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The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is a major pest of citrus due to its role as the vector of the bacterium that causes huanglongbing. In commercial citrus, ACP control currently relies on the application of insecticides, which may not be sustainable long-term, nor practical in urban areas. The sterile insect technique (SIT) is an alternative strategy in which large numbers of pests are reared, sterilized using radiation, and then released into the field to compete with wild individuals for matings, suppressing population growth. As a fundamental step toward the development of SIT for ACP, this study sought to identify the optimum radiation dose required to sterilize ACP without affecting their survival and mating capacity. Virgin adult ACP of both sexes were subjected to doses of X-ray irradiation ranging from 40 to 480 Gy, then paired with a nonirradiated mate and allowed to produce offspring. Fecundity was estimated as the number of eggs laid, and fertility as the proportion of those eggs that hatched. Females were more radio-sensitive than males, exhibiting a major drop in fecundity at even the lowest dose and 100% sterility at 80 Gy. In contrast, a fivefold higher dose (400 Gy) did not achieve complete sterility in males, with around 5% offspring survival. However, F1 progeny of males exposed to 320 Gy or higher were subsequently found to be 100% sterile. This confirmation of inherited sterility suggests that balancing the sterilizing effects of radiation against its mortality-inducing effects may warrant further evaluation.
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Fertilidad , Hemípteros , Animales , Femenino , Fertilidad/efectos de la radiación , Masculino , Control Biológico de Vectores , Rayos X , Longevidad , Citrus , Control de Insectos/métodosRESUMEN
The present study evaluated the reproductive compatibility of Trichogramma pretiosum Riley, 1879, through an integrative approach using biological data and morphometry of three isofemale lines (isolines) collected from two geographical areas. These isolines differed in sequences of mitochondrial DNA and reproductive performance in the laboratory. The wasps used to initiate the isolines were collected in different environments: two lines from a Mediterranean climate in Irvine, California, USA, and one line from a tropical climate in Piracicaba, São Paulo, Brazil. Reproductive compatibility was studied by evaluating the sex ratio and number of adult offspring produced of all mating combinations between adults from these isolines. Morphometry was studied by measuring 26 taxonomically useful characters, followed by a multivariate analysis. For the allopatric matings among Brazilian and North American isolines, a low level of crossing incompatibility was recorded, in only one direction of the crosses; whereas the sympatric North American isolines were incompatible in both directions. Multivariate analysis of the morphometric data indicated no distinct groups, suggesting that despite the genetic and biological differences, the isofemale lines are morphologically similar.
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Reproducción , Avispas , Animales , Brasil , Avispas/genética , ADN Mitocondrial , MitocondriasRESUMEN
Background: Wolbachia bacteria are estimated to occur in more than half of all insect species. In Hymenoptera, Wolbachia often manipulates its host's reproduction to its own advantage. Wolbachia is likely the reason that males are rare in the uniparental Ooencyrtus mirus Triapitsyn & Power (Hymenoptera: Encyrtidae). The likelihood of producing male offspring can be increased by giving mothers a continuous supply of Bagrada hilaris (Burmeister) (Heteroptera: Pentatomidae) host eggs to parasitize for 2-3 weeks, by feeding the parents antibiotics, or by rearing parent wasps at high temperatures; all variables that have been shown to correlate with depleting Wolbachia titers in other organisms. The purpose of the current study was to determine whether thelytoky in O. mirus is due to Wolbachia, and if so, at what time in development the sex change occurs. We also wished to determine if Wolbachia removal results in the production of intersexes, as in some other hymenopterans. Finally, mating behavior was observed to see if and where it breaks down as a result of the species becoming thelytokous. Methods: Females were collected from parental lines of O. mirus reared at 26, 30, 31, 32, 33, 34, and 36 °C. The offspring of these females were reared at 26 °C, and their sex-ratio was determined. In a subsequent experiment, the parental generation was switched between 26 °C and 36 °C during development to narrow down the critical period at which changes occurred that subsequently affected the sex-ratio of their offspring. Results: The sex ratio was male biased in the offspring of O. mirus parents reared at 34 °C and 36 °C (high temperatures), even if the offspring themselves were reared at 26 °C. The constant temperature at which the percentage of males started to increase after two generations was 31 °C (10% males), rising to 39% males at 33 °C, and 100% males at 34 °C and 36 °C. Lasting more than 2 days, the critical period for the change toward a male biased sex ratio was during the second half of the parent's development. Molecular diagnostic assays confirmed that O. mirus females contain Wolbachia and males do not. Examination of preserved males and male-female pairs under a dissecting microscope showed no signs of intersex characters. Observation of the mating behavior of live O. mirus showed that males initiate courtship by drumming their antennae on a female's antennae, but after a few seconds, the females typically turn and walk away. However, a few instances of possible copulation were noted. Conclusions: As hypothesized, the results indicated that thelytoky in O. mirus is likely mediated by Wolbachia bacteria. To maximize the population growth rate without generating males, the best temperature for mass rearing this species is 30 °C.
