Sequence analysis of the internal transcribed spacer 2 (ITS2) region of rDNA for identifying Trichogramma species and evaluating genetic diversity / Análise de sequência do rDNA da região espaçadora do transcrito interno 2 (ITS2) para identificar espécies de Trichogramma e avaliar a diversidade genética

Abstract Species of Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammtidae) are frequently used as biological control agents against Lepidoptera, but practical application of these egg endoparasitoids are complicated because of their complex taxonomy. This study aimed to compare sequences of internal transcribed spacer regions of ribosomal DNA (ITS2-rDNA) of Trichogramma accessions with those deposited in GenBank in order to access the reliability of the ITS2 as a barcode for discriminating species and evaluating the genetic diversity. ITS2-rDNA sequences obtained from seventeen specimens of Trichogramma confirmed previous identifications based on morphological characteristics. Multiple sequence alignment revealed the existence of highly conserved regions in ITS2 sequences while the neighbour-joining dendrogram indicated that the specimens formed three clusters comprising T. manicobai and T. marandobai (group I), T. galloi (group II) and T. pretiosum (group III). The ITS2 marker was shown to be a powerful DNA barcode for discriminating Trichogramma species and could be used to complement the morphological approach.

Resumo Espécies de Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammatidae) são freqüentemente usadas como agentes de controle biológico contra Lepidoptera, esses endoparasitóides de ovos apresentam taxonomia complexa, o que dificulta sua aplicação prática. Este estudo teve como objetivo comparar seqüências de regiões espaçadoras internas transcritas de DNA ribossômico (ITS2-rDNA) de acessos de Trichogramma com aquelas depositadas no GenBank, a fim de avaliar a confiabilidade do ITS2 barcode para discriminar espécies e avaliar a diversidade genética. As seqüências de ITS2-rDNA obtidas de dezessete espécimes de Trichogramma confirmaram identidades anteriores com base em características morfológicas. O alinhamento de múltiplas sequências revelou a existência de regiões altamente conservadas nas sequências ITS2, enquanto o dendrograma indicou que os espécimes formavam três grupos compreendendo T. manicobai e T. marandobai (grupo I), T. galloi (grupo II) e T. pretiosum (grupo III). O marcador ITS2 mostrou ser um poderoso DNA barcode para discriminar espécies de Trichogramma podendo ser usado como complemento da abordagem morfológica.

Sequence analysis of the internal transcribed spacer 2 (ITS2) region of rDNA for identifying Trichogramma species and evaluating genetic diversity.

Species of Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammtidae) are frequently used as biological control agents against Lepidoptera, but practical application of these egg endoparasitoids are complicated because of their complex taxonomy. This study aimed to compare sequences of internal transcribed spacer regions of ribosomal DNA (ITS2-rDNA) of Trichogramma accessions with those deposited in GenBank in order to access the reliability of the ITS2 as a barcode for discriminating species and evaluating the genetic diversity. ITS2-rDNA sequences obtained from seventeen specimens of Trichogramma confirmed previous identifications based on morphological characteristics. Multiple sequence alignment revealed the existence of highly conserved regions in ITS2 sequences while the neighbour-joining dendrogram indicated that the specimens formed three clusters comprising T. manicobai and T. marandobai (group I), T. galloi (group II) and T. pretiosum (group III). The ITS2 marker was shown to be a powerful DNA barcode for discriminating Trichogramma species and could be used to complement the morphological approach.

Which Species of Coconut Moth Occurs in Brazil: Atheloca subrufella vs. Atheloca bondari (Lepidoptera: Pyralidae)?

The moths, Atheloca subrufella (Hulst 1887) and A. bondari, (Heinrich 1956) are species known for their economic impact on coconut production, which Brazil is the fourth largest global producer. The first record of Atheloca in Brazil was reported by Bondar in 1940, where the author registered it being A. subrufella. The studies performed by C. Heinrich in 1956 related the existence of divergence in specimens of Brazilian Atheloca suggesting the presence of morphological differences between the males of A. bondari and A. subrufella. In this study, Atheloca specimens from the five states of northeastern Brazil were used. Samples from Pernambuco state were sent to taxonomist Dr. V. O. Becker (Uiraçu Institute-BA) for identification. Male individuals from the other states were mounted for photographic documentation, highlighting the characteristics that differentiate the two species. A fragment of the cytochrome c oxidase subunit 1 gene was sequenced and then compared with that of the Atheloca spp. deposited in GenBank. An analysis was conducted to evaluate the genetic distance between A. bondari and A. subrufella. The results indicate greater interspecific (0.030-0.034) than intraspecific (0.000-0.002) genetic variation between the groups, reinforcing the hypothesis of two distinct species. A geographic distribution map and a table with the host plants were constructed based on a literature review. This study concluded that the species occurring in Brazil is A. bondari, as suggested by C. Heinrich. Atheloca bondari and A. subrufella have only been reported in plants of the family Arecaceae, but only the coconut tree (Cocos nucifera L.) is shared by the two species.

