge-aliform, winged-aliform, and banded in narrow bands or lines up to three cells wide ...... Paramachaerium gruberi 11a. Ray 1-3 cells in width ...... 12 11b. Axial parenchyma apotracheal diffuse, scanty, vasicentric, and in narrow bands or lines up to three cells wide, with 3-5 or 5-8 cells per parenchyma strand ...... Carydaphnopsis burger 12a. Paratraqueal parenchyma with over eight cells per parenchyma strand ...... Anthodiscus chocoensis 12b. Parenchyma in marginal or in seemingly marginal bands, narrow banded or reticulate; rays and axial elements irregu- larly storied, ray frequency high ...... Cynometra hemitomophylla 13a. Pores solitary, scalariform perforation plates, axial parenchyma vasicentric, lozange-aliform, winged-aliform and confluent and traumatic canals ...... Humiriastrum guianensis 13b. Vessels in dendritic or diagonal and/or radial pattern, rays heterogeneous, 1-3 seriate and parenchyma diffuse and diffuse-in-aggregates ...... Sideroxylon capari 14a. Fluorescence in surface only ...... 15 14b. Fluorescence in ethanol extract only ...... 16 14c. Fluorescence in surface and ethanol extract ...... 31 14d. Fluorescence in water and ethanol extract ...... 35 14e. Fluorescence in surface, water and ethanol extract ...... 37 15a. Parenchyma vasicentric, lozenge-aliform, winged-aliform, confluent and in marginal or in seemingly marginal bands, with 3-4 and 5-8 cells per parenchyma strand ...... Mora oleifera 15b. Ray height over 1 mm, parenchyma diffuse, paratracheal scanty and in marginal or in seemingly marginal bands and radial canals ...... Caryocar costaricense 16a. Pores solitarys ...... 17 16b. Pores solitarys and in groups ...... 18 17a. Ray frequency high, fibers septate and storied, apotraqueal parenchyma diffuse and diffuse-in-aggregates ...... Guaiacum sanctum 17b. Perforation plates scalariform, axial parenchyma diffuse, scanty paratracheal and unilateral . .Vantanea barbourii 18a. Semi-ring porous ...... 19 18b. Diffuse porous ...... 21 19a. Semi-ring porous, parenchyma vasicentric, rays heterogeneous ray and 1-3 seriates in width . . Cedrela odorata 19b. Semi-ring to diffuse porous ...... 20

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1151 20a. Parenchyma aliform, rays heterogeneous and 1-3 seriate ...... Cedrela fissilis 20b. Parenchyma diffuse, paratracheal scanty and vasicentric and in marginal or in seemingly marginal bands ...... Cedrela tonduzii 21a. Rays uniseriate, pits vestured, axial parenchyma vasicentric, vessels with helical thickenings ...... Sclerolobium costaricense 21b. Rays multiseriate ...... 22 22a. Rays 1-10 cells wide, irregularly storied or storied, parenchyma apotracheal diffuse, scanty paratracheal and in mar- ginal or in seemingly marginal bands ...... Swietenia macrophylla or Swietenia humilis 22b. Rays 1 to 3 seriate ...... 23 23a. Axial canals, parenchyma vasicentric, lozange-aliform; confluent, and wide band banded with 2 and 3-4 cells per parenchyma strand ...... Prioria copaifera 23b. Axial canals absent ...... 24 24a. Rays storied ...... 25 24b. Ray not storied ...... 27 25a. Ray homogeneous ...... 26 25b. Ray heterogeneous, 1-3 cells wide, parenchyma apotracheal diffuse and diffuse-in-aggregates, scanty, vasicentric, in narrow bands or lines up to three cells wide and in marginal or in seemingly marginal bands . . Dalbergia retusa 26a. Parenchyma lozenge-aliform, winged-aliform and unilateral parenchyma, and prismatic crystals in chambered cells ...... Dipteryx panamensis 26b. Parenchyma vasicentric, lozenge-aliform, unilateral, and in marginal or in seemingly marginal bands ...... Tabebuia guayacan 27a. Ray heterogeneous ...... 28 27b. Ray homogeneous, parenchyma lozenge-aliform and confluent with 3-5 cells per parenchyma strand ...... Parkia pendula 28a. Ray 1-3 seriate ...... 29 28b. Ray 1-10 seriate, fibers pits common on both radial and tangential walls, two distinct sizes or types in the same ray cell; parenchyma bands more than three cells wide, reticulate and in marginal or in seemingly marginal bands ...... Couratari guianensis 29a. Apotracheal parenchyma absent ...... 