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Research Article
Two new species of Salvia (Lamiaceae) from the dry forests of Dominican Republic
expand article infoItzi Fragoso-Martínez, Gerardo A. Salazar§, Emmanuel Martínez-Ambriz, Martin Reith|
‡ Red de Biodiversidad y Sistemática, Instituto de Ecología, Xalapa, Mexico
§ Universidad Nacional Autónoma de México, México DF, Mexico
| Jardín Botánico Prof. Eugenio de Js Marcano F., Santiago de los Caballeros, Dominican Republic
Open Access

Abstract

We describe two new species of Salvia from the Antillean dry forests, belonging to Salvia section Urbania. These species’ names honor two Latin American botanists who have advanced our understanding either of the Dominican flora or the mint family (Lamiaceae). Salvia claseana is found in the Sierra Martín García. It resembles S. calaminthifolia but differs in having strigose stems, rhombic to trullate leaves with a cuneate, decurrent base, and larger flowers. Salvia martineziana inhabits the Sierra de Bahoruco. It resembles Salvia brachyphylla but differs in having strigose stems with retrorse trichomes and flowers disposed in the axils of the distal leaves. We provide descriptions, photographs, a distribution map and an identification key for the species of Salvia section Urbania from the Domini­can Republic. Additionally, we sequenced three molecular markers (nrITS, trnL-trnF, and trnH-psbA) for the new taxa and other Dominican and Haitian Salvia species to investigate their phylogenetic relationships.

Resumen

Describimos dos nuevas especies de Salvia provenientes de los bosques secos antillanos, pertenecientes a Salvia sección Urbania. Estas especies honran a dos botánicos latinoamericanos que han ampliado nuestro conocimiento ya sea sobre la flora dominicana o la familia de la menta (Lamiaceae). Salvia claseana se encuentra en la Sierra Martín García. Se asemeja a S. calaminthifolia pero se diferencia de ella por tener tallos estrigosos, hojas rómbicas a truladas con una base cuneada, decurrente, y flores más grandes. Salvia martineziana habita en la Sierra de Bahoruco. Es semejante a Salvia brachyphylla pero difiere en tener tallos estrigosos con tricomas retrorsos y las flores dispuestas en las axilas de las hojas distales. Presentamos descripciones, fotografías, mapas de distribución y una clave de identificación para las especies de Salvia sección Urbania de República Dominicana. Además, secuenciamos tres marcadores moleculares (nrITS, trnL-trnF y trnH-psbA) para los nuevos taxones y otras especies de Salvia dominicanas y haitianas, con el fin de explorar sus relaciones filogenéticas.

Key words

Antilles, endemism, Hispaniola, phylogeny, sages

Introduction

The genus Salvia L. (sage) comprises more than 1000 species, which renders it the most diverse group in the mint family or Lamiaceae (Harley et al. 2004). Of these, more than half (580 spp.) belong to Salvia L. subgenus Calosphace (Benth.) Epling, a monophyletic group, that is endemic to the New World, distributed mainly in the Neotropics (González-Gallegos et al. 2020). The Antilles is one of the four diversity centers of Salvia subg. Calosphace, with ca. 45 species (Jenks et al. 2013). From the islands that form the Antilles, Hispaniola harbors the highest number of sages, with 36 species, 86% of them endemic (González-Gallegos et al. 2020).

Salvia subg. Calosphace section Urbania Epling is a group of 10 species endemic to Hispaniola, most of them distributed exclusively in Haiti (Epling 1939). These plants are characterized by a suffrutescent habit with intricate branching, small leaves, usually deltoid-ovate, obovate or flabelliform. The flowers grow in the leaf axils (1–3 per axil) or are rarely arranged in short racemes and are typically blue or violet in color. The calyx has 3-veined posterior lobes; the corolla is ventricose, invaginate and epapillate; the stamens are included in the galea, and the posterior stigmatic branch is longer than the anterior (Epling 1939).

Dominican Republic houses 21 species of Salvia (González-Gallegos et al. 2020), three of them from Salvia section Urbania (Liogier 1994). During November of 2016, a botanical expedition was carried out in Dominican Republic, visiting the montane temperate and arid zones to collect different species of Salvia. Among the collected material, we found specimens of two species from section Urbania that do not match any of the currently described taxa. In this study we describe both and compare them to the taxa that morphologically resemble them the most. Additionally, we provide an identification key for the Dominican species of the section. Finally, using DNA sequences of three molecular markers (nrITS, trnH-psbA intergenic spacer and trnL-trnF region) from most of the sages from Dominican Republic and a few Haitian Urbania species, we evaluate the phylogenetic position of the new taxa and test the monophyly of section Urbania.

Materials and methods

Fieldwork, species identification and conservation status assessment

Herbarium specimens were collected and processed, and the individuals were photographed in situ. Voucher specimens of all the collected taxa were deposited at the herbarium of the Jardín Botánico Nacional Dr. Rafael M. Moscoso (JBSD) and the duplicates (when available) were sent to MEXU and XAL herbaria in Mexico (Thiers 2023). Additionally, for each of the herbarium specimens collected, leaf samples were taken for DNA extraction and flowers were preserved in alcohol for posterior morphological analysis.

