Microlicia crassa and M. maculata spp. nov. (Melastomataceae) from Minas Gerais, Brazil: morphology and leaf anatomy

Rosana Romero, Kleber Resende Silva and Daniela Guimarães Simão

R. Romero (romero@inbio.ufu.br), Inst. de Biologia, Univ. Federal de Uberlândia, Caixa Postal 593, BR-38400-902 Uberlândia, Minas Gerais, Brazil. – K. R. Silva, Programa de Pós Graduação em Biologia Vegetal, Inst. de Biologia, Univ. Federal de Uberlândia, Caixa Postal 593, BR-38400-902 Uberlândia, Minas Gerais, Brazil. – D. G. Simão, Depto de Biologia, Univ. Federal de Santa Maria, Avenida Roraima, 1000, Cidade Univ., BR-97105-900 Santa Maria, Rio Grande do Sul, Brazil.

Two new species of Microlicia from campos rupestres of Minas Gerais are described, illustrated and compared with their relatives. Microlicia crassa sp. nov. is similar to M. formosa, and M. maculata sp. nov. is similar to M. isophylla and M. tetrasticha. The trichomes covering both surfaces of the leaves of the two new species and their relative, M. isophylla may be characterized as sessile glands under a stereomicroscope, but anatomical studies revealed the presence of a short stalk. Microlicia crassa and M. maculata are assessed as ‘Endangered’, according to the IUCN categories and criteria.

In campos rupestres in the surroundings of Diamantina municipality, in Minas Gerais state, Brazil, at least 36 species of Microlicia D. Don are known to occur, of which six are taxonomic novelties (Romero 2013). Recognition of these novelties has been made with caution, because the mor-phological characteristics used to define species are gener-ally very slight, hindering delimitation from closely related species. In Melastomataceae, leaf anatomical characters, such as the morphology and type of trichomes and stomata, and the presence or absence of emergences and position of the palisade parenchyma have provided diagnostic characters for recognizing certain species of Leandra Raddi, Miconia Ruiz & Pav., Acisanthera P. Browne, Tibouchina Aubl. and Microlicia D. Don (Reis et al. 2005). Thus, we believe that leaf anatomical data may also provide support for the characterization of new species of Microlicia.

As a result of ongoing studies on the genus Microlicia in campos rupestres of the Cadeia do Espinhaço, in Minas Gerais state, we here describe two new species, including anatomical data for both, as well as for M. isophylla, a closely related species.

Material and methods

This study was based on literature and on the analysis of Microlicia specimens from the herbaria at BHCB, HUFU, K, MBM, NY, P, RB, SPF, UB, UEC and US (acronyms according to Thiers 2014), and on field observations in Biri-biri State Park, Diamantina, Minas Gerais, Brazil. For the

anatomical study, leaves were fixed in 50% FAA (Johansen 1940) and then preserved in 70% ethyl alcohol. Leaf sam-ples of Microlicia isophylla DC. were removed from speci-mens deposited in the HUFU herbarium (A. P. M. Santos et al. 414), and were hydrated in 5% NaOH (Anderson 1963) for 12 h. Five mature leaves from different nodes of an individual of each of both new species were collected. Young leaves were also studied to confirm the presence and type of trichome and/or emergence, because these structures may be damaged in mature leaves of some Microlicia species. The samples, which were previously dehydrated in an etha-nol series (70, 90 and 100%), were embedded in historesin. Transverse and longitudinal sections were prepared using a rotary microtome with tungsten blades, at the median third of the expanded adult leaf lamina, subsequently stained with toluidine blue (O’Brien et al. 1964) and mounted on per-manent histological slides. Anatomical images were obtained with a Zeiss photomicroscope, using the program DPCon-troller (ver. 2.1.1.183, Olympus). Histochemical tests were performed using Sudan III (Sass 1951) and ferric chloride (Johansen 1940) for lipid and phenolic compound analyses, respectively.

