BIOLOGY Abundance and Richness of Cryptic Species of the willistoni Group of (Diptera: ) in the Biomes Caatinga and Atlantic Forest, Northeastern Brazil

ANA CRISTINA LAUER GARCIA,1,2 DIVA MARIA IZABEL DE OLIVEIRA SILVA,3 AMANDA GABRIELA FELIX MONTEIRO,1 GEO´ RGIA FERNANDA OLIVEIRA,1 4 1 MARTI´N ALEJANDRO MONTES, AND CLA´ UDIA ROHDE Downloaded from https://academic.oup.com/aesa/article/107/5/975/64874 by guest on 27 September 2021

Ann. Entomol. Soc. Am. 107(5): 975Ð982 (2014); DOI: http://dx.doi.org/10.1603/AN14052 ABSTRACT Cryptic species meet the biological deÞnition of species, but are morphologically identical or quite similar. Several ecological studies underestimate richness and neglect important information on cryptic species, as they are rather difÞcult to identify. Among , drosophilids of the willistoni subgroup, which includes Drosophila willistoni Sturtevant, Drosophila paulistorum Dobzhansky & Pavan, Drosophila equinoxialis Dobzhansky, Drosophila tropicalis Burla & da Cunha, Drosophila insularis Dobzhansky, and Drosophila pavlovskiana Kastritsis & Dobzhansky, are good examples of cryptic species. Although several studies have shown that this subgroup is one of the most abundant in the Neotropical region, no identiÞcation to species level has been reported for areas where these individuals live in sympatry. This study evaluates the seasonal oscillations in abundance of this subgroup in biomes with contrasting vegetation and rainfall regimes: the Caatinga and the Atlantic Forest, in northeastern Brazil. Approximately 39,000 drosophilids were captured in 24 collections, of which 18,000 belonged to the willistoni subgroup. The most abundant were D. willistoni, D. pauli- storum, and D. equinoxialis, in this order. D. equinoxialis was recorded in only one of the environments surveyed, represented by few individuals. In all environments, individuals of the willistoni subgroup were more abundant in the rainy season, when richness often was higher. The results underline the importance of humidity and of temperature for the subgroup willistoni and indicate the ecological versatility of some of its species.

KEY WORDS ecology, neotropical region, pernambuco, temperature, humidity

Cryptic species establish morphologically identical or by cryptic species (Salles 1947; Burla et al. 1949; quite similar natural populations that nevertheless re- Breuer and Pavan 1950; Magalha˜es and Bjo¨rnberg main isolated from the reproductive standpoint (Mayr 1957; Vilela 1983, 1992; Vilela and Ba¨chli 1990; Vilela 1963). The notion of cryptic species is one of the most et al. 2002). Examples include the willistoni group of stringent conditions that defy the morphological con- Drosophila, which is composed of three subgroups. cept of species and poses a signiÞcant problem to One of these is the willistoni subgroup, represented by taxonomists. Dobzhansky (1970) underscores the im- six cryptic species. Of these, Drosophila insularis Dob- portance of the identiÞcation of these individuals in zhansky and Drosophila pavlovskiana Kastritsis & evolutionary and ecological studies, and emphasizes Dobzhansky are endemic to the Lesser Antilles and that species are natural entities that exist irrespec- Guiana, respectively. The other four species are tively of our competence to distinguish one from the widely distributed and are often sympatric. Drosophila other. willistoni Sturtevant occurs from the south of the A considerable share of the diversity of small ßies United States to northern Argentina and Uruguay, that comprise the Drosophilidae family is represented Drosophila paulistorum Dobzhansky & Pavan spreads from northern Guatemala to southern Brazil, Drosoph- 1 Universidade Federal de Pernambuco, Centro Acadeˆmico de ila equinoxialis Dobzhansky distributes from southern Vito´ria, Programa de Po´s-Graduac¸a˜oemSau´ de Humana e Meio Am- Mexico and practically reaches central Brazil and biente, Rua do Alto do Reservato´rio, s/n, Bela Vista, 55608-680 Vito´ria Peru, while Drosophila tropicalis Burla & da Cunha is de Santo Anta˜o, PE, Brazil. 2 Corresponding author, e-mail: [email protected]. recorded from northern Guatemala down to central 3 Universidade de Pernambuco, Programa de Po´s-Graduac¸a˜oem Brazil and central Bolivia (Spassky et al. 1971, Dob- Biologia Celular e Molecular Aplicada, Rua Arno´bio Marques, 310, zhansky and Powell 1975, Ehrman and Powell 1982, Bairro Santo Amaro, 50100-130 Recife, PE, Brazil. Santos and Valente 1990). 4 Universidade Federal Rural de Pernambuco, Departamento de Biologia, Campus Dois Irma˜os. Rua Dom Manoel de Medeiros, s/n. In the Neotropical region, the willistoni subgroup is CEP 52171-900 Recife, PE, Brazil. one of the most abundant in numerous environments,