The identification of the specimens was based on the most comprehensive revision of Salvia subgenus Calosphace by Epling (1939), the revision of Salvia of Hispaniola (Liogier 1994) and the revision of sections Ekmania (Torke 2000), Gardoquiiflorae (Zona et al. 2016) and Wrightiana (Zona et al. 2011).

The distribution map was made with QGIS v. 3.30 (QGIS Development Team, 2023), using the distribution data from the herbarium specimen and iNaturalist observations from one of the authors, these occurrences and coordinates are not provided due to the species global conservation status of Critically Endangered. The species’ extent of occurrence (EOO) and species’ area of occupancy (AOO) were calculated using GeoCAT (Bachman et al. 2011). Finally, the conservation status of the new taxa was assessed based on these results and considering the IUCN guidelines (2022).

Taxon sampling, DNA extraction, amplification and sequencing

To test the phylogenetic position of the new taxa, as well as other sage species from the Caribbean, we sampled most of the species from Dominican Republic and a few taxa from Haiti, mainly belonging to section Urbania (Table 1; Suppl. material 1). DNA was extracted either from herbarium specimens or Silica gel-dried tissue, using the 2 × CTAB method (Doyle and Doyle 1987). DNA amplification followed the profiles and primer combinations described by Fragoso-Martínez et al. (2018), the only modification made is that the PCR reactions were carried out in 15 µL volumes. Finally, sequencing was performed at Macrogen, Inc.

Table 1.

Voucher information and GenBank accession codes for the sampled sage species in this study. Detailed specimen information can be found in Suppl. material 1.

Species Voucher information GenBank accession numbers
ITS trnH-psbA trnL-trnF
S. arborescens Urb. & Ekman I. Fragoso-Martínez et al. 459 (JBSD, MEXU) PP905439 PP907529 PP907549
S. arduinervis Urb. & Ekman E. Ekman 3168 (TEX) PP905440 PP907530 PP907550
S. bahorucona Urb. & Ekman I. Fragoso-Martínez et al. 517 (JBSD, MEXU) PP905441 PP907531 PP907551
S. brachyloba Urb. I. Fragoso-Martínez et al. 509 (JBSD, MEXU) PP905442 PP907532 PP907552
*S. claseana Fragoso & Salazar I. Fragoso-Martínez et al. 529 (JBSD, MEXU) PP905459 PP907548 PP907568
*S. calaminthifolia Vahl E. Ekman 9443 (TEX) PP905443 PP907533 PP907553
S. caymanensis Millsp. & Uline I. Fragoso-Martínez & R. Middleton 309, cultivated PP905444 PP907534 PP907554
S. decumbens Alain I. Fragoso-Martínez et al. 519 (JBSD, MEXU) PP905445 PP907535 PP907555
S. foveolata Urb. & Ekman I. Fragoso-Martínez et al. 508 (JBSD, MEXU) PP905446 PP907536 PP907556
S. lachnaioclada Briq. I. Fragoso-Martínez et al. 608 (JBSD, MEXU) PP905447 PP907537 PP907557
S. lavendula Alain T. Clase et al. 1059 (JBSD) PP905448 PP907538 PP907558
*S. martineziana Fragoso & Martínez-Ambr. I. Fragoso-Martínez et al. 497 (JBSD, MEXU) PP905458 PP907547 PP907567
*S. montecristina Urb. & Ekman I. Fragoso-Martínez et al. 527 (JBSD, MEXU) PP905449 PP907539 PP907559
*S. praeterita Epling I. Fragoso-Martínez et al. 607 (JBSD, MEXU) PP905450 PP907540 PP907560
S. selleana Urb. I. Fragoso-Martínez et al. 503 (JBSD, MEXU) PP905451 PP907541 PP907561
S. serotina L. I. Fragoso-Martínez et al. 506 (JBSD, MEXU) PP905452 PP907542 PP907562
*S. subaequalis Epling GenBank BioSample: SAMN22547053 PP905453
S. tenella Sw. T. Clase et al. 8266 (JBSD) PP905454 PP907543 PP907563
S. thormanii Urb. T. Clase et al. 8059 (JBSD) PP905455 PP907544 PP907564
S. tuerckheimii Urb. I. Fragoso-Martínez et al. 607 (JBSD, MEXU) PP905456 PP907545 PP907565
S. uncinata Urb. I. Fragoso-Martínez et al. 575 (JBSD, MEXU) PP905457 PP907546 PP907566

Sequence edition and alignment

The sequences were edited and assembled using Geneious v.10.2.6 (http://www.geneious.com, Kearse et al. 2012). For one species (S. subaequalis Epling), we downloaded the raw data from the GenBank BioSample SAMN22547053 (Rose et al. 2021), and we assembled the ITS region using the default settings for the internal transcribed spacers in GetOrganelle (Jin et al. 2020). The edited and assembled sequences were uploaded to Genbank (www.ncbi.nlm.nih.gov/Genbank) with the accession numbers shown in Table 1. To determine the phylogenetic position of the new taxa, as well as other sage species from the Antilles, we combined the newly sequenced data with the most comprehensive data matrix of Salvia subgenus Calosphace from Fragoso-Martínez et al. (2018). Additionally, we included sequences from posterior studies of the same research group (González-Gallegos et al. 2018; Martínez-Ambriz et al. 2019; Fragoso-Martínez et al. 2021). The matrices of each marker were aligned individually, using MAFFT (Katoh and Standley 2013).

