Updating the checklist of subtribe Hyptidinae (Lamiaceae) from Brazilian Amazonia, with three new taxa and notes on Hyptis sect. Eriosphaeria Raymond

Three new taxa of subtribe Hyptidinae (Lamiaceae) from Amazonian Brazil are here described: two from Pará state Eriope macrostachya var. amazonica , and Hyptis cachimboensis , and Hyptis spathulata from Rondônia state. The diagnostic characters, which distinguish them from their closest relatives are described and discussed, their conservation status is given and a plate is provided for each of the new taxa. The Hyptidinae conspectus an earlier checklist of Lamiaceae from Amazonian Brazil is updated with various name changes since its publication, and with the addition of another two species of Hyptis not previously included. Also included, is a note on Hyptis sect. Eriosphaeria , to which the two new species described here, belong.


Introduction
The subtribe Hyptidinae (Ocimeae tribe, Nepetoidae subfamily) from the Lamiaceae (or Labiatae) is represented by c. 400 species distributed mostly in America, with seven species also found as introductions in the Old World Tropics (Harley & Pastore 2012), perhaps two species occurring naturally in Africa. Hyptis Jacq. still the most species-rich genus in Hyptidinae, even after its dismantling in which several new genera were recognized (Harley & Pastore 2012). In Brazil, this subtribe is represented by all genera, except for Asterohyptis Epling, endemic to Central America and Mexico. Recently, several studies have been published on Brazilian Hyptidinae, describing new species, as well as floristic accounts (e.g., Harley et al. 2019a, b;Harley & Antar 2019;Soares et al. 2019;Soares et al. 2020). The Hyptidinae of the Amazonian region were recently studied by the senior author (Harley 2012), who presented a checklist including records of all species of Lamiaceae (except Aegiphila Jacq., Amasonia L.f. and Clerodendrum L. Vitex L. including Volkameria L., which had previously been treated under the family Verbenaceae) and a key for the Hyptidinae taxa occurring in the region. The objective of the present paper is to update the account of the Hyptidinae of Brazilian Amazonia presented by Harley (2012) and describe the new taxa of the subtribe from to this region.
Results and Discussion 1. Updating the checklist of Hyptidinae of Brazilian Amazonia published by Harley (2012). A generic updating on the Amazonian Hyptidinae.
The checklist of Brazilian Amazonian Hyptidinae (Harley 2012) was published shortly before a major generic revision of Hyptidinae (Harley & Pastore 2012) in which the genus Hyptis was divided into twelve smaller genera, and before a number of other generic adjustments were made, resulting from a molecular phylogenetic study (Pastore et al. 2011;Harley & Pastore 2012). Thus, three new genera and a new combination on Hyptidendron are now added to Brazilian Amazonian Lamiaceae: Hyptidendron Harley (1988).

Taxa newly cited for the Brazilian Amazonian Region
Apart from the generic updating provided above, two species were not included in the first checklist of Hyptidinae from Brazilian Amazonian region (Harley 2012 This section was originally characterized by Bentham (1833: pags. 69 and 88-89) in having the flowers in well-defined pedunculate capitula, clearly hemispherical (rather than globose as seen in the younger stages of several other sections, which are now treated as separate genera), with a straight calyx-tube, and with abundant long usually white trichomes, not only from the base of the flowers, but also in the upper part of the calyx-tube, giving the capitula an extremely hairy appearance. The indumentum is often accompanied by yellowish or orange glands, which seem characteristic of many members of the section. The section possesses another important character, not recorded by Bentham (1833Bentham ( , 1848, which is a gynoecium in which the style is jointed above the level of the ovary, with the basal part (stylopodium) persistent.
The above suite of characters, which is shared by many of the species, which today compose the section, held good for some time until three species from Goiás, with unusual coriaceous subglabrous leaves, dorsiventrally compressed, and sessile, few-flowered capitula, were discovered. Epling (1937: pag. 290) erected a new subsection for this extremely distinctive group: Hyptis subsect. Pachyphyllae Epling. A further species, Hyptis imbricatiformis Harley was subsequently described (Harley 1986b). All four are endemic to the Chapada dos Veadeiros in Goiás state, and share the presence of a stylopodium in the flowers and long silky white hairs extending from their base. Recent findings from molecular DNA studies (Pastore et al. 2011) confirm that this subsection is monophyletic, placing it within sect. Eriosphaeria. Morphological, micromorphological and molecular evidence appear to place the "Pachyphyllae" group as a link between the rest of Hyptis sect. Eriosphaeria and Hyptis sect. Induratae Epling (Harley 1986a;Rudall 1986;Pastore et al. 2011). All three groups are best considered as representing Hyptis sect. Eriosphaeria, with its centre of diversity primarily in the mountains of Goiás, with also a strong presence in similar biomes in Minas Gerais.
