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Alejandro Lizama-Hernández ^a, Ma Ventura Rosas-Echeverría ^a, *, M. Guadalupe del Rio ^b

^a Universidad Autónoma del Estado de Morelos, Escuela de Estudios Superiores del Jicarero, Laboratorio de Sistemática y Evolución de Insectos, Carretera Galeana-Tequesquitengo s/n, Colonia El Jicarero, 62909 Jojutla, Morelos

^b Museo de La Plata, División Entomología- Consejo Nacional de Investigaciones Científicas y Técnicas, Paseo del Bosque s/n, 1900 La Plata, Buenos Aires, Argentina

*Corresponding author: mvrosase@gmail.com (M.V. Rosas-Echeverría)

Received: 01 October 2024; accepted: 27 January 2025

Abstract

The first phylogenetic analysis of the weevil genus Megalostylus endemic to Mexico(Entiminae, Naupactini) is presented, based on a data matrix of 37 morphological characters of adults and 21 terminal taxa. The ingroup comprises 9 species, 4 varieties, and 5 specimens of Megalostylus whose identification at the species level is doubtful. The outgroup includes species representing closely related genera: Pantomorus albosignatus, Naupactus cervinus, and Megalostylodes hirsutus. The objectives were to test the monophyly of Megalostylus, to explore its species relationships, and to determine which synapomorphies allow its identification and differentiation from other genera. The analysis yielded a single cladogram of 61 steps, showing the following phylogenetic sequence: (Pantomorus albosignatus (Naupactus cervinus (Megalostylodes hirsutus (Megalostylus rhodopus (M. morpho 2(M. macrophthalmus (M. tomentosus (M. dilaticollis (M. albicans (M. brevipilis, M. fusiformis (M. splendidus – M. sturmi))))))))))).The results support the monophyly of Megalostylus based on the following synapomorphies: protibia with a prominence opposite to mucro, elytra almost flat, sternite VIII subrhomboidal very elongated, and aedeagus smooth. They also support its sister-group relationship with Megalostylodes.

Keywords: Systematics; Naupactini; Morphology; New species; New varieties; Megalostylodes

Filogenia del género de gorgojos Megalostylus (Coleoptera: Curculionidae: Entiminae), endémico de México

Resumen

Presentamos el primer análisis filogenético del género de gorgojos Megalostylus endémico de México(Entiminae, Naupactini)basado en una matriz de datos de 37 caracteres morfológicos de adultos y 21 taxones terminales. El grupo interno comprende 9 especies, 4 variedades y 5 especímenes de Megalostylus de dudosa identificación a nivel de especie. El grupo externo está formado por 3 especies representantes de géneros relacionados cercanamente: Pantomorus albosignatus, Naupactus cervinus y Megalostylodes hirsutus. Los objetivos fueron poner a prueba la monofilia de Megalostylus, explorar sus relaciones interespecíficas y determinar qué sinapomorfías lo identifican y diferencian de otros géneros. El análisis produjo un solo cladograma de 61 pasos, que muestra la siguiente secuencia filogenética: (Pantomorus albosignatus (Naupactus cervinus (Megalostylodes hirsutus (Megalostylus rhodopus (M. morpho 2(M. macrophthalmus (M. tomentosus (M. dilaticollis (M. albicans (M. brevipilis, M. fusiformis (M. splendidus – M. sturmi))))))))))). Los resultados avalan la monofilia de Megalostylus con base en las siguientes sinapomorfías: protibias con una prominencia opuesta al mucro, élitros casi planos en vista lateral,  esternito VIII subromboidal muy elongado y aedeago liso. También respaldan su estrecha relación con Megalostylodes.

Palabras clave: Sistemática; Naupactini; Morfología; Especie nueva; Variedades nuevas; Megalostylodes

Introduction 

Megalostylus Schoenherr, 1840 (Entiminae, Naupactini) is a genus of broad-nosed weevils distributed in the Mexican states of Durango, Guanajuato, Guerrero, Michoacán, Morelos, Oaxaca, Puebla and Veracruz (Champion, 1911; Muñiz-Vélez et al., 2015; Ordóñez- Reséndiz et al., 2008). It was described by Schoenherr (1840) in his monumental work Genera et species curculionidum, cum synonymia hujus familiae, specie novae aut hactenus minus cognitae based on the type species M. sturmi Boheman, 1840 and has been traditionally characterized by the presence of a short antennal scape comparatively stouter at apex than in other genera (Champion, 1911; Figs. 1-3). The most complete treatment of Megalostylus was conducted by Champion (1911), who recognized 9 species and assigned this genus to “Otiorhynchinaealatae”, group “Cyphina”. He also provided a taxonomic key based on 32 morphological characters of adults and the complete distribution records of the species until then. 

The classification of Megalostylus proposed by Champion (1911) more than a century ago needs a revision of the status of the species, the infraspecific varieties, and a phylogenetic analysis, to test the monophyly, species relationships, and synapomorphies that support the genus.

Materials and methods 

We examined 347 adult specimens obtained from the following entomological collections: IBUNAM, Instituto de Biología, Universidad Nacional Autónoma de México, Mexico City, Mexico; INECOL, Instituto de Ecología, A. C., Xalapa, Veracruz, Mexico; USNM, National Museum of Natural History, Washington DC, USA; NHRS, Swedish Museum of Natural History, Stockholm, Sweden; and BMNH, Natural History Museum, London, England. Moreover, we included material collected by the working team at the Laboratorio de Sistemática y Evolución de Insectos (LabSei) of the ESSJicarero, UAEM.

Taxon sampling. The outgroup comprises 3 species: Megalostylodes hirsutus Champion, 1911, Naupactus cervinus (Boheman, 1840), and Pantomorus albosignatus Boheman, 1840, the former representing the most closely related genus. The ingroup includes 9 species (Figs. 1-3): M. sturmi Boheman,1840, M. rhodopus Boheman, 1840, M. albicans (Lacordaire, 1876), M. splendidus Chevrolat, 1878, M. brevipilis Champion, 1911, M. dilaticollis Champion, 1911, M. fusiformis Champion, 1911, M. macrophthalmus Champion, 1911, and M. tomentosus Champion, 1911 (Table 1). Additionally, 4 described varieties and 5 specimens of doubtful identification at species-level were included as morphospecies1-5.

A list of 37 discrete characters (29 binary and 8 multistate) was recorded from adults, including 33 characters from the external morphology, 2 of the female genitalia, and 2 of the male genitalia (Table 2). The selection of characters was based on previous analyses of the tribe Naupactini (del Río, 2009; Lanteri & del Río, 2017; Rosas et al., 2011). 

For the preparation of genital structures, we followed the methodology described in Rosas et al. (2011) and Lanteri and del Río (2017). A Carl Zeiss Stemi 2000 stereomicroscope, equipped with a reticle eyepiece was used for observations and measurements of the external and internal morphology. Photographs were taken with a Nikon D750 camera equipped with a SIGMA 150 mm 1:2.8 (macro) lens, and drawings were made with Corel Draw (2020 version 22.0.0412). Most characters were illustrated by photographs and drawings, to facilitate the recognition of character states (Figs. 4, 5). They were highlighted with arrows, with indication of character numbers and character states between parentheses. 

A data matrix of 21 terminal groups and 37 morphological characters was compiled (Table 3). Character states that could not be examined (due to insufficient material) were scored with a “?” and character states with inapplicable entries on various terminals were scored with a “–”. Species or varieties showing 2 or more characters were coded as polymorphic characters. To facilitate the coding, Mesquite software version 3.70 was used (Maddison & Maddison, 2021). An implicit enumeration search was performed in the program TNT version 1.5 (Goloboff & Catalano, 2016). The characters were treated as unordered or non-additive, and under equal weights. The species Pantomorus albosignatus was used to root the trees. To evaluate branch support (BS), a standard Bootstrap with 1000 replicates was calculated in TNT. Branch support values greater than 50 were mapped in the cladogram. The resulting cladogram and character state transformations were examined in WINCLADA under fast optimization. All records were georeferenced, and maps created in ArcMap version 10. 4.1 (ESRI, 2015).

Results 

The search for the most parsimonious trees under equal weights yielded one most parsimonious tree (Fig. 6) (L = 63, IC = 0.73, IR = 0.83), showing the following phylogenetic sequence: (Pantomorus albosignatus (Naupactus cervinus (Megalostylodes hirsutus (Megalostylus)))). 

Megalostylodes was found to be sister to the genus Megalostylus based on 12 synapomorphies: epistome scales similar in size, density, and color to those on the rest of the rostrum (0:0), dorsal surface of rostrum slightly to strongly depressed (1:1), absence of pair of dorsolateral carinae (2:0), scape very wide in males and thin in females (3:1), width of scape at apex greater than club width in males (4:1), shape of scape strongly clavate (5:1), color brown to black of the verticillate setae of funicle (9:1), slight lateral projection of pronotum (14:1), absence of a row of denticles on inner margin of protibia (20:0), vestiture of scutellum present, consisting of white scales or seta-like scales (25:1), presence of medial longitudinal depression of ventrites 1 and 2 on males (32:1), and subspherical body shape of spermatheca (33:1). There are also 2 homoplastic characters that support this relationship: pronotum subconical, with sides curved and strongly divergent from apex to base (11:1) and width of base of scutellum wider than interestria 2 (24:2). 

Megalostylus was recovered as monophyletic based on 4 synapomorphies: presence of prominence opposite to mucro in protibia (21:1), elytra in lateral view almost flat (23:1), plate of sternite VIII, subrhomboidal very elongated (34:0), and sculpture of aedeagus smooth (35:1).

The sister genus Megalostylodes is characterized by 3 autapomorphies: pronotum narrower with respect to elytral base (13:0), very long elytral setae, longer than width of elytral interstriae 2 at middle (27:2), and presence of deeply excavated metafemur near apex (31:1), and 3 non-exclusive synapomorphies: presence of glabrous areas in midline of pronotum (12:1), erect disposition of elytral setae (26:1), and slightly convex interestriae (29:1). 

Table 1

List of species included in the cladistic analysis of the genus Megalostylus and their geographic distributions (countries and states).

Species names	Geographic distributions
Pantomorus albosignatus Boheman	Mexico (Aguascalientes, Chihuahua, Coahuila, Mexico City, Durango, Guanajuato, Guerrero, Hidalgo, Monterrey, Oaxaca, Puebla Querétaro, San Luis Potosí, Veracruz, Zacatecas)
Naupactus cervinus (Boheman)	Argentina (Buenos Aires, Catamarca, Córdoba, Corrientes, Entre Ríos, Jujuy, Mendoza, Misiones, Salta, Santa Fe, Tucumán), Brasil (Paraná, Río Grande do Sul, Santa Catarina, São Paulo), México (Distrito federal), Uruguay (Artigas, Canelones, Colina, Durazno, Maldonado, Montevideo, Rio Negro, Treinta y Tres)
Megalostylodes hirsutus Champion	Mexico (Oaxaca)
Megalostylus albicans (Lacordaire)	Mexico (Colima, Estado de México, Guanajuato, Guerrero, Jalisco, Michoacán, Morelos, Nayarit, Oaxaca, Puebla)
Megalostylus brevipilis Champion	Mexico (Colima, Guerrero, Oaxaca)
Megalostylus dilaticollis Champion	Mexico (Guerrero, Michoacán, Morelos, Jalisco)
Megalostylus fusiformis Champion	Mexico (Morelos, Guerrero)
Megalostylus macrophthalmus Champion	Mexico (Oaxaca)
Megalostylus rhodopus Boheman	Mexico (Oaxaca)
Megalostylus splendidus Chevrolat	Mexico (Durango, Guerrero, Morelos, Puebla)
Megalostylus sturmi Boheman	Mexico (Guerrero, Michoacán)
Megalostylus tomentosus Champion	Mexico (Oaxaca)

Table 2 

List of 37 morphological characters, character states and codes. 

Character	Character states
0. Shape, size, density and color of epistome scales compared to those on the rest of the rostrum	Similar in size, density and color (0); smaller, scarcer and generally different in color (1)
1. Dorsal surface of rostrum	Flat (0); slightly to strongly depressed (1)
2. Pair of dorsolateral carinae	Absent (0); present (1)
3. Sexual dimorphism in antennae	Scape width subequal in males and females (0); scape very wide in males and thin in females (1)
4. Width of scape at apex with respect to width of club, in males	Less than club width (0); greater than club width (1)
5. Shape of scape	Capitate (0); strongly clavate (1)
6. Vestiture of scape	Absent to almost absent (0); scales dispersed leaving integument exposed (1); dense, space between scales reduced or absent, not leaving integument exposed (2)
7. Length of funicular antennomeres 1 and 2	Antennomere 2 slightly shorter than 1 or both sub-equal (0); antennomere 2 longer than 1 (1)
8. Antennae, scales on funicular antennomere 2	Absent (0); present (1)
9. Color of verticillate setae of funicle	Whitish or golden (0); brown to black (1)
10. Convexity of eyes	Convex, pronounced curvature, hemispherical shape (0); slightly convex, less pronounced curvature, small elevations (1)
Table 2. Continued
11. Shape of pronotum	Subcylindrical, curved sides and maximum width about half (0); subconical, with sides moderately to strongly curved and strongly divergent from apex to base (1); subconical, with sides slightly curved to straight and strongly divergent from apex to base (2); bell-shaped (3); subtrapezoidal, with sides slightly concave at anterior half and progressively expanding towards lateral sides at posterior half (4)
12. Glabrous areas at midline of pronotum	Absent (0); present (1)
13. Width of pronotum with respect to elytral base	Narrower (0); subequal (1); wider (2)
14. Lateral projection of pronotum	Absent or indistinct (0); slightly projected (1); strongly projected (2)
15. Shape of lateral projections of posterior margin of pronotum	Extended towards sides (0); strongly extended backwards, towards elytral base (1)
16. Sides at basal third of pronotum	Not depressed (0); strongly depressed (1)
17. Shape of posterior margin of pronotum	Straight to slightly bisinuate (0); strongly bisinuate (1)
18. Bulging surface beneath intercoxal granule, at anterior margin of prothorax	Indistinct or absent (0); present (1)
19. Color of legs integument	Light tones, reddish or orange (0); dark tones, black or brown (1)
20. Row of denticles on inner margin of protibia	Absent (0); present (1)
21. Prominence opposite to mucro in protibia	Absent (0); present (1)
22. Elytral anterior margin	Straight (0); bisinuate (1)
23. Shape of elytra in lateral view	Moderately convex (0); almost flat (1)
24. Width of base of scutellum with respect to width of interstria 2	Reduced to indistinct (0); subequal to width of interstria 2 (1); wider than interstria 2 (2)
25. Scutellum vestiture	Scales similar in color to those covering whole surface of elytra (0);  Scales similar, consisted of white scales or seta-like scales (1)
26. Disposition of elytral setae	Recumbent to suberect (0); erect (1)
27. Size of elytral setae	Short (0); long, almost as long as width of interstria 2 at middle (1); very long, longer than width of elytral interestriae 2 at middle (2)
28. Arrangement of elytral setae	Uniform throughout elytral disc (0); only on lateral edges of elytral disc, dorsally without setae (1)
29. Convexity of interstriae	Flat (0); slightly convex (1)
30. Vestiture on legs	Absent or almost absent (0); inconspicuous, scales are scattered leaving integument exposed (1); conspicuous, space between scales is reduced or absent, and covered the integument (2)
31. Deep excavation on metafemur, near apex	Absent (0); present (1)
32. Medial longitudinal depression of ventrites 1 and 2, on males	Indistinct or absent (0); present (1)
33. Spermatheca, body shape	Subcylindrical, slender (almost as wide as base of cornu) (0); subspherical (1)
34. Shape of plate of sternite VIII	Subrhomboidal very elongated, with basal part much longer than apical part (0); subrhomboidal slightly elongated, with basal part as long as apical part (1)
35. Aedeagus, sculpture of median lobe	Granulose (0); smooth (1)
36. Aedeagus, shape of apex of median lobe	Acute to slightly acute (0); truncated (1); arrow shape (2)

Table 3

Data matrix of 21 taxa and 37 morphological characters of Megalostylus and outgroups used for the cladistic analysis. Inapplicable characters are indicated with a dash (-), and missing characters with a question mark (?).

Taxa	Characters
	0-4	5-9	10-14	15-19	20-24	25-29	30-36
Pantomorus albosignatus	101?0	00000	00010	00000	10000	-0100	00?01??
Naupactus cervinus	10100	00100	00010	00000	10100	00000	00001?2
Megalostylodes hirsutus	01011	10001	01101	00000	00102	11201	0111100
Megalostylus albicans	01011	12010	02012	00101	01111	00000	1011010
Megalostylus albicans var. expansus	01011	12010	02022	00101	01111	00000	101??10
Megalostylus albicans var. farinosus	01011	120?0	02012	00101	01111	00000	101????
Megalostylus brevipilis	010?1	120?0	03011	10101	01111	00000	101??10
Megalostylus dilaticollis	01011	12000	04022	01001	01111	00000	10110??
Megalostylus fusiformis	010?1	120?0	03011	10101	01111	00000	10?????
Megalostylus macrophthalmus	01011	100?	12011	00000	01112	10000	00?10??
Megalostylus splendidus	01011	12010	01011	10101	01112	00000	1011010
Megalostylus splendidus var.	01011	12010	01011	10101	01112	00000	1011010
Megalostylus sturmi	01011	12010	0100&11	10111	01112	01100	1011?11
Megalostylus sturmi var. villosus	01011	12010	0100&11	10111	01112	01100	1011011
Megalostylus rhodopus	01011	12001	01111	00000	01112	10010	0011010
Megalostylus tomentosus	01011	110?0	02011	00000	01112	01100	00110??
Morphospecies 1	010?1	12001	01111	10000	0111?	10010	001??10
Morphospecies 2	010?1	11000	01011	10100	01112	10000	001??10
Morphospecies 3	010?1	10001	02011	00000	01112	10011	001??10
Morphospecies 4	010??	11000	02011	00000	01112	01100	00?10??
Morphospecies 5	010?1	12000	04011	00001	01111	00000	101??10

Two main clades were found within Megalostylus: clade I, including Megalostylus rhodopus and morphospecies 1 and 3 (considered belonging to M. rhodopus), is supported by 1 exclusive synapomorphy: elytral setae only present on the lateral edges of elytral disc (28:1). Clade II is supported by 1 exclusive synapomorphy and 1 non-exclusive synapomorphy: scales of scape separate, leaving integument exposed (6:1) and whitish or golden color of the verticillate setae of funicle (9:0), respectively. Within clade II, morphospecies2 is sister to the remaining species, which form a clade supported by 1 non-exclusive synapomorphy: pronotum subconical, sides slightly curved to straight and strongly divergent from apex to base (11:2). Megalostylus macrophthalmus is the sister species of M. tomentosus–M. sturmi var. villosus based on 1 non-exclusive synapomorphy: vestiture of scutellum present, scales similar in color to those covering the whole surface of elytra (25:0). Megalostylus tomentosus (along with morphospecies4) is the sister of a supported clade (BT 58) M. dilaticollis–M. sturmi var. villosus clade, which is supported by 3 exclusive synapomorphies and 1 non-exclusive synapomorphy: integument of legs with dark tones, black or brown (19:1), and width of base of scutellum reduced to indistinct (24:1), vestiture on legs inconspicuous, scales are scattered leaving integument exposed (30:1), and scales separated leaving integument of scape exposed (6:2). Within this clade M. dilaticollis (along with morphospecies 5) is sister to a subclade supported by 1 exclusive synapomorphy and 1 non-exclusive synapomorphy: presence of scales on the second funicular antennomere (8:1) and posterior margin of pronotum strongly bisinuate (17:1) that includes M. albicans and its varieties, M. albicans var. expansus, and var. farinosus sister to M. brevipilis to M. sturmi var. villosus clade supported by 1 exclusive synapomorphy and 1 non-exclusive synapomorphy: pronotum flared or bell-shaped (11:3) and lateral projections of posterior margin of pronotum strongly extended backward, towards elytral base (15:1). Within this clade, M. brevipilis and M. fusiformis are sisters to the M. splendidus–M. sturmi var. villosus clade which is supported by 2 non-exclusive synapomorphies: pronotum subconical, sides curved and strongly divergent from apex to base (11:1), and width of base of scutellum wider than interestria 2 (24:2). The clade M. splendidus–M. sturmi var. villosus also has unresolved relationships between M. splendidus and his variety, both sister to the M. sturmi + M. sturmi var. villosus clade. 

Geographic distribution. The maps (Figs. 7, 8) indicate that the species of Megalostylus are mostly distributed along the Pacific coast, with the greatest richness in the central and southern parts of the country. Megalostylus albicans shows the largest distribution,ranging from Oaxaca to Sinaloa. Six species are distributed in the central states of the country (Guanajuato, Guerrero, Michoacán, Morelos, and Puebla): M. albicans, M. brevipilis, M. dilaticollis, M. fusiformis, M. splendidus, and M. sturmi. The species M. rhodopus, M. macrophthalmus, and M. tomentosus are restricted to Oaxaca, and M. rhodopus reaches the southernmost distribution extending to the Isthmus of Tehuantepec. 

Discussion 

The sister group relationship between Megalostylodes and Megalostylus is well supported by 12 exclusive synapomorphies, and the monophyly of Megalostylus is supported by a combination of 4 exclusive synapomorphies of the protibiae, elytra, and female and male genitalia. Champion (1911) distinguished Megalostylus based on the presence of a prominence opposite to mucro (or toothed tibiae at the external apical angle), and Lanteri and del Río (2017) recovered this character state (car 69[1]) as an apomorphy of Megalostylus.

Within Megalostylus most of the relationships are not well supported nevertheless, it is clear the position of M. rhodopus (including morphospecies 1 and 3) as sister species of the remaining, and it was recovered a well-supported group including M. dilaticollis (= morphospecies 5), M. albicans, M. brevipilis, M. fusiformis, M. splendidus, and M. sturmi. Megalostylus macrophthalmus and M. tomentosus (= morphospecies 4) would be basal regarding this clade. Morphospecies2 is considered a probable new species, however, this status needs further studies. 

The species of Megalostylus are very variable in scale color, size and pronotum shape, and due to that intraspecific variation, some nominal species have been synonymized: M. farinosus Chevrolat, 1878 [junior synonym of M. albicans (Lacordaire, 1876)], M. expansus Pascoe, 1881 [junior synonym of M. albicans (Lacordaire, 1876)] and M. villosus Chevrolat, 1878 [junior synonym of M. sturmi Boheman, 1840]. Our analysis explored the relationships among intraspecific varieties of the genus Megalostylus, and the results support the previously established synonymies. In the case of M. albicans with its varieties (M. albicans var. expansus and M. albicans var. farinosus) there is no reason to consider them as different species. Characters with the greatest contribution to the species relationships are those referring mainly to the vestiture of antennae, legs, elytra, shape, and characteristics of the pronotum, scutellum, and apex of aedeagus.

