Three new cecidogenous species of Palaeomystella Fletcher (Lepidoptera, Momphidae) from the Brazilian Atlantic Rain Forest

Abstract Three new cecidogenous species of Palaeomystella Fletcher (Lepidoptera, Momphidae) from the Brazilian Atlantic Rain Forest are described. Larvae of P. fernandesi Moreira & Becker, sp. n., P. rosaemariae Moreira & Becker, sp. n. and P. tavaresi Becker & Moreira, sp. n. induce galls, respectively, on Tibouchina sellowiana (Cham.) Cogn., T. asperior (Cham.) Cogn. and T. fissinervia (Schrank & Mart. ex DC.) Cogn. (Melastomataceae). Adults, immature stages and galls are illustrated, and data on life history and a preliminary analysis of mitochondrial DNA sequences, including related species, are also provided.


Introduction
Cecidogeny has evolved independently in at least 20 microlepidopteran families, mostly within the Gelechioidea. Although they total only a few hundred species, the majority of these moths have been associated with their gall morphotype only, as most are still awaiting taxonomic description at the specific level (Miller 2005). The Neotropical Melastomataceae host a variety of morphotype galls that are known to be induced primarily by Lepidoptera. In this case, even the family identity of the corresponding gall-inducers remained uncertain for a long time (e.g., Tavares 1917, Houard 1933, Lima 1945 and was only recently associated with the momphine lineage as a subfamily of Coleophoridae (Becker 1999) and is herein treated as Momphidae (sensu van Nieukerken et al. 2011, Heikkilä et al. 2013. Only four of these species have been described, all belonging to Palaeomystella Fletcher, 1940 (Becker 1999, Becker andAdamski 2008). The type-species (P. chalcopeda Meyrick, 1931), for which only the female holotype from Nova Friburgo, Brazil is known, has not yet been associated with any gall morphotype (Becker 1999). The other three species such as P. tibouchinae Adamski, 2008 andP. oligophaga Becker &Adamski, 2008 induce galls on species of Tibouchina Aubl. and Macairea DC. in the Cerrado biome in central Brazil, and P. henriettiphila Becker & Adamski, 2008 induces galls on a species of Henriettea DC. in northeast Brazil (Becker and Adamski 2008). Other similar gall morphotypes occurring in Brazil have been reported for Melastomataceae as induced by unidentified Lepidoptera larvae (e.g., Gonçalves-Alvim et al. 1999, Maia and Fernandes 2004, Carneiro et al. 2009, Santos et al. 2011, Bena and Vanin 2013, Vecchi et al. 2013. Thus, these aspects increase the urgency of alpha-taxonomic work with this specialized moth lineage, together with descriptions of the gall morphotypes that they induce. The majority of the gall morphotypes (ca. 30) that are known to be induced by unidentified lepidopteran larvae in Brazil were originally described from the Atlantic Rain Forest (Rio de Janeiro State), mostly on species of Tibouchina (Tavares 1917, Houard 1933. In fact, this is one of the most diverse genera within the Melastomataceae that occur in this biome, totaling at least 137 species in southern Brazil, where most are endemic (Goldenberg et al. 2012, Guimarães 2014. Although now substantially reduced and fragmented, the Atlantic Rain Forest still supports one of the most diverse communities of plants and animals on earth, with high endemism (for general descriptions and discussions, see Morellato and Haddad 2000, Myers et al. 2000, Carnaval et al. 2009). Thus, as pointed out for this biome by Brito et al. (2013), regarding the expected diversity of leaf miner moths in general, it is expected that several species of cecidogenous momphine moths associated with Melastomataceae await description.
In the course of an ongoing survey on the diversity of microlepidopterans in the Atlantic Rain Forest, Brazil, three momphid species associated with galls induced on three different species of Tibouchina were found recently: one morphotype in Bahia and two others in Rio Grande do Sul. A comparison between their inducers and type material not only revealed the generic affinity of these microlepidopterans with Palaeomystella, but also indicated that they have diagnosable, stable, distinctive characters. Therefore, three new species are proposed here; their last larval instar, pupal and adult stages are described and illustrated, and their life history, including a general description of their galls, is characterized. A preliminary phylogenetic inference based on mitochondrial DNA sequences, including additional members of the genus, is also presented.

