Flora of Espírito Santo: Clusiaceae

Abstract We present here the treatment of Clusiaceae for the flora of the state of Espírito Santo, Brazil. Sixteen species are recognized: seven species of Clusia, four species of Tovomita, two species of Garcinia, two species of Tovomitopsis, and one species of Symphonia. A brief history of the Clusiaceae in Espírito Santo is presented, as well as descriptions, illustrations, and taxonomic commentary on the species.


Introduction
Clusiaceae is a monophyletic group represented by shrubs, trees or hemiepiphytes species, with exudate of different colors, opposite, usually glabrous leaves, without stipules; thyrsoids, terminal or axillary inflorescences, and uni-or bisexual flowers (Stevens 2007;Marinho et al. 2020).The family is inserted in the clusioid clade (Malpighiales), forming a sister group with Bonnetiaceae (Ruhfel et al. 2013).Clusiaceae has a tropical distribution and is composed of three tribes, 15 genera and ~800 species (Stevens 2001 onwards;Marinho et al. 2019).
The Clusiaceae of Brazil is represented by 11 genera and 147 species, of which 49 species are endemic to the country.In the state of Espírito Santo, the family is represented by genera Clusia, Garcinia, Symphonia, Tovomita and Tovomitopsis totaling 16 species (Marinho et al. 2020).

Brief history of Clusiaceae in Espírito Santo
The first species of Clusiaceae based on specimens from Espírito Santo was proposed by Mariz & Weinberg (1982), Clusia spiritu-sanctensis G. Mariz & B. Weinberg (1982: 233).The species was described from specimens collected on rocky outcrops in the region of Vila Velha and resembled the Clusia fluminensis Planchon & Triana (1860: 349), which is cultivated in Brazil.This same region, now in the city of Domingos Martins, was also important for the knowledge of Clusiaceae in the state.The species Clusia aemygdioi Gomes da Silva & B. Weinberg (1985: 162) was described from specimens collected in the high forests of Domingos Martins (Gomes da Silva & Weinberg 1985).Both C. spiritu-sanctensis and C. aemygdioi are accepted species and are described in the present work.Gomes da Silva & Weinberg (1984) Rodriguésia 74: e00552022.2023 also described Clusia marizii Gomes da Silva & B. Weinberg (1984: 22) from specimens collected in riparian forests of Vargem Alta; today, this species is under the synonym of Clusia organensis Planchon & Triana (1860: 349).
The next contribution to the Clusiaceae of Espírito Santo took 27 years to happen.Riguete et al. (2012) published a study on the fruit morphology in the recognition of Clusia species occurring in the state of Espírito Santo, where they presented diagnoses for the fruits, illustrations, photos of living specimens and identification keys for the fruits.Until the publication of this work, the other genera of Clusiaceae that occur in the state only appeared in floristic lists, but without any extensive taxonomic treatment.Four species of Tovomita were included in a review of the genus in the Atlantic Forest (Marinho et al. 2016), and Tovomitopsis was first recorded for the state by Zorzanelli et al. (2015), which also presented a diagnose and photos in the field.In this way, for the first time and almost 40 years after the first species described for the state, a taxonomic treatment of Clusiaceae for Espírito Santo is presented in the present work, including descriptions of the species, identification keys and field images.

Material and Methods
We visited some important collections from the Espírito Santo state (CVRD, MBML, VIES) and also national and international herbaria, considering the most representative collections of Clusiaceae (ESA, INPA, K, NY, RB, SPF, UEC, acronyms following Thiers, continuously updated).Additional specimens from Espírito Santo were consult in online databases such as SpeciesLink (<splink.org.br>) and Reflora (<reflora.jbrj.gov. br>).The names of the authors of taxa follow those used in Flora e Funga do Brasil Project (2022).