The matrix comprising the three molecular markers included 288 taxa, of which 269 belong to Salvia subgenus Calosphace. The length of this matrix was 2,197 bp: 722 bp from the nrITS region, 505 bp from the trnH-psbA intergenic spacer and 970 bp from the trnL-trnF region. Based on the results of the congruence test between plastid and nuclear data performed in previous phylogenetic studies of Salvia subgenus Calosphace (Fragoso-Martínez et al. 2018), and to include the most comprehensive dataset of the subgenus, we decided to concatenate the matrices and analyze them together. However, we are aware that a certain amount of discordance between these kinds of datasets is expected due to different phenomena (e.g., hybridization, introgression, chloroplast capture, etc.) or differences in the inheritance process between the plastid and nuclear DNA. Thus, the trees from the separate analyses (ITS and plastid) are provided as part of Suppl. material 2.

Model selection and phylogenetic analyses

The concatenated matrix included three partitions corresponding to each molecular marker. For each partition, we evaluated 88 molecular substitution models using ModelFinder (Kalyaanamoorthy et al. 2017). The selected substitution models according to the Bayesian Information Criterion (BIC) were: GTR+F+I+G4 for the ITS region, TVM+F+I+G4 for the trnH-psbA IGS and TIM+F+G4 for the trnL-trnF region.

The Maximum likelihood analysis of the concatenated matrix was performed using the IQ-TREE algorithm (Nguyen et al. 2015) in the W-IQ-TREE web server (Trifinopoulos et al. 2016). Due to the length of the resulting tree, only the Angulatae clade is depicted in Fig. 4 and the complete tree is provided as Suppl. material 2. For this figure, the Angulatae clade was extracted from the main tree using the phytools package (Revell 2012) in R (R Core Team 2016) and edited with FigTree version 1.4.4 (Rambaut 2018).

Results

Taxonomic treatment

Salvia claseana Fragoso & Salazar, sp. nov.

Fig. 1

Diagnosis

Similar to Salvia calaminthifolia, but with the stems strigose (vs. cinereous); the leaves rhombic to trullate with the base cuneate, decurrent, the margin crenate-serrate (vs. deltoid-ovate, base truncate, margin subentire); the flowers bigger, with the calyx 5.5–8 mm long and the corolla tube 7.7–9 mm long (vs. 5–5.5 mm long and 5–6 mm long, respectively), and the lower lip reclinate (vs. reflexed).

Figure 1. 

Salvia claseana and the botanist that honors A flowering branch B leaves C flowers D Teodoro Clase at the JBSD herbarium E calyx F corolla G stamens H detail of the connective tooth (Specimen photographs taken from I. Fragoso et al. 529).

Type

Dominican Republic. Azua, Las Charcas • Paraje Boquerón, cañada subiendo por el puente Juana Guayacán, a 4 km de la desviación de la carretera Sánchez (Bani-Azua); 18°21'0.9138"N, 70°31'51.8118"W; 307 m; 28 Nov 2016; I. Fragoso-Martínez, G.A. Salazar & T. Clase 529 (holotype: JBSD 129667; isotypes: MEXU 1512146, XAL 0154233).

Description

Suffruticose herbs , ca. 0.5 m tall; stems strigose with antrorsely appressed trichomes, internodes 1.5–4 cm long. Leaves rhombic to trullate, 1.3–3.6 × 1–3 cm, base cuneate, decurrent, apex acute, margin crenate-serrate, ciliate; upper leaf surface bullate, glabrescent; lower leaf surface pale, densely pubescent, simple trichomes minute, white, with amber spherical glands; petioles 0.5–1.2 cm long. Flowers axillary, 4(–8) per node; pedicels 2.5–6 mm long, trichomes simple, adpressed. Calyx green, tubular-campanulate, 5.5–8 mm long, strigose, with simple and glandular trichomes; tube 4.7–6.5 × 3 mm; lobes deltate, apex apiculate, upper lobe 1.3–1.8 mm long, straight, 3-veined, margin ciliate, lower lobes 1.3–2.5 mm long, straight. Corolla violet with white nectar guides in the lower lobe, ca. 1 cm long, tube 7.7–9 × 2.6–3 mm, ventricose, invaginate, internally epapillate; lobes unequal in length, upper lobe galeate, 3.8–6 mm long, densely pubescent, lower lobe 5–8 × 7.8–8.4 mm, tetralobate, reclinate. Stamens included in the upper corolla lobe, fused close to the corolla opening; filaments 1.5–2 mm long; connective 7–8 mm long, sparsely pilose, with a bilobed tooth close to the insertion with the filament, straight; upper arm of the connective shorter than the lower arm, 3–3.5 mm long, thecae 1–1.5 mm long; lower arm 4.5–5 mm long. Style 9–11 mm long, densely pubescent near the branches, upper branch longer than the lower one, lower branch spathulate. Nectary disc surface with spherical glands near the base of the mericarps, nectary horn ca. 2 mm long, oblong, laterally compressed. Mericarps ovoid, 1.6–2 × 0.7–0.9 mm, smooth.