Materials and methods
Adults used in the study were reared by the first three authors from galls maintained in small plastic vials under controlled abiotic conditions ( (Camacan, Bahia). Immature stages were obtained by dissecting additional galls. Adult specimens were pinned and dry preserved. The immatures were fixed in Kahle-Dietrich´s fluid and preserved in 75% EtOH. For DNA analyses, additional specimens were preserved in 100% EtOH at -20 °C.
For gross morphology studies, the specimens were cleared in a 10% potassium hydroxide (KOH) solution and mounted on slides with either glycerin jelly or Canada balsam. Observations were made with the aid of a Leica® M125 stereomicroscope. Structures selected to be drawn were previously photographed with an attached Sony ® Cyber-shot DSC-H10 digital camera. Then, vectorized line drawings were made with the software CorelPhotoPaint® X4, using the corresponding digitalized images as a guide. At least five specimens were used for the descriptions of each life stage. Measurements were made with an attached ocular micrometer; values are presented as mean ± standard deviation unless noted otherwise.
Specimens used in scanning electron microscope (SEM) analyses were dehydrated in a Bal-tec® CPD030 critical-point dryer, mounted with double-sided tape on metal stubs, and coated with gold in a Bal-tec® SCD050 sputter coater. They were examined and photographed in a JEOL® JSM5800 scanning electron microscope at Centro de Microscopia Eletrônica (CME) of UFRGS.
Molecular phylogeny. Total genomic DNA was purified from larval tissue, using a Qiagen DNA Blood and Tissue Kit, to investigate: (i) monophyly of Palaeomystella fernandesi, P. rosaemariae and P. tavaresi; and (ii) reconstruct phylogenetic relationships within Palaeomystella. For comparison, two pupae of P. oligophaga Becker & Adamski, 2008, from a population of Macairea radula (Bonpl.) DC. located in Brasília, Distrito Federal, were also used for DNA extraction (Table 1). Of the mitochondrial gene cytochrome c oxidase subunit I (CO-I) a piece of 660 base pairs (bp) was amplified using the universal primers LCO1490 (5'-ggtcaacaaatcataaagatattgg-3') and HCO2198 (5'-taaacttcagggtgaccaaaaaatca-3') and following PCR conditions proposed by Folmer et al. (1994). PCR products were treated with Exonuclease I and FastAP™ Thermosensitive Alkaline Phosphatase (Thermo Scientific), sequenced using the BigDye® chemistry, and analyzed on an ABI3730XL DNA analyzer (Applied Biosystems Inc.) at Macrogen (Seoul, Republic of Korea). Sequences were aligned and visually inspected using the algorithm Clustal X in MEGA 5 (Tamura et al. 2011) running in full mode with no manual adjustment. All data generated in this study were deposited in GenBank under the accession numbers KJ188233-KJ188246 (Table 1) and BOLD system, under the project "Momphidae of Brazil" (MOMBR001-14 to 014-14). A phylogenetic tree was reconstructed in order to test the proposed hypothesis of monophyletic status for the three members of Palaeomystella: P. fernandesi, P. rosaemariae and P. tavaresi. The internal relationships of these taxa within Paleomystella and with other species were also investigated. The single currently recognized and named taxon (P. oligophaga) as well as undescribed species (Palaeomystella sp. 1 and Palaeomystella sp. 2) were used in order to cover the widest possible diversity of the genus. Accordingly, variants that match exactly the previously sequenced region in a representative taxon of the sister group of Momphidae (genus Mompha Hübner, 1825) were obtained from GenBank and were incorporated into our analysis as the outgroup (Table 1).
Phylogenetic reconstructions were based on two methods: Bayesian inference (BI), implemented in BEAST 2.0 (Drummond et al. 2012) and maximum likelihood (ML), run in PHYML 3.0 (Guindon et al. 2010). In BI, a relaxed uncorrelated lognormal clock was used together with no fixed mean substitution rate and a Yule prior on Table 1. Specimens used in this study to reconstruct the phylogenetic relationships of the new species of Paleomystella, based on cytochrome oxidase subunit I sequences.
