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Hemiptera: Aphididae) from Costa Rica and Mexico

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Hemiptera: Aphididae) from Costa Rica and Mexico Mier Durante et al.: Ucrimyzus villalobosi, a New Aphid Genus and Species 323 A NEW GENUS AND SPECIES OF ASTERACEAE-INHABITING APHID (HEMIPTERA: APHIDIDAE) FROM COSTA RICA AND MEXICO M. PILAR MIER DURANTE1,*, NICOLÁS PÉREZ HIDALGO1, DAVID MARTÍNEZ-TORRES2, SERGIO GARCÍA-TEJERO1, REBECA PEÑA MARTÍNEZ3 AND JUAN M. NIETO NAFRÍA1 1Departamento de Biodiversidad y Gestión Ambiental, Universidad de León, 24071 León, Spain E-mail: [email protected], [email protected], [email protected], [email protected]. 2Institut Cavanilles de Biodiversitat i Biologia Evolutiva, Universitat de Valencia, Apdo. Correos 22085, 46071 Valencia (Spain) E-mail: [email protected]. 3Prolongación Aldama 188 M1 C55, 16010 México, D.F (Mexico) E-mail: [email protected]. *Corresponding author; E-mail: [email protected] ABSTRACT Ucrimyzus villalobosi Mier Durante & Pérez Hidalgo gen. n., sp. n. (Hemiptera: Aphididae: Macrosiphini) are described from apterous and alate viviparous females collected on species of genera Bidens, Schkuhria, Senecio and Stevia (Asteraceae: Asteroideae) in Costa Rica and Mexico. Principal components analysis (PCA) was done to verify that the studied aphids belong to a single species regardless of their geographical origin or host plant. Molecular analyses were carried out on the sequences of a fragment of the mitochondrial gene encod- ing for cytochrome c oxidase subunit 1 (COI) and of a fragment of the nuclear gene encoding elongation factor 1α (EF1α). The taxonomic discussion takes into account the conclusions of the molecular analyses and the morphologic study compared with other genera of Mac- rosiphini. The identification keys by Blackman & Eastop (2006) to aphids living on each mentioned plant genus are modified to include the new species. Key Words: Ucrimyzus gen. n., aphids, Macrosiphini, morphology, principal components analysis, cytochrome oxidase 1, elongation factor 1α RESUMEN Se describe Ucrimyzus villalobosi Mier Durante & Pérez Hidalgo gen. n., sp. n. (Hemiptera: Aphididae: Macrosiphini) a partir de hembras vivíparas ápteras y aladas recogidas en Costa Rica y México sobre plantas de los géneros Bidens, Schkuhria, Senecio y Stevia (Asteraceae: Asteroideae). El análisis de componentes principales de los especímenes asegura que son de la misma especie cualquiera que sea su origen geográfico o su planta hospedadora. Se han analizado sendos fragmentos del gen mitocondrial que codifica la subunidad 1 de la citocromo c oxidasa (COI) y del gen nuclear que codifica el factor de elongación 1α (EF1α). La discusión taxonómica se ha basado en las conclusiones de los análisis moleculares y en el estudio morfológico comparado con géneros de Macrosiphini. Se modifican las claves de iden- tificación de Blackman & Eastop para los pulgones que viven en los mencionados 4 géneros de plantas para incluir en ellas el nuevo género Ucrimyzus. Palabras Clave: Ucrimyzus gen. n., pulgones, Macrosiphini, morfología, análisis de compo- nentes principales, citocromo oxidasa I, factor de elongación 1α Translation provided by the authors. During expeditions to Costa Rica in 2008, 3 of collected on Bidens, Schkuhria, Senecio and Ste- the authors (M.D., P.H., N.N.) and W. Villalobos via in several Mexican localities, and kept in the (Villalobos Muller et al. 2010) collected apterous collection génerale d’aphides of the Muséum na- and alate viviparous female macrosiphines (Aphi- tional d’Histoire naturelle (Paris, France), were didae, Aphidinae, Macrosiphini) on Bidens pilosa studied; these specimens had been provisionally (Asteraceae, Asteroideae) in the University of identified by the Prof. G. Remaudière asHypero - Costa Rica main campus. Additional individuals myzus sp. and marked as possible new species. 324 Florida Entomologist 96(2) June 2013 Morphological, statistical (PCA) and molecular PCR amplifications of each mentioned frag- (genes COI and EF 1α) studies have been carried ment were done on 3 µL of the extracted DNA. out and have shown that (i) the studied specimens A 710 bp fragment of the 5’ region of the COI belong plausibly to a single species independently gene was amplified using primers LCO1490 and from their geographical origin or host plant, and HCO2198 described by Folmer et al. (1994). PCR (ii) it is a new species, which can not be included conditions for COI amplification were as follows: in any known Macrosiphini genus, and so, a new 94 °C for 1 min, 35 cycles of 94 °C for 30 s, 48 °C genus and its type species are established. for 1 min, and 68 °C for 1 min; a final extension step of 7 min at 68 °C was included after cycling. Amplification of the EF1 fragment was done MATERIAL AND METHODS α using 2 consecutive PCR reactions with primers Ten samples collected in 1 Costa Rican and 6 Efs175 (Moran et al. 1999) and Efr1 (5’GTGTG- Mexican localities have been studied (see “Types” GCAATSCAANACNGGAGT3’) in the first