Phylogenomic analysis provides diagnostic tools for the identification of Anastrepha fraterculus (Diptera: Tephritidae) species complex.

Insect pests cause tremendous impact to agriculture worldwide. Species identification is crucial for implementing appropriate measures of pest control but can be challenging in closely related species. True fruit flies of the genus Anastrepha Schiner (Diptera: Tephritidae) include some of the most serious agricultural pests in the Americas, with the Anastrepha fraterculus (Wiedemann) complex being one of the most important due to its extreme polyphagy and wide distribution across most of the New World tropics and subtropics. The eight morphotypes described for this complex as well as other closely related species are classified in the fraterculus species group, whose evolutionary relationships are unresolved due to incomplete lineage sorting and introgression. We performed multifaceted phylogenomic approaches using thousands of genes to unravel the evolutionary relationships within the A. fraterculus complex to provide a baseline for molecular diagnosis of these pests. We used a methodology that accommodates variable sources of data (transcriptome, genome, and whole-genome shotgun sequencing) and developed a tool to align and filter orthologs, generating reliable datasets for phylogenetic studies. We inferred 3031 gene trees that displayed high levels of discordance. Nevertheless, the topologies of the inferred coalescent species trees were consistent across methods and datasets, except for one lineage in the A. fraterculus complex. Furthermore, network analysis indicated introgression across lineages in the fraterculus group. We present a robust phylogeny of the group that provides insights into the intricate patterns of evolution of the A. fraterculus complex supporting the hypothesis that this complex is an assemblage of closely related cryptic lineages that have evolved under interspecific gene flow. Despite this complex evolutionary scenario, our subsampling analysis revealed that a set of as few as 80 loci has a similar phylogenetic resolution as the genome-scale dataset, offering a foundation to develop more efficient diagnostic tools in this species group.

Sieving Fruit Pulp to Detect Immature Tephritid Fruit Flies in the Field.

Fruit flies of the Tephritidae family are among the most destructive and invasive agricultural pests in the world. Many countries undertake expensive eradication programs to eliminate incipient populations. During eradication programs, a concerted effort is made to detect larvae, as this strongly indicates a breeding population and helps establish the spatial extent of the infestation. The detection of immature life stages triggers additional control and regulatory actions to contain and prevent any further spread of the pest. Traditionally, larval detection is accomplished by cutting individual host fruits and examining them visually. This method is labor intensive, as only a limited number of fruit can be processed, and the probability of missing a larva is high. An extraction technique that combines i) mushing host fruit in a plastic bag, ii) straining pulp through a series of sieves, iii) placing retained pulp in a brown sugar water solution, and iv) collecting larvae that float to the surface was tested. The method was evaluated in Florida with field-collected guava naturally infested by Anastrepha suspensa. To mimic low populations more representative of a fruit fly eradication program, mangos and papaya in Hawaii were infested with a known, low number of Bactrocera dorsalis larvae. The applicability of the method was tested in the field on guava naturally infested by B. dorsalis to evaluate the method under conditions experienced by workers during an emergency fruit fly program. In both field and laboratory trials, mushing and sieving the pulp was more efficient (required less time) and more sensitive (more larvae found) than cutting fruit. Floating the pulp in brown sugar water solution helped detect earlier instar larvae. Mushing and sieving fruit pulp of important tephritid hosts may increase the probability of detecting larvae during emergency programs.

Exceptional larval morphology of nine species of the Anastrephamucronota species group (Diptera, Tephritidae).

Anastrepha is the most diverse and economically important genus of Tephritidae in the American tropics and subtropics. The striking morphology of the third instars of Anastrephacaballeroi Norrbom, Anastrephacrebra Stone, Anastrephahaplacantha Norrbom & Korytkowski, Anastrephakorytkowskii Norrbom, Anastrephanolazcoae Norrbom & Korytkowski, and three newly discovered and as yet formally unnamed species (Anastrepha sp. Peru-82, Anastrephasp.nr.protuberans, and Anastrepha sp. Sur-16), and the more typical morphology of Anastrephaaphelocentema Stone, are described using light and scanning electron microscopy. To contribute to a better understanding of the interspecific and intraspecific variation among species in the mucronota species group and facilitate phylogenetic studies, we integrate molecular and morphological techniques to confirm the identity and describe third instars. Larva-adult associations and the identification of described larvae were confirmed using DNA barcodes. We provide diagnostic characters to distinguish larvae among these nine species of the mucronota group and separate them from those of the 29 other Anastrepha species previously described. We introduce the vertical comb-like processes on the oral margin as a novel character, and the unusual character states, including position and shape of the preoral lobe, and dentate or fringed posterior margins of the oral ridges and accessory plates. Our comparative morphology concurs with most previously inferred phylogenetic relationships within the mucronota group.