Museum collections. Abbreviations of the Brazilian states and institutions from which specimens were examined are:   (Figs 1A-B). Sexes similar in size and color; Forewing length 4.68 to 6.11 mm (n = 7). Head (Fig. 1B): Frons and vertex creamy white; labial palpus mostly dark brown, basal segments angled laterally, terminal segment slightly angled upward; antennae dark brown; proboscis yellowish brown. Thorax: Tegula and mesonotum whitish creamy white with pale-brown scales; legs dark brown. Forewing (Figs 1A, 2A): lanceolate, with 13 veins; L/W index ~ 5.1; dorsally covered mostly by dark-brown scales; with three interconnected white areas that form a longitudinal S-like band; one proximal, rounded, in the anal area, made of pale-creamy white scales, followed by a short stripe aligned in the cubital area, made of creamy white scales, and a third, also rounded and faint, in the cell, made of pale-creamy white scales; a tenuous, U-shaped band of pale-gray scales following the contours of the tornus; three raised tufts of palegray scales, located posteriorly to cubitus, in anal area, in line with mid-cell, and near tornal area respectively; fringes dark brown; ventrally mostly covered by dark-brown scales; retinaculum subcostal; discal cell closed, ~ 0.8× length of forewing, ending near 1/5 of wing margin; Sc ending ca. middle of anterior margin; R 5-branched; R 1 ending near 1/3 of wing margin; R 4 and R 5 stalked ca. 1/2 distance from the cell apex; M 3-branched; CuA 2-branched; CuP weak proximally and not stalked, with well-developed 1A+2A extending more than 1/2 posterior margin. Hindwing (Figs 1A, 2A): strongly lanceolate, with nine veins; L/W index ~ 7.2, ~ 0.8 forewing in length; scales dark brown on both sides; fringes dark brown; frenulum a single acanthus in male, with two distally directed acanthi in female; Sc+R 1 ending ca. 1/2 anterior margin; Rs ending ca. 1/5 anterior margin; M 3-branched, M 1 and M 2 stalked from remnant chorda of cell, from point beyond base of Rs; CuA 2-branched, with CuA 1 stalked to M3; CuP weakly sclerotized, ending 1/3 posterior margin; 1A+2A well developed, ending near basis of posterior margin. Abdomen (not illustrated): pale brown, intermixed with gray scales, with transverse irregular rows of spiniform setae on terga 2-7 in both sexes; eighth sternum ( Fig. 2C) anteriorly expanded medially into a short lobe, associated with a subtriangular sternite.
Distribution. Known only from the type locality, in the Dense Umbrophilous Forest (= Brazilian Atlantic Rain Forest sensu stricto) portions of the CPCN Pró-Mata, São Francisco de Paula, RS, Brazil.
Host Plant. Tibouchina sellowiana (Cham.) Cogn. (Melastomataceae). A small tree (3 to 6 m), endemic to the coastal montane forests of southern Brazil, ranging from Minas Gerais to Rio Grande do Sul, usually flowering in April-May (Souza 1986, Guimarães 2014. Life history (Figs 12A-C). Galls induced by P. fernandesi are common on T. sellowiana at the type locality, during spring (October) and summer (February). They are prosoplasmatic histioid (Küster, in Meyer 1987); fusiform, 6.0 to 18 mm long (n = 12); induced on stem apex (Fig. 12A); without conspicuous projections, bearing a few longitudinal carinae on surface and changing gradually from green to violet as ages; fleshy, without uniformly defined internal chamber; unilocular, unilarval. Most of them house a specialized kleptoparasitic gelechiid moth, whose complex natural history is described in detail elsewhere (Luz et al. 2014). Those that are free from the kleptoparasite fall to the ground in late larval ontogeny and larva complete development on the ground. Pupation occurs inside the gall, within a cylindrical, longitudinally arranged cocoon made of woven white silk (Fig. 12C). The adult emerges presumably after the winter (September), through a circular operculum (with plant epidermis and penellipse white silk frill) on upper half of gall wall (Fig. 12B) constructed by the last instar larva prior to its pupation.