Acknowledgments 

We thank all curators, institutions, collection managers, technicians, and collectors that provided specimens. We also thank Miguel Menéndez-Acuña, Elizabeth Arellano-Arenas, and anonymous reviewers for their helpful suggestions to improve the manuscript. The authors would like to express their sincerest gratitude to Santiago Zaragoza Caballero and María Cristina Mayorga Martínez for their invaluable assistance in providing material for examination. 

References 

Champion, G. C. (1911). Insecta. Coleoptera. Vol. IV, Parte 3. Rhynchophora. Curculionidae. Attelabinae, Pterocolinae, Allocoryninae, Apioninae, Thecesterninae, Otiorhynchinae. In D. Sharp, & G. Champion (Comps.), Biologia Centrali-Americana (pp. 241–246). London: R.H. Porter.

del Río, M. G. (2009). Estudio taxonómico y cladístico de los géneros de la tribu Naupactini (Coleoptera: Curculionidae) distribuidos en la subregión Páramo-Puneña o Zona de Transición Sudamericana (Ph.D. Thesis). Universidad Nacional de la Plata, La Plata, Argentina.

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Lanteri, A. A., & del Río, M. G. (2017). Phylogeny of the tribe Naupactini (Coleoptera: Curculionidae) based on morphological characters. Systematic Entomology, 42, 429–447. https://doi.org/10.1111/syen.12223 

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Ordóñez-Reséndiz, M. M., Muñíz-Vélez R., & Gama-Rojas, F. (2008). Curculiónidos (Coleópteros). In S. Oceguera, & J. Llorente-Bousquets (Coords.), Catálogo taxonómico de especies de México, en Capital natural de México, vol. L. Conocimiento actual de la biodiversidad. CD1. Mexico City. Conabio.

Rosas, M. V., Morrone, J. J., del Río, M. G., & Lanteri, A. A. (2011). Phylogenetic analysis of the Pantomorus-Naupactus complex (Coleoptera: Curculionidae: Entiminae) from North and Central America. Zootaxa, 2780, 1–19. https://doi.org/10.11646/zootaxa.2780.1.1 

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Oscar Iván González-Romero ^a, Xochitl G. Vital ^{a, b,} *

^a Universidad Nacional Autónoma de México, Facultad de Ciencias, Circuito Exterior s/n, Ciudad Universitaria, Coyoacán, 04510 Ciudad de México, México

^b Universidad Nacional Autónoma de México, Posgrado en Ciencias Biológicas, Facultad de Ciencias, Circuito Exterior s/n, Ciudad Universitaria, Coyoacán, 04510 Ciudad de México, México 

*Corresponding autor: vital@ciencias.unam.mx (X.G. Vital)

Received: 18 August 2024; accepted: 5 February 2025

Abstract

While the diversity of sea slugs in the northern area of the Pacific coast of Mexico has been studied thoroughly in the last decades, little is known about the composition of species in the southern states of Mexico. Several field trips were made in 5 localities of Bahías de Huatulco, Oaxaca, where specialized sampling methods focused on sea slugs were carried out. Herein, we documented 49 species of sea slugs, including 37 new records for the state, which increases to 58 species the total sea slug richness known for Oaxaca. This study updates the inventory of sea slugs for the Mexican Pacific coast and contributes to the knowledge of the marine fauna of the Natural Protected Area “Parque Nacional Huatulco”.

Keywords: Bahías de Huatulco; Coral reefs; Molluscs; Nudibranchia; Tropical Eastern Pacific

Babosas marinas (Gastropoda: Heterobranchia) de Huatulco: nuevos registros y ampliaciones de distribución para Oaxaca, México

Resumen

Mientras que la diversidad de babosas marinas en la zona norte de la costa del Pacífico mexicano ha sido estudiada de forma exhaustiva en las últimas décadas, poco se sabe acerca de la composición de especies en los estados del sur de México. Se realizaron diversas visitas a 5 localidades de las bahías de Huatulco, Oaxaca, donde se llevaron a cabo métodos de muestreo especializados con enfoque en babosas marinas. Aquí censamos 49 especies de babosas marinas, con 37 nuevos registros para el estado, lo que incrementa la riqueza de babosas marinas conocida para Oaxaca a 58 especies. Este estudio actualiza el inventario de babosas marinas para la costa del Pacífico mexicano y contribuye al conocimiento de la fauna marina del Área Natural Protegida “Parque Nacional Huatulco”.

Palabras clave: Bahías de Huatulco; Arrecifes de coral; Moluscos; Nudibranchia; Pacífico este tropical

Introduction

Heterobranch sea slugs are gastropods with more than 8,400 described species distributed in all the oceans of the planet, from the intertidal zone to deep waters (Behrens et al., 2022; Camacho-García et al., 2005). Some of these molluscs can do some of the most exceptional processes in the metazoans, such as the incorporation of functional chloroplasts or nematocysts into their tissues (Goodheart & Bely, 2017; Händeler et al., 2009). Furthermore, sea slugs can be important model organisms in several study areas from neuroscience to global climate change (Kandel, 1979; Ziegler et al., 2014); a source of biomedical compounds used for creating novel drugs (Dean & Prinsep, 2017; Fisch et al., 2017) and an attractive target to professional and amateur underwater photographers worldwide (Behrens, 2005). 

More than 370 species of sea slugs have been recorded in the Eastern Pacific from Alaska to Central America (Behrens et al., 2022), of which around 234 have been found on the Mexican Pacific coast (Hermosillo et al., 2006). Although several studies on sea slugs have been performed in this region (e.g., Angulo-Campillo, 2005; Bertsch, 2014; Flores-Rodríguez et al., 2017; Hermosillo, 2009, 2011; Hermosillo & Behrens, 2005; Hermosillo & Gosliner, 2008; Verdín-Padilla et al., 2010), most of them are focused on the northern coast of Mexico, whereas the composition of species of the southern coast, corresponding to the states of Oaxaca and Chiapas, has been poorly recorded in faunal inventories. The current knowledge of Oaxaca’s sea slug fauna comes from 2 checklists of intertidal molluscs on rocky shores of the state (Holguín-Quiñones & González-Pedraza, 1989; Rodríguez-Palacios et al., 1988), a review of the sea slugs preserved at Colección Nacional de Moluscos (CNMO) from Universidad Nacional Autónoma de México (UNAM) (Zamora-Silva & Naranjo-García, 2008), and an ecological analysis of the richness and abundance of molluscs associated with coral ecosystems in the Tropical Eastern Pacific (Barrientos-Luján et al., 2021). Altogether, these studies sum up to 10 known species of Heterobranch sea slugs on the coastal shore of Oaxaca.

Biological inventories are a fundamental base that provides essential information for conservation strategies, such as establishing natural protected areas or monitoring their condition inside them (Alexander et al., 2009; Schejter et al., 2016). Marine molluscs are useful in rapid biodiversity assessments (Benkendorff & Davis, 2002), and they could serve as indicators of the total biological richness in marine reserves (Gladstone, 2002), which possibly extrapolates to different areas and types of habitats. Sea slugs are one of the most diverse groups within the marine molluscs. Nonetheless, they are apparently “rare in space and time” (Schubert & Smith, 2020), as they are not always recorded in the biological inventories due to their small size, their camouflage strategies and inadequate collecting methods applied on the field. This work aims to provide information on the diversity of sea slugs in the southern region of the Mexican Pacific coast, based on surveys conducted in different coral communities in Oaxaca. 

Materials and methods

Five localities of the bay complex known as Bahías de Huatulco, Oaxaca were selected for field studies: San Agustín (hereafter Agustín), La Entrega (hereafter Entrega), El Arrocito (hereafter Arrocito), Isla Montosa (hereafter Montosa) and Playa Conejos (hereafter Conejos). Agustín is located within the Natural Protected Area in the category of National Park “Parque Nacional Huatulco (PNH)” (Conanp, 2003) (Fig. 1, Table 1). This locality has a large and compact homogenous platform of coral colonies dominated by Pocillopora damicornis with patches of dead corals and rocks, similar to Entrega, which additionally has shallow zones of algal turf (López-Pérez & Hernández-Ballesteros, 2004; Ramírez-González, 2005). Arrocito’s substrate is composed of rocks overgrown by algal turf and dead coral fragments mostly in shallow environments, with few coralline formations and a high abundance of sponges. Conejos has big rocks with a sandy bottom and low presence of corals at shallow depths (Ramírez-González, 2005). Finally, Montosa is an island with mixed patches of corals, sand and large boulders as a dominant substrate (López-Pérez & Hernández-Ballesteros, 2004).

A total of 17 field trips were conducted between May 2017 and March 2018, 4 at each locality (except on Montosa, where we searched only once). Using snorkelling and SCUBA diving at maximum depths of 18 m, searches were conducted in subtidal environments on different substrates (Table 1). Most of the specimens were collected in all localities, when necessary, except in Agustín where photos, identification and measuring of the organisms were taken in situ. Additionally, different algal morphotypes where sea slugs may potentially be found were collected in all localities, excluding Agustín. The algae collected were placed in trays with low seawater level, then they were examined after 2 to 3 hours to find specimens attached to the tray walls (Urbano et al., 2019). All organisms were measured and photographed while they were alive and identified according to sea slug identification guides for the Pacific east coast (Behrens et al., 2022; Camacho-García et al., 2005; Hermosillo et al., 2006). Afterwards, the collected specimens were narcotized with magnesium chloride (MgCl₂), preserved with ethanol (96º) (Urbano et al., 2019) and deposited in Colección Nacional de Moluscos (CNMO), Instituto de Biología, UNAM. The nomenclature used in this work follows Bouchet et al. (2017) for supra-family and family categories and World Register of Marine Species (Horton et al., 2024) for genus and species categories. We present the new records for Oaxaca: number of organisms found per locality, their size, their collection number, distribution in the Pacific east coast and remarks of the species, as well as a brief description of undetermined species. 

Table 1

Collecting sites in Bahías de Huatulco: San Agustín, La Entrega, El Arrocito, Isla Montosa, Playa Conejos. Sampling technique: snorkelling (S), scuba diving (SD).

Locality	Latitude (N)	Longitude (W)	Sampling technique	Substrate composition
San Agustín	15°41.185’	96°14.254’	S	Compact coral colonies, rocks and dead coral patches
La Entrega	15°44.642’	96°07.732’	S, SD	Compact coral colonies, rocks, algae and dead coral patches
El Arrocito	15°45.672’	96°06.008’	S, SD	Rocks with algal turf, coral, sponges and dead coral patches
Isla Montosa	15°45.725’	96°05.120’	SD	Boulders, sand and corals
Playa Conejos	15°46.722’	96°03.853’	S	Boulders, sand and corals

Results

A total of 298 specimens belonging to 49 species (38 determined to species level, 10 to genus and 1 to family) were recorded from 5 localities in Bahías de Huatulco; the species were distributed in 22 families and 6 orders/superorders. Nudibranchia had the highest number of species (27), followed by Sacoglossa (10), Aplysiida (6), Pleurobranchida (3), and Cephalaspidea (2), while Umbraculida was represented only by 1 species. A list of the sea slugs recorded from Bahías de Huatulco in this study, complemented with the previous records from Oaxaca, is given in Table 2. 

Class Gastropoda Cuvier, 1795

Subclass Heterobranchia Gray, 1840

Infraclass Euthyneura Spengel, 1881

Cohort Ringipleura Kano, Brenzinger, Nützel, Wilson and Schrödl, 2016

Subcohort Nudipleura Wägele and Willan, 2000

Order Pleurobranchida Pelseneer, 1906

Family Pleurobranchidae Gray, 1827

Berthellina ilisima Ev. Marcus and Er. Marcus, 1967

(Fig. 2A) 

Material examined: 1 organism (18 mm), Entrega (CNMO8025).

Distribution: from Santa Barbara, California to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: animals with nocturnal habits and spongivorous diet (Valdés, 2019).

Berthella sp.

(Fig. 2B)

Material examined: 2 organisms (5, 8 mm), Arrocito (CNMO8274).

Distribution: El Arrocito, Bahías de Huatulco, Oaxaca (this study).

Diagnosis: translucent whitish body with numerous small opaque white dots and low rounded brown tubercles on the dorsum. Small translucent internal shell. The foot protrudes posteriorly from the mantle. The oral velum is triangular and the rolled rhinophores are partially fused. 

Remarks: organisms were found under rocks near white sponges. The observed characters in the specimens allowed their identification only to the genus level. Berthella andromeda, B. strongi and B. martensi are also distributed in the Pacific east coast; however, our specimens had a more translucent and white body than B. strongi and they did not present the opaque white transverse bar of B. andromeda (Ghanimi et al., 2020); the dots on the notum also were darker and more regular compared with B. martensi, which has a band along the edge of the mantle (Behrens et al., 2022).

Pleurobranchus digueti Rochebrune, 1895

(Fig. 2C)

Material examined: 3 organisms (15-20 mm), Entrega (CNMO8020); 1 organism (35 mm), Arrocito (CNMO8006); 1 organism (10 mm), Montosa (CNMO7965).

Distribution: from Santa Barbara, California to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: nocturnal animals hide under rocks during the day (Valdés, 2019).

Order Nudibranchia Cuvier, 1817

Family Dorididae Rafinesque, 1815

Dorididae sp.

(Fig. 2D)

Material examined: 1 organism (6 mm), Conejos (CNMO8261).

Distribution: Playa Conejos, Bahías de Huatulco, Oaxaca (this study).

Diagnosis: yellowish body, with an oval orange-brownish patch covering the dorsum and divided by a cream-whitish band that runs through the middle of the rhinophores all the way to the branchial area. The lamellated rhinophores and the gills are the same colour as the body.

Remarks: the specimen was found under rocks at approximately 3 m depth. The observed characters in the organism did not resemble any recorded species in the literature, but they allowed its identification up to the family level.

Doris sp.(Risbec, 1928)

(Fig. 2E)

Material examined: 1 organism (6 mm), Conejos (CNMO8260).

Distribution: Playa Conejos, Bahías de Huatulco, Oaxaca (this study).

Diagnosis: pale yellowish body with discontinued purple band between the rhinophores and the posterior area of the body. The animal had big tubercles in the central area of the dorsum with visible translucent spicules. Rhinophores were lamellated.

Remarks: specimen found on algae of the genus Caulerpa. This specimen resembles the organism illustrated in Behrens et al. (2022) as Doris immonda; however, the previous authors mention that this identification might not be valid due to its geographical distribution, as D. immonda was originally described for the Indo-Pacific. Also, the diagnosis of D. immonda in Gosliner et al. (2018) does not mention the purple band observed in this individual, these authors described a white opaque marking across the body instead. Therefore, we decided to include this species as Doris sp. until a further review of this taxon is made.

Family Discodorididae Bergh, 1891

Diaulula nayarita (Ortea & Llera, 1981)

(Fig. 2F)

Material examined: 1 organism (4 mm), Arrocito (CNMO7974).

Distribution: from Punta Eugenia, Baja California to Panama (Camacho-García et al., 2005).

Remarks: found under rocks near sponges of the same colour as the specimen.

Discodoris ketos (Marcus & Marcus, 1967)

(Fig. 2G)

Material examined: 1 organism (8 mm), Arrocito (CNMO8028).

Distribution: Gulf of California. Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: it is still unclear whether this species is the same as the circumtropical species Tayuva lilacina (Behrens et al., 2022). Discodoris ketos has a highly specialised diet, feeding on the sponge Haliclona caerulea (Verdín-Padilla et al., 2010).

Geitodoris mavis (Marcus & Marcus, 1967)

(Fig. 2H) 

Material examined: 1 organism (15 mm), Entrega (CNMO7964). 1 organism (13 mm), Arrocito (CNMO8029).

Distribution: from Rosarito, Baja California to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: live specimens darkened their gills when disturbed.

Family Polyceridae Alder and Hancock, 1845

Polycera anae Pola et al., 2014

(Fig. 2I)

Material examined: 2 organisms (8, 10 mm), Conejos (CNMO7981). 

Distribution: from Mexico to Costa Rica (Pola et al., 2014).

Remarks: found on algae of the genus Padina. Specimens found in this work exceed the maximum length of 5 mm reported for the species by Pola et al. (2014).

Polycera cf. hedgpethi Er. Marcus, 1964

(Fig. 2J)

Table 2

Sea slug fauna recorded from Oaxaca state based on this study and the literature. Localities: Puerto Ángel (PA), San Agustín (SA), El Maguey (EM), La Entrega (LE), El Arrocito (EA), Isla Montosa (IM), Playa Conejos (PC), Santa Cruz (SC), Not available data (ND). New records for the state (*). Only determined species in literature were included. References: ¹Holguín-Quiñones and González-Pedraza (1989); ²Rodríguez-Palacios et al. (1988); ³Zamora-Silva and Naranjo-García (2008); ⁴Barrientos-Luján et al. (2021); ⁵This study.

Family	Species	PA	SA	EM	LE	EA	IM	PC	SC	ND	Reference
Pleurobranchidae	Berthellina ilisima*				•						5
	Berthella sp.					•					5
	Pleurobranchus digueti*				•	•	•				5
Dorididae	Dorididae sp.							•			5
	Doris sp.							•			5
Discodorididae	Diaulula nayarita*					•					5
	Diaulula sandiegensis	•									2
	Discodoris ketos*					•					5
	Geitodoris mavis*				•	•					5
Polyceridae	Polycera anae*							•			5
	Polycera cf. hedgpethi*						•				5
	Tambja abdere*						•				5
Chromodorididae	Felimida sphoni*				•	•		•			5
	Chromolaichma dalli*		•				•	•			5
	Chromolaichma sedna*					•	•				5
	Felimare agassizii*		•		•	•	•				5
Cadlinidae	Cadlina sp.				•	•		•			5
Dendrodorididae	Dendrodoris krebsii			•							2
	Doriopsilla janaina*		•		•			•			5
Tritoniidae	Tritonia festiva	•									2
Hancockiidae	Hancockia californica*					•					5
Flabellinidae	Coryphellina marcusorum*						•				5
	Samla telja*		•		•	•	•				5
Cuthonidae	Cuthona divae	•									2
	Cuthona sp. 1					•					5
	Cuthona sp. 2		•								5
Aeolidiidae	Bulbaeolidia sulphurea*					•					5
	Anteaeolidiella chromosoma*					•		•			5
	Anteaeolidiella ireneae*				•						5
	Baeolidia moebii*		•								5
	Limenandra confusa*		•			•					5
	Spurilla braziliana*					•					5
Facelinidae	Favorinus elenalexiarum*				•	•					5
	Phidiana lascrucensis*		•		•	•	•	•			5
Tylodinidae	Tylodina fungina*					•					5
Table 2. Continued
Bullidae	Bulla punctulata									•	1
	Bulla gouldiana									•	4
Haminoeidae	Haminoea sp.		•			•					5
	Aliculastrum exaratum									•	4
Aglajidae	Navanax aenigmaticus*		•		•	•					5
Aplysiidae	Aplysia cf. cedrosensis*							•			5
	Aplysia californica				•						2
	Aplysia hooveri*		•		•	•		•			5
	Dolabella cf. auricularia*		•		•						5
	Dolabella californica				•						2
	Dolabrifera nicaraguana*				•						5
	Phyllaplysia padinae*							•			5
	Stylocheilus rickettsi*				•	•					5
Oxynoidae	Lobiger cf. souverbii*				•			•			5
	Oxynoe aliciae*				•						5
Plakobranchidae	Elysia diomedea		•		•	•		•	•		3, 5
	Elysia cf. pusilla*					•		•			5
	Elysia sp. 1					•	•	•			5
	Elysia sp. 2		•		•	•		•			5
Limapontiidae	Placida cf. dendritica*				•						5
Hermaeidae	Polybranchia mexicana*							•			5
	Caliphylla sp.				•						5
	Hermaea sp.				•						5
	Total	3	14	1	25	27	10	18	1	3

Material examined: 1 organism (14 mm), Montosa. Photographic record only.

Distribution: from Puerto Peñasco, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: found on rocks with bryozoan colonies. Behrens et al. (2022) treat this species as Polycera gnupa; however, it is unclear if this nudibranch is different from the widespread species P. hedgpethi.

Tambja abdere Farmer, 1978

(Fig. 3A) 

Material examined: 1 organism (42 mm), Montosa (CNMO7966).

Distribution: from the Gulf of California, Mexico to Costa Rica (Behrens et al., 2022).

Remarks: the specimen was found on rocks at approximately 13 m depth. According to Hermosillo (2007), T. abdere feeds on the bryozoan Sessibugula translucens which lives in zones with low currents.

Family Chromodorididae Bergh, 1891

Felimida sphoni Ev. Marcus, 1971

(Fig. 3B)

Material examined: 1 organism (5 mm), Entrega (CNMO8270). Three organisms (8-14 mm), Arrocito (CNMO8262, CNMO8272). Three organisms (9-12 mm), Conejos (CNMO8263, CNMO8265). 

Distribution: from the Gulf of California, Mexico to Ecuador (Behrens et al., 2022).

Remarks: animals were found under rocks near to different unidentified sponges.

Chromolaichma dalli (Bergh, 1879)

(Fig. 3C)

Material examined: 5 organisms (17-32 mm), Montosa (CNMO8259). One organism (5 mm), Conejos (CNMO8264).

Distribution: from Islas San Benito, Baja California to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: larger specimens were found at approximately 15 m depth. Matsuda and Gosliner (2018) categorized C. dalli under a temporal nomenclatural name that needs further taxonomic analysis. 

Chromolaichma sedna (Ev. Marcus & Er. Marcus, 1967)

(Fig. 3D)

Material examined: 1 organism (31 mm), Montosa (CNMO8278).

Distribution: from the Gulf of California, Mexico to Ecuador (Behrens et al., 2022).

Remarks: Verdín-Padilla et al. (2010) reported that C. sedna is a polyphagous species that may feed on 16 sponge species. 

Felimare agassizii (Bergh, 1894)

(Fig. 3E)

Material examined: 2 organisms (9, 32 mm), Entrega (CNMO8276, CNMO8281). Four organisms (32-55 mm), Arrocito (CNMO8279, CNMO8280, CNMO8282, CNMO8283). One organism (14 mm), Montosa (CNMO8275).

Distribution: from the Gulf of California, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: Verdín-Padilla et al. (2010) reported that F. agassizii has a polyphagous diet feeding on 9 sponge species.

Family Cadlinidae Bergh, 1891

Cadlina sp.

(Fig. 3F)

Material examined: 5 organisms (4-5 mm), Entrega. One organism (3 mm), Arrocito. Two organisms (4 mm), Conejos. Photographic record only.

Distribution: from Baja California, Mexico to Panama (Behrens et al., 2022).

Diagnosis: oval translucent white body with 4-5 rounded yellow glands around the mantle at each side of the body, simulating an inner semi-oval. The rhinophores are white with a red band in the centre. 