Microlicia crassa R. Romero sp. nov. (Fig. 1)

Type: Brazil, Minas Gerais, estrada Diamantina-Conselheiro Mata, km 172, 20 km do trevo, 18°18′01′′S, 43°48′51′′W, 1325 m a.s.l., 31 Mar 2001 (fl fr), R. Romero and J. N. Nakajima 6088 (holotype: HUFU!, isotypes: BHCB!, K!, P!, RB!, US!).

© 2014 The Authors. Nordic Journal of Botany © 2014 Nordic Society OikosSubject Editor: Bertil Ståhl. Editor-in-Chief: Torbjörn Tyler. Accepted 4 July 2014

Nordic Journal of Botany 000: 001–008, 2014 doi: 10.1111/njb.00635

Early View (EV): 1-EVsee http://tinyurl.com/ctujsze

Figure 1. Microlicia crassa sp. nov. (A) habit, (B) leaf abaxial surface, (C) leaf adaxial surface, (D) floral bud, (E) flower, (F) petal, (G) detail of the petal apex, (H) large stamen, (I) small stamen, (J) gynoecium, (K) immature capsule enclosed by the hypanthium. Drawn by N. Nascimento from the holotype.

2-EV

EtymologyThe specific epithet refers to the coriaceous leaf blades, which are thicker than in most species of Microlicia from Minas Gerais.

DescriptionErect subshrub or shrub, 0.5–1.5 m tall, much branched. Young branches fastigiate, quadrangular, slightly glutinous; young branches, pedicel, hypanthium and calyx lobes with indumentum of glandular trichomes; older branches brown, becoming slightly terete with age, without leaves at the base; nodes light green. Leaves sessile, ascending, imbricate or not, coriaceous, concolorous, green or brownish (dry state); blade 3–8 0.9–4.0 mm, lanceolate or oblong, acute or obtuse at apex, attenuate or rounded at base, with entire margin, often lighter than the rest of the blade, both surfaces with short-stalked glands; 1-nerved or inconspicuously 3-nerved with the central nerve pale on the abaxial surface (dry state) and slightly marked on the adaxial surface, smooth on both surfaces. Flowers solitary, terminal or lateral, 5-merous, zygomorphic resulting from the position of the stamens and style; pedicel 0.8–1.5 mm long. Hypanthium campanulate to oblong, 2.5–3.0 1.5–2.5 mm, green; calyx tube 0.3– 0.5 mm long; calyx lobes foliaceous, triangular-oblong, 3.5–6.0 0.3–0.8 mm, acute at apex. Petals 10.5–13.0 5.5–7.0 mm, magenta, obovate-oblong, glabrous, acute at apex. Stamens 10, dimorphic in size. Large stamens 5; filaments 4.5–5.0 mm long, pink; anthers 3.0–3.5 mm long (including beak), ovate-oblong, tetrasporangiate, vinaceous, their beak 0.5–0.6 mm long, white; connective prolonged, 4.5–5.0 mm, vinaceous, with ventral appendage 1.5– 1.7 mm long, truncate, pink but yellow at apex. Small stamens 5; filaments 4.5–5.0 mm long, pink; anthers 2.5– 3.0 mm long (including the beak), yellow, oblong, tetraspo-rangiate, their beak 0.4–0.6 mm long, yellow; connective 1.0–1.3 mm, prolonged, its ventral appendage 0.5–0.7 mm long, truncate, yellow. Ovary ca 2.5 2.0 mm, superior, 3-locular, glabrous; style 6–7 mm long, pink, terete, slightly curved, glabrous; stigma punctiform. Capsule 3.5–4.5 3.0–3.5 mm, brown, subglobose, dehiscing into 3 valves from the apex; hypanthium covering the entire ovary and peeling off as the fruit matures. Seeds 0.8–1.0 0.4–0.5 mm, slightly curved to one side, pale brown; testa foveolate.

Distribution, habitat and conservationMicrolicia crassa is endemic to Minas Gerais, in the Espinhaço Range, occurring on sandy soils among rocks, in campo rup-estre vegetation from Diamantina and Serro, between 1200 and 1325 m a.s.l. Due to the restricted extent of occurrence and area of occupancy (AOO 20 km2), M. crassa should be considered ‘Endangered’ (EN) B1ab(iii) 2ab(iii) according to the IUCN categories and criteria (2001).