Etymology. Named in honor of Prof. Dr. Geraldo Wilson Fernandes, Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, for his great contributions to the development of cecidology in the Neotropics.

Palaeomystella rosaemariae Moreira & Becker, sp. n.
http://zoobank.org/02056832-C637-4552-8A83-2F896EEE4143 Figs 1C-D, 5-7, 11D-F, 12D-F Diagnosis. Closest to Palaeomystella tavaresi, sharing with this species a valve with a pronounced palmate costa and bladelike signa. These characters distinguish them from all other species of Palaeomystella except P. oligophaga. This, however, has the forewings with R 4 -R 5 fused and the hindwing with M 1 and M 2 stalked from the remnant chorda of the cell (Becker and Adamski 2008). P. rosaemariae differs from P. tavaresi by having: 1) adults, body covered with pale-brown scales interspersed with pale-brown scales tipped with dark brown; 2) males with latero-anterior margin of eighth sternite deeply concave; upper distal section of valva narrower; juxta as long as wide, slightly concave anteriorly; 3) females with signa with inward projection long, thin and curved; 4) pupa with cremaster tubular, dorsally directed, bearing latero-apically a pair of anteriorly curved spines; 5) galls globose, with external surface covered with short spine-like projections, induced on terminal buds of Tibouchina asperior.
Description. Adult (Figs 1C-D). Sexes similar, forewing length 4.81 to 5.59 mm (n = 5). Head (Fig. 1D): Frons pale brown; vertex and labial palpus and antenna with pale-brown scales tipped with dark brown; labial palpus with basal segments angled laterally, terminal segment slightly angled upward; proboscis yellowish brown. Thorax: Tegula and mesonotum with pale-brown scales tipped with dark brown, posterior scales having more pale brown; fore and midlegs dark brown; hindlegs pale brown, tibia and tarsus with intermixed dark-brown scales. Forewing (Figs 1C, 5A): lanceolate, with 13 veins; L/W index ~ 4.5; dorsally covered by pale-brown scales intermixed with scattered, pale-brown scales tipped with dark brown, and with longitudinally aligned groups of brown scales; a narrow, ill-defined, dark-brown streak bisecting the wing longitudinally from base to tornus; 3 raised scale tufts located posterior to cubitus, including 1 wider tuft in anal area, 1 in line with midcell, and 1 near tornal area; fringes pale brown, interspersed with a few pale-brown scales tipped with dark brown; tornal area with two bands of pale-brown scales tipped with blackish brown; ventrally, mostly uniformly covered with dark-brown scales; retinaculum subcostal; discal cell closed, ~ 2/3 length of forewing; ending near 1/5 of wing margin; Sc ending ca. middle of anterior margin; R 5-branched; R 1 ending near 1/3 of wing margin; R 4 and R 5 stalked ca. 1/4 distance from the cell apex; M 3-branched; CuA 2-branched; CuP weak proximally and not stalked, with 1A+2A that is well developed, extending more than half length of posterior margin. Hindwing (Fig. 5A) strongly lanceolate, with 9 veins; L/W index ~ 6.4, ~ 0.8 forewing in length; scales pale brown on both sides; fringes pale brown; frenulum with a single acanthus in male, and with two acanthi in female, proximal acanthus anteriorly divergent, and distal acanthus parallel to wing anterior margin; Sc+R 1 ending at ca. 1/2 anterior margin; Rs ending at ca. 1/5 anterior margin; M 3-branched; CuA 2-branched, with CuA 1 stalked to M3; CuP weakly sclerotized, ending at 1/3 posterior margin; 1A+2A well developed, ending near basis of posterior margin. Abdomen (not illustrated): pale-brown scales intermixed with gray scales, with transverse irregular rows of spiniform setae on terga 2-7 in both sexes; eighth sternum ( Fig. 5C) anteriorly expanded medially into a slender, sharply pointed lobe, associated with a subtrapezoidal sternite.