New species and host plants of Anastrepha (Diptera: Tephritidae) primarily from Suriname and Par, Brazil.

Seventeen new species of Anastrepha, primarily from Suriname, French Guiana and Par, Brazil, are described and illustrated: A. aithogaster Norrbom from Brazil (Par), French Guiana, and Suriname; A. aliesae Norrbom from Suriname; A. brownsbergiensis Norrbom from Suriname; A. crassaculeus Norrbom Rodriguez Clavijo from Colombia (Magdalena, Norte de Santander) and Suriname; A. curvivenis Norrbom from Brazil (Amazonas), Ecuador (Zamora-Chinchipe), Peru (San Martn), and Suriname; A. fuscoalata Norrbom from Brazil (Par), French Guiana, and Suriname; A. gangadini Norrbom from Suriname; A. juxtalanceola Norrbom from Brazil (Par) and Suriname; A. microstrepha Norrbom from Brazil (Bahia) and Suriname; A. mitaraka Norrbom from French Guiana; A. neptis Norrbom from Brazil (Par), Ecuador (Orellana), Peru (Loreto) and Suriname; A. sobrina Norrbom from Brazil (Par), French Guiana, and Suriname; A. surinamensis Norrbom from Suriname; A. tenebrosa Norrbom from Brazil (Par) and Peru (Loreto); A. triangularis Norrbom from Suriname; A. wachiperi Norrbom from French Guiana and Peru (Cusco); and A. wittiensis Norrbom from Suriname. The following host plant records are reported: A. aithogaster from fruit of Parahancornia fasciculata (Poir.) Benoist (Apocynaceae); A. aliesae from fruit of Passiflora coccinea Aubl. and P. glandulosa Cav. (Passifloraceae); A. crassaculeus from fruit of an undetermined species of Pouteria (Sapotaceae); A. fuscoalata from fruit of Trymatococcus oligandrus (Benoist) Lanj. (Moraceae); A. sobrina from fruit of Eugenia lambertiana DC. (Myrtaceae); and A. wittiensis from fruit of Manilkara bidentata (A. DC.) A. Chev. (Sapotaceae).

The Bolivian fauna of the genus Anastrepha Schiner (Diptera: Tephritidae).

The general aim of this study is to contribute to and summarize knowledge of the Bolivian fauna of the genus Anastrepha Schiner (Tephritidae) which includes species of both ecological and economic importance. In addition to compiling data from the literature, we report the results of fruit fly sampling using McPhail or multilure traps in the Tropic of Cochabamba region and at the private natural reserve of Potrerillo del Guendá in Santa Cruz de la Sierra, as well as records from various other sites based on specimens in museum collections. Fifty-seven named species and three unnamed species of Anastrepha are recorded from Bolivia. Distribution maps for all of these species are provided. Numerous new department records are reported as well as the first records for Bolivia of A. castanea Norrbom, A. dissimilis Stone, A. elegans Blanchard, A. haywardi Blanchard, A. macrura Hendel, A. montei Lima, A. punctata Hendel, and A. rosilloi Blanchard. Pacouria boliviensis (Markgr.) A. Chev. (Apocynaceae) is reported as a host plant of A. woodleyi Norrbom Korytkowski, and Myrciaria floribunda (H. West ex Willd.) Berg (Myrtaceae) and Pouteria glomerata (Miq.) Radlk. (Sapotaceae) as host plants of A. fraterculus (Wiedemann). This distribution and host information will be useful to monitor and manage species that damage fruit crops in Bolivia.

Descripción de una nueva especie de Anastrepha grupo curvicauda (Diptera: Tephritidae) / Description of a new species of the Anastrepha curvicauda species group (Diptera: Tephritidae)

RESUMEN Las moscas del género Anastrepha, se consideran plagas de gran importancia económica, no solo por el daño directo que causan a la producción de fruta sino, también, por las restricciones de cuarentena impuestas a los países exportadores de fruta. Por lo tanto, preparar descripciones y generar herramientas de identificación taxonómica, que permitan un reconocimiento adecuado y oportuno de las especies de importancia agrícola, es una actividad básica para la sanidad de las plantas en los países productores y exportadores de frutas. En este trabajo, se describe e ilustra Anastrepha ligiae, una nueva especie del grupo Anastrepha curvicauda, capturada con trampas McPhail, en los municipios de Albán y Anolaima (Cundinamarca) y Vélez (Santander), Colombia y criados en la fruta de la papayuela Vasconcellea pubescens A. DC. (Caricaceae). V. pubescens, se registra por primera vez como una planta hospedante para el género Anastrepha.