PhenologyFlowers were collected in March, June, and October, and fruits in March, June, September and October.

Similar speciesMicrolicia crassa is characterized by its coriaceous, sessile leaves and indumentum of glandular trichomes covering the

branches, hypanthium and calyx lobes, and short-stalked glands on both surfaces of the leaves. These glands possibly have a short pedicel like those found on both sides of the leaves. The hypanthium is campanulate or oblong, the calyx lobes are oblong or triangular-oblong and are equal to or longer than the hypanthium and the petals are glabrous. The gathering G. Hatschbach 69708 has leaves that are strongly imbricate and narrowly lanceolate with an obtuse apex. Furthermore, the leaves (3–4 0.8–1.0 mm), pedicels (ca 0.7 mm long), hypanthium (ca 2.8 2.0 mm) and calyx lobes (ca 2.5 0.5 mm) are smaller than usual.

Among the species that occur in Minas Gerais, M. crassa is most similar to M. formosa Cham. The latter is known only from a few collections, most of them from the 19th century. Currently, M. formosa. is known only from the mountains of Capanema, Caeté and Caraça, in Minas Gerais state, in the region known as ‘Quadrilátero Ferrífero’ (Iron Quad-rangle), located at the southern end of the Espinhaço Range (Harley 1995, Giulietti et al. 1997). Both have small, sessile and ascending leaves, leaf blades lanceolate or oblong, cam-panulate or oblong hypanthia, magenta petals, and dimor-phic stamens with bicolorous anthers. However, M. formosa differs in having leaves with distinctly attenuate bases, petals with an acute-acuminate apex, and triangular calyx lobes that are shorter than the hypanthium (2.0–2.5 mm long) with acute apex and a thickened tip.

Additional specimens examined (paratypes)Brazil, Minas Gerais, Diamantina, estrada Diamantina-Con-selheiro Mata, 20.3 km do asfalto, 18°20′S, 43°53′W, 1200 m a.s.l., 23 Sep 1994 (fr), Splett 624 (HUFU, UB, US); km 172, 18°17′S, 43°37′W, 9 Jun 1998 (fl fr), R. Romero et al. 5423 (CAS, HUFU, UEC); km 184, 18°12′55′′S, 43°35′17′′W, 1295 m a.s.l., 6 Dec 2012 (fr), A. F. A. Ver-siane and K. R. Silva 381 (HUFU). Serro, cabeceira do Rio Jequitinhonha, Cascata Moinho de Esteira, 25 Oct 1999 (fl fr), G. Hatschbach et al. 69708 (HUFU, MBM).

Microlicia maculata R. Romero sp. nov. (Fig. 2)

Type: Brazil, Minas Gerais: São Gonçalo do Rio Preto, Parque Estadual do Rio Preto, 18°06¢54¢¢S, 43°20¢28¢¢W, 20 Feb 2002 (fl fr), J. A. Lombardi 4617 (holotype: BHCB!, isotype: HUFU!).

EtymologyThe specific epithet refers to the brownish–green colour on both sides of the leaf in dry material.

DescriptionErect subshrub, 0.3–0.5 m tall, much branched. Stem terete; young branches fastigiate, quadrangular, glutinous; young branches, pedicel, hypanthium and calyx lobes with indu-mentum of sessile golden glands; older branches brown, becoming slightly terete with age, without leaves at the base. Leaves sessile, ascending, not imbricate, coriaceous, concol-orous, with some dark green blotches on both surfaces (dry state); blade lanceolate, elliptical, ovate or ovate-lanceolate, 1.8–3.0 0.8–2.0 mm, acute to acute-acuminate at apex, at base rounded, cordate, or sometimes attenuate; margins

3-EV

Figure 2. Microlicia maculata sp. nov. (A) flowering branch, (B) leaf abaxial surface, (C) leaf adaxial surface, (D) flower, (E) hypanthium and calyx lobes, (F) petal, (G) small stamen, (H) large stamen, (I) gynoecium, (J) immature capsule enclosed by hypanthium with ovary exposed. Drawn by N. Nascimento from F. N. A. Mello et al. 74a.