Remarks: the specimens were found on white sponges. Specimens resemble Cadlina sp. in Camacho-García et al. (2005), Bertsch and Aguilar Rosas (2016) and Behrens et al. (2022). This undescribed species has been reported in several locations for the Pacific east coast such as Islas Tres Marías (Hermosillo, 2009), Revillagigedo (Hermosillo & Gosliner, 2008), Bahía de Banderas (Hermosillo, 2011), Acapulco (Flores-Rodríguez et al., 2017) and Costa Rica (Camacho-García et al., 2005).

Family Dendrodorididae O’Donoghue, 1924 (1864)

Doriopsilla janaina Er. Marcus and Ev. Marcus, 1967

(Fig. 3G)

Material examined: 1 organism (18 mm), Entrega (CNMO8022). Two organisms (13, 28 mm), Conejos (CNMO8014, CNMO7985). 

Distribution: from Baja California, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: 1 specimen was found on brown algae. Although there are reports of the gregarious behaviour of these animals (Hermosillo et al., 2006), they were found individually in our surveys. 

Suborder Cladobranchia

Family Hancockiidae MacFarland, 1923

Hancockia californica MacFarland, 1923

(Fig. 3H)

Material examined: 3 organisms (5-11 mm), Arrocito (CNMO7968).

Distribution: from Big Lagoon, California to Costa Rica (Behrens et al., 2022).

Remarks: the specimens were found on green algae. Even though the 3 specimens were collected on the same algae, they presented colour variations in their bodies, from transparent brown to reddish-brown. 

Family Flabellinidae Bergh, 1889

Coryphellina marcusorum (Gosliner & Kuzirian, 1990)

(Fig. 3I)

Material examined: 6 organisms (6-18 mm), Montosa (CNMO7976).

Distribution: from Isla San Diego, Baja California to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: specimens were observed at more than 15 m depth. Animals feed on hydroids of the genus Eudendrium (Camacho-García et al., 2005).

Samla telja (Ev. Marcus & Er. Marcus, 1967)

(Fig. 4A)

Material examined: 1 organism (9 mm), Arrocito (CNMO7971). One organism (8 mm), Montosa (CNMO7967).

Distribution: from Puerto Peñasco, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: organisms found on hydroids during daytime as reported by Behrens (2022).

Family: Cuthonidae Odhner, 1934

Cuthona sp. 1

(Fig. 4B)

Material examined: 1 organism (3 mm), Arrocito. Photographic record only.

Distribution: from Puerto Vallarta, Mexico to Islas Catalinas, Costa Rica (Camacho-García et al., 2005).

Diagnosis: translucent whitish body with several opaque white spots. The cerata are globose and have longitudinal yellowish lines with a reddish colour on the base. The smooth rhinophores and the oral tentacles are white coloured on the tips and have a reddish-brown band on the base.

Remarks: juvenile specimen found on hydroids with S. telja individuals. Our species diagnosis match Cuthona sp. 3 in Camacho-García et al. (2005). Before this work, this undescribed species had been reported only in 2 localities that correspond to its geographic distribution limits. 

Cuthona sp. 2

(Fig. 4C)

Material examined: 1 organism (5 mm), Agustín. Photographic record only.

Distribution: San Agustín, Bahías de Huatulco, Oaxaca (this study).

Diagnosis: light orange body with multiple little white spots. The pericardial area is swollen and has a distinctive white patch. The cerata colour base is brown with several little yellow dots. The smooth rhinophores have orange freckles and an orange band near the tips. The oral tentacles are shorter than the rhinophores and have 2 distinctive orange bands, 1 on the base and the other near the centre.

Remarks: the animal was found near egg masses, possibly just laid by the observed specimen. The observed characters in the organism allowed its identification only to the genus level. Specimen diagnosis did not match any recorded species in the literature.

Family Aeolidiidae Gray, 1827

Bulbaeolidia sulphurea Caballer and Ortea, 2015

(Fig. 4D)

Material examined: 4 organisms (4-12 mm), Agustín (CNMO7975, CNMO7995, CNMO8001).

Distribution: from Puerto Vallarta, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: 1 specimen was found on green algae. This species feeds on anemones (Behrens et al., 2022).

Anteaeolidiella chromosoma (Cockerell & Eliot, 1905)

(Fig. 4E)

Material examined: 8 organisms (15-22 mm), Agustín (CNMO7973, CNMO8002, CNMO8005).

Distribution: from Morro Bay, California to Islas Galapagos, Ecuador (Camacho-García et al., 2005).

Remarks: specimens were found under rocks and near coral polyps.

Anteaeolidiella ireneae Carmona et al., 2014

(Fig. 4F)

Material examined: 1 organism (21 mm), Entrega (CNMO8017).

Distribution: from Isla Socorro, Mexico to Panama (Carmona et al., 2014a).

Remarks: feeds on anemones (Behrens et al., 2022).

Baeolidia moebii Bergh, 1888

(Fig. 4G)

Material examined: 1 organism (27 mm), Agustín. Photographic record only.

Distribution: from the Gulf of California, Mexico to Panama (Hermosillo et al., 2006).

Remarks: found under bivalve shells in shallow water (2 m).

Limenandra confusa Carmona et al., 2014

(Fig. 4H)

Material examined: 2 organisms (16, 18 mm), Arrocito (CNMO8033).

Distribution: from the Gulf of California, Mexico to Costa Rica (Carmona et al., 2014c).

Remarks: Carmona et al. (2014c) report that this species feeds on small anemones.

Spurilla braziliana MacFarland, 1909

(Fig. 4I)

Material examined: 1 organism (24 mm), Arrocito. Photographic record only.

Distribution: from Baja California Sur, Mexico to Colombia (Behrens et al., 2022).

Remarks: Carmona et al. (2014b) confirmed that S. braziliana is a widespread species and its presence in the Pacific east coast is probably due to human introduction. 

Family Facelinidae Bergh, 1889

Favorinus elenalexiarum García and Troncoso, 2001

(Fig. 5A)

Material examined: 1 organism (4 mm), Entrega. 1 organism (12 mm), Arrocito. Photographic record only.

Distribution: from the Gulf of California, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: specimens found near Aplysia egg masses.

Phidiana lascrucensis Bertsch and Ferreira, 1974

(Fig. 5B)

Material examined: 1 organism (9 mm), Entrega (CNMO7998). Two organisms (8, 12 mm), Arrocito (CNMO7969, CNMO8003). One organism (24 mm), Montosa (CNMO8036). Five organisms (10-21 mm), Conejos (CNMO7980, CNMO8035). 

Distribution: from Baja California, Mexico to Panama (Behrens et al., 2022).

Remarks: organisms found under rocks, often observed near specimens of Chiton albolineatus. 

Cohort Tectipleura Schrödl, Jörger, Klussmann-Kolb and Wilson, 2011

Subcohort Euopisthobranchia Jörger, Stöger, Kano, Fukuda, Knebelsberger and Schrödl, 2010

Order Umbraculida, Odhner, 1939

Family Tylodinidae Gray, 1847

Tylodina fungina Gabb, 1865

(Fig. 5C)

Material examined: 2 organisms (6, 8 mm), Arrocito (CNMO7994, CNMO8030).

Distribution: from Baja California, Mexico to Islas Galapagos, Ecuador (Hermosillo et al., 2006).

Remarks: Tylodina fungina has a highly specialist diet, feeding on the sponge Aiolochroia thiona and Aplysina gerardogreeni (Behrens et al., 2022; Verdín-Padilla et al., 2010).

Order Cephalaspidea Fischer, 1883

Family Haminoeidae Pilsbry, 1895

Haminoea sp. 

(Fig. 5D)

Material examined: 2 organisms (8, 10 mm), Arrocito (CNMO7958). 

Distribution: from Bahía de Banderas, Mexico to Peru (Behrens et al., 2022).

Diagnosis: body oval. White cream body, with irregular dark brown patches. The animals have an inverted “V” stain in the cephalic area between the eyes. The body is surrounded by multiple orange and light brown dots.

Remarks: specimens found on marine brown cyanobacteria. This species was firstly identified as the Indo-Pacific species Lamprohaminoea ovalis by Valdés and Camacho-García (2004). However, recently phylogenetic analysis has shown that the specimens in the Pacific east coast are an undescribed species more related to the Atlantic and Pacific species of the genus Haminoea (Oskars & Malaquias, 2019, 2020).

Family Aglajidae Pilsbry, 1895

Navanax aenigmaticus (Bergh, 1893)

(Fig. 5E)

Material examined: 5 organisms (30-44 mm), Entrega (CNMO7996, CNMO7999, CNMO8007). Two organisms (21, 33 mm), Arrocito (CNMO8032). 

Distribution: from Baja California, Mexico to Chile (Behrens et al., 2022).

Remarks: collected specimens presented different body colour variations from black, brown, and pink with cream or whitish spots. According to Ornelas-Gatdula et al. (2012), colouration differences on N. aenigmaticus could be probably influenced by environmental factors.

Order Aplysiida

Family Aplysiidae Lamarck, 1809

Aplysia cf. cedrosensis

(Fig. 5F)

Material examined: 1 organism (22 mm), Conejos (CNMO7978).

Distribution: from Bahía de los Angeles, Baja California to Playa Conejos, Bahías de Huatulco, Oaxaca.

Remarks: juvenile specimen found under rocks near red algae. Our diagnosis matched the species A. cedrosensis in Hermosillo et al. (2006). However, Behrens et al. (2022) state that this species might be a synonym of the California black sea hare Aplysia vaccaria. Before this work, the southern distribution of A cedrosensis in the Pacific east coast was reported for Parque de la Reina, Acapulco (Flores-Rodríguez et al., 2017), approximately 440 km northeast from Playa Conejos, Oaxaca.

Aplysia hooveri Golestani et al., 2019

(Fig. 5G)

Material examined: 72 organisms (3-16 mm), Entrega (CNMO7961-7963, CNMO7986, CNMO8010, CNMO8021, CNMO8026, CNMO8034). Two organisms (6, 7 mm), Arrocito (CNMO7972). Five organisms (5-12 mm), Conejos (CNMO7982, CNMO7993). 

Distribution: from Baja California, Mexico to Islas Galapagos, Ecuador (Valdés, 2019).

Remarks: this species was highly abundant in some localities of the study area; it was usually associated with red and brown algae. 

Dolabella cf. auricularia (Lightfoot, 1786)

(Fig. 5H)

Material examined: 1 organism (180 mm), Agustín. One organism (210 mm), Entrega. Photographic record only.

Distribution: from the Gulf of California, Mexico to Ecuador (Zamora-Silva & Naranjo-García, 2008).

Remarks: found on algae at approximately 10 m depth. According to Behrens et al. (2022) there is molecular evidence that confirms that Dolabella auricularia is a species complex.

Dolabrifera nicaraguana Pilsbry, 1896

(Fig. 5I)

Material examined: 1 organism (14 mm), Entrega (CNMO7960).

Distribution: from Bahía de las Cruces, Baja California to Tumbes, Peru (Valdés et al., 2018).

Remarks: cryptic specimen found on rhodolith beds. 

Phyllaplysia padinae Williams and Gosliner, 1973

(Fig. 6A) 

Material examined: 4 organisms (8-18 mm), Conejos (CNMO8277, CNMO8268, CNMO8269).

Distribution: from the Gulf of California, Mexico to Islas Galapagos, Ecuador (Camacho-García et al., 2005).

Remarks: the specimens were found attached to algae of the genera Padina and Caulerpa.

Stylocheilus rickettsi (MacFarland, 1966)

(Fig. 6B)

Material examined: 29 organisms (5-29 mm), Entrega (CNMO7959, CNMO7997, CNMO8008, CNMO8018, CNMO8023, CNMO8024, CNMO8027). Five organisms (6-38 mm), Arrocito (CNMO7957, CNMO7970). 

Distribution: from Baja California, Mexico to Islas Galapagos, Ecuador (Bazzicalupo et al., 2020).

Remarks: the specimens were found on rocks and on different unidentified green, red, and brown algae. 

Subcohort Panpulmonata Jörge et al., 2010

Superorder Sacoglossa Ihering, 1876

Family Oxynoidae Stoliczka, 1868 (1847)

Lobiger cf. souverbii Fischer, 1857

(Fig. 6C)

Material examined: 1 organism (6 mm), Entrega (CNMO8015). One organism (9 mm), Conejos (CNMO7983). 

Distribution: Baja California Sur, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022). 

Remarks: specimens were found associated with algae of the genus Caulerpa as mentioned in the literature (Behrens et al., 2022; Camacho-García et al., 2005). Due to the original description of L. souverbii in the Caribbean region, this sacoglossan is suspected to be a different species.

Oxynoe aliciae Krug et al., 2018

(Fig. 6D)

Material examined: 6 organisms (4-9 mm), Entrega (CNMO8009, CNMO8016).

Distribution: from Baja California Sur, Mexico to Islas Galapagos, Ecuador (Behrens et al., 2022).

Remarks: specimens found as hosts on the algae Caulerpa as mentioned by Krug et al. (2018). 

Family Plakobranchidae Gray, 1840

Elysia cf. pusilla (Bergh, 1871)

(Fig. 6E)

Material examined: 3 organisms (8-11 mm), Arrocito (CNMO8004). Ten organisms (8-10 mm), Conejos (CNMO7977, CNMO8000, CNMO8013). 

Distribution: from Mexico to Costa Rica (Behrens et al., 2022).

Remarks: cryptic specimens were found attached to algae of the genus Halimeda as mentioned in the literature (Behrens et al., 2022; Camacho-García et al., 2005). This species is presumed to be different from E. pusilla, which was originally described in the Indo-Pacific region (Behrens et al., 2022).

Elysia sp. 1

(Fig. 6F)

Material examined: 4 organisms (6-10 mm), Arrocito. Twenty-seven organisms (5-14 mm), Conejos. Photographic record only.

Distribution: from Bahía de Banderas, Mexico to Panama (Hermosillo et al., 2006).

Diagnosis: elongated olive-greenish body. The rolled rhinophores are yellow whitish with light brown patches. The parapodia are strongly folded with an opening in the centre. Some specimens have a white spot in the base of the rhinophores. 

Remarks: specimens were found associated with algae of the genus Halimeda. Our diagnosis resembles the species Elysia sp. 1 in Camacho-García et al. (2005) and Hermosillo et al. (2006). There are numerous records of this undescribed species in the Pacific east coast: Bahía de Banderas (Hermosillo, 2011), Ixtapa, Guerrero (Hermosillo & Behrens, 2005), Papagayo and Parque de la Reina, Acapulco (Flores-Rodríguez et al., 2017), Playa Avellanas and San Pedrillo, Costa Rica (Camacho-García et al., 2005) and Panama (Hermosillo et al., 2006). 

Elysia sp. 2

(Fig. 6G)

Material examined: 4 organisms (13-24 mm), Entrega. Two organisms (19, 22 mm), Arrocito. 8 organisms (12-24 mm), Conejos. Photographic record only.

Distribution: from Islas Revillagigedo, Mexico to Costa Rica (Behrens et al., 2022).

Diagnosis: elongated light-greenish body with several white and dark green specks. The rhinophores are smooth, large, and rolled. The convoluted parapodia are folded with several rounded whitish papillae on the edges. Adult specimens have a purple-pinkish colouration along the margin of the parapodia and in the basis of the rhinophores.

Remarks: some specimens were found on red and green algae of the genus Halimeda and Caulerpa. Our diagnosis matched the species Elysia sp. 2 in Camacho-García et al. (2005) and Elysia sp. in Behrens et al. (2022). This undescribed species was previously documented in multiple locations in the Pacific east coast: Bahía de Banderas (Hermosillo, 2011), Islas Revillagigedo (Hermosillo & Gosliner, 2008), Isla Clipperton (Kaiser, 2007), Ixtapa, Guerrero (Hermosillo & Behrens, 2005) and Costa Rica (Camacho-García et al., 2005). 

Family Limapontiidae Gray, 1847

Placida cf. dendritica (Alder and Hancock, 1843)

(Fig. 6H)

Material examined: 3 organisms (3-5 mm), Entrega (CNMO8019).

Distribution: from the Gulf of California to La Entrega, Bahías de Huatulco, Oaxaca.

Remarks: specimens were found on algae of the genus Bryopsis. Before this work, the southern distribution of P. dendritica in the Pacific east coast was known for Bahía de Banderas, Nayarit (Hermosillo, 2011), approximately 1,200 km northeast from La Entrega, Oaxaca. It is suspected that P. dendritica might be a species complex that encompasses 2 different species in the region (Behrens et al., 2022).

Family Hermaeidae H. Adams and A. Adams, 1854

Polybranchia mexicana Medrano et al., 2018

(Fig. 6I)

Material examined: 1 organism (48 mm), Conejos (CNMO7992).

Distribution: from Baja California, Mexico to Islas Galapagos, Ecuador (Medrano et al., 2018).

Remarks: specimen found under rocks at daylight supporting the reports of the nocturnal habits of the species (Behrens et al., 2022).

Caliphylla sp.

(Fig. 6J)

Material examined: 4 organisms (5-14 mm), Entrega. Photographic record only.

Distribution: from La Entrega, Mexico to Islas Galapagos, Ecuador.

Diagnosis: elongated translucent green body with multiple little dark green and white dots. The bifid rhinophores, the head and the cerata have visible dark green ramified digestive branches. The elongated cerata are flattened and pointed. Some specimens may have a white speck between the eyes and in the posterior part of the head.

Remarks: some specimens were found on algae of the genus Bryopsis. Our diagnosis coincides with the undescribed species Caliphylla sp. in Camacho-García et al. (2005), which has been previously reported in a few localities from Costa Rica and Ecuador.

Hermaea sp.

(Fig. 6K)

Material examined: 1 organism (5 mm), Entrega. Photographic record only.

Distribution: from La Entrega, Oaxaca, Mexico to Playa Real, Guanacaste, Costa Rica.

Diagnosis: cream coloured body with multiple dark green flecks. Rhinophores are auriculate. The arrow-head shape cerata are covered with several white dots and red-brownish ramified digestive branches are visible throughout.

Remarks: the specimen was found on filamentous red algae. Our diagnosis resembles the species Hermaea sp. 3 in Camacho-García et al. (2005), which has been previously reported in Costa Rica.

Discussion

In this study we added 48 sea slug records, increasing by 83% the knowledge of sea slugs’ diversity for Oaxaca, from 10 to 58 species (Table 2). Also, the records presented in this study represent almost 11% of the total sea slug species previously known for the Eastern Pacific, from Alaska to Peru (Behrens et al., 2022). Among the sea slug fauna from Oaxaca, the order Nudibranchia is the most diverse encompassing more than half of the registered species, which is a general trend observed worldwide and in other localities from the Tropical Eastern Pacific (TEP) (García-Méndez & Camacho-García, 2016; Gosliner, 1991; Hermosillo, 2004; Spalding et al., 2007), and might be explained due to phylogenetic, historical and functional variables within the group (Bertsch, 2010). In contrast with other localities on the Pacific coast of Mexico (Table 3), the total number of species recorded in Oaxaca is similar to those reported for Isla Tres Marías, Nayarit (52 spp.), which is a relatively lower species richness compared with other works that involve higher sampling effort and/or more extensive study areas (Angulo-Campillo, 2005; Bertsch, 2014; Hermosillo, 2011; Hermosillo & Behrens, 2005). Moreover, the percentage of shared species between Revillagigedo, Colima and Oaxaca is higher (57.1%) than in other localities; however, this amount could be inaccurate due to the underestimated sea slug diversity in Islas Revillagigedo pointed out by Hermosillo and Gosliner (2008).

Overall, almost all the new sea slug records in this study are endemic to the Panamic biogeographic province (Briggs & Bowen, 2012), with some exceptions previously remarked that are also distributed on the Western Atlantic and/or the Indo-Pacific regions. Most of these exceptions belong to species complex that have not been resolved yet (Behrens et al., 2022), but others have a widespread natural distribution, or they have been introduced possibly by humans’ influence, as it has been suggested for S. braziliana (Carmona et al., 2014b). Interestingly, all the species previously reported for Oaxaca by Rodríguez-Palacios et al. (1988) (see Table 2) are not distributed in the Panamic province nor any warm waters of the Pacific east coast. Therefore, these records should be treated with caution as these species might have been misidentified due to the lack of specific field guides and accessible literature related with the sea slug fauna for this region in the past. 

Coral reef communities, including those inhabiting the TEP, are one of the most thriving habitats for sea slugs, as they encompass a net of biological associations that increase their diversity and inherent productivity (Sanvicente-Añorve et al., 2012; Sreeraj et al., 2013). In general, we found a higher species richness in localities with greater heterogeneity in their substrate composition (Table 1), possibly providing more habitats for sea slugs to succeed in this area. San Agustín, which is inside a Natural Protected Area, did not show a higher species richness compared to other localities. However, the number of species in this locality is underestimated, as we could not perform algae collection as an indirect search method; additionally, other variables need to be analysed to determine whether there is a significant difference in the sea slug diversity between protected and non-protected areas.

The continuous discovery of undescribed sea slug species in the TEP, such as the ones reported in this work: Berthella sp., Dorididae sp., Doris sp., and Cuthona sp. 2 has been a common issue, even in recent years. Phylogenetic and systematic studies have helped to elucidate the status of certain sea slug taxa (e.g., Bazzicalupo et al., 2020; Golestani et al., 2019; Krug et al., 2018; Medrano et al., 2018; Valdés et al., 2018), and represent important efforts to better understand the diversity of the sea slug fauna in the TEP. Nonetheless, there are species recorded more than a decade ago, such as Cadlina sp., Cuthona sp. 1, Elysia sp. 1, Elysia sp. 2, Caliphylla sp. and Hermaea sp. that remain undescribed. In the same way, some studies have found uncertainties in described species related to their geographic distribution. For instance, Behrens et al. (2022) state that Aplysia cf. cedrosensis and Placida cf. dendritica are given names that belong to 2 or more undescribed species that inhabit different biogeographic provinces. The extension of the distribution for those species reported in this work could confirm that they are different species indeed, and further taxonomic studies regarding the description and distribution of each species need to be done.

Table 3

Studies on sea slug diversity from the Pacific coast of Mexico. Shared species refer to those species present in other localities and this study (Huatulco).

Locality	Number of species	Number of shared species (%)	Reference
Bahía de los Angeles, Baja California	117	26 (22.2)	Bertsch (2014)
Baja California Sur	117	31 (26.4)	Angulo-Campillo (2005)
Bahía de Banderas, Nayarit-Jalisco	146	44 (30.1)	Hermosillo (2011)
Islas Tres Marías, Nayarit	52	27 (51.9)	Hermosillo (2009)
Revillagigedo, Colima	42	24 (57.1)	Hermosillo and Gosliner (2008)
Colima, Michoacán and Guerrero	76	38 (50)	Hermosillo and Behrens (2005)
Acapulco, Guerrero	63	27 (42.8)	Flores-Rodríguez et al. (2017)

This study updates the knowledge of the sea slug fauna of Oaxaca and the southern Pacific coast of Mexico; however, many unexplored localities in this region still need to be studied. Since this region could be a potential hotspot of marine biodiversity (Bastida-Zavala et al., 2013), further efforts to find sea slugs are needed. Future samplings involving SCUBA diving on different habitats such as lagoons, mangroves, and rocky shores at different times of the day may help to increase this inventory. This work also contributes to the biological inventory of Parque Nacional Huatulco, which is essential to determine future perspectives in the conservation planning and management of this and other Natural Protected Areas (Bezaury-Creel & Gutiérrez, 2009).