30 29b. Apotracheal parenchyma diffuse, vasicentric, lozenge-aliform winged-aliform and confluent, vessels with 2 distinct diameters, ray height over 1mm ...... Qualea paraensis 30a. Rays homogeneous and heterogeneous, parenchyma in bands 2 cells wide, reticulate and in marginal or in seemingly marginal bands, reticulate bands 2 cells in width ...... Couratari scottmorii 30b. Parenchyma in marginal or in seemingly marginal bands, narrow banded or reticulate with 5-8 cells per parenchyma strand ...... Oreomunnea pterocarpa 31 a. Ray storied ...... 32 31b Rays not storied ...... 34 32a. Rays 1 to 3 seriate ...... 33 32b. Larger rays commonly 4-10 seriate, ray height over 1mm, pits vestured; parenchyma lozenge-aliform, confluent, unilateral, in marginal or in seemingly marginal bands ...... Peltogyne purpurea 33a. Parenchyma confluent and banded, more than three cells wide ...... Dussia macroprophyllata 33b. Parenchyma scanty paratracheal, confluent vasicentric and unilateral ...... Myroxylon balsamum 34a. Pits vestured, axial paratracheal scanty and vasicentric with 2 and 3-5 cells per parenchyma strand ...... Tachigali versicolor 34b. Fibers with distinctly bordered pits, parenchyma diffuse and in bands more than three cells wide, reticulate and in marginal or in seemingly marginal bands ...... Lecythis ampla 35a. Rays exclusively uniseriate; pits vestured ...... 36 35b. Rays 4-10 seriate, semi-ring or diffuse porous, parenchyma apotracheal diffuse, rays heterogeneous with sheath cells, higher than 1mm ...... Cordia gerascanthus 36a. Rays irregularly storied, parenchyma scanty, unilateral paratracheal, winged-aliform, marginal banded ...... Platymiscium pinnatum var polystachyum 36b. Rays storied, parenchyma lozenge-aliform with long wings, and confluent ...... Platymiscium pinnatum 36c. Parenchyma lozenge-aliform and confluent ...... Platymiscium curuense 36d. Parenchyma lozenge-aliform and confluent, traumatic canals sometimes present . . . .Platymiscium parviflorum 37a. Axial canals present ...... 38 37b. Axial canals absent ...... 39

1152 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 38a. Parenchyma vasicentric, lozenge-aliform, confluent and in seemingly marginal bands . . . . Copaifera camibar 38b. Parenchyma vasicentric 1-2 cells in width, and in seemingly marginal bands ...... Copaifera aromatica 39a. Fibers septate; parenchyma diffuse, paratracheal scanty; vasicentric and in marginal or in seemingly marginal bands ...... Astronium graveolens 39b. Ray 1-10 cells in width, storied, axial parenchyma lozenge-aliform, confluent, vasicentric and in bands more than three cells wide ...... Hymenolobium mesoamericanum

Prumnopitys standleyi are similar in shape and timber species, although surface and water frequency to the rays of P. macrostachys. No extract fluorescence can separate only six and differences were seen among species in cross- eight timber species, respectively. Although, field pit apertures. ethanol extract fluorescence was present in many timber species, the color of fluores- cence was sometimes the same. However, DISCUSSION three species were atypical in ethanol extract Forty-five Costa Rican timber species fluorescence color, the purple color found in are considered endangered or threatened. The G. sanctum, yellow in P. purpurea and bluish Costa Rican government has decreed that in T. versicolor, make them easy to identify. 51% of these species are endangered and Guzmán et al. (2008) established that for Mexi- can timber identification it is necessary to use 49% of them are considered to be threatened. mixture of fluorescence tests and other charac- The CITES Appendices includes only eight teristics, such as color or anatomical features. of these species (S. humilis, S. macrophylla, Likewise, identification of the endangered or D. panamensis, G. sanctum, C. costaricense, threatened timber species from Costa Rica also O. pterocarpa, C. odorata and D. retusa) requires anatomical studies. Species of Cedrela (CITES 2002). All gymnosperms growing are easily separated from each other based on in the Costa Rican tropics are cataloged as pore arrangement and axial parenchyma types. endangered or threatened. However, most According to White & Gasson (2008), C. odo- of the gymnosperm species in tropical areas rata is more ring-porous with more aliform around the world are in same situation; they parenchyma than C. fissilis. Cedrela tonduzii is are in decline