R epResentativity of the genus A sphondyliA L oew , 1850 (D ipteRa , C eCiDomyiiDae ) in B RaziL

Representativity of the genus Asphondylia Loew, 1850 (Diptera, Cecidomyiidae) in Brazil. This cosmopolitan genus includes 272 galling species described, of which about 100 occur in the Neotropical Region. The present study goal to evaluate the richness of Asphondylia in Brazil, to provide an updated list of the host plant species, to determine the plant organs where galls are induced, to update the geographic distribution of the genus, to verify it distribution in the Brazilian biomes, and to the list the associated fauna. The survey data was carried out by consulting the Cecidomyiidae collection of the Museu Nacional/UFRJ, the database “Web of Science” using Asphondylia and Brasil/Brazil as keywords, 51 Brazilian inventories and two catalogues. Asphondylia is represented by 58 species in Brazil, among them, twenty are already known and 38 are still undetermined. The Brazilian described species represent 8% of the total of known species of Asphondylia in the world and 21% of species of the Neotropical fauna. This genus is associated with 51 plant species and 20 plant families in Brazil. Astera-ceae comprise the greatest richness of Asphondylia species. This genus was found in five biomes, among them the Atlantic forest has the highest species richness. The associated fauna comprises parasitoids (Hymenoptera) and inquilines (Lepidoptera, Coleoptera and Diptera).


INTRODUCTION
Asphondylia Loew, 1850 belongs to the tribe Asphondyliini, currently with 505 described species, subtribe Asphondyliina, which is the most diverse in the Neotropics (Gagné & Jaschhof, 2014).This cosmopolitan genus includes 272 described galling species, of which about 100 occur in the Neotropical Region (Gagné, 2004).Carneiro et al., (2009a) reports the importance of genera as one of the domi-adult morphology differs little among species.All species share the neckless, cylindrical antennal flagellomeres in both sexes, the needle-like ovipositor and conspicuously enlarged seventh sternite in the female, and the compact male genitalia with the spherical gonostyli positioned dorsally rather than apically (Gagné, 1994;Dorchin et al., 2015).The third-instar larvae possess a well-developed, usually four-toothed spatula on the first thoracic segment.The pupation in all Asphondylia species takes place inside the gall rather than in the soil (Gagné, 1989(Gagné, , 1994)).The pupae are characterized by well-developed horn-like antennal bases, a varying number of facial horns, and transverse dorsal rows of spines on the abdominal segments (Gagné, 1989(Gagné, , 1994) ) that assist in breaking out of the galls just before adults' emergence.
Constant assessment of biodiversity knowledge is essential to guide future efforts inventories in poorly studied areas, so that they can conserve endangered species (Toma & Maia, 2012).Moreover, considering the great diversity in the Neotropics, little is known about the fauna of Cecidomyiidae in this region, especially compared to other parts of the world, getting to be around six times smaller than the Palaearctic fauna (Gagné, 2007;Toma & Maia, 2012).
The present study goal to evaluate the richness of Asphondylia in Brazil, to determine the plant organs where galls are induced, to give an updated list of the host plant species, highlighting the their endemism and categories of conservational status, as well as to verify the species richness in each Brazilian biome, to update the geographic distribution of the genus and the respective associated fauna.

MATERIAL AND METHODS
The survey data on the species of Asphondylia was carried out by consulting the collection of Cecidomyiidae the Museu Nacional/UFRJ (the only reference collection to the family in Brazil) and, concomitantly, a literature review was performed on the database "Web of Science" using Asphondylia and Brasil/Brazil as keywords.Furthermore, the catalog of Cecidomyiidae of the world (Gagné & Jaschhof, 2014) was used to check all described Brazilian Asphondylia species until 2014.Besides, all insect galls Brazilian inventories were examined, as well as the catalog of galls of the Central and South Americas (Houard, 1933), which comprises data on Brazilian galls.
Information about the host plant (botanical species and family), galled plant organs, shape and type of gall, and geographical distribution for each species were provided, whenever possible.Biomes were defined from the collection locations.Data on origin and categories of the conservational status of the host plants were found in the Lista da Flora do Brasil 2016 website (Flora do Brasil, 2020).
According to Carneiro et al., (2009a) the gall morphology associated with the identification of host-plant species is a reliable richness indicator of the gall-inducing insects.About 95% of described species of cecidomyiids from Brazil can be identified based on these two data together (Coelho et al., 2013a).In the present study, we adopted both information to evaluate the richness of the genus Asphondylia.
Considering the high host specificity of the gall midges, the undetermined species probably belong to new species, as they are associated with new records of plants.Based on this specificity, we included also the undetermined species of Asphondylia in the following discussion (Maia & Silva, 2013).