Acknowledgements

We thank Parque Nacional Huatulco for allowing us the entrance to perform the surveys in the locality of San Agustín; OGR acknowledges Comisión Nacional de Becas de Educación Superior, SEP for the financial support as an undergraduate student; XGV acknowledges Consejo Nacional de Ciencia y Tecnología (Conacyt) her PhD scholarship (CVU: 564148). We thank A. Valdés, P. Krug and S. Medrano, who helped with the identification of some organisms, and E. Naranjo-García for providing a space to work in her laboratory. We thank F. Pérez, M. Pérez, and M. A. Arriaga for their hospitality and support in performing this study; to the staff of “Buceo Anfibios Huatulco” and all the people who helped us in the surveys, especially E. Molina, L. Jiménez, A. García and A. Barrera.

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José Alan Herrera-García ^a, Mahinda Martinez ^{a, b,} *, Pilar Zamora-Tavares ^c, Ofelia Vargas ^c, Luis Hernández-Sandoval ^{a, b}

^a Universidad Autónoma de Querétaro, Facultad de Ciencias Naturales, Av. de las Ciencias, s/n, 76230 Juriquilla, Querétaro, Mexico 

^b Universidad Autónoma de Querétaro, Facultad de Ciencias Naturales, Biología, Laboratorio Nacional de Identificación y Caracterización Vegetal, Av. de las Ciencias, s/n, 76230 Juriquilla, Querétaro, Mexico 

^c Universidad de Guadalajara, Centro Universitario de Ciencias Biológicas y Agropecuarias, Instituto de Botánica, Departamento de Botánica y Zoología, Laboratorio Nacional de Identificación y Caracterización Vegetal, Av. Ing. Ramón Padilla Sánchez, 45200 Zapopan, Jalisco, Mexico 

*Corresponding author: mahinda@uaq.mx (M. Martínez)

Received: 20 May 2024; accepted: 19 February 2025

Abstract

Some bromeliads form a compact rosette that accumulates detritus and water, known as phytotelma. The phytotelma is a lentic ephemeral aquatic environment that forms diverse communities with complex trophic levels. Pseudalcantarea grandis, a saxicolous plant, forms a phytotelma. To understand the importance of P. grandis as a eukaryotic diversity reservoir in arid zones, we collected water samples from 5 plants growing in a dry canyon in Zimapán, Hidalgo, Mexico. We analyzed them through metabarcoding of the ITS1 (Internal Transcribed Spacer) and the partial 5.8S gene. We used the Ion Torrent PGM platform for the sequencing, and the taxonomic assignation for the amplicons was made with BLAST in Genbank at NCBI. We found 26 phyla and 543 genera, 80% of which belonged to Ascomycota, Basidiomycota, Blastocladiomycota Chytridiomycota, Glomeromycota, Mucoromycota, and Zoopagomycota phyla. The remaining 20% was composed of 19 phyla belonging to other kingdoms. Photosynthetic organisms were represented by the phyla Bacillariophyta, Charophyta, Chlorophyta, and Ochrophyta. The vascular plants do not live in the tank but constitute the debris sustaining the large number of decomposers. The trophic levels in the tank were detritus, micro- and macro-decomposers, filter feeders, photosynthesizers, micro-predators, aquatic volume predators, surface predators, and parasites.

Keywords: Aquatic; Ephemeral; Arid zone; Phytotelma

Diversidad de eucariotes y niveles tróficos dentro de la bromelia tanque Pseudalcantarea grandis en una zona árida detectados por metabarcoding de ADN ambiental

Resumen

Algunas bromelias forman rosetas compactas que acumulan detritus y agua. Esta acumulación se conoce como fitotelma, un hábitat acuático léntico y efímero con comunidades diversas y niveles tróficos complejos. Psedalcantarea grandis es una planta saxícola y forma un fitotelma. Para entender la importancia de P. grandis como reservorio de diversidad acuática en una zona árida, colectamos muestras de agua de 5 plantas en un cañón de Zimapán, Hidalgo, México y las analizamos por metabarcoding del ITS1 (Internal Transcribed Spacer) y una región parcial del gen 5.8S. La secuenciación se hizo en la plataforma Ion Torrent PGM. Asignamos la identidad taxonómica de los amplicones utilizando BLAST de Genbank. Encontramos 26 phyla y 543 géneros, 80% pertenecen a los phyla fúngicos Ascomycota, Basidiomycota, Blastocladiomycota Chytridiomycota, Glomeromycota, Mucoromycota, y Zoopagomycota. El 20% restante está compuesto por 19 phyla de otros reinos. Los organismos fotosintéticos estuvieron representados por los phyla Bacillariophyta, Charophyta, Chlorophyta y Ochrophyta. Otros organismos fotosintéticos que corresponden a plantas vasculares no viven dentro del tanque, pero forman la hojarasca que mantiene a los descomponedores. Los niveles tróficos en el tanque fueron detritus, micro y macrodescomponedores, filtradores, fotosintetizadores, microdepredadores, depredadores del volumen de agua, depredadores de superficie y parásitos.

Palabras clave: Acuático; Efímero; Zona árida; Fitotelma

Introduction

The Bromeliaceae family is comprised of almost 3,700 species distributed mostly in tropical areas of the Americas (Gouda et al., 2024). They are herbaceous perennial monocots with leaves arranged in rosettes, many of which are epiphytes (Ramírez-Murillo et al., 2004; Rzedowski, 2006). There are 442 species in Mexico (Espejo-Serna & López-Ferrari, 2018) that frequently grow in nutrient and mineral-deficient environments (Bernal et al., 2006; Ramírez-Murillo et al., 2004). Some species form compact rosettes with absorbent trichomes in their interior that allow the accumulation of solids rich in nutrients and water, known as phytotelma (Benzing, 2000; Goffredi et al., 2011).

Phytotelmata are lentic aquatic environments, mostly ephemeral that last less than 3 months (Mogi, 2004). They are freshwater habitats for diverse communities including viruses, Archaea, and bacteria (Brouard et al., 2013, Goffredi et al., 2011). Among the eukaryotes, aquatic mosses, green algae, diatoms, protists, fungi, insects, amphibians, and crustaceans have been documented (Benzing, 2000; Brandt et al., 2017; Kitching, 2001; Ramos & do Nascimento Moura, 2019; Rodríguez-Núñez et al., 2018; Simão et al., 2020). 

Bromeliads that form phytotelma accumulate essential mineral elements, which are the main nitrogen source for the plant (Kitching, 2001). The presence of detritivores and predators is related to the nitrogen concentration in the leaves. Predators increase nutrient flux from the leaf litter of nearby plants to the bromeliad (Benzing & Renfrow, 1974; Ngai & Srivastava, 2006; Nievola et al., 2001; Takahashi & Mercier, 2011).

Unlike terrestrial and aquatic communities in which plants and algae are the main nutrient resources, in the phytotelma leaf litter and invertebrate remains play that role. In trophic networks inside the phytotelma, protists and rotifers are considered as micro-predators that also consume organic particles. Macroinvertebrates can consume the detritus, filter feeders, aquatic predators, and surface predators, whereas bacteria and fungi are the main decomposers that obtain energy directly from the detritus (Brouard et al., 2012; Mogi, 2004).

Gomes et al. (2015) characterized the enzymatic activity of the fungal community inside the bromeliad tank of Vriesea minarum in Brazil. Using cultivation techniques they identified 36 species, 22 of which were Basidiomycota and 14 were Ascomycota. The most relevant genera were Cryptococcus, Candida, and Aureobasidium. These organisms contain enzymes that degrade vegetal material. 

Nutrient intake for the plant is further facilitated by insects (Ngai & Srivastava, 2006). For example, odonatan larvae ingest detritivores, contributing to the nitrogen cycle within the bromeliad by defecating. Leachates from defecation release nitrogen in a form available to the bromeliad and create a suitable niche for other microorganisms providing substrata (Benzing & Renfrow, 1974).

Pseudalcantarea grandis (Schltdl.) Pinzón & Barfuss (Fig. 1A) is a saxicolous tank bromeliad that reaches 2.5 m in height and forms a highly ramified inflorescence in March and April. It is distributed from Central Mexico (Guanajuato, Querétaro, Hidalgo) toward the south (Puebla, Oaxaca, and Chiapas) and into Honduras. It is considered a MegaMexico II endemic species (Espejo-Serna et al., 2010). At the locality of Las Adjuntas, in Hidalgo, the plant is known as “tinaja”, “jarilla”, and “soluche de agua” because its tank can store water. The species grows in crags of the main rivers in the northeastern portion of the region known as Bajío. It grows at elevations ranging from 400 to 1,600 m asl (Espejo-Serna et al., 2010; Rzedowski, 2006). 

Metabarcoding studies describing the communities associated with phytotelma have mostly focused on specific groups such as vertebrates (Brozio et al., 2017), ciliates (Simão et al., 2017), or bacteria (Louca et al., 2017; Rodríguez-Núñez et al., 2018). In Pseudalcantarea grandis, 297 bacteria genera were found, with Proteobacteria (37%), Actinobacteria (19%), and Firmicutes (15%) comprising the highest percentage (71%). The main metabolic functions were aerobic chemoheterotrophy and fermentation. However, rare biosphere bacteria were also found, which could favor micro-ecosystem resilience and resistance (Herrera-García et al., 2022). Comprehensive sequencing of the eukaryotic diversity inside tank bromeliads has been performed in tropical zones (Simão et al., 2020), but not arid areas. The objectives of our study were to describe the eukaryotic diversity in Pseudalcantarea grandis phytotelma to understand which vascular plants form the litter, and to infer the putative trophic levels of the phytotelmata in an arid zone.

Materials and methods

Water samples were collected during the 2018 rainy season at Las Angosturas Canyon, Zimapán, Hidalgo, Mexico (20°50.933’ N, 99°26.7’ W, 900 m asl, see Herrera-García et al. [2022] for a location map) within the Queretano-Hidalguense arid zone, which has been described as a high-diversity and endemicity area (Hernández-Magaña et al., 2017; Hernández & Bárcenas, 1995; Rojas et al., 2013). This arid zone is considered the southernmost portion of the Chihuahuan Desert floristic province. It consists mostly of arid valleys and depressions surrounded by mountains (Hernández & Gómez-Hinostrosa, 2005). 

Las Angosturas Canyon is approximately 12 km long with a mixture of xerophytic scrub and tropical deciduous forest (Fig. 1B). Bromeliads grow on vertical crags with different amounts of the surrounding vegetation. Therefore, the tanks are frequently filled with leaf litter. We selected individuals that were accessible enough to be collected by a rappel and were more than 50 cm in diameter. The associated plants are listed in Table 1. To collect the water inside the bromeliads we used Nest® cell scrapers to scratch the inside of each tank, and the water in the bromeliads was vigorously shaken to obtain a homogeneous sample. Water volumes of 50 to 100 ml were collected using 10 ml sterile serological pipettes. The samples were stored in 50 ml conical Falcon tubes, transported on dry ice, and stored at -79 °C until processing. Physicochemical water parameters were not determined.

Table 1

Las Angosturas Canyon floristic inventory. Plants directly above the sampled individuals are noted in the second column. Vouchers and photographs are noted in the third column.

Family	Species	Association	Reference at QMEX
Amaryllidaceae	Zephyranthes clintiae Traub		A. Herrera 30
Amaryllidaceae	Zephyranthes concolor (Lindley) Bentham & Hook.		A. Herrera 7
Anacardiaceae	Rhus terebinthifolia Schltdl. & Cham.		A. Herrera 21
Anacardiaceae	Pseudosmodingium virletii (Baill.) Engl.		L. Hernández 5029
Apocynaceaea	Asclepias curassavica L.		A. Herrera 20
Apocynaceaea	Vallesia glabra (Cav.) Link		A. Herrera 4
Arecaceae	Brahea dulcis (Kunth) Mart.		Photographic record
Asparagaceae	Agave xylonacantha Salm-Dyck	surrounding	A. Vega 50
Asparagaceae	Dasylirion longissimum Lem.		L. Hernández 4511
Asparagaceae	Yucca filifera Chab.		L. Hernández 4828
Asparagaceae	Yucca queretaroensis Piña		Photographic record
Asteraceae	Baccharis salicifolia (Ruiz y Pav.) Pers.		M. Martínez 6056
Asteraceae	Cirsium sp.		A. Herrera ND
Asteraceae	Gnaphalium luteo-album L.		A. Herrera ND
Asteraceae	Gochnatia hypoleuca (DC.) A. Gray	surrounding	A. Herrera ND
Bignoniaceae	Tecoma stans (L.) Juss.		Photographic record
Boraginaceae	Cordia boissieri A. DC.		M. Martínez 7144
Bromeliaceae	Hechtia glomerata Zucc.	surrounding	S. Zamudio 14469
Bromeliaceae	Hechtia tillandsioides (André) L. B. Smith	surrounding	Photographic record
Table 1. Continued
Family	Species	Association	Reference at QMEX
Bromeliaceae	Pseudalcantarea grandis (Schltdl.)Pinzón & Barfuss		A. Herrera 1
Bromeliaceae	Tillandsia recurvata L.	surrounding	M. Martínez 8220
Burseraceae	Bursera morelensis Ram.	surrounding	Photographic record
Cactaceae	Astrophytum ornatum (DC.) Britton & Rose		Photographic record
Cactaceae	Cylindropuntia imbricata (Haw.) F. M. Knuth ssp. cardenche (Griffiths) U. Guzmán		A. Herrera 12
Cactaceae	Coryphanta sp.		Photographic record
Cactaceae	Echinocereus pentalophus Lem.		Photographic record
Cactaceae	Echinocactus platyacanthus Link & Otto		M. Figueroa 12
Cactaceae	Ferocacutus histrix (DC.) G.E.Linds.		Photographic record
Cactaceae	Mammilaria elongata DC.	surrounding	Photographic record
Cactaceae	Mammillaria longimamma DC.		Photographic record
Cactaceae	Myrtillocactus geometrizans (Mart. ex Pfeiff.) Console	surrounding	Photographic record
Cactaceae	Neobuxbaumia polylopha (DC.) Backeberg		Photographic record
Cactaceae	Opuntia imbricata (Haw.) F. M. Kunth		Photographic record
Cactaceae	Opuntia microdasys (Lehm.) Pfeiff.		Photographic record
Cactaceae	Opuntia rastrera F.A.C. Weber	surrounding	Photographic record
Cactaceae	Stenocereus queretaroensis (F.A.C.Weber ex Mathes.) Buxb.		Photographic record
Cactaceae	Strombocactus disciformis (DC.) Britton & Rose		Photographic record
Cannabaceae	Celtis pallida Torr.		A. Herrera 2
Capparaceae	Capparis incana Kunth		A. Herrera 5
Convolvulaceae	Ipomoea rzedowskii E. Carranza		A. Herrera 17
Crassulaceae	Echeveria secunda Booth	surrounding	A. Herrera 26
Crassulaceae	Pachyphytum sp.		A. Herrera ND
Crassulaceae	Sedum sp.	surrounding	A. Herrera ND
Euphorbiaceae	Cnidoscolus tubulosus (Muell. Arg.) I.M. Johnst.		A. Herrera 9
Euphorbiaceae	Acalypha monostachya Cav.		A. Herrera 15
Euphorbiaceae	Croton ciliato-glandulifer Ort.		L. Hernández 5029
Euphorbiaceae	Jatropha dioica Sessé ex Cerv.		A. Herrera ND
Euphorbiaceae	Ricinus communis L.		A. Herrera ND
Fabaceae	Acacia berlandieri Benth.	surrounding	Photographic record
Fabaceae	Albizia occidentalis Brandegee		Photographic record
Fabaceae	Bauhinia sp.		Photographic record
Fabaceae	Lysiloma microphylla Bentham		A. Herrera 36
Fabaceae	Mimosa leucaenoides Bentham	surrounding	Photographic record
Fabaceae	Mimosa martindelcampoi F. G. Medrano		Photographic record
Fabaceae	Mimosa puberula Bentham		A. Herrera 3
Fabaceae	Pithecellobium dulce (Roxb.) Bentham		Photographic record
Fabaceae	Neltuma laevigata (Humb. & Bonpl. ex Willd.) Britton & Rose		M. Martínez ND
Fabaceae	Vachellia farnesiana (L.) Willd. & Arn.		A. Herrera 10
Table 1. Continued
Family	Species	Association	Reference at QMEX
Fouquieriaceae	Fouquieria splendens Engelm.	surrounding	Photographic record
Lentibulariaceae	Pinguicula aff. moctezumae Zamudio & R.Z. Ortega		Photographic record
Lythraceae	Heimia salicifolia (Kunth) Link		A. Herrera 13
Onagraceae	Hauya elegans DC.	surrounding	A. Herrera 14
Malpighiaceae	Mascagnia macroptera (Moc. & Sessé ex DC.) Nied.		A. Herrera 8
Malvaceae	Pseudobombax ellipticum (Kunth) Dugand		A. Herrera ND
Malvaceae	Malvaviscus arboreus Cav.		M. Martínez 5317
Myrtaceae	Psidium guajava L.	surrounding	Photographic record
Papaveraceae	Argemone ochroleuca Sweet		M. Martínez 6713
Plantaginaceae	Rusellia polyedra Zucc.		A. Herrera 27
Platanaceae	Platanus mexicana Moric.		A. Herrera 16
Poaceae	Arundinaria sp.		A. Herrera ND
Poaceae	Cenchrus sp.		A. Herrera ND
Poaceae	Cynodon dactylon (L.) Pers.		M. Martínez 3197
Poaceae	Eragrostis sp.		E. Carranza 5251
Primulaceae	Samolus ebracteatus Kunth		A. Herrera 11
Pteridaceae	Argyrochosma formosa (Liebm.) Windham		A. Herrera 23
Pteridaceae	Notholaena affinis (Mett.) T. Moore		A. Herrera 24
Pteridaceae	Notholaena jacalensis Pray		A. Herrera 25
Pteridaceae	Pellaea sp.		A. Herrera 34
Ranunculaceae	Clematis drummondii Torr. & A.Gray		M. Martínez 4434
Rhamnaceae	Karwinskia subcordata Schlecht.		A. Herrera 22
Rubiaceae	Nernstia mexicana (Zucc. & Mart. ex DC.) Urb.		A. Herrera 32
Salicaceae	Neopringlea integrifolia (Hemsl.) S. Watson		A. Herrera 19
Salicaceae	Salix humboldtiana Willd.		Photographic record
Sapindaceae	Dodonaea viscosa (L.) Jacq.		A. Herrera ND
Sapindaceae	Sapindus saponaria L.		A. Herrera ND
Sapindaceae	Serjania sp.		A. Herrera ND
Selaginellaceae	Selaginella lepidophylla (Hook. & Grev.) Spring.	surrounding	Photographic record
Selaginellaceae	Selaginella ribae Valdespino		A. Herrera 29
Selaginellaceae	Selaginella selowii Hieron.		A. Herrera 28
Solanaceae	Datura inoxia Miller		V. Martínez 1
Solanaceae	Nicotiana glauca Graham		Photographic record
Solanaceae	Nicotiana trigonophylla Dunal		O. García 434
Solanaceae	Physalis cinerascens (Dunal) Hitch.		L. Hernández 3769
Solanaceae	Physalis philadelphica Lam.		A. Herrera 38
Solanaceae	Solanum lycopersicon L.		Photographic record
Taxodiaceae	Taxodium mucronatum Ten.		M. Martínez 3237
Urticaceae	Urera sp.		H. Rubio 302
Zygophyllaceae	Morkillia acuminata Rose & Painter	surrounding	A. Herrera 18

DNA extraction

Water samples were homogenized and 100 ml was filtered through a 0.22 µm Millipore® nitrocellulose membrane. The membrane was frozen and macerated in liquid nitrogen. We extracted total DNA using the QIAmp DNA Extraction® kit following the manufacturer’s instructions by duplicate to obtain pseudoreplicates and verify reproducibility. DNA quality and concentration were evaluated using NanoDrop® spectrophotometry. 

Amplicon sequencing

To characterize eukaryotic diversity, we amplified a portion of the 5.8 S Internal Transcibed Spacer (ITS) with the ITS1 and ITS2 primers designated by White et al. (1990). The PCR reaction consisted of a final volume of 25 µl, that contained 2 mM of dNTP´s, 2mM µl of each primer, 0.4% DMSO, 0.4 % BSA, 2.5 mM MgCl2, 1.2 mM Buffer, 1.25 U Platinum Taq, 60ng/µl DNA and H2O. Thermocycler conditions were an initial step at 95 °C for 3 min, followed by 30 cycles at 95 °C for 1 min; 52 °C, 45 s, and 72 °C, 2 min, with a final extension step at 72 °C for 5 min. Amplicons were purified with Agencourt® AMPure® XP.

To construct the libraries, we used the Ion Plus Fragment Library kit. The presence, size, and concentration of the fragment were analyzed using Bioanalyzer 2100 with high-sensitivity DNA assay (Agilent). Libraries were quantified using real-time PCR to obtain an equimolar dilution factor to mix the 3 libraries. The template was prepared using a PCR emulsion in the Ion One Touch 2 System (Life Technologies) and quantified by fluorometry in Qubit® 3.0 (Thermo Fisher Scientific). Finally, the template was loaded onto the PGM 318TM chip using the 400-pair base fragment sequencing kit, according to the Ion PGM™ Hi‑Q™ View Sequencing Kit protocol.

We sampled the vascular plants growing at the canyon, directly above the bromeliad, and also the surrounding vegetation. Voucher specimens were deposited at QMEX herbarium. We compared the similarity of the plant inventory obtained by sequencing against the floristic list obtained by field sampling through a Sorensen coefficient analysis at the family level.