Phenology

Flowers were documented from November to May. Fruits have been observed after this period.

Etymology

The epithet “claseana” honors the Dominican botanist Teodoro Clase, head of the Botany Department of the Jardín Botánico Nacional “Dr. Rafael M. Moscoso”. His botanical expeditions have resulted in ca. 12,000 collected specimens and he has described ca. 12 new species of angiosperms from Hispaniola. These contributions are undoubtedly crucial to the knowledge of the flora of Dominican Republic.

Distribution, habitat and conservation status

Endemic to the dry forests with limestone soils from the Sierra Martín García in Azua, Dominican Republic (Fig. 3). Salvia claseana is represented by three collections, each from a different population. In the GeoCat (Bachman et al. 2011) analysis, the species’ extent of occurrence (EEO) is 22.86 km2 with an area of occupancy (AOO) of 12 km2. Considering these results, combined with the high level of endemism of the region, and taking into account the IUCN criteria (IUCN, 2022), we suggest that S. claseana is placed in the category of Critically Endangered (CR).

Comments

Phylogenetically, this species is closely related to Salvia praeterita (Fig. 4); however, the latter has the flowers arranged into racemes, while the new species produces flowers in the leaf axils. Morphologically, Salvia claseana resembles Salvia calaminthifolia the most but differs from it by a number of characters (Table 2), from which the most conspicuous are: the pubescence, which is strigose in the former and cinereus in the latter, the shape and base of the leaves (rhombic to trullate, cuneate vs. deloid-ovate, truncate) and the size of its flowers (bigger in the new species, Table 2). Additionally, Salvia calaminthifolia is not closely related to the new species (Fig. 4) and it seems to be only distributed in Haiti (Liogier, 1994), despite the locality cited in the type specimen (Santo Domingo). No distribution data or collections from Dominican Republic were found representing this species either in the JBSD herbarium or global databases such as GBIF.org (GBIF Secretariat 2023).

Table 2.

Morphological comparison among the new taxa and their more similar species, based data from type specimens, Epling (1939) and Liogier (1994).

S. claseana Fragoso & Salazar S. calaminthifolia Vahl S. praeterita Epling S. martineziana Fragoso & Martínez-Ambr. S. brachyphylla Urb.
STEM
Pubescence Strigose, trichomes antrorse Cinereous, trichomes straight Strigose, trichomes antrorse Strigose, trichomes retrorse Hispid, trichomes straight
LEAF
Shape Rhombic to trullate Deltoid-ovate Ovate to deltoid-ovate Obovate to flabellate Obovate to flabellate
Size (cm) 1.3–3.6 × 1–3 0.5–1.2 × 0.7–1.5 1–2 × 0.5–1 1–1.5 × 0.5–1 1.5–3 × 1–2.5
Base Cuneate, decurrent Truncate Cuneate, decurrent Cuneate, decurrent Cuneate, decurrent
Margin Crenate-serrate Subentire Crenate-serrate Crenate-serrate Crenate-serrate
Upper surface Bullate Smooth Bullate Bullate Bullate
Lower surface Densely pubescent Cinereous, trichomes adpressed Densely pubescent Tomentulose Hispid
INFLORESCENCE
Presence Absent (axillary flowers) Absent (axillary flowers) Present, terminal racemes Absent (axillary flowers) Present, axillary and terminal racemes
Flowers per node 4(–8) 2–6 2–6 2 3
CALYX
Shape Tubular-campanulate Tubular-campanulate Tubular-campanulate Campanulate Tubular-campanulate
Length (mm) 5.5–8 5–5.5 5–7 5–6.8 5.5–6.5
COROLLA
Tube length (mm) 7.7–9 5–6 8.5–9.5 7–9 7.5–9.5
Upper lip length (mm) 3.8–6 3–4.7 4–4.6 4.5–6.5 5.6–6
Lower lip length (mm) 5–8 3.5–5.5 5–6 7–7.5 ca. 6.6
Lower lip position Reclinate Reflexed Reclinate Reflexed Reclinate

Additional specimens examined

Dominican Republic. Azua, Barreras • Sierra Martín García, en los alrededores de Barreras; 170 m; 11 Sep 1984; M. Mejía et al. 1180 (JBSD). Las Charcas • Parque Nacional Francisco Alberto Caamaño, paraje Boquerón; 18° 21'38.4366"N, 70°31'51.24"W; 298 m; 9 May 2014; T. Clase & R. Ovidio S. 8645 (JBSD, MEXU).

Salvia martineziana Fragoso & Martínez-Ambr., sp. nov.