reac- section), considering a sample as the group of tion and then primers Efs175 and Efr2 (5’TTG- specimens collected on the same species of plant GAAATTTGACCNGGGTGRTT3’) in the second in a locality on a specific date. Specimens were semi-nested reaction. PCR conditions used in the preserved in microscopic slides with a water-sol- first reaction were 94 °C for 1 min; 40 cycles of uble mounting medium (Nieto Nafría & Mier Du- 94 °C for 30 s, 50 °C for 1 min and 68 °C for 1.5 rante 1998). Aphids for molecular analyses were min; a final extension step of 7 min at 68 °C was preserved in 96% ethanol until processing; these included after cycling. The semi-nested PCR was individuals were caught together with the holo- done similarly but using 52 °C for the annealing type on the same plant specimen. step and 1 µL of the first PCR product. Morphological measurements were made ac- PCR products were purified by ammonium cording to Nieto Nafría & Mier Durante (1998). In precipitation and reconstituted in 10 mL of LTE the description, measurements are lengths except buffer (10 mM Tris, 0.1 mM EDTA). Direct se- when indicated otherwise as width or diameter. quencing of the amplified fragments was done The comparative morphological study was con- on both directions using PCR primers (Efr2 was ducted on species (i) whose apterae possess swol- used as reverse primer for sequencing the EF1α len siphunculi and have been recorded in North fragment). Sequencing was conducted using the America or (ii) are known to feed on Asteroideae Big Dye Terminator v3.1 Cycle Sequencing Kit species over the World. Specimens belonging to di- (Applied Biosystems) following manufacturer’s verse genera presumably related with the new one instructions, and samples were loaded into an and also specimens of the most part of American ABI 3700 automated sequencer. species currently classifiedNeonasonovia Hille Ris Chromatograms were revised and sequences Lambers, 1949 (subgenus of Hyperomyzus Börner assembled using the Staden package v1.6.0 (Bon- 1933), in the collections of the Muséum national field et al. 1995). Multiple alignments were done d’Histoire naturelle (Paris, France) and the Uni- with Clustal X v1.81 (Larkin et al. 2007) with gap versidad de León (Leon, Spain) were examined. opening and gap extension penalties of 10.0 and Several taxonomic works, mainly Blackman & Ea- 0.2, respectively. stop (2006), Blackman (2010), Foottit et al. (1993), Molecular phylogenetic analyses were con- Heie (1992, 1994, 1995), Miyazaki (1971), and Ni- ducted using MEGA 5 (Tamura et al. 2011). eto Nafría et al. (in press), were consulted. A Leica DC digital camera with IM 1000 ver- RESULTS AND DISCUSSION sion 1.10 software was used for the photomicro- graphs. Ucrimyzus villalobosi Mier Durante and Pérez Hidalgo, The variables used in the PCA (principal com- gen. n., sp. n. ponents analysis) are the metric and meristic characters mentioned in the description of 39 ap- The combined description of the new species terous viviparous females; all samples are repre- and new genus is made under article 13.4 of the sented. Variables were standardized to zero mean International Code of Zoological Nomenclature and unit standard deviation prior to the analysis (International Commission on Zoological Nomen- to give the same weight to all of them. clature, 1999). Molecular phylogenetic analyses were done (i) on a fragment of the mitochondrial DNA con- Type species of Ucrimyzus gen. n. Ucrimyzus villalobosi taining the 5’ region of the cytochrome c oxidase sp. n. 1 (COI) and (ii) on a fragment of the nuclear gene coding for elongation factor-1 alpha (EF1α) of 3 Types of Ucrimyzus villalobosi sp. n. specimens of the Costa Rican sample. DNA extraction was done following the Hot- HOLOTYPE: Apterous viviparous female num- SHOT (Hot Sodium HidrOxide and Tris) method ber 4 of sample CRI-344: COSTA RICA, San Jose (Truett et al. 2000). province, San Pedro de Montes de Oca, campus Mier Durante et al.: Ucrimyzus villalobosi, a New Aphid Genus and Species 325 de la Universidad de Costa Rica (1214 m), Bidens times the body length. Antennal segments I and pilosa, 1-III-2008, Pérez Hidalgo leg., colección II pale brown and delicately rugous on the in- zoológica de la Universidad de León [CZULE], ner margin. Antennal segment III 0.41-0.70 mm Leon (Spain). Paratypes: 42 alate [al.] and 65 and 1.1-2.1 times IV, pale brown (on a proximal apterous [ap.] viviparous females (CZULE, Mu- portion) to dark brown, with scales (ventrally on séum national d’Histoire naturelle, Paris, France, paler portion) and striae, blunt setae 7-15 µm and and Natural History Museum, London, U. K.): 3 0.2-0.6 times D, and 1-20 secondary sensoria with al. and 17 ap. caught at same time as the holo- double-lined and non-ciliate margin placed in a type; 2 al. and 2 ap., MEXICO, Aguas Calientes ventral line of the basal third of the segment. An- state, Pabellon de Arteaga (1904 m), Schkuhria tennal segments IV to VI brown to dark brown anthemoidea, 7-IX-1981, Adame leg.; 4 al.
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