Sequencing quality was evaluated using the FastQC program. Sequences were filtered by the quality value of Phred > 20. We selected sequences larger than 100 bp in the CLC Genomic Workbench v.11.01 (QIAGEN Bioinformatics, Aarhus, Denmark) platform. In the Microbial Genomics module application, we performed an analysis based on the amplicons to aggregate the sequences in operational taxonomic units (OTUs) considering only 99% similarity among them. We deleted unique reads and chimeras. Taxonomic assignation of the amplicons was performed with BLAST in 2023 via the Genbank at NCBI database (Altschul et al., 1990). Over 90% of the OTUs had identity percentages higher than 95% and only 54 OTUs had lower percentages, ranging from 75 to 80%. We manually reviewed them and corroborated the genus of each one. Since BLAST provides determinations at the genus and species levels, we used MEGAN Community Edition V. 6.24.4 to assign kingdom, phylum, class, order, and family. We loaded the BLAST results using the lowest basal common ancestor assignation algorithm (LCA). The analysis is based on the taxonomic hierarchies recognized by NCBI and the results are displayed as a phylogenetic tree that allows simple observation of taxonomic diversity (Huson et al., 2016). We manually reviewed the classifications and to corroborate the taxonomic assignations we used the classification proposed by Simpson (2006) for plants, and Tree of Life (2022) for the other eukaryotic marine taxa (such as Cnidaria). The OTUs at the genus level were used to define the total eukaryotic group diversity present in our sample.

We assigned the ecological function of each genus following the criteria of Mogi (2004) and Brouard et al. (2012). We considered 9 categories: 1) detritus formed by leaf litter and vegetal matter that serves as the main resource for the trophic network; 2) micro decomposers integrated by bacteria (Herrera-García et al., 2022); 3) macro decomposers formed by saprobiotic fungi; 4) filter feeders that use small particles including microorganisms that are in turn consumed by aquatic and surface predators; 5) photosynthetic organisms that require sunlight and serve as food for predators; 6) micro predators that feed on photosynthetic organisms, filter feeders, and micro-decomposers; 7) aquatic predators which are macroinvertebrates that live in the water column and feed mostly on algae and bacteria; 8) surface predators which are restricted to the uppermost portion of the water column and feed on protists, bacteria, and algae; and 9) parasites which are obligate vertebrate parasites that use arthropods as vectors.

Results

The 5 sampled plants had volumes that varied from 50 to 100 ml. The 2 pseudoreplicates were compared and considered as a single pool due to the similarity of the resulting OTUs. We obtained a total of 3,276,538 lectures. After quality and size filtration, the number of useful lectures was reduced to 1,284,998 representing a total of 23,948 OTUs, 762 of which could not be assigned to the species or genus taxonomic category provided by BLAST. 

The diversity of organisms living in the tank consisted of 26 phyla and 543 genera. See Supplementary material T1 for a list of assigned taxa. Fungi were dominant, as 80% of the genera belonged to Ascomycota, Basidiomycota, Blastocladiomycota Chytridiomycota, Glomeromycota, Mucoromycota, and Zoopagomycota phyla. The remaining 20% was composed of 19 phyla: Apicomplexa, Apusozoa, Arthropoda, Bacillariophyta, Bryophyta, Cercozoa, Charophyta, Chlorophyta, Ciliophora, Cnidaria, Colponemidia, Heterolobosea, Hyphochytriomycota, Metamonada, Myxomycota, Ochrophyta, Oomycota, Platyhelminthes, and Tracheophyta (Fig. 2). We identified 25 genera of photosynthetic algae from the phyla Bacillariophyta (with the genera Pseudo-nitzschia, Navicula, and Stephanodiscus), Charophyta (Staurastrum), Chlorophyta (Leskea, Didymogenes, Meyerella, Dolichomastix, Bathycoccus, Mychonastes, Trebouxia, Chamaetrichon, Hazenia, Chloroidium, and Pleurastrum), and Ochrophyta (Nannochloropsis). The other photosynthetic organisms were Bryophyta (Fontinalis, Leskea, and Thuidium).

Protists were represented by 44 genera, 4 of which are relevant to human health: Plasmodium (Apicomplexa) which causes paludism, Giardia (Metamonada) which is responsible for giardiasis, Neobalantidium (Cilliophora) that causes balantidiosis, and Spirometra (Plathelmyntes) which is responsible for sparganosis. 

Tracheophyta (vascular plants) do not live in the phytotelma but do constitute the debris that accumulates in the bromeliad. They comprised 11% of the identified genera. We found a Sorensen coefficient of 45% similarity among the methods in which 14 taxa at the family level were shared (Supplementary material F1). Arthropoda were represented by 13 genera of the Coleoptera, Diptera, Hymenoptera, Lepidoptera, Odonata, and Pocopodia orders. 

Of the eukaryotic organisms, 79% were classified as decomposers, and 7% were classified as micro-predators. In the tank, 3% were photosynthetic organisms, 2% were parasites, and the remaining 9% corresponded to the vascular plants that constitute the detritus. Tracheophyta and Bryophyta constituted the vegetal resources available to micro- and macro-decomposers, and filter feeders. Cercozoa, Apusozoa, Heterolobosea, and Colponemidia were considered micro-predators because they consume some photosynthetic organisms, filter feeders, and micro-decomposers. Ciliophora and 2 Arthropoda genera (Cyprideis and Notodromas) were some of the filter feeders. Apicomplexa, Chytridiomycota, Metamonada, Myxomycota, and Zoopagomycota were the parasites. Aquatic predators were mostly metazoans (Cnidaria and Platyhelminthes) that use filter feeders, photosynthesizers, and macro decomposers as resources. The arthropods Coleoptera, Diptera, Hymenoptera, Lepidoptera, and Odonata were part of the uppermost categories of the trophic network (surface predators). However, their exoskeletons and/or excretions become part of the tank resource or contribute to the nutrient cycling of the microecosystem (Figs. 3, 4). 

Discussion

Eukaryotic composition of the Pseudalcantarea grandis community

The abundance of fungi in the Pseudalcantarea tank appeared to correlate with their function in the trophic network. Fungi are the most important degrading group, responsible for organic decomposition and nutrient recycling in forests, aquatic ecosystems, and the phytotelma (Costa & Gusmão, 2015; Grossart et al., 2019; Grothjan et al., 2019). Fungi also have multiple functions in aquatic environment interactions that favor antagonistic and symbiotic members of the community. They can be predators, parasites, or food for heterotrophic protists. Some can use organic matter, pollen, or zooplankton exoskeletons (Zoopagomycota, Chytridiomycota) (Grossart et al., 2019). Vegetal matter decomposition enhances detritus quality for detritivores degrading vegetal polysaccharides into monosaccharides through enzymatic reactions which are then easily digested by microorganisms (Krauss et al., 2011). Fungi and protist interactions for vegetal matter transformation are poorly documented. A symbiotic relationship between them is unknown and difficult to study because of the microscopic scale at which they occur (Grossart et al., 2019).

The fungal diversity found in the Pseudalcantarea grandis tank was high compared to that reported in previous studies of other Bromeliaceae species. We found 436 genera, whereas Gomes et al. (2015) identified 36 genera using cultivation techniques. Other papers already pointed out that metagenomic studies detect higher diversity levels than other techniques (Simão et al., 2020). The 36 genera found by Gomes et al. (2015) were also found in P. grandis. Cryptococcus, Candida, and Aureobasidium have specific enzymatic activity in plant material degradation, which suggests that degradation reactions by these organisms are frequent in the phytotelma. The primers used in our study were developed for fungi (White et al., 1990), therefore they might be overrepresented.

We found 44 protist genera in the phytotelma. Some have mixotrophic nutrition, in that they obtain their energy through photo- and heterotrophy depending on the environmental conditions in which they grow (Jones, 2000). In aquatic environments where light is available but dissolved organic carbon (DOC) is scarce, photosynthetic organisms are better represented. Low light and high DOC favor heterotrophic organisms (Jones, 2000). The latter condition is what was present in the sampled tanks. Therefore, the development of photosynthetic protists is not favored because of the large and abundant bacterial community that competes for elements such as phosphorus (Brouard et al., 2012; Herrera-García et al., 2022). In the rainy season, the tank of P. grandis is surrounded by vegetation that intercepts light and deposits leaf litter, therefore favoring the conditions for fungi and decomposers (Grossart et al., 2019; Herrera-García et al., 2022, Kitching, 2000).

Direct observations and sampling of the phytotelma of tropical zones have revealed Diptera, Odonata, Oligochaeta, Ostracoda, beetles, copepods, pseudoscorpions, scorpions, isopods, Lepidoptera, hemipterans, homopterans, orthopterans, and arachnids in tank bromeliads (Cutz-Pool et al., 2016; Marino et al., 2013). Using environmental DNA with specific primers, amphibians (Brozio et al., 2017) and ciliates (Simão et al., 2017) have been found in tank bromeliads with high water availability. These results suggest that aridity and strong water seasonality in our study area were responsible for the lack of amphibians and the low arthropod and ciliate diversity we found.

Phytotelma seasonality is an important factor. In a rainy forest, the annual precipitation is 3,000 mm and rain is present for 280 days, therefore the water in the tank lasts longer (Brouard et al., 2012). In contrast, in our location, the annual precipitation is 391 mm and the phytotelma is available only through the rainy season from May to June (INFAED, 2012). The presence of Plasmodium is relevant since its most common vector is Aedes, suggesting that at some point during the phytotelma duration, the mosquito is in contact with the water, completing the parasite life cycle (Williams, 2007).

The absence of vertebrates is characteristic of phytotelma communities (Mogi, 2004). One exception is in rainforest bromeliad, where bromeliad tadpoles can be found. We did not find vertebrates. Strong seasonality was probably the reason for their absence. Not all OTUs could be assigned to the species or genus taxonomic category at 99% identity level we used. We did not find an identity for 762 of the 23,984 sequences, possibly because the sequences of these organisms are not available in the NCBI database, or because the organisms have not yet been described.

The tracheophytes found in the phytotelma could not be identified at the generic level. There are 2 possible explanations: the reads were short (150 bp) and therefore insufficient, or the genera growing at the canyon are not in the GenBank NCBI (National Center for Biotechnology Information) database. However, 30 families of vascular plants were detected, 16 of which correspond to the families found by field collection.

Trophic structure of the Pseudalcantarea grandis tank

We propose 9 trophic levels for tank bromeliads in arid zones, —2 more than those suggested by Mogi (2004), and 3 more than those suggested by Brouard et al. (2012). Detritus is the main nutrient source. Macrodecomposers process leaf litter into small organic matter particles, including their waste. The particles are then stored in the phytotelma where filterers and invertebrates process them. Dead organisms, feces, and leaf litter stored at the bottom of the tank are used by bacteria and other microorganisms such as fungi to assimilate nutrients (Brouard et al., 2012). We found that 48 plant genera (45 Tracheophyta, and 3 Bryophyta) constitute the detritus, and although Brouard et al. (2012) recognized organic litter as a resource, they did not identify the organisms that provided it. The large amount of detritus was due to the type of surrounding vegetation, which was a tropical deciduous forest in this study. In P. grandis macro decomposers are fungi of the Ascomycota, Basidiomycota, and Myxomycota phyla. We concur with Brouard et al. (2012) that ciliates are filterers. Mogi (2004) did not consider microorganisms to be autotrophs, but Brouard et al. (2012) included that category, and in P. grandis algae constitute this level. Insects were categorized as surface predators in their diagram; they include the odonate family Coenagrionidae, which includes the 2 genera we found in P. grandis, Ischnura, and Agriocnemis (Brouard et al., 2012). Neither Mogi (2004) nor Brouard et al. (2012) included parasites at a trophic level, but some of the genera found in P. grandis are obligate parasites, mostly from the Alveolata kingdom, which has birds and mammals as hosts but uses arthropods as vectors.

We found 26 phyla with 543 genera in the phytotelma of P. grandis growing in an arid zone. The identified diversity suggested that the organisms that inhabit these small ephemeral water bodies are adapted to prolonged dry spells and develop quickly when the phytotelma has water. The biota was mostly composed of fungi (over 80% of the diversity) that specialize in plant detritus degradation. Water bodies shelter aquatic groups that cannot exist in areas outside the P. grandis phytotelma. It is easier to assess the diversity of organisms within a tank than to comprehend their interactions. The trophic network proposed for eukaryotes indicates that they fulfill different functions.

As final considerations, we conclude that the analyzed phytotelma had a large detritus accumulation, and water was present briefly. Most of the diversity belonged to fungi (80%) because of the large amount of plant detritus in the tank. Photosynthesizers were scarce but included 25 algal genera and 3 Bryophyta. We found 45% Sorensen coefficient similarity between the plant detritus and the specimens collected with herbarium specimens. We also found a low arthropod and ciliate diversity, and the tank also harbors protist genera, some of which have medical implications. We found 9 trophic levels in the tank. Unlike tropical areas, in which algal production can support non-detrital food webs, in our arid zone system, detritus degradation was the main energy source. 

Acknowledgments

Funding was provided by Conahcyt through grant 293833 for the Laboratorio Nacional de Identificación y Caracterización Vegetal. Diana Velázquez designed and executed Figure 3. Two anonymous reviewers helped to improve the manuscript with their comments.

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César Marco Aurelio Jurado-Vargas ^a, *, José Cruz-de León ^b, José Tulio Mendez-Montiel ^c

^a Universidad Michoacana de San Nicolás de Hidalgo, Facultad de Biología, Laboratorio de Investigación en Invertebrados, Ciudad Universitaria, Av. Fco. J. Mujica s/n, 58030 Morelia, Michoacán, México

^b Universidad Michoacana de San Nicolás de Hidalgo, Facultad de Ingeniería en Tecnología de la Madera, Laboratorio de Conservación y Preservación de la Madera, Ciudad Universitaria, Av. Fco. J. Mujica s/n, 58030 Morelia, Michoacán, México 

^c Universidad Autónoma Chapingo, Dirección de Ciencias Forestales, Carretera Federal México-Texcoco Km 38.5, 56230 Texcoco, Estado de México, México

*Autor para correspondencia: cjurado@umich.mx (C.M.A. Jurado-Vargas)

Recibido: 02 abril 2024; aceptado: 11 marzo 2025

http://zoobank.org/urn:lsid:zoobank.org:pub:4737516A-ED56-4106-B714-86B23F67BCF4 

Resumen

A partir de un análisis morfológico y taxonómico de adultos del género Calymmaderus Solier, 1849 (Ptinidae: Dorcatominae), se describen y proponen 2 nuevas especies para el Neotrópico en México: C. robustus sp. nov. y C. semioblongus sp. nov. Ellas se distinguen de las 7 especies reportadas y reconocidas actualmente para México con base en su coloración, pubescencia y estrías elitrales laterales. Se describe la estructura genital de machos.

Palabras clave: Carcoma; Edificios históricos; Xilófagos

Two new Neotropical species of the genus Calymmaderus (Coleoptera: Ptinidae), associated to structural wood in Taretan, Michoacán, Mexico

Abstract

Through morphological and taxonomic analysis of adults of the genus Calymmaderus Solier, 1849 (Ptinidae: Dorcatominae), 2 new Neotropical species in Mexico are described and proposed: C. robustus sp. nov. and C. semioblongus sp. nov. They are distinguished from the 7 species currently reported and recognized for Mexico based on their coloration, pubescence and lateral elitral striae. The genital structure of males is described.

Keywords: Woodworm; Historic buildings; Xylophages

Introducción

El género Calymmaderus incluye especies de escarabajos xilófagos de distribución mundial, para el cual se reconocen 89 especies americanas (Blackwelder, 1945; Lüer y Honour, 2017; Toskina, 2000; White, 1974, 1982, 1983, 1984), de las que 7 especies se encuentran en México: C. dejeani (Pic, 1905), C. donckieri (Pic, 1904), C. oblongus (Gorham, 1883), C. pupatus (Gorham, 1883), C. semirufus (Champion, 1913), C. sharpi (Gorham, 1886) y C. subvestitus (Champion, 1913) (White, 1983; Zaragoza et al., 2016). El estudio de las especies de este género es complicado debido a la falta de revisiones taxonómicas, lo limitado de sus descripciones diagnósticas y la carencia, en la mayoría de los casos de ilustraciones de los órganos genitales, estudio que le da mayor robustez a la determinación de especies (Bercedo et al., 2008; Lüer y Honour, 2017; Viñolas, 2018). Los miembros de Calymmaderus se distinguen de las demás especies de Ptinidae por su cuerpo alargado y oblongo, antenas con los 2 últimos segmentos de la clava estrechamente unidos, coxas protorácicas expuestas ventralmente, un lóbulo metasternal bifurcado que aloja al último segmento antenal cuando el cuerpo está en reposo y el número de suturas elitrales laterales (Arango, 2012; Español, 1992; White, 1971). Las especies de este género son de importancia económica al dañar la madera estructural de edificios históricos y bienes culturales. La especie más conocida en México es C. oblongus (Gorham, 1883), a la que varios autores señalan como asociada al deterioro de la madera (Bercedo et al., 2008; Cibrián et al., 1995; Jurado-Vargas, 2020; Jurado-Vargas y Cruz, 2010, 2020; Jurado-Vargas et al., 2003; Pichardo et al., 2017; White, 1974).

El presente estudio tiene como objetivo describir 2 especies nuevas de Calymmaderus, que fueron encontradas en la localidad de Taretan, Michoacán, México, ubicada en la Zona de Transición Mexicana, que es un área de alta diversidad y endemismos de artrópodos y presenta la mayor mezcla biótica entre elementos neárticos y neotropicales (Halffter, 2017). Ambas especies representan taxones morfológicamente diferentes a las especies descritas en la literatura conocida para este grupo. White fue el último investigador que publicó datos sobre especies de Calymmaderus y otros géneros de escarabajos de la familia Ptinidae americanas entre 1981 y 1984, dejando pendientes algunos ejemplares con estatus incierto aún por resolver. En este trabajo presentamos la descripción y propuesta de las 2 nuevas especies de Calymmaderus referidas.

Materiales y métodos

El área de estudio corresponde a la localidad de Taretan, Michoacán, México (1,140 m snm, 19°20’3.32” N, 101°55’3.02” O) (fig. 1), que presenta un clima cálido A (w), con precipitación media anual de 1,240 mm y temperatura media anual 25° C (INEGI, 2010).

Se recolectaron 463 ejemplares del género Calymmaderus de la techumbre de madera del templo de San Ildefonso por medio de 2 trampas de luz blanca, durante el periodo de emergencia de los adultos en los meses de junio-agosto del 2017 y 2018. Los ejemplares se fijaron en una solución de alcohol al 80%. La determinación de los ejemplares se realizó usando descripciones y claves de identificación para el género y especies de Calymmaderus (Arango, 2012; White,1971, 1974, 1982, 1983, 1984). En el proceso, la preparación de los órganos se realizó mediante la técnica de montaje permanente en laminillas con resina sintética (Gaviño et al., 2004). 

La descripción de los caracteres morfológicos de los ejemplares adultos se realizó mediante un microscopio estereoscópico Nikon a 45 aumentos. Se obtuvieron imágenes de los ejemplares en posición dorsal, lateral y ventral, además de metasterno, abdomen y antenas, mediante un microscopio estereoscópico Carl Zeiss (modelo Axio Zoom V16). Las imágenes de las preparaciones de la estructura genital se lograron a través de un microscopio compuesto Marca Leica a 100 aumentos y una cámara Panasonic adaptada a la distancia focal del ocular. Para obtener detalles adicionales de la morfología externa de los ejemplares, se tomaron imágenes por medio del microscopio electrónico de barrido modelo JSM-6400.

Se determinaron las medidas de largo y ancho de 30 ejemplares de cada una de las especies que se describen con un vernier digital, con las que se realizó un análisis estadístico descriptivo básico de los parámetros utilizados para su descripción. La fenología de emergencia de adultos se determinó mediante gráficos de los registros mensuales de captura de los ejemplares adultos en los periodos de trabajo. Los holotipos y paratipos de las 2 especies se depositaron en la Colección de Invertebrados (Insectos Xilófagos) de la Facultad de Biología de la Universidad Michoacana (CIFBUM-XIL).

Resultados

Descripciones

Calymmaderus robustus Jurado-Vargas sp. nov. (figs. 2-7, 14)

http://zoobank.org/urn:lsid:zoobank.org:act:1C158DC5-F92E-4A6C-A089-09DC362DFC6E 

Diagnosis. Cuerpo robusto en vista lateral, 1.9 veces más largo que ancho, color negro uniforme mate, pubescencia blanquecina decumbente y abundante, separada menos de su longitud, élitros paralelos después de la base hasta 3/4 partes; antenas desde la base de café rojizo a café negruzco en los últimos segmentos; metasterno con menos pubescencia que resto del cuerpo. Lóbulo mestasternal con muesca profunda en forma característica de U.

Holotipo ♂, largo 3.8 mm, ancho 2.1 mm. Cuerpo robusto en vista lateral, 1.9 veces más largo que ancho (figs. 2, 3), color negro mate, excepto en palpos maxilares y labiales, que presentan el segmento basal café oscuro y los otros segmentos café rojizo. Tarsos de color café rojizo. Pubescencia decumbente blanquecina, abundante, uniforme en todo el cuerpo. Puntuaciones elitrales de la superficie ampliamente distribuidas, no alineadas e intercaladas con otras puntuaciones diminutas en todo el cuerpo y separadas 1.0 veces su diámetro. Cabeza: ojos grandes y abultados, distancia interocular 1.1 veces el diámetro vertical de un ojo. Clava antenal ligeramente mayor (1.1 veces) que el resto de los segmentos; 9º segmento de la clava antenal ligeramente más largo que los segmentos 10º y 11º (fig. 6). Pronoto: acampanado, 1.9 veces más ancho que largo; borde anterior curvo y márgenes laterales redondeados. Superficie densamente punteada y pilosa, con las puntuaciones laterales equidistantes y separadas en general el equivalente a su diámetro; en el disco las puntuaciones son más separadas (1-2 veces su diámetro). Élitro: puntuaciones del disco y laterales separadas de 1 a 2 veces su diámetro (algunas veces un poco más separadas y alargadas); puntuaciones más cercanas a la base elitral más aglutinadas y puntuaciones laterales hacia la parte distal del élitro más alargadas; ranura lateral visible después de la base del tercer esternito hasta el ápice (fig. 4), abarcando menos de la mitad del élitro. Metasterno: convexo, más ancho que largo, carinas bien delimitadas; puntuaciones cercanas al proceso metasternal más pequeñas, separadas 1.0 veces su diámetro; puntuaciones se agrandan más hacia la parte media y a los lados, separadas de 1.0 a 1.5 veces su diámetro; en la parte media y hasta el borde posterior son más pequeñas y menos densas; surco longitudinal corto y no muy marcado (fig. 5). Abdomen: primero, segundo y quinto esternitos más o menos del mismo largo; segundo con borde posterior ligeramente curvo en centro; tercero un poco más corto que anteriores, y cuarto más corto que el resto (fig. 3). Edeago: longitud: 1.1 mm; anchura 0.48 mm. Parámeros bifurcados apicalmente, con borde apical redondeado; piezas accesorias de los parámeros 4.2 veces más largos que su ancho, con pilosidad larga desde la mitad hasta su extremo. Endófalo: ápice más esbelto que la base, terminando en punta con un diente curvo en forma de gancho, saco interno provisto de por lo menos doce espinas transversales a lo largo; las 2 más cercanas a la base 2 veces más largas que las demás y de posición paralela; 2 espinas al centro más pequeñas y entrecruzadas con las espinas basales más largas (fig. 14).