Distribution. P. rosaemariae is known only from the type locality, the fragments of lowland Dense Umbrophilous Atlantic Forest of Coxilha das Lombas, Santo Antônio da Patrulha, RS, Brazil.
Host plant. Tibouchina asperior (Cham.) Cogn. (Melastomataceae), a shrub (0.5 to 1.0 m), in humid grassland areas, endemic to Santa Catarina and Rio Grande do Sul (Souza 1986, Guimarães 2014. At Coxilha das Lombas, where the southernmost portions of lowland Dense Umbrophilous Atlantic Forest occurs, these shrubs are common along the borders of forest fragments located in poorly drained, swampy areas, associated with the formation of lagoons and also influenced by sand dunes. (Figs 12D-F). Galls induced by P. rosaemariae are located at distal axillary buds of the host. At the type locality, they occur in low numbers per plant. Galls are prosoplasmatic histioid (Küster, in Meyer 1987); small, delicate, globoid (5.2 to 7.28 mm long; n = 7), green to reddish, covered with several short spine-like projections (Fig. 12D). Unilocular, unilarval, pupates away from the gall. Little is known about the life history of this species. In laboratory, mature last instar larva invariably made a lateral orifice by chewing the gall wall (Fig. 12E) and moved directly to the bottom of the plastic pot. There, they promptly began to construct a cocoon by tying together small pieces of dried leaves with silk, where the pupation occurred (Fig. 12F). The adult emerged through a slit made at the terminal end of the cocoon. Specimens that pupated in the laboratory during the summer emerged as adults in the following autumn (May).

Life history
Etymology. Named in honor of Prof. Dr. Rosy Mary dos Santos Isaias, an anatomist of the Departamento de Botânica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, for her great contributions to the development of cecidology in the Neotropics.  Lima, 1945: 303-305, figs 180, 183, 184] misidentification.

Palaeomystella tavaresi
Diagnosis. Closest to Palaeomystella rosaemariae, sharing with this species a pronounced palmate costa of the valve and a bladelike signa. As already mentioned, these characteristics differentiate them from all other species of Palaeomystella except P. oligophaga. This species, however, has the forewings with R 4 -R 5 fused and the hindwing with M 1 and M 2 stalked from the remnant chorda of the cell (Becker and Adamski 2008). P. tavaresi differs from P. rosaemariae by having: 1) adults with body covered with pale-brown scales tipped with brown, and brown scales; 2) males with lateroanterior margin of eighth sternum anteriorly expanded medially into a stout, rounded lobe; valva with distal portion of costa wider; juxta longer than wide, anteriorly convex; 3) females with signa having inward projections shorter, straight and stout; 4) pupa with cremaster slightly bifurcated and posteriorly directed, with a latero-apically located pair of blunt spines; 5) galls of rosette type, induced on apical/terminal buds of Tibouchina fissinervia shoots, causing growth of clustered short leaves with a cylindrical gall chamber.
Distribution. Palaeomystella tavaresi is known only from the type locality, in preserved fragments of the Atlantic Rain Forest at the Serra Bonita Reserve, Camacan, Bahia, Brazil.
Host plant. Tibouchina fissinervia (Schrank & Mart. ex DC.) Cogn.(Melastomataceae), a pioneer tree species that grows up to 20 m tall in the Atlantic Rain Forest, where it is endemic, ranging from Bahia to São Paulo (Freitas 2011). In the Serra Bonita Reserve, these trees are relatively common at higher altitudes, above 600 m, growing spontaneously in areas that were formerly cleared for agriculture and along trails and in clearings in pristine forests, resulting from the fall of other trees.
Life history (Figs 12G-I). Gall prosoplasmatic histioid (Küster, in Meyer 1987), of the rosette type (internal length from 18 to 31 mm; n = 6), induced on growing shoots causing growth of clustered short leaves (Fig. 12G); green, progressively darkening during senescence, after emergence of the moth; unilocular, unilarval. A longitudinal, narrow, cylindrical chamber is formed in the middle (Fig. 12H), where the larva develops and pupate. The mature last instar larva constructed a brown silk plug/gate near the middle of the chamber consisting of two convex hatches that open horizontally (Fig. 12I). Then it constructed a flimsy silk cocoon in the proximal sector of the chamber, where pupation occurred. To exit the cocoon the adult pushed the hatches open, and emerged through the terminal leaflets of the gall. At the type locality, the galls occur in small numbers on T. fissinervia trees, occasionally in groups of a few per plant. Under laboratory conditions, mature galls collected in the spring (October) had the adults emerging ca. 15 days later.