4-EV

pore (0.2–0.4 mm diameter) compared to other Microlicia species. In fruit the apex of the ovary is completely exposed (Fig. 2J) and the hypanthium falls off only in mature capsules.

The affinities of M. maculata appear to be with M. tetrasticha Cogn. They share an indumentum of glan-dular trichomes, ascending, sessile leaves, short pedicel (ca 1 mm long), campanulate hypanthium, pink to pink– magenta petals, and the dimorphic stamens with bicolor-ous anthers (Romero 2013). Microlicia maculata differs in lacking imbricate leaves and lanceolate, elliptical, ovate or ovate-lanceolate leaf blades. Microlicia isophylla, like M. maculata, has ascending and small (3–7 1–2 mm) leaves, often of the same length of the internode, but dif-fers by having lanceolate to elliptic-lanceolate leaves, with attenuate, rounded, rarely cuneate bases and acute to short-acuminate apices, and young branches, leaves, hypanthium, and calyx lobes are covered with sessile, golden glands, that are occasionally mixed with an indumentum of short, pale trichomes (Romero 2013).

Additional specimens examined (paratypes)Brazil, Minas Gerais: Augusto Lima, Serra do Cabral, 18°00′40′′S, 44°19′41′′W, 20 Mar 1994 (fl fr), C. M. Sakuragui et al. CFCR 15257 (K, SPF); Diamantina, ca 27 km de Serro on road (MG 2) to Diamantina, 1200 m a.s.l., 26 Feb 1968 (fl fr), H. S. Irwin et al. 20932 (CAS, HUFU, NY, US); estrada Diamantina-Extração, 28 Jan 1986 (fl fr), D. C. Zappi et al. CFCR 9311 (K 2 sheets, SPF); estrada Diamantina-Conselheiro Mata, km 185, próximo à grande inselberg, 23 Feb 1986 (fl fr), J. Semir et al. CFCR 9489 (K, SPF); 5 km west of Diamantina, estrada para Gouveia, 18°13′S, 43°38′W, 16 Feb 1991 (fl fr), M. N. Arbo et al. 5228 (K, SPF); estrada para Biribiri, 18°09′S, 43°36′W, 1063 m a.s.l., 19 Oct 2007 (fr), F. N. A. Mello et al. 74a (HUFU); trilha Casa dos Ventos, 18°10′50′′S, 43°53′41′′W, 5 Dec 2012 (fr), A. F. A. Versiane and K. R. Silva 370 (HUFU).

Leaf anatomyAll three analyzed species displayed depressions on both surfaces of the leaves (Fig. 3A–C), the most prominent being found in M. isophylla (Fig. 3B). The epidermis, seen in frontal view, possesses fundamental cells with straight to slightly sinuous walls, as well as anomocytic stomata on both surfaces (Fig. 3D–E). The epidermis is uniseriate with similar sized cells on both surfaces in transverse section (Fig. 3A–C, G), covered by a thin cuticle (Fig. 3G). The stomata are slightly projected in relation to the remaining epidermal cells (Fig. 3G). The mesophyll is homogeneous, composed of palisade parenchyma (Fig. 3A–C, G), the num-ber of layers varying among species, with five in M. isophylla (Fig. 3B, G), six to seven in M. maculata (Fig. 3C) and nine in M. crassa (Fig. 3A). This tissue is also found surrounding the leaf margin of all three species (Fig. 3A–C).