Paratipo ♀. Aspecto general del cuerpo similar al macho. Largo 4.1 ancho 2.3 mm, ojos evidentemente más grandes y abultados que el macho, distancia interocular 1.3 veces el diámetro vertical de un ojo. Tercera y cuarta sutura de los esternitos curvada a los lados.

Resumen taxonómico

Etimología. El epíteto de la especie robustus deriva de la forma robusta del tórax en vista lateral (mucho más alta que el abdomen), que caracteriza y diferencia a los ejemplares de esta especie en comparación con las especies conocidas.

Material examinado. Holotipo ♂: CIFBUM-XIL Núm. 224. México: Michoacán. Taretan. 19.VII.2017. C. Jurado Col. Trampa de luz blanca, madera estructural de pino. 19°20’3.32” N. 101°55’3.02” O. 1,140 m snm. Paratipos: 28; 13 ♂, 17 ♀. CIFBUM-XIL Núms. 226-252, con los mismos datos del holotipo. 

Comentarios taxonómicos

Variabilidad: largo 3.1-4.2 mm (media 3.72) ancho 1.7-2.3 mm (media 2.08). Macho más pequeño que la hembra (de 3.1 a 3.7 mm; tabla 1). Ojos más pequeños, distancia interocular de 1.0 a 1.1 veces del diámetro vertical del ojo; último esternito normal, con sutura recta. Hembra: 3.8 a 4.2 mm, ojos más grandes y abultados que el macho, distancia interocular de 1.2 a 1.3 veces el diámetro vertical de un ojo; último esternito con sutura ligeramente curvada en el centro. Especie con morfología poco variable; color general negruzco mate, pubescencia blanquecina uniforme, algunas veces con pubescencia menos abundante en el metasterno.

Especie de hábitos nocturnos asociada con madera estructural de Pinus sp.; los ejemplares se capturaron en el interior de la techumbre de madera del templo de “San Ildefonso” Taretan, con trampa de luz. La emergencia de adultos inicia la última semana de junio, con máximo a mediados de julio y disminuye la última semana de julio; la emergencia es rara en agosto. Cohabita con otras especies de Ptinidae y termes, reinfestando durante muchos años la madera del mismo sitio (fig. 7). 

Tabla 1

Medidas y variación de largo y ancho de machos y hembras adultos de Calymmaderus robustus sp. nov. 

Medidas	Promedio (mm)	Error estándar	Rango (mm)
N (hembras) = 13 N (machos) = 17	Machos	Hembras	Machos	Hembras	Machos	Hembras
Longitud	3.438	3.947	0.055	0.032	3.1 – 3.7	3.8 – 4.2
Ancho	1.931	2.194	0.036	0.020	1.2 – 2.1	2.1 – 2.3

De acuerdo con la clave de identificación usada para especies de Calymmaderus (White, 1983), los ejemplares son definitivamente diferentes a las descripciones de las especies mexicanas descritas, considerando la variación en el conjunto de los siguientes caracteres. Todas las especies citadas para México presentan cuerpo oblongo o semiesférico (C. sharpi) en vista lateral; en cuanto al color, los ejemplares descritos de todas las especies van de café rojizo o café oscuro a casi negro, mientras que la coloración del integumento de la especie que se describe es negro mate uniforme. En cuanto a la pubescencia, a diferencia de las especies conocidas en las que tiene tonalidades amarillentas a grisáceas, la especie propuesta presenta pubescencia blanquecina, que contrasta fuertemente con el color del cuerpo. La mayoría de las poblaciones de las especies comparten un tamaño de longitud y ancho similares, excepto C. donckieri y C. subvestitus, que son notablemente más pequeñas. La carencia de información acerca de la armadura genital de las especies no permite hacer comparaciones al respecto. Sin embargo, se presenta la comparación de la armadura genital del macho de las 2 especies propuestas en este documento y C. oblongus, especie mejor conocida de amplia distribución en México (figs. 14-16), lo que respalda la diferenciación entre ellas. 

Calymmaderus semioblongus Jurado-Vargas sp. nov. (figs. 8-13, 15)

http://zoobank.org/urn:lsid:zoobank.org:act:7170D451-739D-4507-96C3-A826C5BA4E13 

Diagnosis. Forma oblonga, esbelto en vista dorsal, 2.1 veces más largo que ancho, élitros paralelos después de la base hasta 3/4 partes de su extensión; superficie del cuerpo de coloración homogénea de café oscuro a casi negro; pubescencia amarillenta densa y decumbente, separada menos del largo de su longitud; puntuaciones laterales de élitros más agrandadas en la base que en el resto del élitro.

Holotipo ♂, largo 3.8 mm, ancho 1.8 mm. Coloración de café oscuro a casi negro; superficie brillante; antenas y tarsos con la misma coloración del cuerpo; palpos labiales y palpos maxilares café rojizo. Cuerpo oblongo y alargado, 2.1 veces más largo que ancho (figs. 8, 9). Vestidura: pubescencia amarillenta, densa, decumbente y abundante en todo el cuerpo, puntuaciones uniformes en todo el cuerpo, excepto las puntuaciones laterales del élitro que se presentan de forma aglutinada e irregular (fig. 10). Cabeza: ojos pequeños separados 1.4 veces el diámetro vertical de un ojo. Clava antenal 1.3 veces más larga que el resto de los segmentos, noveno segmento de la clava antenal del mismo largo que el décimo y undécimo (fig. 12). Pronoto: acampanado 1.6 veces más largo que ancho, convexo dorsalmente; margen lateralmente redondeado, con un reborde lateral y borde anterior regularmente curvo, no proyectado; disco no prominente; pubescencia uniforme, superficie densamente punteada. Élitro: puntuaciones elitrales irregulares, no alineadas en hileras, separadas de 1 a 2 veces su diámetro, en el disco elitral también a la misma distancia; puntuaciones laterales de la base un poco más grandes y algunas están separadas el equivalente a su diámetro; en el tercio posterior del élitro las puntuaciones son más pequeñas; puntuaciones basales laterales aglutinadas formando una estría a la altura del segundo esternito, que continua hasta el ápice ya bien marcada como ranura basal; por encima otra línea no muy marcada forma otra estría también a nivel del segundo esternito, la ranura ocupa ligeramente más de la mitad del élitro hasta el ápice. Metasterno: ancho del lóbulo metasternal, escasamente mayor que su longitud, muesca profunda, ápice del lóbulo muy arqueado en forma de V; puntuaciones más grandes cerca del lóbulo y a lo largo del borde anterior del metasterno, separadas el equivalente a su diámetro; a los lados las puntuaciones son más espaciadas de 1.0 a 1.5 veces su diámetro, algunas separadas hasta 2 veces su diámetro; del centro hasta el borde posterior, las puntuaciones son más pequeñas y escasas, separadas de 1.0 a 2.0 veces su diámetro; surco longitudinal ligeramente marcado y extendido hasta la mitad del metasterno (fig. 11). Abdomen: segundo ventrito más largo que el resto; el primero y tercero de largo similar, el cuarto es el más estrecho, el quinto presenta una concavidad paralela al ventrito cerca del borde posterior (fig. 9). Edeago: longitud 0.65 mm; anchura 0.41 mm, parámeros bifurcados apicalmente, el más grande de apariencia bilobulada en el extremo; piezas accesorias de los parámeros 4.5 veces más largos que anchos, con pilosidades desde la mitad hasta el ápice. Endofalo: ápice redondeado, saco interno con un par de dientes paralelos al endofalo en la base; sobre estos dientes, hay 2 dientes adicionales curvos con aspecto de media luna (fig. 15).

Paratipo ♀. Aspecto general del cuerpo similar al macho. Largo 4.1 mm. Ancho 1.9 mm. Distancia interocular 1.4 veces el diámetro vertical del ojo, segunda y tercera suturas abdominales curvadas en el centro, quinta sutura ligeramente curvada a los lados.

Resumen taxonómico

Etimología. El epíteto semioblongus es aplicado por la similitud morfológica externa con la especie oblongus, conocida para varias localidades de México.

Material examinado. Holotipo ♂: CIFBUM-XIL No. 253, Michoacán, Taretan. 18.VIII.2017. C. Jurado Col. Trampa de luz blanca, madera estructural de pino. Col. C. Jurado. 19°20’3.32” N. 101°55’3.02” 0. 1,150 m.snm. Paratipos: 29 individuos CIFBUM-XIL Núms. 255 a 282 (14 ♂ y 14 ♀), mismos datos que el holotipo.

Comentarios taxonómicos

Variabilidad: largo: 3.0-4.8 mm; ancho 1.4- 2.1 mm. Especie muy homogénea. Los machos notablemente menores que las hembras, con tamaño que varía de 3.0 a 3.7 mm (a veces hasta 3.8 mm), con ojos más pequeños y menos separados que en las hembras (distancia interocular de 0.8 a 1.2 el diámetro vertical de un ojo); área frontal algo más aplanada que la hembra, surco longitudinal del metasterno más largo que en la hembra.

Tabla 2

Medidas y variación de largo y ancho de machos y hembras adultos de Calymmaderus semioblongus sp. nov. 

Medidas	Promedio (mm)	Error estándar	Rango (mm)
N (machos) = 15  N (hembras) = 15	Machos	Hembras	Machos	Hembras	Machos	Hembras
Longitud	3.507	4.133	0.059	0.070	3.0-3.8	3.8-4.8
Ancho	1.627	1.886	0.025	0.026	1.4-1.7	1.7-2.1

Tabla 3. Medidas y variación de largo y ancho de machos y hembras adultos de Calymmaderus oblongus de la localidad de Tacícuaro, Michoacán.

Medidas	Medidas	Promedio (mm)	Error estándar	Rango (mm)
N (machos) = 15  N (hembras) = 15	n♂= 15 n♀= 15	♂	♀	♂	♀	♂	♀
Longitud	Longitud	3.507	4.133	0.060	0.070	3.0-3.8	3.8-4.8
Ancho	Ancho	1.627	1.887	0.025	0.026	1.4-1.7	1.7-2.1

Hembra: cuerpo más grande que el macho, de 3.9 a 4.8 mm, ojos más grandes y separados que el macho (distancia interocular de 1.4 a 1.6 veces el diámetro vertical del ojo), última sutura esternal más curva a los lados que en el macho. 

Especie de hábitos nocturnos asociada con madera estructural de Pinus sp. Se capturó en el interior de la techumbre de madera del templo de San Ildefonso en Taretan, Michoacán, con trampa de luz. La emergencia de los adultos inicia la última semana del mes de junio, en el mes de julio se registra la máxima emergencia con el inicio del verano, hasta disminuir en agosto con pocos ejemplares y en septiembre su emergencia es rara. Cohabita con otras especies xilófagas de Ptinidae y de termitas. Reinfesta las estructuras de madera por muchos años en el mismo sitio (fig. 13).

Calymmaderus semioblongus sp. nov. es una especie diferente a las otras especies mexicanas del género (White, 1983). Muestra similitud morfológica con la especie oblongus, pero con diferencias evidentes como la coloración del integumento (café oscuro en la especie descrita, a diferencia de oblongus que tiende más a café rojizo; sus medidas de longitud y ancho del cuerpo son menores (3.0 a 4.5 mm de longitud y ancho 1.4 a 2.1 mm, tabla 2), en comparación con C. oblongus (de 3.3 a 5.2 mm de largo y ancho de 1.6 a 2.4 mm, tabla 3). Ambas especies presentan de 2 a 3 hileras de puntuaciones laterales desde la base elitral que forman estrías; en oblongus, la estría más externa y cercana a la ranura elitral es más corta y las otras 2 estrías son menos marcadas que en la especie que se describe. La comparación de la estructura genital de machos de C. oblongus y C. semioblongus sp. nov. también mostró diferencias claras en el número y posición de espinas en el endofalo de estas 2 especies; C. oblongus presenta 2 espinas en forma de cornamenta en la parte más alejada de la base (figs. 14-16). 

Agradecimientos

Al Laboratorio de Investigación en Invertebrados de la Facultad de Biología y a la División de Estudios de Posgrado de la Facultad de Ingeniería en Tecnología de la Madera de la UMSNH, por apoyar el proyecto de insectos xilófagos en madera estructural en Michoacán. A Mauricio Quesada, jefe del laboratorio Nacional de Síntesis Ecológica, ENES, UNAM, Unidad Morelia, por facilitar el uso del microscopio Axiostar Zoom V16, para la toma de imágenes de ejemplares. A las autoridades eclesiásticas del Templo de la “Asunción” de Taretan (en especial al párroco Francisco Javier Valencia Durón), por permitir el trabajo de campo en el inmueble. A Roberto Sibaja por la elaboración y ubicación del mapa del sitio. Finalmente agradecer a José Fernando Villaseñor Gómez, de la Facultad de Biología, quien colaboró en la última revisión del texto, con sugerencias acertadas y positivas.

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Francisco Lorea-Hernández*

Instituto de Ecología, A.C., Carretera antigua a Coatepec 351, Col. El Haya, 91073 Xalapa, Veracruz, Mexico 

*Corresponding author: francisco.lorea@inecol.mx (F. Lorea-Hernández)

Received: 27 February 2024; accepted: 21 January 2025

Abstract

Following a detailed morphological survey of the Licaria collections in several herbaria, various taxonomic entities not recognized before were detected. Here, 11 new species of Licaria from Mesoamerica are described and illustrated. Possible relations to other species in the genus are commented.

Keywords: Lauraceae of Central America; Lauraceae of Mexico; Licaria of Central America; Licaria of Mexico

Nuevas especies de Licaria (Lauraceae) de Mesoamérica

Resumen

Como resultado de un análisis morfológico detallado de las colecciones del género Licaria en diferentes herbarios, se detectaron varias entidades taxonómicas no reconocidas previamente. Aquí se describen e ilustran 11 especies nuevas del género Licaria de la región mesoamericana. Se comentan además las posibles relaciones con otras especies del género.

Palabras clave: Lauraceae de Centroamérica; Lauraceae de México; Licaria de Centroamérica; Licaria de México

Introduction

Licaria is an endemic genus to the Americas represented mostly by medium-sized to large tree species that grow principally in the intertropical region of this part of the world. The hermaphrodite, perigynous flowers with only the third whorl of stamens fertile, whose anthers are bisporangiate, as well as the fruit often seated on a double rimmed cupule, constitute the combination of characters that distinguishes the genus among the Lauraceae. Licaria was last revised by Kurz (2000), who recognized 38 species, including 12 in the Mesoamerican area. That work is an adaptation with minor changes of Kurz doctoral dissertation (Kurz, 1983). Since then, in the lapse of 40 years, subtracting new combinations and synonyms, 31 new species have been described, adding 9 to the flora of Mesoamerica (Burger & van der Werff, 1990; Gómez-Laurito & Cascante, 1999; Gómez-Laurito & Estrada, 2002; Hammel, 1986; van der Werff, 1988, 2009). The impulse given to field work with the onset of the Flora Mesoamericana project resulted in a significant increment of herbarium specimens during the last 2 decades of the past century, but Kurz did not see most of them; those specimens have been frequently the source of new species described afterwards in Licaria. Furthermore, the expanded collections have improved our knowledge about variation of morphology within species and consequently have led to better circumscriptions of them. On the other hand, it is important to say that Kurz missed part of the diversity of Licaria because he relied mainly on what the big museums of Europe and the USA have, but did not search in several of the smaller regional herbaria; some of which preserve important specimens that support the recognition of several species that have elsewhere been synonymized or validate larger geographic distributions of taxa.

Contrary to what is found in most American genera of the Lauracae, Licaria presents a rather wide variation in flower morphology, e.g., in orientation of tepals, extent of stamens fusion, presence and extent of fusion of glands, presence of staminodes, and shape and position of anther openings. These features, together with the presence and type of hairiness on flower parts, as well as position and structure of inflorescence, constitute the basis for the recognition of species in the genus (Kostermans, 1937; Kurz, 2000; Mez, 1889). In the course of the revision of Licaria for the Flora Mesoamericana project, a number of herbarium specimens with distinctive character combinations were found, which had not been recorded previously in the genus. Differences are mainly in flower morphology, but complementary vegetative features or other data emphasize their singularity. In several cases, there are no additional specimens, but the collection on which the description is based. This situation has not been considered a drawback since, as is discussed in every case, the peculiarity of the plants is so patent that there is little chance to think they represent part of the variability of any other known species. The case strengthens the idea that we have as yet an incomplete picture of the extent of diversity and distribution in many taxa. Field work is still important and necessary.

Materials and methods

Specimens identified as Licaria (as well as unidentified, putative lauraceous material) collected in the Mesoamerican area from different herbaria (A, CAS, DS, ENCB, F, GH, HEM, K, MEXU, MICH, MO, NY, P, TEX, US, XAL) were carefully analyzed, both for vegetative and reproductive morphology. Particular attention was paid to floral characters; flowers were dissected using a Zeiss, Stemi DV4 stereomicroscope. Information of vegetative characters was collected directly from dry specimens, while floral morphology was surveyed in rehydrated material. Groups of specimens sharing similar morphology were matched with keys and descriptions of currently accepted species, in order to apply the correct names to them. To prevent misinterpretations, all available type specimens (either from herbaria cited above or accesible on Global Plants database (http://plants.jstor.org/search?plantNam) were also considered during the process of identification. Specimens that did not fit any of the known species were evaluated to determine their singularity; those whose floral and vegetative features combined do not overlap with that of accepted taxa are here proposed as new species. Following the methodology mentioned above, description of vegetative characters is based on dry specimens, while floral characters are described as they look in rehydrated material. In order to apply a standardized nomenclature to characters, applicable information in Radford et al. (1974) was used; particularly, base and apex of leaves, pubescence, tridimensional form of flowers, and shape of tepals were described according to that text.

Descriptions

Licaria breedlovei Lorea-Hern. sp. nov. (Figs. 1, 3)

Diagnosis. Trees similar to L. alata Miranda, but different because of the convex flanks of the base of the leaves, sparsely sericeous lower leaf surface, obloid flowers, densely tomentose tepals abaxially, partially pubescent staminal filaments abaxially and adaxially, densely tomentose hypanthium outside, densely hirsute-tomentose inside, and sparsely puberulent style.

Trees up to 15 m tall; twigs sericeous, soon glabrate, brownish gray or dark brown, lenticellate, conspicuously ridged, the ridges narrow, wing-like (at least when dry), each ridge originates at one extreme of petiole insertion, acropetal. Buds densely sericeous-tomentose. Leaves alternate, petioles 9-10.5(-12) mm long, bi-marginate above, glabrous, round below, sericeous, glabrescent, blades (17.5-)23-28 × (3.5-)4.5-7 cm, narrowly elliptic, pinninerved, secondary veins 11-13 pairs, upper leaf surface glabrous, lower surface sparsely sericeous, glabrescent, leaf apex acuminate, leaf base obtuse, somewhat conduplicate, the basal flanks of the blade convex and projected above the midvein. Inflorescences (7.5-)10-16.5 cm long, axillary to tiny, deciduous bracts, on the proximal section of new twigs, paniculate, sericeous-tomentose, peduncle (2.5-)4-6.5 cm long, sometimes glabrescent, pedicels 0.5-1.2(-2.5) mm long, densely tomentose, hairs yellowish-gray to yellowish-brown. Flowers obloid, tepals conspicuously inflexed, outer tepals 0.8-1 × 1.7-2 mm, very widely ovate, densely tomentose abaxially, slightly papillose toward the apex, glabrous or sometimes with scattered sericeous hairs at the base adaxially, inner tepals 0.6 × 1-1.1 mm, ovate or elliptic, tomentose abaxially, glabrous adaxially, staminodes of whorls I and II absent, stamens of whorl III 1-1.1 mm long, fused throughout, filaments pubescent at the base abaxially, glabrous adaxially or pubescent on the upper section, anthers 0.2-0.3 mm long, glabrous, with apical openings, glands ca. 0.2 mm, free, compressed, glabrous, sometimes reduced or absent, hypanthium ca. 0.8 mm deep, densely tomentose outside, hirsute-tomentose inside, pistil 1.3-1.4 mm long, ovary 0.8-1 mm long, glabrous, style sparsely puberulent. Fruits unknown.

Taxonomic summary

Type. Mexico. Chiapas: municipality of Petalcingo (actually municipality of Tila), steep slope of Ahk’ulbal Nab above Petalcingo, 1,700 m, 28 March 1981, D. E. Breedlove 50394 (holotype CAS; isotype MO 6485834).

Etymology. This species is dedicated to Dr. Dennis E. Breedlove, who spent so much effort in field work documenting the flora of Chiapas, pursuing the aim of elaborating a plant species compendium for that region of Mexico.

Distribution and habitat. So far the species is known only from the type collection, in an area covered by montane rain forest which, according to Breedlove (data from label of type specimen), had a canopy layer reaching 25 – 35 m, and species of the genera Alfaroa, Brunellia, Calatola, Hedyosmum, Matudea, Meliosma, Nectandra, Oreopanax, Quercus, and Turpinia, among others. 

Phenology. Regarding the date when the plant was collected, it must have flowers around the end of winter and early days of spring; fruit season is not known, but possibly occurs during winter, for it has been observed that maturation of fruits in most Lauraceae takes around a year after flowering. It seems that the species is not deciduous.

Conservation status. Given the deep environmental degradation that currently prevails in the region where the species is only known, it is suspected that it is critically endangered.

Remarks

The presence of erect, concave tepals, and stamens with extrorse sporangia places L. breedlovei in subgenus Licaria. It is one element in the group (here called Licaria excelsa species group) constituted by L. alata, L. excelsa Kosterm., L. minutiflora (here described), L. pergamentacea W. C. Burger, L. sarukhanii (here described), and L. tomentulosa (here described), as it shares with them morphological features like ridged twigs, large narrow-elliptic leaves (frequently reaching 28-30 cm long), and anthers with sporangia opening apically. Within the group, L. breedlovei is distinguished from the other species by the combination of somewhat conduplicate leaf-base, lower leaf surface persistently sericeous, stamens fused throughout, and sparsely pubescent style. In addition, it differs particularly from L. alata for the obloid flowers (vs. ellipsoid), tepals densely tomentose abaxially (vs. tepals glabrous abaxially), filaments pubescent abaxially and adaxially (vs. filaments glabrous), and hypanthium densely tomentose outside and inside (vs. hypanthium glabrous on both faces).

Licaria dolichopoda Lorea-Hern. sp. nov. (Figs. 2, 3)

Diagnosis. Trees, similar to L. misantlae (Brandegee) Kosterm., glabrous throughout, leaves mostly caudate, inflorescences botryoid, flowers long-pedicellate, without staminodes, and fruit cupule thick, deeply crateriform, its outer rim slightly lobed.