or are restricted to isolated areas similar to C. odorata and C. fissilis, although (Farjon et al. 1993). C. tonduzii is less ring porous. Aliform paren- The development of identification keys, chyma is well defined in C. tonduzii, but not like the one presented in this study, requires in C. odorata, C. salvadorensis or C. fissilis. knowledge of wood characteristics and struc- C. odorata is considered to be easy to identify ture, as described in IAWA standards (IAWA by its reddish color and distinct odor. Another 1989). Fluorescence provides a quick test for important difference among C. tonduzii and wood identification and it had been utilized other Cedrela species is that C. tonduzii has by several authors for species separation. For lower wood density than the other four species. example, Miller & Wiemann (2006) found C. salvadorensis has distinctive anatomical fea- differences in water and ethanol fluorescence tures that facilitate its identification; its rays are between Dalbergia nigra and D. spruceana. larger, commonly 4-10 seriate, than the rays of Guzmán et al. (2008) found fluorescence spe- other Cedrela species, which are 1-3 seriate in cies in the Anacardiaceae, Leguminose and width. Furthermore, its rays are heterogeneous Rubiaceae from and South Africa. Fluo- and homogenous, but the rays of the other rescence is one of the important distinguishing Cedrela species are not. However, Bonilla et al. characteristics of endangered or threatened (2004) reported that C. salvadorensis has 1-3

Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 1153 seriate rays, similar to those of other Cedrela. separating it from other Podocarpus species. Therefore, we attribute the wider rays that we Abundant axial parenchyma also separates P. found in C. salvadorensis in Costa Rica to macrostachys from P. costaricensis and P. gua- regional differences. temalensis. Patel (1967) agreed with this result: In species of Platymiscium, traumatic he found that P. dacrydioides can be separated canals were seen in P. parviflorum, but not in from P. totara, P. hallii and P. acutifolius by other species. Traumatic canals have not been the abundance of axial parenchyma. Bauch et previously reported in Platymiscium (Espi- al. (2006) mentioned that P. costarricensis has noza & León 2002). Storied rays have been more axial parenchyma than the commercially reported for most Platymiscium, for example important species P. salignus growing in Chile P. lasiucarpium, P. duckei, P. pinnatum and and Argentina. P. yucatanum (Pérez 1993, Espinoza & León Many of the Costa Rican species included 2002). However, Detienne & Jacquet (1983) in this study, are also present in other tropical reported that rays are irregularly storied in P. regions, and our results, both as wood descrip- ulei, similar to P. pinnatum var polystachyum. tions and the identification key, and are appli- The differences found in Couratari spe- cable to the wider region. Species conservation cies agreed with previous research, which is a goal for many countries, especially in has also reported differences. For example countries where deforestation of natural forests Leon (2008) separated C. guianensis from C. has increased in the last few years. There is a multiflora by ray width, and Richter (1982) strong interest to protect timber species which maintained that parenchyma distribution as have been over-exploited for many years. This well as type and configuration of inorgan- identification key and wood descriptions will ic contents can be employed for separating assist in the protection of species categorized species of Lecythidaceae. as endangered or threatened, and will promote Canessa (1989) agreed with our results reliable conservation plans. in Copaifera species; he found that the axial parenchyma in C. camibar was different than that of C. officinalis and C. pubiflora. Paren- ACKNOWLEDGMENT chyma is reported to be vasicentric, aliform to The authors wish to thank The Council confluent, and aliform of the lozenge type in for International Exchange of Scholars of the these species (Melandri & Espinoza de Pernía Department of Scholar and Professional Pro- 2009). The ray dimensions and axial parenchy- grams of USA, Premios Ford de Conservación ma of some Copaifera species