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A New Species of Neotropical Drosophila (Diptera, Drosophilidae)

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A New Species of Neotropical Drosophila (Diptera, Drosophilidae) Revista Brasileira de Entomologia 61 (2017) 232–238 REVISTA BRASILEIRA DE Entomologia A Journal on Insect Diversity and Evolution www.rbentomologia.com Systematics, Morphology and Biogeography A new species of Neotropical Drosophila (Diptera, Drosophilidae) belonging to the guarani group a a,∗ b Vilma Ratcov , Carlos R. Vilela , Beatriz Goni˜ a Universidade de São Paulo, Instituto de Biociências, Departamento de Genética e Biologia Evolutiva, São Paulo, SP, Brazil b Universidad de la República, Facultad de Ciencias, Instituto de Biología, Sección Genética Evolutiva, Montevideo, Uruguay a r a b s t r a c t t i c l e i n f o Article history: Drosophila butantan sp. nov., a species belonging to the guarani group and closely related to Drosophila Received 2 March 2017 nigrifemur from Bolivia, is described based on a female, and some of its offspring, collected at the forest Accepted 8 June 2017 reserve of the Instituto de Biociências da Universidade de São Paulo, Cidade Universitária “Armando de Salles Available online 22 June 2017 Oliveira”, São Paulo City, state of São Paulo, Brazil. Although externally similar, the two apparently forest- Associate Editor: Sarah Oliveira dwelling species can be told apart by having distinct oviscapt valves and spermathecal introverts and tips. Accordingly, a proposal is made to also include D. nigrifemur, a previously unassigned species, in the Keywords: guarani group. The two species seem to be also related to Drosophila alexandrei and Drosophila guaraja Atlantic forest as indicated by their external morphology, their elongate spermathecae and the not so sharply pointed Brazil oviscapt valves. The karyotypes of the new species differ from those described for D. alexandrei and D. Female terminalia Karyotypes guaraja, while those of D. nigrifemur remain still unknown. Photomicrographs of the male and female Male terminalia imagines, in addition to drawings and photos of their terminalia, are also included. © 2017 Sociedade Brasileira de Entomologia. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Introduction identify them to the species level, we suspected they were conspe- cific. After having established an isofemale line (coded I48F71) we Most species of Drosophila belonging to the Neotropical guarani could confirm the previous male/female association and analyze group are readily recognized by their dark brown body and by their male terminalia, especially the aedeagus and the inner sper- having wings with clouded main crossveins. The guarani species mathecal capsules. At first sight, based on the shape of the latter group was proposed by Dobzhansky and Pavan (1943: 66) and structures, we suspected they could belong to Drosophila alexan- is currently (Bächli, 2017) composed by 16 species: Drosophila drei, a species collected in Rio Grande do Sul state (Cordeiro, 1951), alexandrei Cordeiro, 1951, D. araucana Brncic, 1957, D. cuscungu only known from its original description and whose type specimens Vela and Rafael, 2005, D. ecuatoriana Vela and Rafael, 2004, are thought to be lost. The analysis of the male terminalia was of D. griseolineata Duda, 1927, D. guaraja King, 1947, D. guaru no help as they were neither depicted nor illustrated in the original Dobzhansky and Pavan, 1943, D. huilliche Brncic, 1957, D. limbin- description of the candidate species. ervis Duda, 1925, D. maculifrons Duda, 1927, D. ornatifrons Duda, Ten years later, in December 2006, 10 specimens (4 males: 6 1927, D. pichinchana Vela and Rafael, 2004, D. quitensis Vela and females) of this undescribed species were identified among 495 Rafael, 2004, D. subbadia Patterson and Mainland in Patterson, drosophilids aspirated from living inflorescences of Goeppertia 1943, D. tucumana Vilela and Pereira, 1985, and D. urubamba monophylla (Vell.) Borchs. & S. Suárez (Borchsenius et al., 2012) Vilela and Pereira, 1993. (Marantaceae), in the same forest reserve (Vaz et al., 2014). During a two-year Drosophilid fauna survey conducted from More recently, analyses of the mitotic metaphase chromo- September, 1995 through August, 1997 (Ratcov, 2002 unpub- somes obtained from the larval brain sampled from isofemale lished thesis) in an urban forest reserve in São Paulo City, we line I48F71 and described below suggest the unknown species collected some small unknown dark brown male and female ima- we have collected and Drosophila alexandrei belong to different gines belonging to the guarani group. Although we were unable to species. The purpose of the present paper is to describe this unknown forest-dwelling Neotropical species of the Drosophila guarani group, which is attracted to but apparently does not breed on inflore- ∗ Corresponding author. scences of Goeppertia monophylla, a Marantaceae species locally E-mail: [email protected] (C.R. Vilela). known as caeté. http://dx.doi.org/10.1016/j.rbe.2017.06.002 0085-5626/© 2017 Sociedade Brasileira de Entomologia. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/). V. Ratcov et al. / Revista Brasileira de Entomologia 61 (2017) 232–238 233 1 2 5 3 4 6 Figs. 1–6. Drosophila butantan sp. nov., paratypes and pupa from isofemale line I48F71: (1, 2) male paratype, and (3, 4) female paratype, in dorsal and right lateral views, respectively; (5, 6) ordinary puparium + pupa, dorsal and oblique dorsal views. Scale bar = 1 mm. Material and methods the Combine ZP software (open source software CombineZP at http://combinezp.software.informer.com). One female collected with an entomological net over a banana- Initially, the isofemale line I48F71 was kept in a short-lasting baited trap in late August 1996, including her first-generation powdered milk-agar medium (Bächli et al., 2000) for about 10 years. offspring (5 males, 8 females), plus two specimens (1 male, 1 However, during the past 11 years, the cited strain has been also female) of a much later generation of the same isofemale line successfully maintained in a long-lasting modified banana-agar (I48F71) yielded in the laboratory in a modified banana-agar culture medium (Goni˜ and Vilela, 2016). The imagines are kept in medium (Goni˜ and Vilela, 2016) were used in the description of conventional cylindrical glass tubes (20 × 100 mm), containing ca. ◦ the new species. The trap was set in the forest reserve of the 3 mL of modified banana-agar medium for 3 weeks, at 18 ± 1 C Instituto de Biociências da Universidade de São Paulo, Cidade Uni- and 13:11 h (L:D) photoperiod. During this period, they are trans- versitária “Armando de Salles Oliveira”, São Paulo city, state of São ferred once a week to new vials to reach sexual maturation. A tiny Paulo, an urban fragment of the montane Atlantic forest of south- ball (ca. 1 mm diameter) of solid and fresh baker’s yeast is added eastern Brazil. Label data accompanying each type specimen are after transferring the flies. The flies are discarded three weeks after recorded as given; a comma indicates a line change, and a slash, emergence and, one week later, two V-shaped filter paper strips a label change. Our own notes or interpretations are included in are inserted into the medium and the vials are placed inside one brackets. All specimens are deposited in the Museu de Zoologia ¼ l flask containing wet sand at the bottom. Emerged flies of the da Universidade de São Paulo, São Paulo, Brazil (MZUSP). Dissec- new generation are then aspirated once a week and transferred tions of reproductive structures follow Wheeler and Kambysellis again to a modified banana-agar medium, restarting the cycle. This (1966), as modified by Kaneshiro (1969) and Bächli et al. (2004). method has also been successfully used in our laboratory in São For morphological terminology, measurements, indices, prepara- Paulo city to keep strains of some species belonging to the cardini tions of microscope slides as well as illustrations refer to Vilela and and tripunctata groups. Since early February 2017 the wet sand has Bächli (1990, 2000) and Bächli et al. (2004). Whenever in the same successfully been replaced by exfoliated vermiculite. It is worth plate, all line drawings were drawn to the same scale, and all pho- mentioning the calm behavior of the imagines of the new species tomicrographs were taken and enlarged to the same magnification. inside the culture flasks containing sand or vermiculite, facilitating Photomicrographs of imagoes and pupa (4× objective) were the aspiration and transfer of emerged flies to new vials. taken with an Olympus camera (PM2) loaded with an analog 35 mm Male and female mitotic chromosomes were obtained from Fujichrome Professional 64T film and attached to an Olympus ste- cytological preparations of cerebral ganglia of third instar larvae reomicroscope (SZ11) with a ring illuminator. The analog images of both sexes of the new species sampled from the isofemale were later digitized with an Epson scanner (Perfection 4180 photo). line I48F71, according to the method of Imai et al. (1977, 1988). Photomicrographs of wing (4× objective, optovar 1×) and termi- All cytological preparations were made from single individuals ◦ nalia structures (10× objective, optovar 1×) were taken with an cultured with banana-agar-yeast media at 18–20 C. Cytological ® Olympus Q-Color 5 digital camera attached to an Olympus BX60 preparations were observed in an Olympus BX60 microscope ® microscope. A set of 20–50 pictures was taken by manually focus- equipped with an Olympus U-MAD-3 camera. Suitable mitotic ® ing each structure at different depths. This set of photomicrographs cells were selected and then digitalized using the Image-Pro Plus was then digitally stacked to create an all-in-focus composite using version 5.1 software. Furthermore, mitotic chromosomes from 234 V. Ratcov et al. / Revista Brasileira de Entomologia 61 (2017) 232–238 sulcate.
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