ABSTRACT Fruit flies of the genus Anastrepha are considered pests of great economic importance because they do not only cause direct damage to the fruit production, but also due to the quarantine restrictions imposed to fruit exporting countries. Therefore, preparing descriptions and generating taxonomic identification tools that allow an appropriate and accurate recognition of the species of agricultural importance are basic plant health activities in the fruit producing and exporting countries. In this work, we describe and illustrate Anastrepha ligiae, a new species of the Anastrepha curvicauda group captured with McPhail traps in the municipalities of Albán and Anolaima (Cundinamarca) and Vélez (Santander), Colombia, and reared from papaya fruit Vasconcellea pubescens A. DC. (Caricaceae). V. pubescens is the first time recorded as a host plant of the genus Anastrepha.

New records of Anastrepha (Diptera: Tephritidae) primarily from Colombia.

New distribution information, primarily from Colombia, is provided for 60 species of Anastrepha, including the first records of 33 species from Colombia: A. acca Norrbom, A. acris Stone, A. amita Zucchi, A. amplidentata Norrbom, A. atrox (Aldrich), A. barbiellinii Lima, A. bezzii Lima, A. canalis Stone, A. cocorae Norrbom Korytkowski, A. compressa Stone, A. cordata Aldrich, A. crebra Stone, A. cryptostrepha Hendel, A. cryptostrephoides Norrbom Korytkowski, A. furcata Lima, A. fuscicauda Norrbom Korytkowski, A. galbina Stone, A. grandicarina Norrbom Korytkowski, A. hamata (Loew), A. katiyari Norrbom, A. macrura Hendel, A. margarita Caraballo, A. minuta Stone, A. nigripalpis Hendel, A. normalis Norrbom, A. pastranai Blanchard, A. pseudanomala Norrbom, A. pulchra Stone, A. similis Greene, A. speciosa Stone, A. urichi Greene, A. willei Korytkowski, and A. zuelaniae Stone. In addition, A. acris is reported from Costa Rica, A. alveata Stone from El Salvador, A. antunesi Lima and A. bahiensis Lima from Bolivia, A. barbiellinii from Trinidad Tobago, A. bezzii, A. canalis, A. coronilli Carrejo González, A. cryptostrepha and A. minuta from Bolivia, and A. willei from Argentina and Bolivia. A list of the 83 species of Anastrepha now known to occur in Colombia is provided, as well as illustrations of at least the wing and aculeus tip or male genitalia for 60 species.

Phylogenetic relationships of the tribe Toxotrypanini (Diptera: Tephritidae) based on molecular characters.

Current hypotheses of relationship among the species of the fruit fly genera Anastrepha and Toxotrypana are tested using sequence data from six DNA regions: the mitochondrial regions 16S, CAD, and COI, and the nuclear regions EF1a, PER, and PGD. DNA sequences were obtained from 146 species of Anastrepha, representing 19 of the 21 species groups as well as five of the six clades of the robusta group, and four species of Toxotrypana in addition to species of Hexachaeta, Pseudophorellia, Alujamyia, and 13 other tephritid genera used as outgroups. The results indicate that Hexachaeta is more closely related to the Molynocoelia group than to Toxotrypana and Anastrepha, and it is removed from the tribe Toxotrypanini. The group Anastrepha+Toxotrypana and the genus Toxotrypana are strongly supported as monophyletic, consistent with previous studies, but Toxotrypana arises within Anastrepha, confirming that Anastrepha as currently defined is paraphyletic. The placement of Toxotrypana within Anastrepha is clearly defined for the first time with high support, as the sister group to the cryptostrepha clade of the robusta group of Anastrepha. Within Anastrepha, the daciformis, dentata, leptozona, raveni, and striata species groups are highly supported clades. The serpentina group is recognized with lower support, and the fraterculus and pseudoparallela groups are supported with minor alterations. The robusta group is resolved as polyphyletic, but four of the six species clades within it are recovered monophyletic (one clade is not represented and another is represented by one species). The punctata and panamensis groups are resolved together in a clade. At least some species of the mucronota group are related, however this group requires further study. The benjamini, grandis, and spatulata groups appear to be polyphyletic. Relationships among the species groups are generally poorly resolved, with the following exceptions: (1) the lineage including Toxotrypana, the cryptostrepha clade, and the tripunctata group; (2) the sister group relationship of the daciformis+dentata groups; (3) a clade comprising the punctata and panamensis groups; and (4) the large clade comprising the pseudoparallela+spatulata+ramosa+grandis+serpentina+striata+fraterculus groups.