Results and Discussion
Clusiaceae Lindl.
Clusiaceae plants can be shrubs, trees, or hemiepiphytes and secrete an exudate that can be translucent, white, yellow, greenish, orange, or red.The leaves are opposite, usually decussate, entire, and glabrous, without stipules.Secretory ducts can be conspicuous on the leaf blade or absent.The inflorescences are terminal or axillary, solitary flowers rarely occur, bracts and bracteoles can be present or absent.The flowers are uni-or bisexual, actinomorphic, usually 4-5(-8)-merous.The sepals are free or connate, usually decussate.The petals are free, decussate, or imbricate.The stamens are free or fused in synandrium or staminal tube, resin secretors or not, the anthers are longitudinal or poricidal, staminodes can be present or absent in staminate flowers, with resin secretion or not, and a pistillode can be present or absent in staminate flowers.The gynoecium is composed of a 2-5(-21)-locular ovary, style usually short, generally with large stigma; nectariferous disc can be present or absent, staminodes can be present or absent in pistillate flowers, with resin secretion or not.The fruits can be dehiscent or indehiscent, berries or carnose capsules, septicidal or septifragal.The seeds can be surrounded by an aril or not, be vascularized or not.The aril is usually orange, occasionally white, yellow, or red (Stevens 2001 onwards;Stevens 2007;Marinho et al. 2020).The species of Clusia are trees, shrubs, or hemiepiphytes, dioecious, rarely hermaphrodite, with eperulate terminal buds and a white or yellowish exudate.The leaves are opposite, glabrous, petiolate or sessile, with coriaceous or subcoriaceous leaf blades, secondary veins barely conspicuous or not, and secretory ducts conspicuous or not on the leaf blade.The inflorescences are terminal, thyrsoid or solitary flowers.The flowers are showy, often resinous.The calyx is free, 4-5 (-17) sepals.The corolla is free, 4-8(-10) petals, usually opposed to sepals.The staminate flowers with 4-∞ stamens, free or fused in a synandrium, with resin secretors or not, anthers are longitudinal or poricidal, staminodes can be present or not, with resin secretors or not, and pistillode can be present or not.The pistillate flowers have of a 4-21-locular ovary, with short or absent styles, rarely conspicuous, terminal or subterminal stigma, papillose or smooth, staminodes can be present or not, with resin secretion or not.The fruits are dehiscent, carnose, septicidal capsules, persistent stigmas, sepals, petals, and staminodes, persistent or not.The seeds have a non-vascularized aril, orange or red.