Glandular trichomes, on both surfaces, have a spherical head and short, uniseriate peduncle (Fig. 3F), appearing sessile when examined under a stereomicro-scope. These structures are associated with foliar depressions (Fig. 3A–B); these depressions are shallower on the leaves

entire, sometimes slightly crenulate; both surfaces with incon-spicuous short-stalked glands on the central the nerve only, inconspicuously visible. Flowers solitary, terminal, some-times lateral, 5-merous, zygomorphic as a result of the position of stamens and style; pedicel 0.5–1.0 mm long. Hypanthium terete, campanulate, 1.8–2.0 1.5–2.0 mm, cream or cream and dark pink, smooth or 10-costate; calyx tube 0.3–0.9 mm long; calyx lobes foliaceous, triangular to oblong-triangular, 1.5–4.0 1.0–1.5 mm, acute or obtuse at apex. Petals 4.3–6.0 2.0–3.5 mm, oblong, pink-magenta, glabrous, acute-acuminate at apex. Stamens 10, dimorphic in size. Large stamens 5; filaments 2.5–3.0 mm long, pink; anthers 1.8–2.0 mm long (including beak), oblong, tetrasporangiate, pink, their beak 0.5–0.7 mm long, white or pale pink, with large pore 0.2–0.4 mm in diameter; connective prolonged 1.5–2.0 mm, pink, with ventral appendage 1.0–1.5 mm long, truncate, expanded, pink but yellow at apex. Small stamens 5; filaments 3.0–3.5 mm long, pink; anthers 1.4–1.6 mm long (including the beak), yellow, oblong, tetrasporangiate, their beak 0.3–0.5 mm long, yellow; connective 0.6–0.7 mm, prolonged, its ventral append-age ca 0.2 mm long, obtuse, yellow. Ovary ca 2 1 mm, superior, 3-locular, glabrous; style 2–3 mm long, pink, terete, slightly curved, glabrous; stigma punctiform. Capsule 3.5–4.0 2.5–3.0 mm, brown, globose, dehiscing into 3 valves from the apex; hypanthium covering more than 2/3 of the ovary and peeling off as the fruit matures. Seeds 0.5–0.6 0.3–0.4 mm, slightly curved to one side, pale brown; testa foveolate.

Distribution, habitat and conservationMicrolicia maculata is endemic to the Espinhaço mountain range in Minas Gerais, occurring in campo rupestre vegeta-tion, at 1000–1200 m a.s.l. Due to the restricted extent of occur-rence and area of occupancy (AOO 20 km2), M. maculata should be considered ‘Endangered’ (EN) B1ab(iii) 2ab(iii) according to the IUCN categories and criteria (2001). Speci-mens were found around Diamantina, in Biribiri State Park, in Rio Preto State Park, and Serra do Cabral State Park; all are natural reserves maintained by the state government. Despite the high vulnerability of campos rupestres in Minas Gerais, the occurrence of M. maculata in three conservation units guarantees some protection of its populations.

PhenologyFlowers were collected in January, February, and March, and fruits in January, February, March, October and December.

Similar speciesMicrolicia maculata is characterized by its small, ascending, sessile leaves. The leaves are brownish green in dry mate-rial, the same length as the internodes, and are covered by short-stalked glands. The branches, hypanthium and calyx lobes are covered by glandular trichomes. As in M. crassa, we believe that these glands have a short stalk. The hypanthium is campanulate, the calyx lobes are foliaceous, and equal to or slightly longer than the hypanthium. The connective of the smaller stamens has a ventral appendage that does not exceed 0.2 mm in length, and the style is short (2–3 mm long). Furthermore, the anthers of the larger stamens have a large

5-EV

Figure 3. Leaf anatomy of Microlicia species. (A), (D), (H) M. crassa sp. nov.; (B), (E), (G) M. isophylla; (C), (F), (I) M. maculata sp. nov. (A)–(C) general aspect in cross-section, (D)–(E) epidermis in frontal view, abaxial and adaxial surfaces, respectively, showing anomocytic stomata, (F) cross-section with a short-stalked trichome, (G) cross-section showing leaf tissues (arrow pointing to stoma slightly elevated), (H) detail of midvein in cross-section, (I) group of sclereids in longitudinal section. Scale bars 200 mm (A)–(C); 50 mm (D), (F), (H), (I); 20 mm (E); 100 mm (G).

6-EV

Table 1. Presence (1) and absence (0) of leaf anatomical characteristics in Microlicia crassa sp. nov., M. isophylla and M. maculata sp. nov.