Diagnosis

Similar to Salvia brachyphylla, but differing from it by the pubescence of the plant (strigose with retrorse trichomes vs. hispid with erect trichomes); having smaller leaves (1–1.5 × 0.5–1 cm vs. 1.5–3 × 1–2.5) with a tomentulose lower leaf side (vs. hispid); flowers axillary (vs. forming racemes), calyx campanulate (vs. tubular-campanulate) and lower lobe of the corolla reflexed (vs. reclinate).

Figure 2. 

Salvia martineziana and the botanist that honors A flowering branch B leaves C flowers D Martha Martínez-Gordillo conducting fieldwork in Mexico E calyx F corolla G stamens H detail of the connective tooth (Specimen photographs taken from I. Fragoso et al. 497).

Type

Dominican Republic. Independencia, Duvergé • Parque Nacional Sie­rra de Bahoruco, 6.8 km al S de Puerto Escondido por el camino a la Caseta 1; 18°17'10.4994"N, 71°34'6.3978"W; 965 m; 24 Nov 2016; I. Fragoso-Martínez, G.A. Salazar & T. Clase 497 (holotype: JBSD 129573; isotypes: MEXU 1512142, XAL 0154234).

Description

Suffruticose herbs , 0.3–0.5 m tall; stems strigose with retrorsely appressed trichomes, internodes 0.8–2.4 cm long. Leaves obovate to flabellate, 1–1.5 × 0.5–1 cm, base cuneate, decurrent, apex acute to obtuse, margin crenate-serrate, ciliate; upper leaf surface bullate, densely hirsute; lower leaf surface whitish, tomentulose simple trichomes minute, curved, white, with yellow spherical glands; petioles 4–8 mm long. Flowers axillary, 2 per node; pedicels 1.5–4.5 mm long, hirsute. Calyx green, campanulate, 5–6.8 mm long, densely hirsute, with simple and glandular trichomes; tube 3–5 × 3 mm; lobes ovate-deltate, apex apiculate, upper lobe 2.5–4 mm long, curved backwards, 3-veined, margin ciliate, lower lobes 1.6–2.5 mm long, straight. Corolla violet with white nectar guides in the lower lobe, 1.2–1.4 cm long, tube 7–9 × 2.9 mm, ventricose, invaginate, internally epapillate, lobes unequal in length, upper lobe galeate, 4.5–6.5 mm long, densely pubescent, lower lobe 7–7.5 × 8.4 mm, tetralobate, reflexed. Stamens included in the upper corolla lobe, fused close to the corolla opening; filaments 1.5–2 mm long; connective 7.5–8.5 mm long, sparsely pilose, with an entire tooth close to the insertion with the filament, retrorse; upper arm of the connective slightly longer than the lower arm, 4–4.5 mm long, thecae 1–1.5 mm long; lower arm 3.5–4 mm long. Style 13–15 mm long, densely pubescent near the branches, with simple and capitate glandular trichomes, upper branch longer than the lower one, lower branch spathulate. Nectary disc surface with spherical glands near the base of the mericarps, nectary horn ca. 1 mm long, oblong, laterally compressed. Mericarps ovoid, 1.5–2 × 0.4–0.6 mm, smooth.

Phenology

Flowers were documented from November to May. Fruits have been observed after this period.

Etymology

The epithet “martineziana” honors the Mexican botanist Martha Martínez Gordillo, specialist of the Euphorbiaceae and Lamiaceae families. Dr. Martínez works at the FCME herbarium at the Universidad Nacional Autónoma de México. She has conducted fieldwork mainly in the states of Chiapas, Guerrero, Mexico and Oaxaca. Her study of the Mexican flora, particularly that of Gue­rrero, has led to the description of more than 30 species of angiosperms, many of them from the genus Salvia. Dr. Martínez has taught botany to numerous generations of Mexican biologists, and her exemplary professional and academic ethics, determination, kindness and generosity are an inspiration to her students.

Distribution, habitat and conservation status

Endemic to the dry forests with limestone soils from Sierra de Bahoruco in Independencia, Dominical Republic (Fig. 3). Salvia martineziana is represented by two collections, each seemingly belonging to different populations, both from the southern region of Puerto Escondido in the Sierra de Barohuco. In the GeoCat (Bachman et al. 2011) analysis, the species’ extent of occurrence (EEO) is 0.09 km2 with an area of occupancy (AOO) of 8 km2. Considering these results, combined with the high level of endemism of the region, and taking into account the IUCN criteria (IUCN 2022), we suggest that S. martineziana should be placed in the category of Critically Endangered (CR).

Figure 3. 

Distribution map of the four taxa of Salvia section Urbania from Dominican Republic. Salvia calaminthifolia was excluded from the map due to the lack of distribution data in the country aside from the type specimen.

Comments

This species is sister to a clade formed by two other Dominican species of Salvia section Urbania (Fig. 4). It differs from S. praeterita by lacking flowers in racemes and from S. claseana by the size of the leaves (smaller in S. martineziana: 1–1.5 × 0.5–1 cm vs. 1.3–3.6 × 1–3 cm) and the pubescence of the lower side of the blades (tomentulose vs. densely pubescent). Morphologically, S. martineziana resembles S. brachyphylla the most, a Haitian species. However, it differs mainly by the lack of racemes, having the flowers distributed in the axils of the upper portion of the branches.