Trees up to 25 m; twigs glabrous, smooth or slightly ribbed, reddish-brown or grayish-brown, sparsely lenticellate. Buds glabrous. Leaves alternate, petioles (3.5-)6.5-9(-11) mm long, slightly sulcate above, rounded and smooth or slightly ribbed below, glabrous, blades (4.5-)7-11(-13) × (1.5-)2.5-4(-5) cm, elliptic or narrowly elliptic, pinninerved, secondary veins 6-11 pairs, both upper and lower leaf surfaces glabrous, leaf apex caudate, sometimes acuminate, base cuneate to obtuse. Inflorescences 2.5-5 cm long, axillary to tiny, decidous bracts, disposed on very short shoots axillary to leaves, botryoid, or apparently paniculate, due to suppression of terminal bud of the floriferous branchlet, glabrous throughout, peduncle 0.2-1.7 cm long, pedicels (6-)10-13(-16) mm long, glabrous. Flowers widely obovoid to turbinate, greenish-yellow, tepals concave, conspicuously inflexed, outer tepals 0.9-1.3 × 1.1-1.7 mm, widely ovate, blunt cuspidate, glabrous abaxially and adaxially, short-ciliate, inner tepals 0.75-1.1 × 1.1-1.2 mm, ovate to widely ovate, glabrous abaxially and adaxially, short-ciliate, staminodes of whorls I and II absent, stamens of whorl III 0.6-1 mm long, fused throughout, filaments glabrous abaxially and adaxially, sometimes with a few hairs at the base abaxially, anthers ca. 0.1 mm, glabrous, with apical openings, glands ca. 0.4 mm, free, oblong, sometimes almost square, obtuse or sub-acute, glabrous, hypanthium 0.6-0.9 mm deep, glabrous outside and inside, pistil ca. 1.5 mm long, glabrous throughout, ovary 0.7-0.9 mm long. Fruits ca. 27 × 21 mm, ovoid or ellipsoid, cupule ca. 15 × 20 mm, crateriform, clearly bimarginate, inner margin ca. 2 mm tall, erect, outer margin ca. 3.5 mm tall, oblique to slightly reflexed, shallowly lobed, thick, pedicel ca. 5.5 × 4.5 (at the base) and 7 (at the apex) mm, obconic, continuous with the cupule.

Taxonomic summary

Type. Costa Rica. Puntarenas: Reserva Forestal Golfo Dulce, Osa Peninsula, Rancho Quemado, 8°44’ N, 83°36’ W, 200-300 m, 2 May 1988, B. Hammel et al. 16790 (holotype MO; isotypes MEXU, TEX).

Paratypes. Costa Rica. Guanacaste: Cantón de Tilarán, San Gerardo Abajo, río Caño Negro, Fincas Quesada y Arce, 10°18’40’’ N, 84°50’02’’ W, 1,100-1,200 m, 5 December 1991, E. Bello & E. Cruz 4262 (XAL). Puntarenas: Cantón de Osa, Rincón, filas al margen izquierdo de Quebrada Vaquedano, 8°38’45’’ N, 83°35’25’’ W, 400 m, 21 July 1990, G. Herrera 4000 (XAL); Cantón de Osa, Aguabuena, cuenca media y superior de Quebrada Orito, Rincón, 8°42’40’’ N, 83°31’40’’ W, 400 m, 25 October 1990, G. Herrera 4510 (MEXU); Cantón de Osa, Rancho Quemado, sector oeste, Sierpe, 8°41’00’’ N, 83°35’40’’ W, 350 m, 25 August 1982, J. Marín & G. Marín 499 (MO, XAL).

Etymology. The name of this species alludes to the distinctive long pedicels that bear the flowers.

Distribution and habitat. Currently the species is known from 2 rather distant areas in Costa Rica that differ in ecological conditions. One of them, in the province of Guanacaste, is located in the Pacific foothills of the southern end of the Sierra de Guanacaste, while the other, in the province of Puntarenas, is in the lowlands of the Peninsula de Osa toward the southeastern extreme of the country. The first one is covered by pre-montane humid forest, and the second by humid tropical forest. 

Phenology. Flowering is apparently distributed in 2 peaks: May-July and October-December. Fruits are only known from October.

Conservation status. There is no information about the abundance of the species in the places where it has been collected, neither about the ecological conditions of the vegetation there. However, the distribution range in the southeast is embedded in the Reserva Forestal Golfo Dulce, by the boundaries with the Parque Nacional Corcovado. Thus, there might not be problems with the persistence of the species in that region. The other point of the species distribution is, according to satellite images, within a region with deeply transformed vegetation, although not too far from the Parque Nacional Volcán Arenal, which could hold the species in its flora.

Remarks

Because of the erect, concave tepals, and extrorse sporangia of stamens in the flowers, L. dolichopoda must be also considered in the subgenus Licaria. The glabrous condition found in every structure of the plant body (stem, leaves, and flowers), as well as the apical openings of stamens, and the lack of staminodes found in this species places it beside L. eurypaniculata (here described), from which it differs in the botryoid architecture of the inflorescence, and the conspicuous caudate leaves. Collection duplicates of L. dolichopoda have been previously distributed as L. cufodontisii Kosterm. (Herrera 4000), and as L. misantlae (Herrera 4510).

Licaria eurypaniculata Lorea-Hern. sp. nov. (Figs. 4, 6)

Diagnosis. Among the species with glabrous leaves, inflexed tepals, lacking staminodes, as well as apical anther openings, this species is distinguished by paniculate inflorescences where the flowers are not aggregate on the floriferous axes, orbicular flowers, tepal surface glabrous abaxially and adaxially, fused stamens throughout, glabrous hypanthium outside and inside, and glabrous pistil.

Trees 6-7 m tall; twigs glabrous, smooth or slightly ribbed, reddish-brown, sparsely lenticellate. Buds glabrous. Leaves alternate, petioles (5.5-)7-9(-10.5) mm long, slightly sulcate above, glabrous, blades (7-)11-16(-19.5) × (2-)3-4.5(-6) cm, narrowly elliptic to lanceolate or elliptic, pinninerved, secondary veins 7-9 pairs, both upper and lower leaf surfaces glabrous, leaf apex acuminate to short caudate, base obtuse to cuneate. Inflorescences 10-11 cm long, apparently terminal, if it is axillary to a bract, this one is inconspicuous, paniculate, basal axes longer than the peduncle, glabrous throughout or sparsely puberulent toward the end of the secondary axes, flowers spaced, not aggregate on the axes, peduncle ca. 3 cm long, pedicels 2-3.2 mm long, glabrous or sparsely puberulent. Flowers spheroidal, greenish, tepals conspicuously inflexed, outer tepals ca. 0.5 × 0.9-1.0 mm, widely ovate to depressed ovate, glabrous abaxially and adaxially, inner tepals ca. 0.4 × 0.6-0.7 mm, ovate or almost orbicular, glabrous abaxially and adaxially, staminodes of whorls I and II absent, stamens of whorl III 0.5-0.6 mm long, fused throughout, filaments glabrous outside, sparsely tomentose inside, anthers ca. 0.1 mm, glabrous, with apical openings, glands ca. 0.1 mm, free, widely elliptic or orbicular, glabrous, hypanthium ca. 0.7 mm deep, glabrous outside, glabrous or sparsely pubescent on the distal section inside, pistil ca. 1.1-1.2 mm long, glabrous throughout, ovary ca. 0.6-0.7 mm long. Fruits (not wholly ripe) 15-17 × 12-12.5 mm, ovoid or ellipsoid, cupule 10-11 × 14-15 mm, crateriform, clearly bimarginate, inner margin 1.1-1.3 mm tall, erect, outer margin 0.5-0.7 mm tall, perpendicular to the inner one, pedicel 3-3.6 × 2-2.2 (at the base) and 3.6-4 (at the apex) mm, obconic.

Taxonomic summary

Type. Panama. Bocas del Toro: along road to Chiriquí Grande, c. 10 road miles from continental divide and about 2 miles along road east of highway, 8°45’ N, 82°15’ W, 300 m, 15 April 1987, G. McPherson 10830 (holotype MO; isotype XAL 124596).

Paratype. Panama. Bocas del Toro: along road to Chiriquí Grande, 10 road-miles from continental divide, ca. 2 road-miles along road east of highway, 8°55’04’’ N, 82°10’04’’ W, 300 m, 9 February 1987, G. McPherson 10453 (MO 5048286).

Etymology. The name of this species alludes to the very long basal secondary axes of the inflorescence, which give a broad triangular profile to the panicle.

Distribution and habitat. The only 2 known collections of the species come from the same area in the Atlantic lowlands of western Panama. No information about the vegetation found at the site was recorded, but considering the geographical factors of the place, it is expected to be tropical rain forest. 

Phenology. Flowers in spring, and fruits mature in winter or early spring. The plant is not deciduous.

Conservation status. It seems that the species is not frequent in the area where it was found, for it has been collected only twice. On the other hand, the view of the area from satellite images shows that most of the original vegetation has been cleared; therefore, the species might be endangered.

Remarks

There are no species that come close morphologically to L. eurypaniculata. The other species with a general glabrous condition almost throughout the plant body, L. dolichopoda, is very different, in the shape of the leaves and the structure of the inflorescence. Given the erect, concave tepals, and stamens with extrorse sporangia that L. eurypaniculata presents, it is also a member of the subgenus Licaria. 

Licaria gibbitepala Lorea-Hern. sp. nov. (Figs. 5, 6)

Diagnosis. Trees similar to L. excelsa, but distinguished by the obovate to oblanceolate leaves, densely yellowish to orange-brown puberulent inflorescences, tepals with basal half conspicuously swollen, puberulent abaxial surface, sericeous adaxial surface at the base, apically pubescent ovary, and pubescent style.

Trees 15-20 m tall, trunk ca. 25 cm DBH, bark smooth, fragrant; twigs hollow, inhabited by ants, glabrous, sparsely lenticellate, slightly ridged, each ridge originating at one extreme of petiole insertion, acropetal. Buds glabrous or partially pubescent. Leaves alternate, petioles 10-12(-14) mm long, bimarginate above, glabrous, blades (20-)24-30 × (9-)11-13 cm, obovate or oblanceolate, sometimes elliptic, pinninerved, secondary veins (12)14-16 pairs, leaf surface glabrous above and below, leaf apex apiculate, sometimes apiculate-acuminate, leaf base obtuse or rounded. Inflorescences 9-15 cm, seemingly terminal, but actually axillary to tiny, deciduous bracts, on the proximal section of new twigs, paniculate, conspicuously puberulent along all axes, hairs yellowish to orange-brown, peduncle (1-)3.5-4.5 cm, sparsely puberulent, glabrescent, pedicels 2.5-5.5(-7) mm, densely puberulent. Flowers widely obovoid, yellowish-green, fragrant, perianth thick, coriaceous, tepals clearly inflexed, their base conspicuously swollen, outer tepals 1.2-1.4 × 1.6-1.9 mm, widely ovate, puberulent abaxially, sericeous-tomentose at the base and toward the margins adaxially, hairs orange-brown, inner tepals 0.8-1 × 1-1.2 mm, ovate, puberulent abaxially, sericeous-tomentose at the base and tomentose at the middle adaxially, staminodes of whorls I and II absent, stamens of whorl III 0.7-0.8 mm long, fused along their filaments, filaments tomentose at the base outside, tomentose throughout inside, hairs orange-brown, anthers ca. 0.2 mm, free or fused just at the base, glabrous, with apical openings, glands ca. 0.3 mm, only 3 given the fusion of adjoining glands, widely oblong, glabrous, hypanthium 1-1.2 mm deep, obconic, densely puberulent outside, hairs yellowish to orange-brown, sericeous-tomentose inside, hairs reddish-brown, pistil 1.6-1.8 mm long, top of the ovary and style pubescent, ovary 0.8-1.2 mm long. Fruits (nearly ripe) 19-23 × 15.5-16.5 mm, ellipsoid, cupule ca. 16.5 × 18.5 mm, cotyliform, sparsely lenticellate, seemingly tri-margined, the 2 regular margins plus the swollen projections of the tepals, inner margin ca. 1.6 mm tall, erect, outer margin 2.2-2.5 mm tall, erect, pedicel 4.5-7 mm long, continuous with the base of the cupule, 3.5 mm diam. at the base.

Taxonomic summary

Type. Costa Rica. Limón: Reserva Biológica Hitoy Cerere, 300 m aguas abajo de la confluencia del río Hitoy con el río Cerere, margen izquierda por la fila que lleva al cerro Bobócora, 9°39’00’’ N, 83°02’45’’ W, 200 m, 20 February 1989, G. Herrera & A. Chacón 2427 (holotype MO, isotype XAL 124445).

Etymology. The name is derived from the conspicuous swollen condition of the base of the tepals; the feature is distinctive. 

Distribution and habitat. So far, this species is known only from the southwestern end of Costa Rica. Although no information about the habitat was recorded, there is no doubt that the place lies within the tropical rain forest territory.

Phenology. With flowers and ripe fruits around the end of winter. The species has perennial leaves.

Conservation status. There is no information about the abundance or extent of distribution of the species, but as it is known to grow in the grounds of a nature reserve, it can be expected that it is not under high risk of extinction but certainly endangered.

Remarks

Licaria gibbitepala seems to be closely related to L. tomentulosa and, at the same time, to the group of species around L. excelsa. All of them have ridged twigs, large leaves, flowers with erect, concave tepals, no staminodes, and sporangia with apical openings. The hollow twigs, along with the obovate to oblanceolate leaves, and the conspicuous gibbous base of the tepals distinguish L. gibbitepala.

Licaria gracilis Lorea-Hern. sp. nov. (Figs. 7, 9)

Diagnosis. Small trees with glabrous leaves, botryoid, few-flowered inflorescences, glabrous, erect tepals, lacking staminodes, free stamens, anther openings elliptic, lateral, strongly oblique, glands present, glabrous hypanthium outside and inside, glabrous pistil.

Trees to 6 m tall; twigs smooth, dark brown to reddish brown, pruinose, puberulent, glabrescent, sparsely lenticellate. Buds glabrous. Leaves alternate, petioles (4-)8-11 mm long, puberulent, soon glabrous, canaliculate above, blades 8.5-13 × 3.5-5.5 cm, elliptic or narrowly elliptic, pinninerved, secondary veins 5-7 pairs, leaf surface glabrous above and below, but the lower surface puberulent in the beginning, leaf apex caudate, sometimes just acuminate, base obtuse to cuneate. Inflorescences 2.5-3.5 cm long, axillary to leaves and to tiny, deciduous bracts, on the proximal section of new twigs, botryoid, racemiform, few-flowered (less than 10 flowers), glabrous throughout or with some hairs toward the end of the peduncle, peduncle 1.8-2 cm long, pedicels 3-4.5 mm long, glabrous. Flowers obovoid, pale yellow, tepals erect to slightly inflexed, concave, outer tepals ca. 0.8 × 1.3 mm, very widely ovate to depressed ovate, glabrous abaxially, with some long, appressed hairs ascending from the base adaxially, inner tepals ca. 0.5 × 0.8 mm, ovate or widely ovate, glabrous abaxially, adaxially like the outer tepals, staminodes of whorls I and II absent, stamens of whorl III ca. 0.7 mm long, free, but very close one to each other, widely ovate in outline, filaments sparsely tomentulose outside and inside, anthers ca. 0.4 mm, tomentulose at the base outside, tomentulose inside along the central line, openings lateral, oblique, glands ca. 0.4 mm, rounded, glabrous, hypanthium ca. 0.7 mm deep, obconic, glabrous inside and outside, pistil ca. 1.7 mm long, glabrous, ovary ca. 0.8 mm long. Fruit unknown.

Taxonomic summary

Type. Panama. Chiriquí: Punta Burica, El Chorogo, alrededores de la finca de Fernando Chavarría, adyacente al límite fronterizo, cabecera del río San Bartolo, 8°17’07’’ N, 82°58’56’’ W, 395 m, 15 May 2007, J. E. Aranda et al. 3912 (holotype MO 6456198).

Etymology. The name of the species is derived from the attractive view that the slender inflorescences give to the plant.

Distribution and habitat. The species is known only from the location where it was first and last collected. No information about the habitat is mentioned in the data-label of the specimen, except that was collected nearby a ranch. The original vegetation must have been tropical evergreen forest or semi-evergreen forest.

Phenology. Flowers during spring; ripe fruits expected during winter or early spring, since maturation of fruits in most Lauraceae takes around a year after flowering.

Conservation status. The species might be (critically) endangered, for most of the land in the area where it was collected has been transformed for diverse agricultural purposes.

Remarks

The few-flowered, botryoid inflorescence, tiny flowers, and very oblique, lateral openings of the sporangia distinguish this species straightaway. There is no species whose general morphology indicates association with L. gracilis. The singular way that the openings of the stamens are displayed resemble that found in Licaria cogolloi van der Werff, and L. caribaea Gómez-Lau. & Cascante, but besides this, there is no other feature that might suggest a relationship to those species. For the features of its flowers, Licaria gracilis belongs to subgenus Licaria.

Licaria minutiflora Lorea-Hern. sp. nov. (Figs. 8, 9)

Diagnosis. Trees similar to L. pergamentacea, but distinct by the presence of hollow twigs, glabrous buds, abaxially densely tomentulose tepals, internally pubescent hypanthium, apically pubescent ovary, and pubescent style.

Trees up to 20 m tall, trunk ca. 30 cm DBH; twigs hollow, inhabited by ants, glabrous, smooth or slightly ridged, each ridge originates at one extreme of petiole insertion, acropetal, sparsely lenticellate. Buds glabrous. Leaves alternate, petioles (7-)15-20(-30) mm, glabrous, bimarginate above, blades (11-)20-28(-34.5) × (3-)5-9(-13) cm, narrowly elliptic or narrowly ovate, pinninerved, secondary veins 9-12 pairs, leaf surface glabrous on both sides, leaf apex acute, slightly apiculate, base acute or obtuse. Inflorescences (6-)9-12(-15) cm long, axillary to tiny, deciduous bracts, on the proximal section of new twigs or axillary to leaves, paniculate, tomentulose, peduncle (0.5-)1.5-3(-4) cm long, pedicels (1.2-)2.5-4(-6) mm long, densely tomentulose. Flowers obovoid or ellipsoid, yellowish-green, tepals inflexed, outer tepals 0.5-0.8 × 0.8-1 mm, widely ovate, densely tomentulose abaxially, tepal surface concealed or almost so by hairs, glabrous adaxially, inner tepals 0.5-0.7 × 0.5-0.6 mm, ovate, densely tomentulose abaxially, glabrous adaxially, staminodes of whorls I and II absent, stamens of whorl III 0.5-0.6 mm long, free or barely united by filament base, filaments tomentose at base outside, glabrous or sparsely pubescent along the medial line inside, anthers ca. 0.2 mm, glabrous, openings apical, glands ca. 0.2 mm, free, rounded, hypanthium 0.6-0.8 mm deep, obconic, densely tomentulose outside, sericeous on upper half inside, hairs yellowish or reddish, pistil ca. 1.4 mm long, top of the ovary and style pubescent, ovary 0.8-1 mm long. Fruits ca. 19.5 × 15-16.5 mm, ellipsoid, cupule 15-16 × 18-20 mm, urceolate, lenticellate, conspicuously bimarginate, inner margin 1.2-1.5 mm tall, outer margin 1-1.2 mm, extended, pedicel 6.5-9 mm long, continuous with the cupule, 2.8-3.6 mm diameter at base.

Taxonomic summary

Type. Costa Rica. Alajuela: camino entre la estación de la Reserva Forestal de San Ramón y el camino a la colonia Palmareña, finca de don Bolívar Ruiz, margen derecha río San Lorencito, 10°12’53’’ N, 84°36’28’’ W, February 1987, G. Herrera 500 (holotype MO 3587621; isotypes MEXU 638804, TEX).

Paratypes. Costa Rica. Alajuela: Reserva Biológica Monteverde, Poco Sol, La Cutacha de San Bosco, 10°22’ N, 84°40’ W, 900 m, 1 April 1989, E. Bello 784 (MEXU 1304162; MO 6142952; XAL 124442); Bosque Eterno de los Niños, Reserva de Arenal, río Peñas Blancas, Quebrada Agua Gata, Finca Villalobos, 10°23’ N, 84°42’ W, 1,000 m, 20 April 1990, E. Bello 2208 (MO 6130824; XAL 124446); Cantón de Upala, Colonia La Libertad, 10°52’ N, 85°17’ W, 300 m, 3 August 1991, Q. Jiménez & G. Rivera 1011 (MO 6117397; XAL 124597). Guanacaste: Parque Nacional Guanacaste, Estación Pitilla, 10°00’15’’ N, 85°25.2’ W, 500 m, 27 May 1989, G. Herrera et al. 2942 (MO; XAL 124428).

Etymology. The very small flowers, whose tepals are less than 1 mm long, is the feature on which the species name is based.

Distribution and habitat. This species is known from the northern hills of the Cordillera de Guanacaste (Guanacaste Mountain Range) and the western part of the Cordillera Central (Central Mountain Range), between 200 and1,000 m asl. Prevailing vegetation in the region is tropical evergreen forest and montane rain forest. The species has been collected also in pasture fields. There is a fruiting specimen from the Osa Peninsula (Hammel et al. 16984) that seems to belong to L. minutiflora but until confirmed with a flowering specimen, the presence of the species in this part of the country remains uncertain.

Phenology. Flowers toward the end of winter and early spring; fruits must be ripe around the end of autumn or early winter.

Conservation status. Most specimens of this species have been collected within nature reserves. Therefore, it is considered not threatened, even though its abundance is still unknown.

Remarks

As has been mentioned elsewhere in this paper, Licaria minutiflora is part of the L. excelsa species group. Its general appearance resembles that of L. pergamentacea, for the size and shape of the leaves, structure and hairiness of the inflorescence, and for having small oblong-ellipsoid, not coriaceous flowers. However, besides the presence of hollow twigs, it differs by having flowers with the upper part of the hypanthium homogeneously pubescent inside, and a pubescent pistil.

Licaria ochracea Lorea-Hern. sp. nov. (Figs. 10, 12)

Diagnosis. Trees similar to L. multinervis H. W. Kurz, but differing by lower surface indument of leaves composed by 2 types of hairs, the most numerous tomentulose, the fewer sericeous, long inflorescences, terminal or axillary to leaves, with rachis up to 15 cm, fully exserted anthers, clearly stalked glands, conspicuous hypanthium tube projected beyond the insertion point of stamens. 