Epling also created three other new subsections (Epling 1936(Epling , 1937 to accommodate four species, which did not appear to fall within those of Bentham, but a more detailed molecular study is required to elucidate their relationships. At the present time, 32 species are recognized in the section. The two subsections, Hyptis subsect. Sessilifoliae and subsect. Velutinae, are relevant to this paper, as each contains a new species from Amazonian Brazil. The two subsections are separated largely on the basis of leaf shape, although both typically have sessile or subsessile leaves. In Hyptis subsect. Velutinae, the leaves taper gradually towards the slender, often rounded, cuneate or acuminate base, while in Hyptis sect. Sessilifoliae, the base of the leaves is broader and cordate to truncate, sometimes almost clasping the stem. There also seems to be a difference in the indumentum of the two sections, although this is not clearly expressed in the literature. That of Hyptis sect. Sessilifoliae is composed of more delicate, softer trichomes, while those of Hyptis sect. Velutinae are more rigid and sometimes slightly silvery in appearance.
As shown by the molecular analysis (Pastore et al. 2011), Hyptis sect. Eriosphaeria subsect. Sessilifoliae forms a well-supported clade, based on the five species analysed, and the morphological features support this. Eight species were recognized (Epling 1949). These eight species include a complex array of forms, and some collections appear to be intermediate between the species recognized. This is especially true with the extremely variable Hyptis crenata and two other widely collected species, Hyptis dilatata Benth. and Hyptis goyazensis A.St.-Hil. ex Benth. Although, based on empiric analyses, we suspect that this may be due to the long history of habitat destruction, especially of forest, and the resultant expansion of more open savanna habitats, including pasture, where the species occur. Originally, forest may have acted as a barrier to gene flow, allowing a number of species to evolve in isolation. Subsequently, with the merging of populations, unrestricted gene flow may have allowed a whole range of intermediate forms to appear. Judging by present-day distribution, Hyptis dilatata probably evolved in the northern part of South America, particularly Amazonia, while H. goyazensis arose in the Western savannas south of the Amazon, in Mato Grosso and Goiás. The taxonomy of the Hyptis crenata complex has still not been elucidated, and the species shows a great deal of variation, which seems to be partly correlated with geography. This entire group is urgently requiring a detailed biosystematic and molecular study.
However, a number of taxa are sufficiently distinct, and merit recognition. These mostly have very narrow distributional ranges in montane areas. Examples include Hyptis alpestris A.St.-Hil. ex Benth., from mountains in SW Minas Gerais and neighbouring northern São Paulo, Hyptis hamatidens Epling & Játiva from northern Mato Grosso, and Hyptis obtecta Benth. from Pará, Tocantins and Goiás. This latter species, for which Epling created a new monotypic subsection (Hyptis subsect. Obtectae Epling 1936: 289), has now been analysed for its DNA (Pastore et al. in press), and is shown to form a clade with other members of Hyptis subsect. Sessilifoliae. This group is characterized by a mainly herbaceous habit, with sessile, often rugose and villous leaves, truncate to cordate at base and with a terminal, usually sub-corymbose inflorescence of capitula. Hyptis obtecta is unusual in having very small erect, conduplicate, imbricate leaves, under 1 cm long, which are closely applied to the stem. Leaf indumentum is silky, with short adpressed trichomes. However, a number of taxa have recently been discovered in which the leaves are also small, imbricate, sometimes conduplicate, as in H. obtecta, and with similar leaves. The new species here described, Hyptis cachimboensis, certainly has the habit and leaf shape and indumentum of a species belonging to Hyptis subsect. Sessilifoliae, but differs in its very small leaves, which are imbricate and erect, and often conduplicate, thus bridging the morphological gap between Hyptis obtecta and other members of subsect. Sessilifoliae.  Fig. 1 The new variety described in this paper is very similar to the more widespread E. macrostachya var. macrostachya, but differs in its subglabrous leaves, often with the base unequal, and rounded to weakly cordate, the unusual setose trichomes with a thickened, dark, ± conical base, persistent, especially on the stems and petioles, and with flowers that are slightly larger than in the typical variety.