are characterized of Ford Motor Company, and Vicerrectoría de by high anatomy variation (Regina et al. 2002), Investigación y Extensión del Instituto Tec- and our results confirm that. nológico de Costa Rica (ITCR) for financial On the other hand, Patel (1967) found sim- support of this research. ilar results in Podocarpaceae species when he evaluated axial parenchyma. They mentioned that numerous species of the Podocarpus RESUMEN are characterized by diffuse axial parenchyma Un total de 45 especies de árboles de Costa Rica and a considerable variation of trached cross- se catalogaron como amenazadas o en peligro de extin- field pit apertures (size, form, number per ción, de las cuales, CITES (Convention on International cross-field). Scanty axial parenchyma in P. Trade Endangered Species) incluye solamente ocho en sus guatemalensis can be used for separating this Apéndices. Sin embargo, la identificación de las especies species from other Podocarpaceae in Costa basadas en su anatomía es muy limitada. El presente estu- dio tiene el objetivo describir y comparar la anatomía y la Rica. This is in agreements with P. spicatus fluorescencia de las especies amenazadas o en peligro de growing in New Zealand, in which the lack extinción de Costa Rica. Muestras de madera de las espe- of axial parenchyma is the main feature to cies en peligro de extinción o amenazadas de la xiloteca

1154 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 61 (3): 1133-1156, September 2013 del Instituto Tecnológico de Costa Rica y del Laboratorio Costa Rica. 1996b. Se mantiene la restricción a la corta o de Productos Forestales de los Estados Unidos en Wiscon- aprovechamiento del árbol conocido como Almendro sin se examinaron, se describió su anatomía, se evaluó su nombre científico Dipteryx panamensis) (25663). actividad fluorescente y se midió su densidad. La superficie MINAE, San José, Costa Rica. de la madera fue fluorescente en ocho especies, el extracto Costa Rica. 1997. Declara en veda total aprovechamiento en agua fue fluorescente en seis especies y el extracto en de árboles en peligro extinción indicados en el pre- etanol fue positivo en 24 especies. Muchas de las especies sente decreto (25700). MINAE, San José, Costa Rica. presentaban porosidad difusa, excepto algunas Cedrela Detienne, P. & P. Jacque. 1983. Atlas d’identification des odorata, C. fissilis, Cordia gerascanthus C. salvadoren- bois de l’amazonie et des regions voiseines. Cen- sis y C. tonduzii que presentaban porosidad semi-anular. tre Technique Forestier Tropical, Nogent s/Marne, Vasos con distribución déndrica se encontró en Sideroxylon France. capari y poros solitarios en Guaiacum sanctum y Vantanea Espinoza, N. & W. León. 2002. Estudio anatómico del barbourii. Los vasos más cortos se encontraron en Guaia- leño de 56 especies de la subfamilia Papilionoideae cum sanctum y los vasos más largos en Humiriastrum (Leguminosae) en . Rev. For. Venezolana guianensis, Minquartia guianensis y Vantanea barbourii. 46: 59-71. Finalmente, la información de la anatomía y de su fluores- Farjon, A., C. Page & N. Schellevis. 1993. A preliminary cencia se utilizó para construir una clave de identificación, word list of threatened conifer taxa. Biodivers. Con- donde la actividad de fluorescencia juega un papel impor- serv. 2: 304-326. tante en la identificación. Gasson, P., R. Miller, D.J. Stekel, F. Whinder & K. Zie- minska. 2010. Wood identification of Dalbergia Palabras clave: fluorescencia, madera de Costa Rica, nigra (CITES Appendix I) using quantitative wood maderas tropicales, clave de identificación. anatomy, principal components analysis and naive Bayes classification. Ann. Bot. 105: 45-56. REFERENCES Gasson, P. 2011. How precise can wood identification be? Wood anatomy role in support of the legal timber Álvarez. D. 1986. Deforestación, causas y soluciones, p. trade, especially CITES. IAWA J. 32: 137-154. 17-20. In COSTA RICA Ministerio de Gobernación y Gasson, P., P. Baas & E. Wheeler. 2011. Wood anatomy Policía. La verdadera emergencia nacional. San José, of CITES-listed tree species. IAWA J. 32: 155-198. Costa Rica. Guzmán, J.A.S., H.G. Richter, R.R. Anda & F.J. Talavera. Bauch, J., G. Koch, J. Puls, T. Schwarz & S. 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