Figure 4. 

Phylogenetic relationships of the Angulatae clade based on three molecular markers (nrITS, trnH-psbA and trnL-trnF) with an increased taxon sampling including the new species and other Hispaniolan taxa. Bootstrap values ≥80% are shown above the branches. The names of the new species are written in purple, while the names in gray correspond to the sections of the taxa from the Hispaniolan subclade. An extended version of the phylogenetic tree is provided in Suppl. material 2.

Additional specimens examined

Dominican Republic. Independencia, Duvergé • 5.2 km al S de Puerto Escondido en camino a Acetillar, Sierra de Bahoruco; 170 m; 9 May 1985; T. Zanoni et al. 34648 (JBSD).

Identification key for the species of Salvia section Urbania

1a Flowers in racemes 2
2a Leaves and calyces densely hispid S. brachyphylla
2b Leaves and calyces strigose to glabrate 3
3a Bracts persistent 4
4a Bracts flabellate, surrounding the flowers almost completely S. saccifera
4b Bracts ovate, not surrounding the flowers completely S. subglabra
3b Bracts deciduous 5
5a Lower leaf surface greenish, glabrous; flowers 6–12 per node S. hotteana
5b Lower leaf surface whitish, strigose; flowers 2–6 per node S. praeterita
1b Flowers axillary, not forming racemes 6
6a Leaf margin subentire, entire or sinuate 7
7a Leaves coriaceous S. montecristina
7b Leaves membranaceus 8
8a Lower leaf surface incanous, grayish S. mornicola
8b Lower leaf surface glabrate, greenish 9
9a Leaves oblong-elliptical S. tortuensis
9b Leaves deltoid-ovate S. subaequalis
6b Leaf margin crenate-serrate 10
10a Leaf base truncate S. calaminthifolia
10b Leaf base cuneate 11
11a Leaves rhombic to trullate, 1–3 cm wide, lower leaf surface densely pubescent S. claseana
11b Leaves obovate to flabellate, 0.5–1 cm wide, lower leaf surface tomentulose S. martineziana

Phylogenetic relationships

The Hispaniolan species included in the phylogenetic analysis are distributed in four different clades. The species from Salvia section Micranthae (Benth.) Epling — Salvia serotina L. and S. tenella Sw.—, are part of a clade that also contains taxa from sections Bracteata Epling and Subrotundae (Epling) Epling (99% BS; Suppl. material 1). Salvia decumbens Alain is included in the Flocculosae clade, along with species from the Andean region such as S. discolor Kunth and S. leucocephala Kunth (100% BS; Suppl. material 1). Salvia occidentalis Sw. is part of the Uliginosae clade and it is closely related to other species of section Microsphace (Briq.) Benth. (100% BS; Suppl. material 1). Most of the taxa surveyed are found in the Angulatae clade (Fig. 4), forming a Hispaniolan subclade (100% Bootstrap Support; BS). The species from this subclade belong to four different sections (Table 3) and one is unassigned (Salvia lavendula Alain). From these, only sections Urbania Epling and Gardoquiiflorae Epling are monophyletic (both with 100% BS). However, the latter is nested in a clade formed by species of two sections: the paraphyletic Ekmania Epling and the polyphyletic Wrightiana (Fig. 4).

Table 3.

Sections sampled and the percentage of taxa sequenced in this study.

Epling’s sections sampled Taxa sampled from the total
Ekmania Epling 8 spp. (100%)
*Gardoquiiflorae Epling 2 spp. (66%)
Micranthae 4 spp. (57%)
*Urbania Epling 6 spp. (50%)
Wrightiana Epling 2 spp. (66%)

The new species described in the present study belong to the monophyletic section Urbania. Within this clade Salvia montecristina Urb. & Ekman, endemic to Dominican Republic, is sister to the remaining species (Fig. 4). These taxa are arranged into two subclades, one containing Haitian species (100% BS) and another one formed by Dominican taxa (99% BS). The latter subclade comprises the new species and Salvia praeterita Epling.

Discussion

The new taxa described here are an addition to the sage species of Dominican Republic, increasing its diversity to 23 species (González-Gallegos et al. 2020), five of them belonging to Salvia section Urbania. As depicted in Fig. 3, these taxa are microendemic and their range of distribution is very narrow. Consequently, the conservation status assessment for the new taxa placed them both in the critically endangered (CR) category, following the IUCN criteria (IUCN 2022). Future studies should address the state and characteristics of the populations, to provide further insight into their dynamics and expand information about their conservation status.

Both species described here fit the circumscription of Salvia section Urbania provided by Epling (1939). Moreover, this section was proved here to be monophyletic, rendering its phylogenetic relationships congruent with the morphology of the group. The clade formed by this section is related to other Hispaniolan taxa belonging to three other sections, only one of them monophyletic (Table 3). These results are consistent with the findings of Fragoso-Martínez et al. (2018), where many of the clades of the Neotropical sages show strong geographical structure instead of morphological congruence. Nevertheless, section Urbania shows both morphological and geographic structures. Only a few of the 41 non-monotypic sections sampled to date show the same pattern (e.g., Lavanduloideae Epling, Membranaceae (Benth.) Epling and Sigmoideae Epling); however, less than half of the species of Salvia subgenus Calosphace have been sequenced. Thus, there are still relationships to be explored or defined.