New taxa from the Brazilian Amazonian Region
Shrub to 4 m tall with older stems woody, pale brown < 1.4 cm diam., branched from the base, leafless below; upper stems more or less terete, often densely covered by long, rigid, ± setose and patent uniseriate trichomes, broad-based and 4-5 mm long, these mixed with very small, slender, few-celled uniseriate trichomes, patent at base and strongly curled above (often almost hooked, but not rigid). Many setose trichomes possess a stout, often conical, very rigid and dark-coloured base, persisting as a rigid projection, after the upper more slender, hyaline part has been eroded. Cauline leaves aromatic, with petiole (0.7-)1.2-2.5(-3.7) cm long, indumentum as on stems, but sparser and with few small curved trichomes, and broad-based setose trichomes frequent and often eroded, leaving dark, rigid projections; lamina (3.9-)6.7-8.2 × (1.9 -)3-3.9 cm long, lanceolate to broadly ovate-lanceolate, usually at least twice as long as broad, (the leaves on short, non-flowering, side-branches smaller with lamina c. 1.8 × 0.8 cm, and sometimes narrowly ovate), membranous to cartaceous, discolorous, adaxial surface bright green when fresh, glabrous or nearly so, rugulose, with occasional usually very short, thick, conical trichomes, or very few short, setose trichomes, and numerous small, sessile, sunken glands, abaxial surface pale green, glabrous except along midrib and veins, midrib with some setose, patent and thick-based, uniseriate trichomes, veins with few, smaller, more slender trichomes, and very numerous small, sunken glands; base rounded to weakly cordate, often unequal, apex acute, margin somewhat irregularly and shallowly serrate, with teeth thickened along margin and especially at apex; older leaves often turning vinaceous.  Eriope macrostachya var. amazonica is at present only known from the type locality, but should be looked for in other similar campinas in Pará, and perhaps elsewhere in Amazonia (Fig.  2). The collection Miranda 606 from Pará state, Tucurui, Campina de Santa Rosa deposited in INPA, recently determined as Eriope macrostachya Mart. ex Benth., which was described as "more glabrous than typical", and should be re-examined. It is the only other known collection of this species from Amazonia, and could well prove to be a further record of this new variety. The area where this variety occurs is composed of deep sand, varying in colour from almost white to a pale brownish orange, and has been subject in the past to the wholesale removal of sand for building. This has left a much-disturbed vegetation with occasional clumps of secondary woodland, and hollows as a result of the sand removal. In the rainy season these form temporary pools, although no aquatic species were observed. The open areas are dominated with a cover of Chamaecrista ramosa (Vog.) Irwin & Barneby var. ramosa, suggesting a much disturbed habitat. Eriope was so far found in only two main areas, although much of the site was not visited. In all, less than 50 plants were observed, although this probably represents a small part of the total population. It seems to occur on slightly raised areas and well away from standing water, and often nearby taller vegetation. Very young, dwarf plants, only about 20 cm tall, were observed growing below the Chamaecrista.
The varietal name was given in reference to the region where this variety is distributed.
The variety was flowering and fruiting during February to July, but further observations needed.
Eriope macrostachya var. amazonica is mainly threatened by the destruction of the habitat, due to sand removal. This did not seem to be happening during our visit, but we noticed enormous mounds of excavated sand near where we entered the area. When this supply is exhausted, excavation will probably restart, unless steps are taken to preserve the area and control access.
All the material was collected from the same area. This species falls into the criterion b-III of IUCN (2017), as the area of occurrence is less than 10 km from the city of Belém (capital of the state of Pará), thus one can anticipate a steady decline in the quality of the habitat due to urban expansion. Even if the site eventually becomes a conservation area, it is still likely to be affected by increased visits by local people for leisure activities.
Apart from the typical variety of Eriope macrostachya, three other varieties are known (Harley 1976): E. macrostachya var. hypoleuca Benth, E. macrostachya var. platyantha (Epling & Mathias) Harley and E. macrostachya var. grandiflora (Epling) Harley. The first of these, E. macrostachya var. hypoleuca, grows principally on canga (iron-rich) soils in Minas Gerais and has † These technically mark the point where the peduncle -below -joins the true pedicel -above, but in this description, for convenience, the whole structure is referred to as the pedicel.