Identification key for Clusiaceae genera in Espírito Santo
Clusia has ~400 species.Seventy-nine species can be found in Brazil, of which 30 are endemic (Marinho et al. 2020).Most Clusia species are found in the northern region of Brazil, mainly in the Amazon phytogeographic domain.In the Southeast Region there are 17 species, distributed mainly in regions of Cerrado and Atlantic Forest.The six species of Clusia found in the state of Espírito Santo occur mainly in areas of rocky outcrops (inselbergs), high elevation in Atlantic Forest and restingas.
Clusia aemygdioi is similar to Clusia hilariana, mainly regarding its flowers and fruits.These species differ in that Clusia aemygdioi presents subsessile petiole (up to 5 mm long) and elliptical or obovate leaves with a rounded or obtuse base and rounded and slightly obtuse apex, while Clusia hilariana has a larger petiole (9-35 mm long) and obovate leaves with acute, cuneate, or attenuate base and rounded, occasionally truncate apex.Schltdl., Linnaea 8: 181. 1833.
Clusia hilariana is morphologically similar to Clusia aemygdioi.These species have large and showy flowers, staminate flowers with stamens organized in a conical synandrium without pistillode, with resinous staminodes forming a ring at the base of the synandrium, and pistillate flowers with many carpels, large ovary, with resinous staminodes at the base of the ovary.The fruits are large and the persistent subapical stigmas in the fruit form a crown-like structure.The diagnostic characters of C. hilariana were discussed in the comments of C. aemygdioi.Leaf blades 4-12 × 3-7 cm, green on adaxial side in vivo, light green on abaxial side in vivo, brown to light brown on both sides in sicco, coriaceous, obovate, base acute, cuneate, or decurrent, apex rounded or truncate, margin revolute.Midrib proeminent on both sides, visible up to the apex of Rodriguésia 74: e00552022.2023 the leaf blade; secondary veins 1.5-3 mm apart from each other, slightly prominent or inconspicuous on both sides, forming an angle of 40°-60° with the midrib; inframarginal vein slightly conspicuous or inconspicuous on the adaxial side, crenate.Secretory ducts inconspicuous.Flowers grouped in terminal thyrsoid inflorescences, with numerous flowers; peduncle 25-35 mm long; bracts 2, ovate to triangular, carenate, early caducous; bracteoles 4, ovate to triangular, carenate, persistent on fruit.Floral buds globose, green.Sepals 4, 3.8-4.5 × 3.7-4 mm, decussate, triangular-subrotund, subcoriaceous with hyaline margin, green; petals 4, 3.5-4.5 × 2.5-3.5 mm, obovate, fleshy, cream to yellowish green.Staminate flowers with numerous free stamens, yellowish green in vivo, not resiniferous, 2.5-4 mm long; anthers apical, caducous by longitudinal slits; pistillode absent.Pistillate flowers with 4 short non-resiniferous staminodes, linear, anantherous, early caducous; ovary 4-locular, 3-4 mm long, oblong, styles erect, short, connivant, stigma apical, connivent.Capsules submature with 10-15 × 5-8 mm, sepals persistent, petals and staminodes early caducous, stigmas apical.Seed aril orange.Clusia melchiorii has disjunct distribution in the Guyana Shield (Brazil, Colombia, Guyana and Suriname) and in eastern Brazil, in the states of Bahia and Espírito Santo, occurring in montane to upper montane elfin forests in the Guyana Shield and in rocky outcrop areas, restingas and Tabuleiro forests in Bahia and Espírito Santo.Clusia melchiorii is assessed as a species of Least Concern (LC) according to the IUCN Red List (BGCI & IUCN 2019).

Clusia nemorosa
Clusia organensis has staminate flowers formed by free and claviform stamens, covered with resin and with a central pistillode, similar to the staminate flowers of Clusia spiritu-sanctensis.These species differ in that Clusia organensis has subcoriaceous and oblanceolate leaf blades, and flowers with vinaceous petals, or red with white margin, while Clusia spiritu-sanctensis has coriaceous and obovate leaf blades, and flowers with petals that are white with red internal base.Some populations of C. organensis that occur in Rio de Janeiro may have flowers with white or cream petals.Fig. 1i Shrubs, trees, or hemiepiphytes, up to 12 m tall; exudate white or cream.Petiole 5-9 × 1-2 mm, tightly carenate.Leaf blades 2.5-16 × 1.5-12 cm, green on adaxial side in vivo, light green on abaxial side in vivo, brown to light brown on both sides in sicco, coriaceous, obovate, base acute, cuneate, or decurrent, apex rounded or truncate, margin revolute.Midrib proeminent on both sides, visible up to the apex of the leaf blade; secondary veins 1.5-2.5 mm apart from each other, slightly prominent or inconspicuous on both sides, forming an angle of 30°-50° with the midrib; inframarginal vein slightly conspicuous or inconspicuous on the adaxial side, crenate.Secretory ducts inconspicuous.Flowers grouped in thyrsoid inflorescences, terminal: ♂ 5-9 flowers, ♀ 3 flowers; peduncle 15-25 mm long; bracts 2, ovate to triangular, carenate, early Clusia spiritu-sanctensis occurs throughout the state of Espírito Santo and in the northern region of the state of Rio de Janeiro.This species occurs in restingas, tropical rain forest, and rocky outcrops.Clusia spiritu-sanctensis was assessed as Data Deficient (DD) in the red list of Espírito Santo (Fraga et al. 2019).
Clusia spiritu-sanctensis is very similar to Clusia fluminensis Planch.& Triana, a species that occurs naturally in Rio de Janeiro and is also cultivated as ornamental.These species differ mainly in their distribution and in that C. spiritusanctensis presents staminate flowers with 30-40 stamens and reduced pistillode, at the same height or less than the stamens, while C. fluminensis presents staminate flowers with 10-15 stamens and a well-developed pistillode, greater than the height of the stamens.Vegetatively, C. spiritu-sanctensis presents strongly carenate (keeled) petiole and midrib, while C. fluminensis presents slightly carenate petiole and midrib.
Clusia spiritu-sanctensis has greater morphological similarity with Clusia organensis; to distinguish these taxa, see the comments under Clusia organensis.