Zona et al. (2016) conducted the first phylogenetic analysis of Hispaniolan sages. Their study, based on five species, suggested that the island’s sage diversity originated from at least two colonization events. Our expanded taxonomic sampling has now revealed two additional colonization events: one involving species from section Micranthae and another from section Microsphace.

Despite a recent surge in the discovery of new Neotropical sages, the Dominican Republic has remained relatively unexplored in this regard. The last new sage species from the country was described almost 40 years ago (Liogier 1988). This underscores the critical need for continued botanical expeditions and taxonomic research to broaden our understanding of plant diversity in this region. Furthermore, incorporating more species of Salvia subgenus Calosphace into phylogenetic analyses will provide valuable insights into the evolutionary history of this lineage, and will allow us to group taxa with their closest relatives.

These newly discovered taxa from Hispaniola, which are critically endangered, are the first species of Salvia section Urbania to be evaluated under the IUCN criteria. However, they mirror a pattern seen in other native sages on the island, where some species are vulnerable (Salvia arborescens Urb. & Ekman, and S. decumbens Alain) and others are critically endangered (Salvia buchii Urb., S. haitiensis Urb., S. lachnaioclada Briq. and S. paryskii Skean & Judd). These results highlight the need for a comprehensive assessment of all Hispaniolan sage species, to develop effective strategies for their protection.

Acknowledgements

IFM is grateful for the support from the Posgrado en Ciencias Biológicas, Universidad Nacional Autónoma de México during her doctoral studies and the scholarship for doctoral studies (grant 324065) from Consejo Nacional de Ciencia y Tecnología (CONACyT, Mexico). The authors thank the Ministerio de Medio Ambiente y Recursos Naturales from the Dominican Republic for the collection permit; the staff of the JBSD herbarium, for managing the exchange and donation of the specimens; Teodoro Clase by providing information on the localities of the Dominican sages and conducting fieldwork with IFM and GAS. We also acknowledge María del Rosario García Peña of the MEXU herbarium, Elizabeth Séptimo from the JBSD herbarium and Israel Acosta Rosado of the XAL herbarium, for the facilities provided to process the type specimens. Finally, the authors are grateful to A.I. Bieler Antolín for the photographs of the floral structures; L.I. Cabrera for guidance with lab work.

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

The Society of Systematic Biologists (SSB) provided a grant to IFM to conduct fieldwork in the Dominican Republic. Additional financial support for the laboratory work and sequencing was provided by the Programa Fronteras de la Ciencia FC-2016 (Project 1867). Publication fees were covered by the SNII-CONAHCYT grant (83797) provided to IFM.

Author contributions

IFM wrote the manuscript, EMA, GAS and MR provided feedback on the draft and made insightful suggestions to improve it. IFM and GAS conducted the field expedition in 2016 and MR provided them with information about the localities of the surveyed taxa. EMA extracted the DNA samples, amplified the molecular markers, made the map, assessed the conservation status of the new taxa and assembled all the figures of the manuscript. IFM processed the molecular data and performed phylogenetic analyses.

Author ORCIDs

Itzi Fragoso-Martínez https://orcid.org/0000-0003-3661-1076

Gerardo A. Salazar https://orcid.org/0000-0002-5203-5374

Emmanuel Martínez-Ambriz https://orcid.org/0000-0002-4770-4174

Martin Reith https://orcid.org/0000-0001-7830-9161

Data availability

All the newly sequenced data was uploaded to the NCBI with the accession numbers provided in Table 1. The complete phylogenetic tree of the combined dataset is provided as Suppl. material 2.