† † The first calyx measurement is taken along the posterior side, and the second along the anterior side of the calyx. The two measurements are separated by " / ".   Branched, erect shrub to 3 m tall, much branched above, with stems terete, bare below, 2−3.5 mm. diam., at least in upper stems, with epidermis densely hairy, with coarse, acute, rigid, silvery, antrorse trichomes strongly appressed to stem. Lower parts of plant not seen. Leaves subsessile, with petiole usually less than 1 mm, crowded along the upper, ultimate branches, densely imbricate, ascending. Lamina of leaf slightly coriaceous, discolorous, 11−16 × 3.6−5 mm, often weakly spathulate, widest just below obtuse and often rotund apex and tapering gradually towards narrowly rounded base, and often slightly concave above towards apex, margin rather indistinctly serrate-crenate below apex and entire in lower half; adaxial surface probably dark green when fresh, covered with strongly appressed, silvery, rigid trichomes and numerous orange-red sessile glands, abaxial surface grey to white, with similar but denser indumentum, especially along prominent, thickened midrib and with sessile glands, lateral nerves usually obscured except sometimes those near base on underside of lamina. Inflorescence small, ± hemispherical capitula, clustered at stem apex, on short, hairy peduncles, c. 5 mm long, arising from the axils of foliaceous bracts, similar to the leaves, but smaller, and with the stem leaves extending upwards to the base of the inflorescence. Capitula c. (4−)7−9 mm in diam., with narrow, linear bracteoles, the outer < 6.5 × 0.5 mm, with subulate apices often overtopping the flowers, ± discolorous and with indumentum similar to the leaves, but with adaxial surface subglabrous, and with margins strongly ciliate, especially towards base. Inner bracteoles often narrower. Flowers very shortly pedicellate, and with long, silky hairs extending upwards from the base and often overtopping them. Calyx at anthesis c. 4.5−5 mm long, tube 2−2.5 mm long, cylindrical, membranous, densely covered on external surface by orange-red sessile glands, and distally covered with antrorse, rigid, long hairs, ± glabrous towards base, internal surface with numerous shorter hairs except towards base, calyx lobes 2−2.5 mm long, subequal, narrowly deltate-lanceolate, densely long-ciliate and, especially on external surface, very densely hairy with long slender ± appressed hairs and sessile glands; calyx in fruit scarcely accrescent, c. 5.5 × 1 mm, tube 3−3.5 mm, broadly cylindrical, thin-walled, lobes 2−2.5 mm long, indumentum as previously described; corolla c. 7 mm long, colour not recorded, tube c. 5 mm long, very slender, c. 0.2−0.3 mm diam., straight for most of its length, slightly up-curved just below throat, externally thinly pilose distally, especially on lower part, and with sessile glands above, glabrous near base, lobes glabrous, with numerous orange-red sessile glands on external surface, anterior lobe concave, not compressed; stamens glabrous, gynoecium with stylopodium overtopping ovary at first, style glabrous with short stigmatic lobes. Nutlets 1.5−2 × 0.9−1 mm, oblong-ellipsoid, very pale brown with dimpled surface, glabrous, not mucilaginous when wet.
Hyptis spathulata is, up to the present, only known from a single collection, made in a campo cerrado area in the state of Amazonia, in the West and bordering on the state of Rondônia (Fig. 2). It was probably collected within the Parque Nacional dos Campos Amazônicos, although label data is not precise. Apart from the recent record of Hyptis velutina from the Eastern Amazon (see above), it is the only other member of the subsection to occur in Amazonia. The remaining seven species of Hyptis subsect. Velutinae all occur further South (Epling 1949   This last species is the most widespread; frequent throughout Central Brazil, it extends north-east into Bahia, north, into Pará and Tocantins states, and westward into Mato Grosso, Mato Grosso do Sul and Eastern Bolivia. From the field data provided, it is assumed that the new species is characteristic of humid soils, in open areas such as cerrado, probably similar to the habitats occupied by H. velutina.
The name was given in reference to the spathulate leaves shape.
The limited information available indicates that the species can be found in flower and fruit in July.
Hyptis saxatilis is a frequent component of the cerrado vegetation in Goiás state and in the Distrito Federal, to which it is restricted. The only other member of subsection Velutinae recorded from Amazonian Brazil is Hyptis velutina Pohl ex Benth. This is a fairly low perennial herb, frequent in cerrado in the Centre-West region of Brazil (states of Goiás, Mato Grosso and Mato Grosso do Sul), extending into Bolivia, with a few records in the Northeast region, and a few records from Tocantins state and a single record from Pará. In appearance it is very different from H. spathulata., both in habit and leaf shape.