0013-8746/14/0975Ð0982$04.00/0 ᭧ 2014 Entomological Society of America 976 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 107, no. 5 Downloaded from https://academic.oup.com/aesa/article/107/5/975/64874 by guest on 27 September 2021

Fig. 1. Collection sites of drosophilids in the state of Pernambuco, northeastern Brazil, in the Caatinga biome (black squares) and Atlantic Forest biome, biogeographical subregions Brejos de Altitude (triangles) and Pernambuco (circles). Inset shows the map of South America, Brazil (light gray), and the northeastern region of the country (dark gray). Site codes: SET-1 (Serra Talhada, 07Њ 57Ј S, 38Њ 18Ј W); SET-2 (Serra Talhada, 07Њ 56Ј S, 38Њ 17Ј W); CAT-1 (Buõ´que, 08Њ 24Ј S, 37Њ 09Ј W); CAT-2 (Buõ´que, 08Њ 36Ј S, 37Њ 14Ј W); TRI (Triunfo, 07Њ 50Ј S, 38Њ 06Ј W); MAD (Brejo da Madre de Deus, 08Њ 12Ј S, 36Њ 24Ј W); BEZ (Bezerros, 08Њ 53ЈS, 36Њ 29Ј W); BON (Bonito, 08Њ 30Ј S, 35Њ 42Ј W); TAP (Sa˜o Lourenc¸o da Mata, 08Њ 07Ј S, 35Њ 17Ј W); POM (Pombos, 08Њ 08Ј S, 35Њ 23Ј W); ZOO (Recife, 08Њ 03Ј S, 34Њ 52Ј W); SAL (Tamandare´ ,08Њ 45Ј S, 35Њ 06Ј W). and sometimes accounts for the largest proportion of gimes: the Caatinga and the Atlantic Forest in north- individuals collected (Dobzhansky and Pavan 1950, eastern Brazil. In the latter, the biogeographic subre- Martins 2001, Medeiros and Klaczko 2004, Tidon 2006, gions Brejos de Altitude and Pernambuco were Do¨ge et al. 2008, Acurio and Rafael 2009, Rohde et al. surveyed. 2010). Nevertheless, in several studies carried out in areas where more than one species of this group has Materials and Methods been recorded, its representatives are not identiÞed to species level. This situation is observed in Brazilian Study Sites. Adult drosophilids were collected in biomes like the Atlantic Forest (Pavan 1950, Sene et 12 sites in the state of Pernambuco, northeastern Bra- al. 1980, Tidon-Sklorz and Sene 1992, De Toni et al. zil. Of these, four are located in the Caatinga biome 2001, Silva et al. 2005, Mateus et al. 2006, Torres and and eight are in the Atlantic Forest biome. In the Madi-Ravazzi 2006, De Toni et al. 2007, Cavasini et al. latter, four are in the Brejos de Altitude subregion and 2008, Do¨ge et al. 2008, Gottschalk et al. 2009, Bizzo et four are in the Pernambuco subregion (Fig. 1). In each al. 2010), the Cerrado (Sene et al. 1980, Torres and site, two collections were conducted, totaling 24 sam- Madi-Ravazzi 2006, Tidon 2006, Blauth and Gottschalk plings, at periods characterized by contrasting rainfall 2007, Mata et al. 2008), the Amazon Forest (Martins levels (the dry and the rainy season). 1989, 2001), and the Caatinga (Sene et al. 1980). The vegetal formations in Caatinga are character- In light of the importance and of the paucity of ized by xerophilous steppic savanna, and the dry sea- information about species of the willistoni subgroup, son is long (Eiten 1982). Mean annual rainfall varies the current study assessed the seasonal oscillations in between 400 and 600 mm, most of which in three willistoni representativesÕ richness and abundance in consecutive months between November and June biomes with contrasting vegetation and rainfall re- (AbÕSaber 1977). The Caatinga is the only exclusively September 2014 GARCIA ET AL.: CRYPTIC SPECIES OF THE willistoni GROUP OF Drosophila 977