Garcinia L.
Garcinia is composed of shrubs, treelets and medium-sized trees with eperulate terminal buds.The species secrete a white to yellowish exudate and can be monoecious, dioecious, gynodioecious or andromonoecious (Leal et al. 2013).The leaves are opposite, glabrous, occasionally membranaceous or more often subcoriaceous to coriaceous, with conspicuous primary, secondary and tertiary veins.The inflorescences are axillary fascicles.Flowers can be unisexual or bisexual, with 2-4(-5) sepals fused at the base and 2-4(-6) free petals.Staminate flowers have numerous free stamens, in fascicles or not, inserted over a central disc; the anthers are longitudinal or rarely poricidal, with an inconspicuous connective; some species have pistillode.Bisexual flowers have fewer stamens than staminate flowers, and in some species these stamens are sterile, and the flowers are functionally pistillate.The ovary is (1-2-)3-locular, syncarpic, with short or absent style.Fruits are yellow to orange berries when mature, often edible for humans.Seeds are only 1-4 per fruit, cylindrical, without aril.

Identification key for Garcinia species in Espírito Santo
The two species of Garcinia that are native to Espírito Santo can be easily differentiated through the surface of young branches, which in G. brasiliensis are rough or even papillose, and smooth in G. gardneriana.Furthermore, the species can be distinguished by the shape of the fruit, which is ellipsoid in G. brasiliensis and globose to ellipsoid in G. gardneriana, and by the rostrum of the fruit, absent or short in G. brasiliensis and well developed (2-6 mm) in G. gardneriana.However, rostrum morphology may be variable between species (Bittrich 2003), and therefore should be used in combination with other characters.Additionally, G. brasiliensis is apparently the only Brazilian species of Garcinia that has perfumed flowers (Van den Berg 1979), a fact that can be useful for identifying species from Espírito Santo during fieldwork.Vernacular names: bacupari, guanandi, guanandi-da-areia.

Symphonia L.f.
The species of Symphonia are trees, hermaphroditic, with perulate terminal buds, and secrete a yellow exudate.The leaves are opposite, glabrous, petiolate, leaf blade subcoriaceous, intersecondary and tertiary veins conspicuous.The inflorescences are terminal or axillary, cymose or solitary flowers.The flowers are bisexual.The sepals are free, 5-merous.The petals are free, 5-merous, usually alternate to sepals.The androecium is organized in fascicles, each formed by five fused stamens, united in the base, and forming a staminal tube surrounding the style.The anthers are long with glandular connective.The ovary is 5-locular, syncarpous.The fruits are globose indehiscent, with style and stigmas persistent.The seeds are exarillate.