RESULTS
Asphondylia is represented by 58 species in Brazil (21 from the papers retrieved from the Web of Sci-  Gagné & Jaschhof, 2014, 9 exclusively from the MNRJ, and 47 simultaneously from the MNRJ and literature).The MNRJ comprehended data on 11 determined and 36 undetermined species, all mounted on slide.The later comprised 14 new records.Among them, 20 are known and 38 are still undetermined.The last ones comprise 15 species found in the MNRJ and 23 retrieved from the literature (Maia, 2013a;Maia & Fernandes, 2004;Maia & Souza, 2013;Maia et al., 2008a;Maia et al., 2014;Mendonça et al., 2014;Pamplona et al., 2000;Rodrigues et al., 2014;Urso-Guimarães & Scareli-Santos, 2006.)The described species represent 8% of the known species of Asphondylia in the world and 21% of Neotropical species.
Asphondylia is associated with 51 plant species and 20 plant families in Brazil.Asteraceae comprise the greatest richness of Asphondylia species (n = 23).Data on other families are shown in Table 1.Baccharis L. and Mikania Willd.(Asteraceae) were the plant genera that host the greatest number of Asphondylia species, both with five.Mikania glomerata (Asteraceae) was the unique plant species that comprised more than one species of Asphondylia.Seventeen species of host plants are endemic in Brazil, thirty four are native, and one is exotic.The endemic plants host Asphondylia maricensis Maia & Couri, 1992, A. peploniae Maia, 2001, A. serrata Maia, 2004, and 13 undetermined species.Concerning the conservational status, Baccharis concinna was the single species categorized as a vulnerable, seven were categorized as less concerning (C.serrata, G. polymorpha, M. glomerata, P. cf. cinerea, P. notata, Z. montana, A. fasciculata), and the remaining have not been yet evaluated for threat of extinction.Baccharis concinna host a single species of Asphondylia, The galls of Asphondylia were found on leaves, stems, flower buds, buds, flowers, fruits, inflorescences and simultaneously on leaves/stems.Leaves comprised the greatest richness of Asphondylia species (n = 14) (Table 2).Many gall shapes were found: globose, ovoid, rosette, spherical, elliptic, swollen, cylindrical, conical, drop-shaped, amorphous, aggregate and fusiform (the terminology used by each author was repeated in the present work).The majority of the analyzed morphotypes were glabrous and one-chambered.There was an evident predominance of green (n = 18) galls, the color of the most galled plant organ (leaf ).But, yellow, brown, black and cream galls were also found (again, we repeat the color information given by each author) (Table 3).
Asphondylia obtained.The Cerrado was second biome in richness of species, followed by Pampa, Caatinga and Amazonian Forest, with 10, 5, 3 and 1 species, respectively.All species were found in a single biome, excepting Asphondylia moehni Skuhravá, 1989, with occurrence in the Pampa and Atlantic forest.The species were collected in Para (PA), Sergipe (SE), Ceará (CE), Pernambuco (PE), Bahia (BA), Espírito Santo (ES), Minas Gerais (MG), Rio de Janeiro (RJ), São Paulo (SP), and Rio Grande do Sul (RS) (Fig. 2).The majority of the species were found in RJ, SP, MG, and RS, the best surveyed states.The other states have been less surveyed.All species were recorded in a single Brazilian state, with four exception, A. moehni (in RJ, SP and RS), A. glomerata (in RJ and SP) A. cfr.cordiae (in RJ and SP) and Asphondylia sp.(in ES and SP) (Table 3).As the geographical distribution of all host plant species is wider than that of the galler (Table 4), we expect that researches in less sampled states can result in an expansion of the galling species distribution.
Only five species from Brazil are known in the larval stage, pupa and adult of both sexes (Table 5).Asphondylia ulei, characterized only in the larval stage, is the described species that most lacks morphological information.The number of undescribed species to Brazil indicates that Asphondylia is more diverse than it seems.
The associated arthropod fauna comprises parasitoids and inquilines.The former are represented by three families of Hymenoptera: Eurytomidae, Braconidae and Pteromalidae, and the later by three insects orders: Lepidoptera, Coleoptera and Diptera.The beetles are represented by a single family, Curculionidae, whereas the flies are represented by three, Tephritidae, Sciaridae and Cecidomyiidae (Table 6).The taxonomical knowledge of the associated fauna is superficial, with most records at order or family level and only two in genus level.
Nine species of Asphondylia were attacked by Hymenoptera, the most important natural enemy of these midges in the world (Gagné, 1994).In Brazil, Eurytomidae and Pteromalidae are very frequent parasitoids in Cecidomyiidae galls; while Braconidae are less common (Maia & Azevedo, 2009).Five species of Asphondylia induce galls which are invaded by inquilines.Lepidoptera, Curculionidae, Tephritidae, Sciaridae, and Cecidomyiidae are frequent inquilines of Cecidomyiidae galls in Brazil.