Table 1. Absolute abundance of species of the willistoni subgroup of Drosophila and of the other drosophilid species captured in different collection efforts in each of the three environments surveyed in the dry and rainy seasons

Locality willistoni Subgroup Other Biome Environment Date Season Total code D. willistoni D. paulistorum D. equinoxialis Total species Caatinga SET-1 15 Feb. 2009 Dry 3 Ð Ð 3 275 278 23 June 2009 Rainy 1,611 133 Ð 1,744 771 2,515 SET-2 15 Feb. 2009 Dry 1 Ð Ð 1 783 784 23 June 2009 Rainy 1,975 154 Ð 2,129 1,069 3,198 CAT-1 13 Mar. 2009 Dry Ð Ð Ð Ð 273 273 25 June 2008 Rainy 312 Ð Ð 312 630 942 CAT-2 13 Mar. 2009 Dry Ð Ð Ð Ð 350 350 25 June 2008 Rainy 149 55 Ð 204 1,011 1,215 Total 4,051 342 Ð 4,393 5,162 9,555

Atlantic Forest Brejos de altitude TRI 30 Jan. 2009 Dry 8 7 Ð 15 284 299 Downloaded from https://academic.oup.com/aesa/article/107/5/975/64874 by guest on 27 September 2021 23 June 2009 Rainy 774 298 Ð 1,072 602 1,674 MAD 16 Dec. 2010 Dry 31 4 Ð 35 35 70 10 Nov. 2010 Rainy 274 5 3 282 315 597 BEZ 8 Dec. 2009 Dry 44 Ð Ð 44 395 439 12 July 2008 Rainy 117 11 Ð 128 4,064 4,192 BON 6 Feb. 2010 Dry 994 11 Ð 1,005 862 1,867 17 May 2010 Rainy 3,028 262 Ð 3,290 894 4,184 Total 5,270 597 3 5,871 7,451 13,322 Pernambuco TAP 5 Mar. 2011 Dry 682 42 Ð 724 2,960 3,684 14 Oct. 2010 Rainy 746 73 Ð 819 1,152 1,971 POM 5 Jan. 2009 Dry 215 25 Ð 240 201 441 12 July 2008 Rainy 1,528 292 Ð 1,820 681 2,501 ZOO 23 Feb. 2009 Dry 338 284 Ð 622 1,097 1,719 4 July 2008 Rainy 1,356 867 Ð 2,223 1,532 3,755 SAL 10 Dec. 2009 Dry 323 11 Ð 334 390 724 28 July 2009 Rainy 733 449 Ð 1,182 76 1,258 Total 5,921 2,044 7,964 8,089 16,053 Total geral 15,242 2,983 3 18,228 20,702 38,930

Collection site codes are described in Fig. 1.