Developmental Studies of Maize-Infesting Picture-Winged Flies (Diptera: Ulidiidae).

Eleven species of picture-winged flies (Diptera: Ulidiidae) attack maize (Zea mays L.) in the Americas. Field and laboratory studies were used to determine developmental times on sweet corn ears for the three most common species attacking the crop in the United States, Chaetopsis massyla (Walker), Euxesta eluta Loew, and Euxesta stigmatias Loew. Egg plus larval stage developmental times were evaluated in early Spring and late Fall 2009, and late Spring 2010, by placing newly deposited eggs in protected ears in the field. Newly formed puparia were removed daily from cages and held in the laboratory to determine pupal developmental times. Developmental times were compared with flies reared on artificial diet in the laboratory. Ear- and diet-reared adults were held until their death to determine adult longevity. Developmental times, including for pupae from ear-reared larvae, were significantly affected by species and season. All three species required nearly twice as long to complete development in the late Fall compared to late Spring studies. Flies required 3-13 d longer to complete development on artificial diet than on ears. Euxesta eluta adults lived two to three times longer than the other species, and females of all species lived 10-15% longer than males. Species and seasonal developmental times are discussed in relation to ear developmental times and control strategies. It is estimated that 16-19 generations per year of all three fly species can develop in the field in the sweet corn production area of southern Florida.

New species and host plants of Anastrepha (Diptera: Tephritidae) primarily from Peru and Bolivia.

Twenty-eight new species of Anastrepha are described and illustrated: A. acca (Bolivia, Peru), A. adami (Peru), A. amplidentata (Bolivia, Peru), A. annonae (Peru), A. breviapex (Peru), A. caballeroi (Peru), A. camba (Bolivia, Peru), A. cicra (Bolivia, Peru), A. disjuncta (Peru), A. durantae (Peru), A. echaratiensis (Peru), A. eminens (Peru), A. ericki (Peru), A. gonzalezi (Bolivia, Peru), A. guevarai (Peru), A. gusi (Peru), A. kimi (Colombia, Peru), A. korytkowskii (Bolivia, Peru), A. latilanceola (Bolivia, Peru), A. melanoptera (Peru), A. mollyae (Bolivia, Peru), A. perezi (Peru), A. psidivora (Peru), A. robynae (Peru), A. rondoniensis (Brazil, Peru), A. tunariensis (Bolivia, Peru), A. villosa (Bolivia), and A. zacharyi (Peru). The following host plant records are reported: A. amplidentata from Spondias mombin L. (Anacardiaceae); A. caballeroi from Quararibea malacocalyx A. Robyns & S. Nilsson (Malvaceae); A. annonae from Annona mucosa Jacq. and Annona sp. (Annonaceae); A. durantae from Duranta peruviana Moldenke (Verbenaceae); and A. psidivora from Psidium guajava L. (Myrtaceae).

Nuclear ribosomal internal transcribed spacer 1 (ITS1) variation in the Anastrepha fraterculus cryptic species complex (Diptera, Tephritidae) of the Andean region.

The nuclear ribosomal internal transcribed spacer 1 (ITS1) was sequenced for Anastrepha fraterculus (Wiedemann, 1830) originating from 85 collections from the northern and central Andean countries of South America including Argentina (Tucumán), Bolivia, Perú, Ecuador, Colombia, and Venezuela. The ITS1 regions of additional specimens (17 collections) from Central America (México, Guatemala, Costa Rica, and Panamá), Brazil, Caribbean Colombia, and coastal Venezuela were sequenced and together with published sequences (Paraguay) provided context for interpretation. A total of six ITS1 sequence variants were recognized in the Andean region comprising four groups. Type I predominates in the southernmost range of Anastrepha fraterculus. Type II predominates in its northernmost range. In the central and northern Andes, the geographic distributions overlap and interdigitate with a strong elevational effect. A discussion of relationships between observed ITS1 types and morphometric types is included.

An integrative approach to unravel the Ceratitis FAR (Diptera, Tephritidae) cryptic species complex: a review.