DISCUSSION
This genus is one of the most frequent in Brazil, represented by 20 described species (Carneiro et al., 2009a).All determined species reported in the present study were cited by Carneiro et al., (2009a), except Asphondylia gochnatiae Maia, 2008.According to Maia et al. (2009), Asphondylia is associated with 66 plant families in the world, being more frequent on Asteraceae, Fabaceae and Chenopodiaceae.In the present study, Asteraceae are also pointed out as an important host plant family, but this is not the case of Fabaceae and Chenopodiaceae.Another difference is the indication of Rubiaceae as an important host family.
Eighteen Asphondylia species are associated with these endemic hosts.Among them, three are identified at species level, Asphondylia maricensis, Baccharis concinna was categorized as a vulnerable species, so the undetermined galler which is associated with this host can be also vulnerable, if it occurs exclusively on it.It is important to highlight this situation, where a probably new species can be considered vulnerable.So, it can become extinct before being formally described, implying silent extinction.
The highest incidence of galling species on leaves confirmed the global pattern pointed out by Felt (1940), probably because the leaves represent an abundant and frequent resource (Maia et al., 2008a).
The highest number of species (39 spp.) was recorded in the Atlantic forest, what corresponds to about 67% of the total, but this result does not necessarily indicate that the Atlantic forest comprises the greatest richness of Asphondylia species, as the highest number of inventories developed by taxonomists was carried out in this biome.In addition to that, there are more available specimens of the Atlantic forest for identification.
In spite of being one of the most surveyed biomes, the Cerrado comprises only 10 Asphondylia species.This can be explained by the low number of available specimens for identification.Differing from the multivoltine species of the restingas, the gall midges of the Cerrado are generally univoltine, with longer life cycles and consequently lower success of rearing.The other biomes have been little studied.
Considering all species (58 spp.), the genus in Brazil is quite representative of the Neotropical fauna, although the morphological knowledge of many species is partial due to the lack of knowledge of the life cycle phases or both sexes.The species described in the larval stage, pupal and adult male and female correspond to the more recently described, and indicate the current tendency to describe a new taxon only when all stages are shown (except egg, which contributes little to the taxonomy).
The records of inquilines exemplify some ecological interactions between Asphondylia species and other insects and suggest the role of these gall midges as ecosystem engineers.The importance of the parasitoid wasps as natural enemies of the cecidomyiids are confirmed.These can be evaluated as more data on the associated fauna are known.

CONCLUSIONS
The taxonomical knowledge of the Brazilian species of Asphondylia is still incipient.Asphondylia maricensis, A. peploniae and A. serrata are endemic based on their occurrence exclusively on endemic host plants.Asteraceae is confirmed as the botanical family which hosts the greatest richness of Asphondylia species.The geographic distribution of the Brazilian Asphondylia species is poorly known.The genus exhibits genetic plasticity to different environmental conditions, as it has been recorded in five Brazilian biomes.The associated fauna is little studied.

TABLE 2 :
Distribution of the number of Asphondylia species (Diptera, Cecidomyiidae) per plant organs in Brazil.
C. & Maia, V.C.: The genus AsphondyliA Loew, 1850 (Diptera, Cecidomyiidae) in Brazil whereas the less concerning plants host nine species and the others 48.

TABLE 3 :
Host plants, gall characterization and geographic distribution of Asphondylia species (Diptera Cecidomyiidae) in Brazil (AF: Atlantic Forest).Note: Three botanical names were updated.

TABLE 4 :
Species of host plants, their distribution in Brazil and the galling species distribution.

TABLE 5 :
Brazilian species of Asphondylia (Diptera, Cecidomyiidae) and described life cycle phases.The letter "x" indicates known phase.

TABLE 6 :
Asphondylia species (Diptera, Cecidomyiidae) and associated fauna in Brazil. .peploniae and A. serrata.As they induce gall exclusively on endemic plants, they are proposed here as endemic too.The other 15 species, although still undetermined, probably are endemic based on the high specificity between the gallers and the host plants, but more comparative morphological studies among them are necessary before proposing it.Unfortunately, the available material is not enough, as the species are represented by different life cycle stages. A