Brazilian biome, and its biodiversity has been poorly pended 1.5 m above the ground and kept in the col- studied. Even though it has undergone expressive en- lection sites for three consecutive days. vironmental changes because of anthropogenic inßu- The individuals of the willistoni group were Þrst ence, the Caatinga is one of the Brazilian biomes least sorted from the other specimens based on an analysis protected by conservation units (Leal et al. 2003). of external morphology. Whenever the number of The Atlantic Forest is formed by trees that reach individuals collected was suitably high, 100 ßies of 35 m, bushes, and epiphytical species (Klein 1978). It willistoni subgroup of each gender were identiÞed per is one of the richest biomes in terms of biodiversity, sampling. Females were characterized by electropho- and one of the most seriously threatened in the planet resis of the enzyme acid phosphatase 1 (Acph-1), ac- (Ribeiro et al. 2009). The biome is particularly dev- cording to Garcia et al. (2006). Males were identiÞed astated in northeastern Brazil, north of the Sa˜o Fran- based on the analysis of the hypandrium (Wheeler and scisco River, and today it covers Ͻ7% of its original area Kambysellis 1966), according to Rohde et al. (2010). (Siqueira-Filho and Leme 2006). There, the biome splits When collections provided Ͼ200 individuals of the into the biogeographic subregions called Brejos de Al- willistoni group, the number of representatives of each titude and Pernambuco (Da Silva and Casteleti 2005). In species was estimated considering the proportion of those areas, mean annual rainfall varies between 1,300 individuals identiÞed. and 2,400 mm (Veloso et al. 1991). Voucher specimens were deposited in the droso- Brejos de Altitude is characterized by humid forest philid collection of the Laborato´rio de Gene´tica of the “islands” surrounded by typical Caatinga vegetation. Centro Acadeˆmico de Vito´ria of the Universidade These areas have been associated with the existence of Federal de Pernambuco. plateaus between 500 and 1,000 m above sea level Data Analysis. The chi-square test was used to (a.s.l.; Andrade-Lima 1960). The Pernambuco subre- assess the differences in abundance of species of the gion is located in the coastal region of northeastern willistoni subgroup between the three environments Brazil. Altitudes are Ͻ500 m a.s.l. This subregion in- surveyed and seasonality in the software BioEstat 5.0 cludes the states of Alagoas, Pernambuco, Paraõ´ba, and (Ayres et al. 2007). Rio Grande do Norte (Ribeiro et al. 2009). Capture and Identification of Drosophilids. Results were captured using traps with banana baits, Musa acuminata Colla, prepared according to the model Approximately 39,000 drosophilids were captured developed by Tidon and Sene (1988). Ten traps were in the 24 collection efforts in the sites surveyed in the used in each collection effort. All traps were sus- Caatinga and Atlantic Forest (Brejos de Altitude and 978 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 107, no. 5 Downloaded from https://academic.oup.com/aesa/article/107/5/975/64874 by guest on 27 September 2021

Fig. 2. Absolute abundance of the willistoni subgroup of Drosophila (A) and of D. willistoni and D. paulistorum (B) compared with the other drosophilids collected in the Caatinga and in the Atlantic Forest (subregions Brejos de Altitude and Pernambuco), northeastern Brazil.

Pernambuco subregions). Of these ßies, Ͼ18,000 be- (Fig. 2A). In the dry season, the abundance of this longed to the willistoni subgroup (Table 1). In all subgroup was lower than the value recorded for the environments surveyed, abundance of these individ- other species in the three environments surveyed, and uals was higher in collections carried out during the the Caatinga was the environment where the sub- rainy season (Fig. 2A). In this season, abundance of group was least represented (Fig. 2A). individuals of the willistoni subgroup in some envi- Relative abundance of the willistoni subgroup was ronments exceeded that of the other species captured similar in the three environments surveyed (49.61, September 2014 GARCIA ET AL.: CRYPTIC SPECIES OF THE willistoni GROUP OF Drosophila 979