Etymology. P. tavaresi is named in memory of the Jesuit priest Joaquim da Silva Tavares, a Portuguese naturalist and a pioneer in the study and description of Brazilian cecidology (Tavares 1917).
Remarks. Lima (1945: 304, 305, Figs 180, 183, 184) illustrated the gall, wing venation and male genitalia of a species that he identified as a member of Walshia Clemens (Cosmopterigidae), reared from galls on branches of a species of Tibouchina collected in Petrópolis, Rio de Janeiro. The gall, genitalia and wing venation appear almost identical to those of P. tavaresi and very likely represent the same species. Tavares (1917: 31, pl . 1, Figs 1, 2) described a stem gall, also from a Tibouchina sp., collected at Tijuca and Petrópolis, Rio de Janeiro, which exactly resembles the galls of P. tavaresi. However, his description of the moths as "shiny, brunneous, with several golden spots on the upper side of forewings" does not match the one described here.  100, 20, 200, 100, 20, 200, 100, 20, 200 µm, respectively. Molecular phylogeny. A total of 660 nucleotide sites were analyzed for species of Palaeomystella from different host plants, and 211 (32%) of these were variable. According to the phylogenetic hypothesis proposed here, all species were recovered Figure 12. Galls induced by Palaeomystella species: A-C P. fernandesi D-F P. rosaemariae G-I P. tavaresi A on Tibouchina sellowiana, general view B operculum made by last-instar larva on gall surface before pupation; C pupal cocoon in a dissected gall (arrow indicates the operculum shown in B) D on Tibouchina asperior, general view E exit hole made by last larval instar on gall surface F pupal cocoon constructed between two leaves, uncovered by pulling them apart (direction indicated by arrows) G on Tibouchina fissinervia, general view H longitudinally dissected gall, showing gall chamber (arrow indicates position of exit orifice on cocoon) I internal chamber in detail, showing the exit orifice on cocoon (asterisk). Scale bars = 10, 2, 4, 10, 5, 4, 10, 10, 2 mm, respectively. as monophyletic lineages within the Palaeomystella group of Momphidae, in both methods of inference (BI and ML) with full branch support (Fig. 13). Regarding internal relationships, P. rosaemariae was placed as a sister of P. tavaresi with strong posterior probability (= 1) and bootstrap (=100). P. fernandesi was more distantly related, although with low branch support (< 0.8, posterior probability; < 70, bootstrap). Despite the strong internal statistical branch support of the three new lineages of momphids, the external relationships for Palaeomystella were poorly resolved (i.e., position of clades), and even the monophyly of the genus lacks statistical support. Mompha was used to root the tree, but its position as a sister clade of Palaeomystella was not supported (Fig. 13). The evolutionary divergence observed between comparisons of pairs of species was markedly high, showing greater genetic variation in this group of momphids (Table 2), particularly between clades (Fig.  13). An average of 18% (± 3%) of K2P differences was found between species of Palaeomystella, ranging from 14 (± 2%) to 24% (± 3%). The maximum divergence observed among clades was 20%, found between P. fernandesi and the clade formed by P. rosaemariae + P. tavaresi + Palaeomystella sp. 1 (Fig. 13). The genetic divergence within Palaeomystella (ca. 18%) was greater than between this genus and Mompha (16%). sampling, and then to sample at least a few more loci in order to better understand the relationships within this group of momphids.