Molecular, Morphological, and Biological Differentiation between Anagrus virlai sp. n., an Egg Parasitoid of the Corn Leafhopper Dalbulus maidis (Hemiptera: Cicadellidae) in the New World, and Anagrus incarnatus from the Palaearctic Region (Hymenoptera: Mymaridae).

The common New World egg parasitoid of the corn leafhopper Dalbulus maidis (DeLong) (Hemiptera: Cicadellidae), an economically important pest of maize from Argentina to southern USA, has long been misidentified as the Palaearctic species Anagrus incarnatus Haliday or its synonym A. breviphragma Soyka (Hymenoptera: Mymaridae). Using a combination of genetic and morphometric methods, and available biological information, specimens reared from eggs of D. maidis in Argentina and Mexico, described and illustrated here as Anagrus (Anagrus) virlai Triapitsyn sp. n., are shown to be different from those of A. incarnatus from the Palaearctic region. Mitochondrial and nuclear ribosomal DNA sequence data provide clear evidence for the separation of the two species. Anagrus virlai is also known from Brazil, Colombia, Guadeloupe (France), and Guyana.

First Occurrence of Trichogramma bruni Nagaraja (Hymenoptera: Trichogrammatidae) Parasitizing Eggs of Anticarsia gemmatalis Hübner (Lepidoptera: Erebidae) in Brazil.

The occurrence of Trichogramma bruni Nagaraja (Hymenoptera: Trichogrammatidae) is described for the first time parasitizing eggs of the velvetbean caterpillar Anticarsia gemmatalis Hübner (Lepidoptera: Erebidae) in Lapa, Paraná, Brazil. This parasitoid was successfully kept in the laboratory on eggs of the velvetbean caterpillar; previously, this species was mainly associated with forest and fruit trees environments, although snap beans and other annual crops were also recorded as hosting lepidopteran eggs parasitized by T. bruni. This is the sixth Trichogramma species recorded parasitizing eggs of A. gemmatalis on soybeans in Brazil.

Sequence analysis of the internal transcribed spacer 2 (ITS2) region of rDNA for identifying Trichogramma species and evaluating genetic diversity

Abstract Species of Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammtidae) are frequently used as biological control agents against Lepidoptera, but practical application of these egg endoparasitoids are complicated because of their complex taxonomy. This study aimed to compare sequences of internal transcribed spacer regions of ribosomal DNA (ITS2-rDNA) of Trichogramma accessions with those deposited in GenBank in order to access the reliability of the ITS2 as a barcode for discriminating species and evaluating the genetic diversity. ITS2-rDNA sequences obtained from seventeen specimens of Trichogramma confirmed previous identifications based on morphological characteristics. Multiple sequence alignment revealed the existence of highly conserved regions in ITS2 sequences while the neighbour-joining dendrogram indicated that the specimens formed three clusters comprising T. manicobai and T. marandobai (group I), T. galloi (group II) and T. pretiosum (group III). The ITS2 marker was shown to be a powerful DNA barcode for discriminating Trichogramma species and could be used to complement the morphological approach.

Resumo Espécies de Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammatidae) são freqüentemente usadas como agentes de controle biológico contra Lepidoptera, esses endoparasitóides de ovos apresentam taxonomia complexa, o que dificulta sua aplicação prática. Este estudo teve como objetivo comparar seqüências de regiões espaçadoras internas transcritas de DNA ribossômico (ITS2-rDNA) de acessos de Trichogramma com aquelas depositadas no GenBank, a fim de avaliar a confiabilidade do ITS2 barcode para discriminar espécies e avaliar a diversidade genética. As seqüências de ITS2-rDNA obtidas de dezessete espécimes de Trichogramma confirmaram identidades anteriores com base em características morfológicas. O alinhamento de múltiplas sequências revelou a existência de regiões altamente conservadas nas sequências ITS2, enquanto o dendrograma indicou que os espécimes formavam três grupos compreendendo T. manicobai e T. marandobai (grupo I), T. galloi (grupo II) e T. pretiosum (grupo III). O marcador ITS2 mostrou ser um poderoso DNA barcode para discriminar espécies de Trichogramma podendo ser usado como complemento da abordagem morfológica.