45.97, and 44.06% in the Pernambuco subregion, fact that we compared abundance of the willistoni Caatinga and Brejos de Altitude subregion, respec- subgroup in other sites, considering the spatial varia- tively). tion in the parameter across the Caatinga biome. The analyses of hypandrium and of the electropho- In the Atlantic Forest, relative abundances of the retic pattern of the enzyme Acph-1 were carried out willistoni subgroup varied, from Ϸ44% in the Brejo de for Ϸ3,000 individuals. The proportions obtained in Altitude subregion to Ϸ50% in the Pernambuco sub- these analyses were used to infer the total number of region. Our relative abundance values and those pub- individuals of each species of the willistoni subgroup lished by Sene et al. (1980) are higher than the num- in each sampling. Only two species of the subgroup bers reported for other subregions of the biome (D. willistoni and D. paulistorum) were recorded in characterized by lower mean temperatures in the win- the Pernambuco subregion and in the Caatinga. Low ter, like the subregions Arauca´ria (Franck and Valente abundance of D. equinoxialis was observed only in the 1985, Saavedra et al. 1995, Cavasini et al. 2008), Interior Brejos de Altitude subregion (Table 1). (Dobzhansky and Pavan 1950, Sene et al. 1980, Tidon- Downloaded from https://academic.oup.com/aesa/article/107/5/975/64874 by guest on 27 September 2021 In all environments and sites surveyed, the most Sklorz and Sene 1992, Silva et al. 2005, Torres and abundant species of the subgroup was D. willistoni, Madi-Ravazzi 2006), and Serra do Mar (Dobzhansky followed by D. paulistorum (Table 1; Fig. 2B). The and Pavan 1950, Sene et al. 1980, Bizzo et al. 2010). The highest abundance of these two species was observed comparison of these results underscores the prefer- in the Pernambuco subregion, while the lowest was ence of this subgroup for warmer regions, as originally recorded in Caatinga (Table 1; Fig. 2B). Higher abun- pointed out by Dobzhansky and Pavan (1950). dance values of D. willistoni and of D. paulistorum In the three environments surveyed here, abun- were always observed in the rainy season. D. pauli- dance of the willistoni subgroup was higher in collec- storum was not collected in the Caatinga in the dry tions conducted in the rainy season. A similar Þnding season (Table 1; Fig. 2B). was reported in Brazil, in the Cerrado biome (Tidon The chi-square test revealed signiÞcant differences 2006, Mata et al. 2008), and in three subregions of the in abundance of D. willistoni between the Caatinga Atlantic Forest, namely Interior, Serra do Mar, and and the Brejos de Altitude subregion (␹2 ϭ 159.421; Arauca´ria (Dobzhansky and Pavan 1950, Franck and df ϭ 1; P Ͻ 0.0001), between the Caatinga and the Valente 1985, Tidon-Sklorz and Sene 1992, Saavedra et Pernambuco subregion (␹2 ϭ 350.672; df ϭ 1; P Ͻ al. 1995, De Toni et al. 2007, Garcia et al. 2012). These 0.0001), and between the subregions Brejos de Alti- results conÞrm the importance of humidity for these tude and Pernambuco (␹2 ϭ 37.87; df ϭ 1; P Ͻ 0.0001). drosophilids, previously discussed in the pioneering Abundance of D. paulistorum varied signiÞcantly be- studies by Dobzhansky and Pavan (1950), Dobzhan- tween the Caatinga and Brejos de Altitude subregion sky (1957), and Spassky et al. (1971). (␹2 ϭ 69.249; df ϭ 1; P Ͻ 0.0001), the Caatinga and This preference of the willistoni subgroup for pe- Pernambuco subregion (␹2 ϭ 1,213.166; df ϭ 1; P Ͻ riods of higher humidity raises the question of whether 0.0001), and the subregions Brejos de Altitude and the frequency of these individuals should indeed be Pernambuco (␹2 ϭ 792.014; df ϭ 1; P Ͻ 0.0001). lower in environments like the Caatinga, which are In the comparison between the dry and the rainy subjected to periods of intense water deÞcit, com- season, signiÞcant differences were observed in abun- pared with more humid environments, such as the dances of D. willistoni and D. paulistorum in the Caat- Atlantic Forest. However, the Þndings we report here inga (␹2 ϭ 4,035.016, df ϭ 1, P Ͻ 0.0001 and ␹2 ϭ contradict this expectation, as the relative abundance 342.000, df ϭ 1, P Ͻ 0.0001, respectively). The same of the willistoni subgroup in the Caatinga was actually was noted for the Brejo de Altitude (␹2 ϭ 1,842.402, higher than the value recorded in the forested envi- df ϭ 1, P Ͻ 0.0001 and ␹2 ϭ 512.243, df ϭ 1, P Ͻ 0.0001, ronments of Brejos de Altitude. In this sense, in spite respectively) and in the Pernambuco subregions of the preference of the willistoni subgroup for periods (␹2 ϭ 1,328.834, df ϭ 1, P Ͻ 0.0001 and ␹2 ϭ 851.572, of higher rainfall, it still is ßexible as to inhabiting arid df ϭ 1, P Ͻ 0.0001, respectively). and forest environments alike. D. willistoni was the most abundant species of the subgroup, in all environments surveyed. This has been Discussion observed in some survey efforts in other Brazilian This study investigated abundance and richness of regions (Spassky et al. 1971; Medeiros and Klaczko the willistoni subgroup of Drosophila in contrasting 2004; Schmitz et al. 2007; Garcia et al. 2008, 2012; rainfall regimes. The environments studied were the Rohde et al. 2010). Higher abundance of the species Caatinga and Atlantic Forest (subregions Brejo de has also been recorded in some locations in Mexico, Altitude and Pernambuco) in northeastern Brazil. The Panama, Guatemala, Cuba, Haiti, Jamaica, Puerto region is one of the least surveyed in terms of diversity Rico, Colombia, Venezuela, Guiana, Peru, Bolivia, and of the Drosophilidae family (Gottschalk et al. 2008). Ecuador (Dobzhansky 1957, Spassky et al. 1971, Acu- The relative abundance of the willistoni subgroup rio and Rafael 2009). was Ϸ46% in the Caatinga, a value that is much higher D. paulistorum was the second species of the willi- than that recorded for the same biome in a previous stoni subgroup in abundance in the environments sur- study by Sene et al. (1980), in Rio Grande do Norte, veyed in the current study. This pattern had also been Paraiba, and Bahia. This discrepancy may be explained recorded in some locations in the Atlantic Forest, based on differences in collection periods, and on the Caatinga, Amazon, and in the Cerrado in Brazil 980 ANNALS OF THE ENTOMOLOGICAL SOCIETY OF AMERICA Vol. 107, no. 5