Anatomical characters M. crassa M. isophylla M. maculata

Depressions on both surfaces 1 1 1Epidermis with cell walls straight to slightly sinuous on both surfaces (frontal view) 1 1 1Epidermis uniseriate with similar cell size on both surfaces 1 1 1Cuticle thin 1 1 1Leaves amphistomatic 1 1 1Stomata anomocytic, slightly projected 1 1 1Trichomes glandular and uniseriate with short stalk 1 1 1Mesophyll homogeneous 1 1 1Palisade parenchyma surrounding margin 1 1 1Midrib sunken in mesophyll 1 1 1Collateral vascular bundle circular in midrib 1 1 1Collenchyma angular on adaxial surface on midrib 1 0 1Sclereids on adaxial surface of midrib 1 0 0Sclereids on abaxial surface of midrib 0 0 1Phenolic substances on epidermis 0 1 1Mesophyll with phenolic substances only on adaxial layer of palisade parenchyma 0 1 0Mesophyll with phenolic substances only on adaxial and abaxial layers of palisade parenchyma 0 0 1Phenolic substances on all layers of the mesophyll 1 0 0Phenolic substances on sheath of vascular bundle 1 1 1Phenolic substances on collenchyma of midrib 1 0 0Druses scattered on mesophyll 1 1 1

of M. maculata, and consequently the trichomes are more exposed (Fig. 3C). Therefore, in older leaves trichomes occur less frequently or are damaged. Species of Microlicia frequently have some type of trichome covering the vegeta-tive and reproductive structures. Based on scanning electron microscopy, Wurdack (1986) described at least four dif-ferent types of trichomes for the genus: elongated smooth hairs, conical hairs without enations, long-stalked glands with thin-walled heads and unfurrowed sessile glands, the latter occurring in the majority of species. However, ana-tomical data of the three analyzed species revealed that, at least regarding the leaf blade, the unfurrowed sessile glands displayed a short peduncle, and therefore should be charac-terized as short-stalked glands.

The midrib, which is level with the mesophyll (Fig. 3A–C, G), displays a circular-shaped collateral vas-cular bundle in all three species, as observed in M. crassa (Fig. 3H) and M. isophylla (Fig. 3G). An angular collenc-hyma can also be observed in this region, facing the adaxial surface of M. crassa (Fig. 3H) and of M. maculata (Fig. 3C). Isolated columnar sclereids were observed associated with the collenchyma of M. crassa (Fig. 3H) or in other groups, facing the abaxial surface in M. maculata (Fig. 3C, I). Idioblasts containing phenolic substances occur more frequently in the chlorenchyma, as compared with the lining and support-ing tissues (Fig. 3A–C, G), differing, however, in the posi-tion between the layers of palisade parenchyma (Table 1). These compounds can also be observed in the epidermis of M. isophylla (Fig. 3G) and M. maculata, on the sheath of the vascular bundle (Fig. 3G–H) and the collenchyma of M. crassa (Fig. 3H). Calcium oxalate druses occur scat-tered throughout the mesophyll of the analyzed species, as observed in M. isophylla (Fig. 3G).

Although few species of Microlicia have been studied, our results indicate that some foliar anatomical features may be considered diagnostic, such as: foliar depressions, which are more conspicuous in M. isophylla; number of

mesophyll layers, with five in M. isophylla, six to seven in M. maculata and nine in M. crassa; collenchyma on adaxial surface in M. crassa and M. maculata; as well as the distri-bution of sclereids on the adaxial and abaxial surfaces of M. crassa and M. maculata, respectively, and of phenolic substances in different tissues (Table 1). The remaining characteristics, such as uniseriate epidermis, amphistom-atic leaves with slightly projected anomocytic stomata and short, glandular and uniseriate trichomes, reveal no variation among the species, suggesting that they are com-monly found in other representatives of the genus, because this has already been recorded for leaves of M. polystemma Naudin (Reis et al. 2005) and M. hatschbachii Wurdack (Cassiano et al. 2010). Therefore, new anatomical studies in Microlicia can contribute to the characterization and recognition of its representatives, the phylogeny of which is as yet uncertain (Fritsch et al. 2004, Simon et al. 2009), providing a possible data set to help differentiate closely related genera, such as Lavoisiera and Trembleya (Almeda and Martins 2001).