This paper reviews all information gathered from different disciplines and studies to resolve the species status within the Ceratitis FAR (Ceratitis fasciventris, Ceratitis anonae, Ceratitis rosa) complex, a group of polyphagous fruit fly pest species (Diptera, Tephritidae) from Africa. It includes information on larval and adult morphology, wing morphometrics, cuticular hydrocarbons, pheromones, microsatellites, developmental physiology and geographic distribution. The general consensus is that the FAR complex comprises Ceratitis anonae, two species within Ceratitis rosa (so-called R1 and R2) and two putatitve species under Ceratitis fasciventris. The information regarding the latter is, however, too limited to draw final conclusions on specific status. Evidence for this recognition is discussed with reference to publications providing further details.

Description of third instar larvae of Ceratitis fasciventris, C. anonae, C. rosa (FAR complex) and C. capitata (Diptera, Tephritidae).

Third instar larvae of members of the Ceratitis FAR complex, including Ceratitis fasciventris (Bezzi), Ceratitis anonae Graham, and Ceratitis rosa Karsch are described and compared with those of Ceratitis capitata (Wiedemann). Diagnostic characters, such as presence vs. absence of a secondary tooth on the mandibles, previously used to separate Ceratitis capitata from Ceratitis rosa, are shown to vary in each species. Significant variation in diagnostic morphological characters among populations of Ceratitis rosa from east and south Africa is documented; however, the differences are not simply congruent with the R1 and R2 designations based on other studies. Quantitative measures of numerous morphological characters are consistently smaller in the larvae of Ceratitis fasciventris and distinguish them from other species of the FAR complex. Larvae of Ceratitis capitata can be distinguished from those of the FAR complex by characters such as absence of accessory plates of the oral ridges, the shape of the anterior spiracle, and the pattern of dorsal spinules. Previous studies indicated that absence of accessory lobes separate the genus Ceratitis from Bactrocera, but this is shown to be incorrect, as accessory lobes are in fact present in several species of Ceratitis.

Examination of the pest status of corn-infesting Ulidiidae (Diptera).

Larvae of 11 species of picture-winged flies (Diptera: Ulididae) are known to feed on corn plants (Zea mays L.) in the western hemisphere. Larvae emerge from eggs deposited in leaf axils and corn silk to feed mostly within ears, but the primary versus secondary nature (i.e., pest status) of their infestation is not known for all of these species. Choice and no-choice tests by using a split-plot design were conducted in greenhouse and field trials to determine the pest status on sweet corn of three of these species found in Florida: Chaetopsis massyla (Walker), Euxesta eluta Loew, and E. stigmatias Loew. The main treatments (uninfested ears and ears experimentally infested with either Spodoptera frugiperda [Lepidoptera: Noctuidae] or E. eluta larvae) were applied at first silk. The subtreatments (C. massyla, E. eluta, or E. stigmatias adults caged on ears) were applied 7 d later and maintained for 10 d. All three fly species were reared from uninfested and experimentally infested ears in both choice and no-choice tests in greenhouse and field trials confirming both primary and secondary modes of ear infestation. More flies of all three species emerged from ears that were preinfested with S. frugiperda compared with uninfested ears suggesting either preference for or greater survival within ears previously infested by S. frugiperda. Fewer E. eluta and E. stigmatias emerged from ears preinfested with E. eluta in no-choice field tests, suggesting that previous infestation by this fly may negatively affect oviposition or that older fly larvae affect survival of neonate larvae. All three species studied here should be considered primary pests that can render unprotected sweet corn ears unmarketable.

Recent Mediterranean fruit fly (Diptera: Tephritidae) infestations in Florida--a genetic perspective.

Microsatellite and mitochondrial DNA (mtDNA) variability data were used to study infestations of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) in Florida in 1997 and 1998. A total of 132 flies collected in monitoring traps or as larvae removed from fruit were examined at three polymorphic mtDNA restriction sites and two microsatellite loci. All of the flies sampled in Florida in 1997 displayed the mitochondrial AAB haplotype and represent a novel introduction of Mediterranean fruit flies into the state. All flies collected in central Florida in 1998 also displayed the AAB haplotype. Microsatellite analysis of these specimens from 1998 detected only alleles that were present in 1997. These results strongly indicate that the 1998 Florida outbreaks were derived from the Florida populations from the previous year. According to our analyses, the Mediterranean region is the most likely source for the 1997 Florida infestation. Flies from a small outbreak in Miami Springs, Dade County, FL, early in 1998 had a different mtDNA haplotype, characterized by the AAC restriction pattern. Microsatellites of these specimens showed significant differences in their allelic distribution from AAB flies, indicating an origin from a separate source population. South America is the most likely source for the Miami Springs flies.