(Spassky et al. 1971; Medeiros and Klaczko 2004; References Cited Schmitz et al. 2007; Garcia et al. 2008, 2012; Rohde et Ab’Saber, A. N. 1977. Os domõnios morfoclimaticos da al. 2010). D. willistoni and D. paulistorum were col- ´ ´ Ame´rica do Sul: primeira aproximac¸a˜o. Geomorfologia 52: lected in the Pernambuco subregion of the Atlantic 1Ð22. Forest and in the Caatinga (Rohde et al. 2010). These Acurio, A., and V. Rafael. 2009. Taxonomic survey of Droso- species have been reported as the only representatives philidae (Diptera) in the Yasuni National Park, Ecuador- of the subgroup in some locations in the Atlantic ian Amazon. Acta Amazonica 39: 713Ð718. Forest, to the south of Sa˜o Francisco River (Spassky et Andrade-Lima, D. 1960. Estudos Þtogeogra´Þcos de Per- al. 1971; Medeiros and Klaczko 2004; Schmitz et al. nambuco. Arquivo do Instituto de Pesquisas Agronoˆmicas 2007; Garcia et al. 2008, 2012), and in the Caatinga de Pernambuco 5: 305Ð341. (Rohde et al. 2010). Ayala, F., and J. R. Powell. 1972. Enzyme variability in the Dobzhansky (1957) and Spassky et al. (1971) Drosophila willistoni group. VI. Levels of polymorphism stressed that humidity is a limiting factor for these two and physiological function of enzymes. Biochem. Genet. Downloaded from https://academic.oup.com/aesa/article/107/5/975/64874 by guest on 27 September 2021 7: 331Ð345. species in South America, especially for D. paulisto- Ayres, M., M. Ayres, Jr., D. L. Ayres, and A. A. Santos. 2007. rum. The chi-square tests carried out in this study BioEstat Ð Aplicac¸o˜es estatõ´sticas nas a´reas das cieˆncias conÞrm this pattern, as the signiÞcant differences in biome´dicas. Sociedade Civil Mamiraua´, Bele´m, Brazil. abundance of both species, between the rainy and the Bizzo, L., M. S. Gottschalk, D. C. De Toni, and P.R.P. Hof- dry season. In all collections in the driest times, pop- mann. 2010. Seasonal dynamics of a drosophilid (Dip- ulation sizes of the two species dropped. In those tera) assemblage and its potencial as bioindicator in open periods, the number of D. paulistorum individuals cap- environments. Iheringia, Se´rie Zoologia 100: 185Ð191. tured was smaller than that of D. willistoni. The re- Blauth, M. L., and M. S. Gottschalk. 2007. A novel record of markable dependence of these species on humidity is Drosophilidae species in the Cerrado biome of the state evident in the Caatinga, the most arid of the environ- of Mato Grosso, west-central Brazil. Drosoph. Inf. Serv. ments surveyed in the current study. There, in the 90: 90Ð96. Breuer, M. E., and C. Pavan. 1950. Genitalia masculina de rainy season, the abundance of the two species was ´ “Drosophila” (Diptera): Grupo “annulimana”. Revista higher than the sum of the abundances of all other Brasileira de Biologia 10: 469Ð488. species captured, while in the dry season D. willistoni Burla, H., A. B. Da Cunha, A. R. Cordeiro, T. Dobzhansky, was represented by a few individuals and D. paulisto- C. Malogolowkin, and C. Pavan. 1949. The willistoni rum was not recorded at all. In the Atlantic Forest group of sibling species of Drosophila. Evolution 3: 300Ð environments, where humidity is more consistently 314. retained in periods when rains are sparse (Por et al. Cavasini, R., R. P. Mateus, and M.L.T. Buschini. 