As mentioned above, the diversity of moth-induced melastome galls is in fact much greater that presented here (e.g. Hanson et al. 2014). Several species belonging to this group, in addition to the two that we included in the molecular analyses, are present in our collections and still await description. Furthermore, in contrast to our expectations the species described here are not only similar from a gross morphology perspective, but share several fine-scale morphological characteristics with those belonging to Mompha Hübner. For example, the divided male valva into two sections and the female bursa with a bladelike signa are also found within the latter genus (e.g., Hodges 1998, Wagner et al. 2004. A pupal cremaster bearing spines, similar to those described in this study, is also found among the Palearctic species of Mompha (e.g., Patočka and Turčani 2005). Contrary to what is known for the larvae of two species of Palaeomystella that were proposed by Adamski and Becker (2008), all species described here are bisetose regarding the prothoracic L-group setae, a characteristic of Mompha species (Stehr 1987, Wagner et al. 2004. Also in contrast to their findings, the present species do not show a reduction in the number of setae on the anal plate. Thus, the generic status of the species described in this study may change in the future, pending descriptions of additional taxa, further studies of phylogeny, and taxonomic revision of the family. A decision concerning this matter would be precipitate at present, as Mompha is similarly a poorly known genus, with species that are difficult to collect and have a wide variation of feeding habits (including cecidogeny). Therefore, several species either remain undescribed or lack taxonomic descriptions that are sufficiently detailed to allow comparisons. Furthermore, DNA sequences for only a few species are available, and as with Palaeomystella, show a wide range of evolutionary divergence (4-14%) (for discussion, see Emery et al. 2009).
This study illustrates further variation in gall morphotypes. Such variation has long been known to exist among Melastomataceae galls (Tavares 1917, Houard 1933, confirming that they are associated with different species of lepidopteran inducers. At least two of the galls described here may have appeared before in the literature, but none of them has been accurately associated with any cecidogenous species. The fusiform type induced by P. fernandesi on T. sellowiana was reported by Toma and Mendonça (2013), in a gall survey that they carried out at the type locality, as being induced by an unidentified member of Gelechioidea. As already mentioned, depending upon the time of year, most such galls are associated in the field with a specialized kleptoparasitic gelechiid moth, with which the cecidogenous species may be confounded (Luz et al. 2014). Galls similar to the rosette type induced by P. tavaresi in T. fissinervia were illustrated by Tavares (1917) and Houard (1933) for Tibouchina sp. in Rio de Janeiro State. Lima (1945) showed the same type of gall for an unidentified species of Tibouchina, also in Rio de Janeiro, but as mentioned above, he erroneously associated it with an unidentified species of Walshia (Cosmopterigidae). Furthermore, galls of the same rosette type as those of P. tavaresi, apparently induced by unidentified species of Lepidoptera, were shown by Maia and Fernandes (2004) on Miconia theizans (Bonpl.) Congn. occurring in Minas Gerais State, and by Santos et al. (2011) and Isaias et al. (2013) on Henriettea succosa (Aubl.) DC. in Pernambuco State. Further studies should be conducted to determine whether such galls are induced by P. tavaresi or by closely related species.
The present results also demonstrate the existence of considerable variation in lifehistory styles for the pupal stage of Palaeomystella species, which should be taken into account in future studies. That is, last-instar larvae may remain endophylous until pupation in either sessile (P. tavaresi) or dehiscent (P. fernandesi) galls, or may leave them to pupate in leaf litter (P. rosaemariae). Although varying little in the general integumentary morphology, their pupae show considerable variation in the size and shape of the cremaster which may provide useful characters for future species identifications. Unfortunately, the other known species of Palaeomystella (Becker and Adamski 2008) were not described under the scanning electron microscope, and thus cannot be closely compared. These structures are supposedly used to anchor the pupa laterally to the cocoon/pupal chamber. They have likely evolved in conjunction with the habit of emerging of the adults in the pupation sites, which apparently first appeared in the Lepidoptera evolution within the Gellechioidea (e.g. Powell 1973, Becker 1982. Variation in chaetotaxy among species may also prove to be useful to identify different lineages within Palaeomystella. For example, in addition to the variation in number of setae described by Adamski and Becker (2008) for the prothoracic L-group and anal plate, a numerical variation in chaetotaxy was also detected on the head (AF1 seta), thorax and abdomen (SD setae).