Hymenopteran parasitoids associated with frugivorous larvae in a Brazilian caatinga-cerrado ecotone.

The purpose of this study was to investigate native species of parasitoids of frugivorous larvae and their associations with host plants in commercial guava orchards and in typical native dry forests of a caatinga-cerrado ecotone in the State of Minas Gerais, Brazil. Nine species of parasitoids were associated with larvae of Anastrepha (Tephritidae) and Neosilba (Lonchaeidae) in fruit of Psidium guajava L. (Myrtaceae), Ziziphus joazeiro Mart. (Rhamnaceae), Spondias tuberosa Arruda (Anacardiaceae), Spondias dulcis Forst. (Anacardiaceae), Syzygium cumini (L.) Skeels (Myrtaceae), and Randia armata (Sw.) DC. (Rubiaceae). Doryctobracon areolatus was the most abundant species, obtained from puparia of Anastrepha zenildae, An. sororcula, An. fraterculus, An. obliqua, and An. turpiniae. This is the first report of Asobara obliqua in Brazil and of As. anastrephae and Tropideucoila weldi in dry forests of Minas Gerais State. The number of species of parasitoids was higher in areas with greater diversity of cultivated species and lower pesticide use. The forest fragments adjacent to the orchards served as shelter for parasitoids of frugivorous larvae.

Geographic pattern of morphological variation of the coconut mite, Aceria guerreronis Keifer (Acari: Eriophyidae), using multivariate morphometry.

The coconut mite, Aceria guerreronis Keifer, has become one of the most important pests of coconut in the Americas and Africa and recently in Southeast Asia. Despite the great economic importance of this mite, there is a lack of information on its origin and invasion history that are important to guide the search of biological control agents as well as the adoption of quarantine procedures. This study evaluates morphometric variation among A. guerreronis populations throughout its occurrence area, relates this variation with historical sequence of records, looking for information on its biogeography. Samples of 27 populations from the Americas, Africa and Asia were analysed using Principal Component Analysis and Canonical Discriminant Analysis. Results showed significant morphometric variability of A. uerreronis throughout its distribution area, with a high variability among American populations and otherwise a high similarity among African and Asian populations. The geographic pattern of variation of mite populations observed supports the hypothesis that A. guerreronis originated in the Americas and was introduced into Africa and Asia. Some inferences related to taxonomy of Eriophyoidea mites were included.

Parasitoid diversity (Hymenoptera: Braconidae and Figitidae) on frugivorous larvae (Diptera: Tephritidae and Lonchaeidae) at Adolpho Ducke Forest reserve, central amazon region, Manaus, Brazil.

This study aimed to identify parasitoid species of frugivorous larvae and to describe the tritrophic interactions involving wild fruits, frugivorous insects and their natural enemies at Adolpho Ducke Forest Reserve (RFAD) (Manaus, AM, Brazil). Collections were performed in four 1 km2 quadrants in the corners of the RFAD. The wild fruits were collected inside the forest in access trails leading to each collection area and in trails that surrounded the quadrants, up to five metres from the trail on each side. The fruits were placed in plastic containers covered with thin fabric, with a vermiculite layer on the base to allow the emergence of flies or parasitoids. Seven Braconidae species were collected, distributed among Opiinae: Doryctobracon areolatus (Szépligeti, 1911), Utetes anastrephae (Viereck, 1913), and Opius sp., and Alysiinae: Asobara anastrephae (Muesebeck, 1958), Phaenocarpa pericarpa Wharton and Carrejo, 1999, Idiasta delicata Papp, 1969, and Asobara sp. Parasitism rates by braconids and figitids are presented. Doryctobracon areolatus was the most frequent, parasitizing the highest number of fly species, and showing the highest parasitism percentage in larvae feeding on Micropholis williamii fruits. The collected figitids belong to Aganaspis nordlanderi Wharton, 1998 and A. pelleranoi (Brethes, 1924). All 15 tritrophic associations are new records for the Brazilian Amazon region. The RFAD is an important natural reservoir of frugivorous larvae parasitoids.