2008. Car- 2005), the reduction in population sizes of these spe- acterizac¸a˜o da assembleia de Drosophila (Diptera, Droso- cies was not so dramatic in the dry season as compared philidae) em dois fragmentos ßorestais na regia˜o Sul do with the numbers recorded in the Caatinga. Brasil. Revista Eletroˆnica Lato Sensu 3: 1Ð14. This study also reports the Þrst record of D. equi- Chaves, N. B., and R. Tidon. 2008. Biogeographical aspects of drosophilids (Diptera, Drosophilidae) of the Brazilian noxialis in the Brejos de Altitude subregion. In Brazil, savanna. Revista Brasileira de Entomologia 52: 340Ð348. this species was recently observed in the Atlantic Da Silva, J.M.C., and C.H.M. Casteleti. 2005. Estado da bio- Forest, Pernambuco subregion (Rohde et al. 2010), diversidade da Mata Atlaˆntica brasileira, pp. 43Ð59. In C. and formerly in Cerrado (Burla et al. 1949, Spassky et Galindo-Leal and I. Gusma˜o-Caˆmara (eds.), Mata Atlaˆn- al. 1971, Chaves and Tidon 2008), and in the Amazon tica: biodiversidade, ameac¸as e perspectivas. Fundac¸a˜o Forest (Burla et al. 1949, Spassky et al. 1971, Ayala and SOS Mata Atlaˆntica and Conservac¸a˜o Internacional, Belo Powell 1972). Horizonte, Brazil. In sum, the current study proves the importance of De Toni, D. C., P.R.P. Hofmann, and V.L.S. Valente. 2001. the taxonomic identiÞcation of the willistoni subgroup First record of Zaprionus indianus (Diptera, Drosophili- in the face of the practical difÞculty to characterize its dae) in the State of Santa Catarina, Brazil. Biotemas 14: representatives at species level. Our results under- 71Ð85. De Toni, D. C., M. S. Gottschalk, J. Cordeiro, P.R.P. Hof- score the signiÞcance of humidity and of temperature mann, and V.L.S. Valente. 2007. Study of the Drosophila as factors that inßuence the distribution of this im- (Diptera, Drosophilidae) assemblages on Atlantic Forest portant group of Neotropical drosophilids, and point Islands of Santa Catarina State. Neotrop. Entomol. 36: to the interesting ecological versatility of some of the 356Ð375. species it contains. Dobzhansky, T. 1957. Chromosomal variability in Islands and continental populations of Drosophila willistoni from Central America and the West Indies. Evolution 11: 280Ð Acknowledgments 293. Dobzhansky, T. 1970. Genetics of the evolutionary process. We are grateful to the funding agencies Science and Tech- Columbia University Press, New York, NY. nology Support Foundation of the State of Pernambuco Dobzhansky, T., and C. Pavan. 1950. Local and seasonal (FACEPE), Pro-Rectory for Research and Postgraduate variations in the relative frequencies of species of Dro- (PROPESQ) of the Federal University of Pernambuco (UFPE), sophila in Brazil. J. Anim. Ecol. 19: 1Ð14. Pro-Rectory for Research and Post-Graduation (PRPPG) of Dobzhansky, T., and J. R. Powell. 1975. The willistoni group the Federal Rural University of Pernambuco (UFRPE), and of sibling species of Drosophila, pp. 589Ð622. In R. C. King National Council for ScientiÞc and Technological Develop- (ed.), Handbook of genetics, vol. 3. Plenum Publishing ment (CNPq). Corporation, New York, NY. September 2014 GARCIA ET AL.: CRYPTIC SPECIES OF THE willistoni GROUP OF Drosophila 981

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