References

  • Bachman SP, Moat J, Hill A, de la Torre J, Scott B (2011) Supporting Red List threat assessments with GeoCAT: Geospatial conservation assessment tool. ZooKeys 28: 117–126. https://doi.org/10.3897/zookeys.150.2109
  • Doyle J, Doyle J (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19: 11–15.
  • Epling C (1939) A revision of Salvia subgenus Calosphace. Repertorium Specierum Novarum Regni Vegetabilis 110: 380.
  • Fragoso-Martínez I, Martínez-Gordillo M, Salazar GA, Sazatornil F, Jenks AA, García Peña MR, Barrera-Aveleida G, Benitez-Vieyra S, Magallón S, Cornejo-Tenorio G, Granados Mendoza C (2018) Phylogeny of the Neotropical sages (Salvia subg. Calosphace; Lamiaceae) and insights into pollinator and area shifts. Plant Systematics and Evolution 304(1): 43–55. https://doi.org/10.1007/s00606-017-1445-4
  • González-Gallegos JG, Fragoso-Martínez I, González-Adame G, Martínez-Ambriz E, López-Enríquez I (2018) Salvia ozolotepecensis, S. patriciae and S. sirenis (Lamiaceae), three new species from Miahuatlán district, Oaxaca, Mexico. Phytotaxa 352(2): 143–159. https://doi.org/10.11646/phytotaxa.362.2.2
  • González-Gallegos JG, Bedolla-García BY, Cornejo-Tenorio G, Fernández-Alonso JL, Fragoso-Martínez I, García-Peña MR, Harley RM, Klitgaard B, Martínez-Gordillo MJ, Wood JRI, Zamudio S, Zona S, Xifreda CC (2020) Richness and distribution of Salvia subgenus Calosphace (Lamiaceae). International Journal of Plant Sciences 181(8): 831–856. https://doi.org/10.1086/709133
  • Harley RM, Atkins S, Budantsev AL, Cantino PD, Conn BJ, Grayer R, Harley MM, de Kok R, Krestovskaja T, Morales R, Paton AJ, Ryding O, Upson T (2004) Lamiaceae. In: Kubitzki K, Kadereit JW (Eds) The families and genera of Vascular Plants VII. Dicotyledons: Lamiales (except Acanthaceae including Avicenniaceae). Springer, Berlin, 167−275. https://doi.org/10.1007/978-3-642-18617-2_11
  • Jenks AA, Walker JB, Kim S-C (2013) Phylogeny of New World Salvia subgenus Calosphace (Lamiaceae) based on cpDNA (psbA-trnH) and nrDNA (ITS) sequence data. Journal of Plant Research 126(4): 483–496. https://doi.org/10.1007/s10265-012-0543-1
  • Jin J-J, Yu W-B, Yang J-B, Song Y, dePamphilis CW, Yi T-S, Li D-Z (2020) GetOrganelle: A fast and versatile toolkit for accurate de novo assembly of organelle genomes. Genome Biology 21(1): 241. https://doi.org/10.1186/s13059-020-02154-5
  • Kalyaanamoorthy S, Minh BQ, Wong TKF, Von Haeseler A, Jermiin LS (2017) ModelFinder: Fast model selection for accurate phylogenetic estimates. Nature Methods 14(6): 587–589. https://doi.org/10.1038/nmeth.4285
  • Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30(4): 772–780. https://doi.org/10.1093/molbev/mst010
  • Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A (2012) Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics (Oxford, England) 28(12): 1647–1649. https://doi.org/10.1093/bioinformatics/bts199
  • Liogier AH (1988) Novitates Antillanae XIV. Phytologia 64(5): 345–348.
  • Liogier AH (1994) Salvia (Labiatae). In: Liogier AH (Ed.) La Flora de La Española VI. Universidad Central del Este, San Pedro de Macorís, 293–314.
  • Martínez-Ambriz E, Fragoso-Martínez I, Martínez-Gordillo M (2019) A new species of Salvia from the Fulgentes clade (Lamiaceae), from Puebla, Mexico. Phytotaxa 409(1): 29–38. https://doi.org/10.11646/phytotaxa.409.1.4
  • Nguyen L-T, Schmidt HA, von Haeseler A, Minh BQ (2015) IQ-TREE: A fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology and Evolution 32(1): 268–274. https://doi.org/10.1093/molbev/msu300
  • QGIS Development Team (2023) QGIS Geographic Information System Ver. 3.30. Open Source Geospatial Foundation Project. http://qgis.osgeo.org [accessed: 01.08.2024]
  • R Core Team (2016) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna. https://www.R-project.org/
  • Rose JP, Kriebel R, Kahan L, DiNicola A, González-Gallegos JG, Celep F, Lemmon EM, Lemmon AR, Sytsma KJ, Drew BT (2021) Sage insights into the phylogeny of Salvia: Dealing with sources of discordance within and across genomes. Frontiers in Plant Science 12: 1–14. https://doi.org/10.3389/fpls.2021.767478
  • Thiers B (2023) Index Herbariorum: a global directory of public herbaria and associated staff. New York Botanical Garden’s Virtual Herbarium. http://sweetgum.nybg.org/ih [accessed: 20.07.2024]
  • Trifinopoulos J, Nguyen L-T, von Haeseler A, Minh BQ (2016) W-IQ-TREE: A fast online phylogenetic tool for maximum likelihood analysis. Nucleic Acids Research 1–4(W1): W232–W235. https://doi.org/10.1093/nar/gkw256
  • Zona S, Finch K, Clase T, Jestrow B (2016) A synopsis of Salvia sect. Gardoquiiflorae (Lamiaceae), with a note on the origins of Caribbean Salvia species. Phytotaxa 255(3): 214–226. https://doi.org/10.11646/phytotaxa.255.3.3

Supplementary materials

Supplementary material 1 

Additional information of the sampled sage species in the study

Itzi Fragoso-Martínez, Gerardo A. Salazar, Emmanuel Martínez-Ambriz, Martin Reith

Data type: pdf

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (196.53 kb)
Supplementary material 2 

Complete phylogenetic trees of the plastid, nuclear and combined dataset

Itzi Fragoso-Martínez, Gerardo A. Salazar, Emmanuel Martínez-Ambriz, Martin Reith

Data type: pdf

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Download file (8.92 MB)
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