Geographic pattern of morphological variation of the coconut mite, Aceria guerreronis Keifer (Acari: Eriophyidae), using multivariate morphometry / Padrão geográfico de variação morfológica do ácaro do coqueiro, Aceria guerreronis Keifer (Acari: Eriophyidae), utilizando morfometria multivariada

The coconut mite, Aceria guerreronis Keifer, has become one of the most important pests of coconut in the Americas and Africa and recently in Southeast Asia. Despite the great economic importance of this mite, there is a lack of information on its origin and invasion history that are important to guide the search of biological control agents as well as the adoption of quarantine procedures. This study evaluates morphometric variation among A. guerreronis populations throughout its occurrence area, relates this variation with historical sequence of records, looking for information on its biogeography. Samples of 27 populations from the Americas, Africa and Asia were analysed using Principal Component Analysis and Canonical Discriminant Analysis. Results showed significant morphometric variability of A. uerreronis throughout its distribution area, with a high variability among American populations and otherwise a high similarity among African and Asian populations. The geographic pattern of variation of mite populations observed supports the hypothesis that A. guerreronis originated in the Americas and was introduced into Africa and Asia. Some inferences related to taxonomy of Eriophyoidea mites were included.

O ácaro do coqueiro, Aceria guerreronis Keifer, se tornou uma das pragas mais importantes do coqueiro nas Américas e África e, recentemente, no sudeste da Ásia. Apesar da importância econômica deste ácaro, faltam informações sobre sua origem e seu histórico de invasão, o que é importante para orientar a busca de agentes de controle biológico assim como a adoção de medidas quarentenárias. Este estudo teve como objetivo avaliar a variação morfométrica entre populações de A. guerreronis ao longo de sua área de ocorrência, relacionar esta variação com a sequência histórica de registros, buscando informações sobre sua biogeografia. Amostras de 27 populações das Américas, África e Ásia foram analisadas utilizando-se Análise dos Componentes Principais e Análise Discriminante Canônica. Os resultados mostraram uma significante variabilidade de A. guerreronis ao longo de sua área de distribuição, com uma alta variabilidade entre as populações americanas e, por outro lado, uma alta similaridade entre populações africanas e asiáticas. O padrão geográfico de variação de populações do ácaro observado suporta a hipótese de que A. guerreronis é originário da América e foi introduzido na África e Ásia. Algumas inferências relacionadas à taxonomia de ácaros Eriophyoidea foram incluídas.

Parasitoid diversity (Hymenoptera: Braconidae and Figitidae) on frugivorous larvae (Diptera: Tephritidae and Lonchaeidae) at Adolpho Ducke Forest Reserve, Central Amazon Region, Manaus, Brazil / Diversidade de Parasitóides (Hymenoptera: Braconidae e Figitidae) de larvas frugívoras (Diptera: Tephritidae e Lonchaeidae) na Reserva Florestal Adolpho Ducke, Amazônia Ocidental, Manaus, Brasil

This study aimed to identify parasitoid species of frugivorous larvae and to describe the tritrophic interactions involving wild fruits, frugivorous insects and their natural enemies at Adolpho Ducke Forest Reserve (RFAD) (Manaus, AM, Brazil). Collections were performed in four 1 km² quadrants in the corners of the RFAD. The wild fruits were collected inside the forest in access trails leading to each collection area and in trails that surrounded the quadrants, up to five metres from the trail on each side. The fruits were placed in plastic containers covered with thin fabric, with a vermiculite layer on the base to allow the emergence of flies or parasitoids. Seven Braconidae species were collected, distributed among Opiinae: Doryctobracon areolatus (Szépligeti, 1911), Utetes anastrephae (Viereck, 1913), and Opius sp., and Alysiinae: Asobara anastrephae (Muesebeck, 1958), Phaenocarpa pericarpa Wharton and Carrejo, 1999, Idiasta delicata Papp, 1969, and Asobara sp. Parasitism rates by braconids and figitids are presented. Doryctobracon areolatus was the most frequent, parasitizing the highest number of fly species, and showing the highest parasitism percentage in larvae feeding on Micropholis williamii fruits. The collected figitids belong to Aganaspis nordlanderi Wharton, 1998 and A. pelleranoi (Brethes, 1924). All 15 tritrophic associations are new records for the Brazilian Amazon region. The RFAD is an important natural reservoir of frugivorous larvae parasitoids.