Conservation status. Species assessment: EOO: 4 km 2 AOO: 4 km 2 IUCN Assessment: Critically Endangered (CR).Its conservation status has been classified as Critically Endangered (CR) according to criteria B: B1 and B2 a, b(iii). The only known specimen of this species was collected within or near the area that was later to become part of the Parque Nacional dos Campos Amazônicos. However, Hyptis spathulata falls within the criterion b-III da IUCN (2017), as the region suffers frequently from illegal mining and disputes regarding land-ownership, which threaten to seriously damage the habitat. Harley, sp. nov Slender, erect, aromatic perennial herb or subshrub 35-70 cm tall, upper stems slightly branched, ± quadrangular, slender, c. 2.5 mm diam., villous with long, straight, patent uniseriate trichomes, mixed with very short patent hairs, stems often bare below due to leaf-fall. Leaves sessile, erect and imbricate, sometimes slightly conduplicate and partially obscuring abaxial surfaces, upper and middle cauline leaves with lamina membranous, 8-12 × 6-9 mm, lowest < 15 × 9 mm, ± oblong with strongly cordate to almost sagittate base, apex ± acute to obtuse, margin strongly revolute or recurved, crenate, adaxial surface green, bullate between sunken midrib and primary veins, with long, whitish uniseriate trichomes, weakly antrorsely adpressed and with numerous very minute, patent trichomes, often with a small apical gland, abaxial surface pale green, with long, delicate, whitish trichomes mostly restricted to midrib and primary veins, and with a continuous indumentum of minute, often  Hyptis cachimboensis is at present known from two areas. The majority of records (in fact all, but one) occur in the Serra do Cachimbo in southern Pará state. The single record from Mato Grosso state, in Parque Estadual Cristalino, Novo Mundo, is a few hundred km to the South of Cachimbo (Fig.  4). The intervening terrain, with only one small, unpaved road, the BR-163 which connects them, crosses what must be some of the least botanically known regions in Brazil. Information on habitat is somewhat sparse, but in both localities the plants occur in open grassland, between about 380 to 485 m altitude.

Hyptis cachimboensis
Conservation status. Species assessment: EOO: 236.6 km 2 AOO: 16 km 2 IUCN Assessment: Its conservation status has been classified as Endangered (EN) according to criteria B: (B1) and B2 a, b(iii). This species occurs within the boundaries of the Parque Estadual do Cristalino and almost certainly within the Reserva Biológica Nascentes da Serra do Cachimbo. However, it is considered here to be endangered due to the rapid expansion of agriculture in the North of Mato Grosso state, which implies a decline in as yet undisturbed areas, where this species is likely to occur.
Hyptis cachimboensis was apparently first collected by L. de C. Soares in September 1952, in the Serra do Cachimbo, followed by a few other collections in the same decade, and then in the succeeding years almost up to the present. Previously specimens have often been assigned to the widespread Hyptis crenata Pohl ex Benth., to which it is obviously related. Hyptis cachimboensis is an erect herb, with narrowly deltate, not aristate, calyx lobes and the capitula with bracteoles 6-7 mm long, not rigid. The small leaves are imbricate, erect, and often conduplicate, thus bridging the morphological gap between Hyptis obtecta and other members of subsect. Sessilifoliae. The description of the flowers at anthesisis has been taken from Silva et al. 89: Pará state: Itaituba (mun.): Serra do Cachimbo, 25 Apr. 1983 (NY). The new species here described, Hyptis cachimboensis, certainly has the habit and leaf shape and indumentum of a species belonging to Hyptis subsect. Sessilifoliae, but differs from other members of the section by the conspicuous smaller leaves in H. cachimboensis.

Conclusion
It is hoped that this presented paper, by adding several taxa not previously recorded for this state, will provide further stimulus for a future Flora of Pará, the feasibility of which is now under discussion. This important and very large area of Brazil is one of those which, due in part to lack of resources of both specialist personpower and infrastructure and due to the high logistical costs required for exploration, is still very under-collected and poorly supplied with detailed floristic studies; but see Viana & Giulietti-Harley (2016, 2018a for the Flora of the canga vegetation of the Serra dos Carajás.