Trees up to 22 m tall, twigs smooth, densely tomentose, hairs initially yellowish-brown, then greysh, sparsely and inconspicuously lenticellate. Buds tomentose. Leaves alternate, petioles (10-)15-20 mm long, tomentose, glabrescent, channeled above, blades 12.5-18.5 × (2.5)3.5-4.5 cm, narrowly elliptic, sometimes narrowly oblanceolate, pinninerved, secondary veins (9)10-13 pairs, leaf surface glabrous above, tomentose below, most hairs sinuous, rather appressed, or patent, fewer hairs straight, appressed, leaf apex acuminate, base narrowly cuneate. Inflorescences (7-)10-18 cm long, axillary to leaves, less frequently terminal, paniculate, densely tomentose, hairs like on twigs, flowers agglomerate at the end of terminal axes, peduncle 0.2-1.5(-3.5) cm long, pedicels (1-)1.5-2 mm long, tomentose. Flowers narrowly oblong, tepals erect, concave, outer tepals, 1-1.3(-1.5) × (1-)1.1-1.3 mm, widely ovate, tomentose abaxially, glabrous adaxially, except for a few long, appressed hairs, coming from the base, inner tepals 1-1.1(-1.4) × 0.8-0.9 mm, ovate, with a pattern of pubescence similar to that of outer tepals, staminodes of whorls I and II absent, stamens of whorl III 3-3.3(-3.8) mm long, coherent along their filaments or even at the base of the anthers, easily separable, filaments tomentose on both faces, anthers 0.9-1.1 mm long, free or coherent at the base, glabrous, completely exserted, openings dorsolateral, glands 0.7-0.8 mm, free, oblanceolate, glabrous, clearly stalked, stalk pubescent, hypanthium 1.2-1.3(-1.6) mm deep, extended 0.3 mm beyond the insertion point of stamens, tomentose outside, densely hirsute-tomentose inside, hairs golden-brown, pistil 3.6-3.9 mm long, sparsely pubescent, at least some hairs on the upper half of the ovary and lower half of the style, ovary 0.9-1.1 mm long. Fruits unknown.

Taxonomic summary

Type. Nicaragua. Matagalpa (according to current maps it should be Jinotega): pasture and small woods, tropical premontane forest, Hacienda Santa María del Ostuma, 10 km N of Matagalpa, 1,300 m, 17 July 1978, P. C. Vincelli 756 (holotype MO 2984723; isotypes LL, MEXU 691292).

Etymology. The name refers to the conspicuous orange-brown pubescence of the species on young twigs and main axes of inflorescences.

Distribution and habitat. Only known from the place of type collection, where the Cordillera Isabelia and Cordillera Dariense meet.

Phenology. Flowers in summer; fruit season is not known, but since it has been observed that maturation of fruits in most Lauraceae takes around a year after flowering, possibly ripe fruits are present during spring or early summer.

Conservation status. There is no certainty on the conditions that might be affecting the survival of the species. The type was collected in a private property that used to be preserved for ecotourism activities, but its current situation is unknown. However, since the species has not been collected again, it is suspected to be endangered.

Remarks

The collection on which the description of this species is based was initially identified by Kurz (2000) as L. multinervis; actually it is cited as a paratype of this species. But the impression of being conspecific with this taxon disappears with a detailed survey of the morphology. Important differences are the pubescence on the lower surface of the leaves (simple in L. multinervis vs. made of 2 types of hairs in L. ochracea), position of the inflorescences (axillary to small bracts in the proximal section of new branches in L. multinervis vs. axillary to leaves or terminal in L. ochracea), length of inflorescences (up to 3.5 cm long in L. multinervis vs. 5-7 times longer in L. ochracea), protrusion of anthers (partially exserted in L. multinervis vs. fully exerted in L. ochracea), and projection of hypanthium tube beyond the insertion point of stamens (short in L. multinervis vs. conspicuous in L. ochracea). However, L. multinervis and L. ochracea seem to be more closely related to each other than to the rest of species in the area that present fully exserted stamens, namely L. agglomerata van der Werff, L. capitata (Schltdl. et Cham.) Kosterm., L. nitida van der Werff, and L. vanderwerffii (here described).

Licaria rufotricha Lorea-Hern. sp. nov. (Figs. 11, 12)

Diagnosis. Small trees, distinct for the conspicuously obovoid flowers, aggregate toward the end of the secondary floriferous axes, with inflexed tepals, glabrous abaxially, reddish-brown hirsute-sericeous adaxially, lacking staminodes, stamens fused throughout, reddish-brown hirsute, anther openings apical, hypanthium mostly glabrous outside, reddish-brown hirsute inside, and ovary distally pubescent. 

Trees to 12 m tall; twigs smooth or inconspicuously ridged, each ridge originating at one extreme of petiole insertion, acropetal, lenticellate, grayish, densely puberulent, glabrescent. Buds glabrous. Leaves alternate, petioles 10-13 (-17) mm long, slightly channeled above, puberulent, blades (14.5-)16.5-20(-23) × 4-5.5(-9) cm, elliptic or narrowly elliptic, pinninerved, secondary veins 6-7 pairs, leaf surface densely puberulent above when young, glabrescent, rather persistently puberulent below, leaf apex acuminate, base cuneate, sometimes obtuse or narrowly cuneate to indistinctly attenuate. Inflorescences (3-)5-8(-10.5) cm, axillary to tiny, deciduous bracts, on the proximal section of new twigs, or axillary to new leaves, paniculate, flowers aggregate at the end of secondary axes, densely puberulent on peduncle and rachis, glabrate toward the ultimate axes, peduncle (2-)3.5-5.5(-7.5) cm long, pedicels 2-3.5(-4.5) mm long, glabrous or sparsely puberulent. Flowers obovoid or obconic, pale yellow, tepals conspicuously inflexed, outer tepals 1-1.2 × 2-2.4 mm, widely ovate, glabrous abaxially, hirsute-sericeous adaxially, hairs reddish-brown, inner tepals 0.8-1 × 0.8-0.9 mm, ovate or elliptic, glabrous abaxially, generally mostly hirsute adaxially, hairs reddish-brown, surpassing the apical margin of the tepals, making them appear fimbriate, staminodes of whorls I and II absent, stamens of whorl III, 0.6-0.7 mm long, fused throughout, filaments hirsute on both faces, hairs reddish-brown, anthers ca. 0.2 mm, pubescent, except around the margin of the openings, hairs like those of the filaments, glands ca. 0.4 mm, free, rounded or oblate, glabrous, hypanthium 0.8-0.9 mm deep, glabrous or sparsely puberulent outside, hirsute inside, hairs like those of the filaments, pistil 1.7-2 mm long, ovary ca. 0.8 mm long, sparsely pubescent distally, hairs whitish. Fruits unknown.

Taxonomic summary

Type. Panama. Panama: 5-10 km NE of Altos de Pacora, on trail at end of road, ca. 750 m, 7 March 1975, S. Mori & J. Kallunki 4977 (holotype MO 2992664).

Etymology. The name given to this species refers to the dense red-brown pubescence in the interior of the flowers, covering the adaxial surface of tepals, stamen filaments, anthers, and hypanthium inside.

Distribution and habitat. Just known from the place where the type was collected (around 25-30 km NE of Panama City), with tropical evergreen forest or semi-evergreen forest as original vegetation.

Phenology. Flowers around the end of winter or early spring; ripe fruits expected during winter, since maturation of fruits in most Lauraceae takes around a year after flowering.

Conservation status. Probably the species is endangered. Most of the vegetation in the surroundings of the place where the species was collected has been cut down for cattle raising or agricultural purposes. But the southern boundary of the Parque Nacional Chagres (Chagres National Park) is very close to that place, and the Nature Reserve Cocobolo is a few kilometers ENE of it as well. So, even it has not been collected again, it is hoped the species is still around there.

Remarks

There is no other species in the Mesoamerican area with the distinctive reddish-brown pubescence found in the interior of the Licaria rufotricha flowers. Yellowish to reddish-brown pubescence is present on the filaments of stamens and/or inside the hypanthium of several species, like L. agglomerata, L. excelsa, L. gibbitepala, and L. tomentulosa, but it is never found on the tepals surface. In fact, its general morphology does not suggest a relation to any species in the region. 

Licaria sarukhanii Lorea-Hern. sp. nov. (Figs. 13, 15)

Diagnosis. Trees similar to L. alata, but differing by terminal, subsessile inflorescences, longer pedicels (up to 10.5 mm), flowers widely ellipsoid, abaxial tepal surface tomentose at the base and papillose toward the apex, hypanthium densely tomentose outside, sericeous-tomentose inside, and style pubescent.

Trees to 20 m tall, twigs glabrous, grayish to grayish-brown, slightly ridged, each ridge originating at one extreme of petiole insertion, acropetal, sparsely lenticellate. Buds glabrous or pubescent at the apex of the bud scales. Leaves alternate, petioles (11-)13-17(-20) mm long, slightly bimarginate, glabrous, blades (12.5-)20-32(-37) × (5-)7-10(11.5) cm, narrowly elliptic, occasionally oblanceolate, pinninerved, secondary veins (12)14-18 pairs, leaf surface glabrous on both sides, leaf apex acuminate, sometimes acute, base obtuse or acute. Inflorescences 12-22 cm long, terminal, paniculate, tomentose, soon glabrescent, hairs patent or oblique, grey or yellowish-grey, peduncle 0.2-0.4 cm long, pedicels (3-)5-8(-10.5) mm long, glabrous or sparsely tomentose. Flowers broadly ellipsoid, pale green, tepals inflexed, outer tepals 0.8-0.9 × 1.8-2 mm, very widely ovate, concave, tomentose over the basal half, and papillose-pubescent over the distal half abaxially, glabrous or with few straight hairs at base adaxially, inner tepals 0.7-0.9 × 0.4-0.8 mm, ovate, pubescence pattern abaxially like that of outer tepals, but less papillose, glabrous adaxially, staminodes of whorls I and II absent, stamens of whorl III 0.7-0.9 mm long, fused along their filaments, filaments tomentose along the central line on both faces, anthers 0.2-0.4 mm long, glabrous, openings apical, glands ca. 0.4 mm, oblong, flattened, frequently reduced or absent, glabrous or tomentose on the base abaxially, hypanthium ca. 1 mm deep, densely tomentose outside, hairs sinuous, sericeous-tomentose inside, pistil 1.3-1.6 mm long, ovary 0.8-1.2 mm long, glabrous, style rather densely pubescent. Fruits 27-30 × 17-18 mm, ellipsoid, cupule 15-17.5 × 18-19.5 mm, urceolate, bimarginate, but the margins barely discernible, ca. 0.5 mm, erect, pedicel 4-5 × 3.5-4.5 mm.

Taxonomic summary

Type. Mexico. Chiapas: municipio de Ángel Albino Corzo, Reserva de la Biosfera El Triunfo, aprox. 1.5 km al E de Campamento El Triunfo, sobre el Sendero Bandera, 15°36’ N, 92°50’ W, 1,850 m, 27 April 1993, F. Lorea 5522 (holotype XAL 151063; isotypes [to be distributed]).

Paratypes. Mexico. Chiapas: municipio de Ángel Albino Corzo, Polígono I de la Reserva de la Biosfera El Triunfo, 15°39’ N, 92°48’ W, 1,900 m, 21 February 1993, S. Solórzano 67 (MEXU 754117); Reserva El Triunfo, camino a Cerro de la Bandera, 25 March 1986, M. L. Ávila and V. H. Hernández s/n (MEXU 877857, TEX 146383); Reserva de la Biosfera El Triunfo, sendero Palo Gordo, 15°40’10’’ N, 92°48’42’’ W, 1,990 m, 22 March 2006, F. González-García s/n (XAL 113841).

Etymology. The species is dedicated to José Sarukhán, a Mexican plant ecologist who has played an important role in making a huge amount of information about Mexico’s biodiversity, ecosystem conservation, and sustainable development accessible to the public.

Distribution and habitat. So far, the species is known only from the central region of the Sierra Madre de Chiapas, from 1,800 to 2,000 m asl where the montane rain forest is the dominant type of vegetation.

Phenology. Flowers toward the end of winter and early spring; ripe fruits were found in the same interval of time.

Conservation status. All the collections of the species come from the nature reserve El Triunfo, located in the Sierra Madre de Chiapas, which has persisted largely undisturbed for its almost forty years of existence. Thus, considering the size of the area covered with montane rain forest in the reserve, the species might be regarded as vulnerable.

Remarks

Licaria sarukhanii is part of the L. excelsa species group, with a closer relation to L. breedlovei, because they share tomentose inflorescences and flowers rather widely ellipsoid to obloid. They differ by the conduplicate leaf bases, sparsely sericeous lower leaf surface, conspicuously pedunculate inflorescences axillary to small bracts or leaves, fused anthers, and style sparsely pubescent present in L. breedlovei, in contrast to flat leaf base, glabrous lower leaf surface, terminal, sub-sessile inflorescences, free anthers, and conspicuously pubescent style in L. sarukhanii. 

Licaria tomentulosa Lorea-Hern. sp. nov. (Figs. 14, 15)

Diagnosis. Trees similar to L. excelsa, but distinguished by presenting hollow twigs, inflorescences yellowish-brown tomentulose, flowers conspicuously obovoid, tepals densely tomentulose abaxially, and apex of the ovary and style conspicuously pubescent.

Trees up to 25 m tall, and trunk 50 cm DBH, twigs hollow, inhabited by ants, smooth or slightly ridged, glabrous, grayish-brown, sparsely lenticellate. Buds glabrous. Leaves alternate, petioles (14-)17-22(-24) mm long, bimarginate above, glabrous, blades (22-)25-30(-34) × (6-)8-10(-12) cm, narrowly elliptic or elliptic, pinninerved, secondary veins (9)11-13 pairs, leaf surface glabrous on both sides, leaf apex acute or short acuminate, base obtuse, rounded or shortly attenuate, particularly in leaves close to the twig tips. Inflorescences 6.5-9.5(13.5) cm long, axillary to tiny, deciduous bracts, on very short branches axillary to leaves, paniculate, tomentulose, hairs yellowish-brown, peduncle 2.5-4 cm long, pedicels (1-)2-3.5 mm long, densely tomentulose. Flowers obovoid, thick, coriaceous, greenish-yellow, tepals inflexed, concave, outer tepals 0.7-1 × 1.4-1.7 mm, widely ovate, densely tomentulose abaxially, hairs almost concealing the surface, marginal tomentose bands and some long, appressed hairs at the base adaxially, inner tepals 0.6-0.9 × 0.8-0.9 mm, ovate, sometimes slightly conduplicate, densely tomentulose abaxially, pattern of pubescence adaxially like in the outer tepals, staminodes of whorls I and II absent, stamens of whorl III 0.8-1.1 mm long, fused throughout, filaments tomentose outside on lower half, densely tomentose all over the inner face, hairs reddish-brown, anthers ca. 0.2 mm, glabrous outside, densely tomentose inside, openings apical, glands 0.3-0.4 mm, frequently fused between adyacent stamens, transversely oblong, oblong when free, hypanthium 1-1.2 mm deep, densely tomentulose outside, tomentulose inside, sometimes only in the upper half, hairs mainly reddish-brown, pistil 2-2.3 mm long, ovary 1.1-1.4 mm long, apex of the ovary and style conspicuously pubescent. Fruits (not fully ripe) ca. 13 × 13 mm, spheroidal, cupule 13-14 × 17-18 mm, crateriform, lenticellate, bimarginate, inner margin 0.7-1 mm tall, erect, outer margin 0.5-0.7 mm, erect to spreading, pedicel 4-6 × 3-4 (apex) and 2 mm (base).

Taxonomic summary

Type. Costa Rica. Limón: Cordillera de Talamanca, Matina, intersección de río Barbilla y quebrada Cañabral, por la fila al norte, 10°01’ N, 83°24’ W, 100-200 m, 11 October 1988, G. Herrera 2165 (holotype MO 3693485; isotypes MEXU 529888, 529889, 717774, and 718334; TEX).

Paratypes. Costa Rica. Alajuela: Upala, Bijagua, El Pilón, Cerro La Carmela, entre río Celeste y cabeceras del río Chimurria, 10°43’15’’ N, 84°59’45’’ W, 1,000 m, 11 July 1988, G. Herrera 2056 (MEXU 529875, 717781; MO 3693486; US 3655515). Limón: Reserva Biológica Hitoy Cerere, Valle de la Estrella, sendero a Cerro Bobócara, 9°41’00’’ N, 83°04’20’’ W, 798 m, 17 August 1990, G. Herrera 4115 (MEXU 1304090; MO 6142953; XAL 124444); Reserva Indígena Talamanca, camino a Soki entre la Quebrada Amubri, margen izquierda de río Lari, 9°29’40’’ N, 82°89’40’’ W, 200 m, 29 June 1989, A. Chacón 25 (MO; XAL 124437).

Etymology. The name is for the abundant tomentulose pubescence that covers the axes of the inflorescence, as well as the abaxial surface of tepals and hypanthium. 

Distribution and habitat. Known only from Costa Rica, along the Atlantic lowlands adjacent to the Sierra de Talamanca, and the Sierra de Guanacaste. Tropical rain forest used to be the predominant vegetation there, but currently a big amount of land has been changed to agricultural activities or other affairs.

Phenology. Flowers in summer and early autumn; ripe fruits in summer.

Conservation status. Given the deterioration of the habitat, the species might be considered vulnerable or endangered, in spite of its wide range of distribution. 

Remarks

Licaria excelsa and L. tomentulosa are much alike. Considering the available material, distinguishing characters seem to work well to separate both species; hollow twigs, as well as type and distribution of pubescence on vegetative and floral parts, are consistently present in all the specimens considered as L. tomentulosa, making it a clear morphological unit, distinct from L. excelsa.

Licaria vanderwerffii Lorea-Hern. sp. nov. (Figs. 16, 17)

Diagnosis. Similar to L. agglomerata, but different by the free stamens, white pubescence of stamens, hypanthium densely white tomentose inside, and pistil glabrous.

Shrubs or small trees 1-3 m tall, twigs hollow, smooth, sparsely lenticellate, glabrous, reddish-brown. Buds glabrous. Leaves alternate, petioles ca. 12.5-15 mm long, slightly channeled and bimarginate above, glabrous, blades ca. 15-23 × 6-7.5 cm, narrowly elliptic, pinninerved, secondary veins 9-11 pairs, leaf surface glabrous on both sides, leaf apex acuminate, base obtuse or cuneate. Inflorescences ca. 1.5-2 cm long, axillary to leaves, axillary to tiny, deciduous bracts, on the proximal sections of new twigs too, paniculate, but strongly condensed, with very short rachis and secondary axes, peduncle ca. 0.5 cm long, glabrous, pedicels (1.2-)1.5-2.5(-3) mm long, glabrous. Flowers ellipsoid, slightly swollen in the middle of the hypanthium, greenish, tepals erect, outer tepals 0.5-0.6 × 0.5-0.6 mm, ovate, glabrous abaxially, glabrous adaxially except for some sericeous hairs at the base, inner tepals 0.4-0.5 × 0.2-0.3 mm, ovate, glabrous abaxially, glabrous adaxially or with some sericeous hairs at the base, staminodes of whorls I and II absent, stamens of whorl III 1-1.2 mm long, free, filaments white-tomentose throughout, denser on inside, anthers 0.3-0.4 mm, wholly exserted, glabrous or sparsely tomentose at the base outside, tomentose on lower half inside, openings dorsal, glands absent, hypanthium 1.1-1.3 mm deep, narrowly rhomboid, extending a little beyond the insertion point of stamens, glabrous outside, densely white-tomentose inside, pistil 2-2.3 mm long, glabrous, ovary 1-1.2 mm. Fruit unknown.

Taxonomic summary

Type. Panama. Darién: Cerro Tacarcuna S slope, 1,250-1,450 m, 26 January 1975, A. Gentry & S. Mori 13920 (holotype MO 2300651; isotype F 1763407).

Etymology. The species is named after Hendrik (Henk) Hessel van der Werff, a prolific scholar of the family Lauraceae in the Neotropics.

Distribution and habitat. So far, only known from the type collection, in the mountains of the southeastern edge of Panama, where premontane rain forest prevails.

Phenology. Flowers in winter; probably ripe fruits expected during autumn or early winter, since maturation of fruits in most Lauraceae takes around a year after flowering.

Conservation status. The place of the type collection is within the boundaries of the nature reserve Parque Nacional del Darién, but the altitudinal range within which the specimen was collected stretches barely for 40 km along the reserve. So, the species should be considered endangered.

Remarks

The collection on which the description of L. vanderwerffii is based was cited by Kurz (2000) as L. triandra, but there are several features that separate the former from this species. Whereas L. triandra presents branchlets solid, paniculate inflorescences (up to 6 cm long), stamens fused throughout (or almost so), glands at the base of filaments, anthers partially exserted, and hypanthium glabrous inside, L. vanderwerffii has hollow branchlets, sub-capitate inflorescences (up to 2 cm long), stamens free, no glands at the base of filaments, anthers wholly exserted, and hypanthium densely white tomentose inside.

This species is closely related to L. agglomerata; both present small, glomerate inflorescences, ellipsoid to narrowly rhomboid flowers with erect tepals, no staminodes, fully exserted stamens, no glands on filaments, pubescent stamens, and hypanthium tomentose within. However, L. vanderwerffii differs by having stamens free throughout, white tomentum on stamens and (densely on) inner face of hypanthium, and glabrous pistil, against stamens fused by their filaments, brownish-orange tomentum on stamens and upper part of hypanthium inside (sometimes completely glabrous), and puberulent style in L. agglomerata. It is worth to mention that they differ in ecological conditions as well; while L. agglomerata grows in lowland habitats (200-700 m), L. vanderwerffii is a mountain dweller above 1,200 m.

Acknowledgements

I would like to thank the curators of the herbaria cited in the Materials and methods section for allowing the study of specimens of Licaria in their collections. I also thank Jens Rohwer and an anonymous reviewer for their comments and calling my attention to several slips and wrong phrasing in some paragraphs. Eva Piedra assisted me with the preparation of figures. Phil Brewster kindly reviewed the final english version of the manuscript. I thank one of the reviewers who suggested to include the following exsiccata as part of the studied material for several of the species described here, however, it was not possible to see the specimens. So even they correspond to duplicates of the studied material, they are cited here just as a reference for researchers who might need to study them.

L. breedlovei; D. Breedlove 50394, isotype TEX 472525.

L. dolichopoda; B. Hammel et al. 16790, isotypes CR 147811 and CR 2918355; E. Bello & E. Cruz 4262, paratype CR 1596155; G. Herrera 4000, paratype CR 1596509; G. Herrera 4510, paratypes CR 159602, CR 176525 and CR 1561583; J. Marín & G. Marín 499, paratype CR 1596363.

L. gibbitepala; G. Herrera & A. Chacón 2427, isotype CR 1596536.

L. minutiflora; G. Herrera 500, isotype CR 151205; E. Bello 784, paratype CR 1513414; E. Bello 2208, paratype CR 1596135; Q. Jiménez & G Rivera 1011, paratypes CR 1561584 and CR 157780; G. Herrera 2942, paratype CR 1596539.

L. tomentulosa; G. Herrera 2165, isotypes CR 146925 and CR 2921047; G. Herrera 2056, paratypes CR 147034, and CR 2920969; G. Herrera 4115, paratypes CR 1596519, and CR 207025; A. Chacón 25, paratype CR 1596195.

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