Acknowledgements – The first author thanks Fundação de Apoio à Pesquisa do Estado de Minas Gerais (FAPEMIG proc. 0703-11) and Univ. Federal de Uberlândia for supporting expeditions to the campos rupestres in Minas Gerais State, to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-REFLORA proc. 563541/2010–5 and PROTAX proc. 562290/2010–9) for supporting research on Melastomataceae in Minas Gerais State, to Coordenação de Apoio a Pessoal de Nível Superior (CAPES) for a post-doctoral fellowship (Ciência sem Fronteiras proc. 9612/12–2), and Elizabeth Woodgyer and Eve Lucas for their warm hospitality during her stay at the Royal Botanic Gardens, Kew. K. R. Silva thanks Programa de Pós Graduação em Biologia Vegetal (UFU) for financial support to collect in Diamantina, to Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for a Masters degree fellowship and to Laboratory of Plant Morphology and Images (LAMOVI) for providing support. The authors thank Natanael Nascimento for preparing the illustrations.

7-EV

IUCN 2001. IUCN red list categories and criteria, ver. 3.1. – IUCN Species Survival Commission.

Johansen, D. A. 1940. Plant microtechnique. – McGraw-Hill.O’Brien, T. P. et al. 1964. Polychromatic staining of plant cells walls

by toluidine blue O. – Protoplasma 59: 368–373.Reis, C. et al. 2005. Anatomia foliar de Melastomataceae do

cerrado do Estado de São Paulo. – Rev. Bras. Bot. 28: 451–466.

Romero, R. 2013. Taxonomic notes in Microlicia (Melastomata-ceae, Microlicieae). – Phytotaxa 110: 48–54.

Sass, J. E. 1951. Botanical microtechnique. – Ames, The Iowa State College Press.

Simon, M. F. et al. 2009. Recent assembly of the Cerrado, a neotropical plant diversity hotspot, by in situ evolution of adaptations to fire. – Proc. Natl Acad. Sci. USA 106: 20359–20364.

Thiers, B. 2014. Index Herbariorum: a global directory of public herbaria and associated staff. – New York Bot. Gard. Virt. Herb., sweetgum.nybg.org, accessed 20 Jan 2014.

Wurdack, J. J. 1986. Atlas of hairs for neotropical Melastomata-ceae. – Smithson. Contrib. Bot. 63: 1–80.

References

Almeda, F. and Martins, A. B. 2001. New combinations and new names in some Brazilian Microlicieae (Melastomataceae), with notes on the delimitation of Lavoisiera, Microlicia and Trem-bleya. – Novon 11: 1–7.

Anderson, L. C. 1963. Studies on Petradoria (Compositae): anatomy, cytology, taxonomy. – Trans. Kans. Acad. Sci. 66: 632–684.

Cassiano, D. S. A. et al. 2010. Caracterização morfoanatômica de folhas e caules de Microlicia hatschbachii Wurdack, Melasto-mataceae. – Rev. Bras. Farmacogn. 20: 529–535.

Fritsch, W. P. et al. 2004. Phylogeny and circumscription of the near endemic Brazilian tribe Microlicieae (Melastomataceae). – Am. J. Bot. 91: 1105–1114.

Giulietti, A. M. et al. 1997. Espinhaço range region. eastern Brazil. – In: Davis, S. D. et al. (eds), Centers of plant diversity. A guide and strategies for the conservation. Vol III. The Ameri-cas. WWF/IUCN, Cambridge, pp. 397–404.

Harley, R. M. 1995. Introdução. – In: Stannard, B. L. (ed.), Flora of the Pico das Almas, Chapada Diamantina, Bahia, Brasil. R. Bot. Gard. Kew, pp. 1–76.

8-EV