Este trabalho teve por objetivo identificar as espécies de parasitóides de larvas frugívoras e descrever as interações tritróficas desses inimigos naturais com seus insetos hospedeiros e frutos silvestres da Reserva Florestal Adolpho Ducke (RFAD). As coletas foram realizadas em quatro quadrantes de 1 km², ocupando áreas próximas aos cantos da RFAD. Os frutos silvestres foram coletados no interior da floresta nas trilhas de acesso a cada área de coleta e nas trilhas que delimitavam os quadrantes, até cinco metros em cada lado. Os frutos foram acondicionados em recipientes plásticos, cobertos com tecido fino, contendo camada de vermiculita, para obtenção das moscas ou dos parasitóides, que eram individualizados, contados, sexados e fixados em álcool 70 por cento. As taxas de parasitismo por braconídeos e figitídeos são apresentadas. Foram coletadas sete espécies de Braconidae, distribuídas em Opiinae: Doryctobracon areolatus (Szépligeti, 1911), Utetes anastrephae (Viereck, 1913), Opius sp. e Alysiinae: Asobara anastrephae (Muesebeck, 1958), Phaenocarpa pericarpa Wharton e Carrejo, 1999, Idiasta delicata Papp, 1969 e Asobara sp. Doryctobracon areolatus foi a espécie mais frequente, parasitando o maior número de espécies de moscas e apresentando a maior percentagem de parasitismo em larvas em frutos de Micropholis williamii. OS figitídeos pertenciam a Aganaspis nordlanderi Wharton, 1998 e A. pelleranoi (Brethes, 1924). Todas as 15 associações tritróficas obtidas representam novos registros para a região Amazônica brasileira.

Intraspecific variation in Trichogramma bruni Nagaraja, 1983 (Hymanoptera: Trichogrammatidae) associated with different hosts

Trichogramma bruni is an insufficienty studied South American species whose limits are still not well defined. Thus, the objective of the present study was to characterize T. bruni taxonomically and to determine the association between morphological variations as well as host and habitat, based on morphological and biological studies. Specimens from the Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ) collection, and from the University of California Riverside (UCR) and specimens collected from the vegetation of forest parks with native areas planted with eucalyptus in Piracicaba and Itatinga, State of Säo Paulo, were also analyzed. The holotype deposited at Univeridade Federal de Minas Gerais (UFMG) collection was also examined. The variability in the genital capsule of T. bruni observed both among individuals of the same progeny and among specimens from different hosts is remarkable and is mainly related to the dorsal lamina. Therefore, an association of diagnostic characters rather than the dorsal lamina alone should be used for the identification of T. bruni and intraspecific variations should be considered. The intraspecific variation observed for T. bruni is a factor that should be considered for its identification, since the influence of the environment (habitat + host) and the variation among individuals itself is responsible for the plasticity observed in the genital capsule. Heliconius erato phyllis, Hamadryas feronia, Erosina hyberniata and Mechanitis lysiminia are new hosts of T. bruni

Intraspecific variation in Trichogramma bruni Nagaraja, 1983 (Hymenoptera: Trichogrammatidae) associated with different hosts.

Trichogramma bruni is an insufficiently studied South American species whose limits are still not well defined. Thus, the objective of the present study was to characterize T. bruni taxonomically and to determine the association between morphological variations as well as host and habitat, based on morphological and biological studies. Specimens from the Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ) collection, and from the University of California Riverside (UCR) and specimens collected from the vegetation of forest parks with native areas planted with eucalyptus in Piracicaba and Itatinga, State of São Paulo, were also analyzed. The holotype deposited at Univeridade Federal de Minas Gerais (UFMG) collection was also examined. The variability in the genital capsule of T. bruni observed both among individuals of the same progeny and among specimens from different hosts is remarkable and is mainly related to the dorsal lamina. Therefore, an association of diagnostic characters rather than the dorsal lamina alone should be used for the identification of T. bruni and intraspecific variations should be considered. The intraspecific variation observed for T. bruni is a factor that should be considered for its identification, since the influence of the environment (habitat + host) and the variation among individuals itself is responsible for the plasticity observed in the genital capsule. Heliconius erato phyllis, Hamadryas feronia, Erosina hyberniata and Mechanitis lysiminia are new hosts of T. bruni.