Tovomita Aubl.
Tovomita species are usually dioecious trees with few underwood shrub species and eperulate terminal buds.They are easily recognized by the presence of prop roots and yellowish exudate.The leaves are chartaceous to coriaceous, always shiny and grouped at the apex of the branches.The flowers are unisexual and organized in terminal inflorescences with a dichasial or thyrsoid arrangement and articulated pedicels.The central flower of the dichasium always opens first and usually does not have an articulated pedicel.The floral bud oblong, globose, or cylindrical, with different apex types.The flowers are usually inconspicuous: white, greenish, or pale yellow, becoming brownish when senescent, without resin.The flowers may have two or four sepals; however, the outer pair always covers the entire floral bud.The staminate flower has free stamens and an inconspicuous pistillode; pistillate flowers have staminodes similar to the stamens and ovary 4-5(-7)-carpellate.The ovary will give rise to a fleshy capsule containing a seed in each valve.The seed has a fleshy aril which can be yellow, orange or red.

Identification key for
Tovomita fructipendula is distinguished from other species in Espírito Santo by its smaller size, usually shrubby, small and narrow leaves (Marinho et al. 2016), in addition to brown and asperous fruits (Fig. 4b), which do not occur in other species of Tovomita in the state.Some specimens from the Duas Bocas Biological Reserve (Fraga 2304) and the Augusto Ruschi Biological Reserve (Rossini 377, Rossini 469, Vervloet 2167) have small and narrow leaves, but the fruits have smooth epicarp, contrasting with the typical asperous fruits of T. fructipendula.For the precise identification of these specimens, it is necessary to collect materials with floral buds or flowers.Tovomita fructipendula has been frequently identified as T. brasiliensis, a synonym, and was assessed as Least Concern (LC) according to the IUCN Red List (BGCI & IUCN 2019).Vernacular name: guanandi-mirim.Fig. 4d-f Trees up to 6 m tall, exudate scarce.Petioles 0.7-1.1 cm long, green.Leaf blades 7.2-10.9× 3-4.8 cm, purplish red in sicco, subcoriaceous, lanceolate to oblong, base convex to decurrent, apex acuminate to slightly apiculate.Venation: secondary veins 10-12 pairs, 4-10 mm apart from each other, immersed adaxially, prominent abaxially, slightly arcuate near the margin; intersecondary veins present, one or two per intercostal area, slightly thinner than the secondary veins; intramarginal vein present.Inflorescences: ♂ a lax 9-flowered thyrsoid, lacking primary flower, ♀ a 3-flowered dichasium.Pedicels 7-17 mm long.Floral buds 4-5 mm long, globose, apex rounded.rounded,apex rounded,green to pale yellow;petals 4,    Tovomita guianensis has a disjunct distribution between the Amazon and Atlantic Forests and is widely distributed in South America, occurring from the coast of Venezuela to the central region of Bolivia (Marinho 2019).In Brazil, T. guianensis occurs in Amapá, Amazonas, Maranhão and Pará, in the Amazon region; Alagoas, Bahia, Pernambuco and Espírito Santo, in the Atlantic Forest (Marinho et al. 2016).In Espírito Santo, the species is known for only three specimens from lowland forests in the north of the state, the southern limit of species.

Tovomita guianensis
Among the species of Espírito Santo, Tovomita guianensis is the only one with intersecondary veins that are slightly thinner than the secondary ones.In a herbarium, the specimens take on a purplishred coloration which is also characteristic of the species.Tovomita guianensis is recognized for its globose floral buds, yellow and subclavate stamens and staminodes (Fig. 4d), in addition to fruits with smooth epicarp (Fig. 4e).Tovomita guianensis had been identified as T. brevistaminea, a synonym (Marinho 2020a).The species was not assessed for its conservation status in the state of Espírito Santo but was assessed as a species of Least Concern due to its wide distribution (Marinho & Beech 2019).(Schltdl.) Planch. & Triana, Ann. Sci. Nat., Bot. 14, série IV: 278. 1860. Fig. 4g-i Trees up to 10 m tall, exudate abundant.Petioles 1.3-2.8cm long, green or reddish.Leaf blades 6.5-17 × 3-8 cm, generally grayish to cooper-coloured in sicco, coriaceous, obovate to oblong, base decurrent, apex rounded.Venation: secondary veins 22-40 pairs, 1.8-3 mm apart from each other, prominent in both surfaces, straight near the margin; intersecondary veins present, similar to the secondary veins; intramarginal vein present.Inflorescences: ♂ a lax 3-9-flowered thyrsoid, lacking primary flower, ♀ a 3-flowered dichasium.Pedicels 12-30 mm long.Floral buds 6-7 mm long, globose, apex rounded.Sepals 4,rounded to oblong,apex rounded,green;[11][12][13][14][15][16][17][18][19][20]  Tovomita leucantha is endemic to the Atlantic Forest of the Southeast region of Brazil, not occurring only in the state of São Paulo (Marinho et al. 2016;Marinho 2020a).In Espírito Santo, the species is known for a single specimen collected on the montane forest of Irupi, south of the state (Fig. 4g).Tovomita leucantha was assessed as Endangered in the red list of Espírito Santo (Fraga et al. 2019), especially due to the low number of localities in which it can be found.

Tovomita leucantha
Tovomita leucantha is recognized by its obovate leaves with obtuse apex, greyish in sicco, coriaceous and with numerous secondary veins (22-40 pairs).The stamens are subclavate, similar to those of T. guianensis, but white, like the petals (Fig. 4g).Some immature specimens of T. riedeliana are similar to T. leucantha but the latter can be distinguished by the subclavate stamens (vs.terete in T. riedeliana) and short anthers (ca.0.5 mm vs. ca. 1 mm in T. riedeliana).

Tovomitopsis
The species of Tovomitopsis are dioecious trees or shrubs, with a white-yellowish exudates, and eperulate terminal buds, and conspicuous prop roots.The leaves are membranaceous, chartaceous, subcoriaceous or coriaceous, grouped at the apex of the branches, and can present black dots or not.The thyrsoid inflorescences are terminal and bearing small, yellowish to greenish flowers, which offer resin to pollinators (Bittrich et al. 2003).The flowers of the genus have small external sepals, not covering the floral bud, which distinguishes it from Tovomita.The flowers also are unisexual with staminodes in the pistillate flowers, and pistillodes in the staminate flowers.Stamens and staminodes are free and covered by the resin.The ovary is 4-carpellate, and will give rise to capsular and fleshy fruits.The seeds have an orange to reddish aril.
The limits between T. saldanhae and T. paniculata are unclear (Marinho et al. 2021b), since the leaf morphology appears to be largely plastic according to the altitudinal variation and distance from the coast.What we know for now is that T. saldanhae occurs in regions with high altitude, while T. paniculata has a preference for lowland forests.

Figure 5 2 .
Figure 5 -a-d.Tovomitopsis paniculata -a.young leaf in adaxial view; b. staminate flower; c. pistillate flowers and buds; d. prop roots.e-g.Tovomitopsis saldanhae -e.branch with mature and immature fruits; f. staminate flower, note the two pairs of petals of different sizes; g. pistillate flower and buds, note the black dots on the abaxial leaf surface.Only a-c are specimens from Espírito Santo.Photos: a. José Elvino Nascimento Jr.; b-c.João Paulo Zorzanelli; d.Gabriel Marcusso; e. Rodrigo Freitas; f.Rodrigo Penati; g.Luciano Pedrosa.
Clusia nemorosa is a species with wide distribution in Brazil, occurring in all regions, except in the Southern Region of Brazil.There may be populations of C. nemorosa with gynodioecious flowers, but these populations have not yet been found in Espírito Santo.In Espírito Santo it is found mainly in restingas, tropical rain forests and rocky outcrops.Clusia nemorosa is assessed as a species Rodriguésia 74: e00552022.2023 of Least Concern (LC) according to theIUCN Red  List (BGCI & IUCN 2019).Vernacular name: clusia-capelinha, clusia-purunga.Clusia nemorosa differs from all species of Clusia of Espírito Santo in that it presents staminate flowers with filiform stamens, with lateral thecae, connective higher than the thecae, and central resinous staminodes.