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Heterópteros , Avispas , Wolbachia , Animales , Masculino , Femenino , Temperatura , Wolbachia/genética , Razón de Masculinidad , Avispas/microbiología , ReproducciónRESUMEN
Girault (1917) very briefly described and diagnosed the encyrtid wasp species Ooencyrtus californicus Girault (Hymenoptera: Encyrtidae) with the following limited data (p. 22): "Sacramento, California, from bug eggs on Pinus sabiniana, September". The first author examined its two syntypes, poorly mounted on a slide, of which only parts of the four antennae and a slightly damaged fore wing remain (Triapitsyn et al. 2020). They concluded that this species was better considered a nomen dubium until fresh specimens could be collected from the same host plant in or near the type locality. In July 2019, an effort was made to re-collect O. californicus from foothill (or gray) pine, Pinus sabiniana (Pinaceae), in the Oakhurst area of Madera County, California, USA, where this pine is abundant, but no specimens were captured. At the same time, a very similar insect was collected from sentinel eggs of the invasive stink bug Bagrada hilaris (Burmeister) in Riverside, California. This insect was cultured in the laboratory as part of a B. hilaris biological control program. Due to the poor preservation of the type specimens and in the absence of genetic evidence, it was impossible to positively and properly attribute specimens of this newly collected parasitoid to O. californicus. Therefore, to provide a much needed scientific name for this native egg parasitoid, it was described as Ooencyrtus lucidus Triapitsyn Ganjisaffar in Triapitsyn et al. (2020). The two nominal species were separated based on minor differences in the proportions of the scape, pedicel, and first funicular segment of the female antenna (Triapitsyn et al. 2020). John S. Noyes, world expert on Encyrtidae and one of the reviewers of Triapitsyn et al. (2020), disagreed with the erection of O. lucidus as a new taxon based on the morphological similarities of the female antenna with that of O. californicus, so the description was published with the understanding that the two might be conspecific. But to settle this one way or another new specimens of O. californicus had to be collected to allow morphological and molecular analyses.
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Heterópteros/parasitología , Avispas/anatomía & histología , Avispas/clasificación , Animales , California , FemeninoRESUMEN
Soyka (1946) described the European fairyfly species Anagrus bakkendorfi Soyka (Hymenoptera: Mymaridae) from a single female collected on the same day, 7.x.1931, together with several non-type specimens incorrectly labeled as paratypes (Chiappini Triapitsyn 1999), on a window at its type locality, the former St. Ignatius Jesuit College in Valkenburg, Limburg, the Netherlands. Soyka (1956) did not mention type specimens of any of his species of Anagrus Haliday. He described Anagrus avalae Soyka (Soyka 1956) based apparently on a specimen collected on Mt. Avala, Belgrade, Serbia, former Yugoslavia, along with several other nominal species (some also from Valkenburg) that were later synonymized with A. avalae and A. bakkendorfi by Chiappini (1989) and Chiappini Triapitsyn (1999). The only, minor, morphological difference between the two nominal species is the length of the ovipositor in females relative to the length of the protibia, being at least 2.6× in A. bakkendorfi and at most 2.3× in A. avalae (Triapitsyn 2015). Specimens identifiable as both species were captured by the second author in 2000 by a Malaise trap in Iratibizkar, Irati Forest (Selva de Irati), Navarra, Spain. Chiappini Triapitsyn (1999) discussed both nominal species but despite recognizing their apparent conspecificity, no formal synonymy was proposed. Triapitsyn Berezovskiy (2004) and Triapitsyn (2015) also did not propose synonymy because at that time genetic evidence for conspecificity was lacking. However, Triapitsyn et al. (2019) obtained DNA and sequenced selected mitochondrial and nuclear ribosomal gene regions (COI and ITS2) from a specimen unambiguously identified by the first author as A. avalae that was collected in Sevenoaks, Kent County, England, United Kingdom. Because of this we decided to collect fresh specimens identifiable as A. bakkendorfi for molecular analysis and genetic comparison with those identifiable as A. avalae. Based on the previous record (Triapitsyn 2015), several females of A. bakkendorfi were successfully captured by the second author on 4.ix.2020 in Irati Forest, preserved in ethanol, and shipped to the first and third authors for morphological and molecular identification, respectively.
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Himenópteros , Animales , Femenino , Himenópteros/genética , MasculinoRESUMEN
The cryptic species that make up the Euwallacea fornicatus species complex can be readily distinguished via their DNA sequences. Until recently, it was believed that the Hawaiian Islands had been invaded by only one of these cryptic species, E. perbrevis (tea shot hole borer; TSHB). However, following the 2016 deposition of a DNA sequence in the public repository GenBank, it became evident that another species, E. fornicatus (polyphagous shot hole borer; PSHB), had been detected in macadamia orchards on Hawai'i Island (the Big Island). We surveyed the two most-populous islands of Hawai'i, Big Island and O'ahu, and herein confirm that populations of TSHB and PSHB are established on both. Beetles were collected using a variety of techniques in macadamia orchards and natural areas. Individual specimens were identified to species using a high-resolution melt assay, described herein and validated by subsequent sequencing of specimens. It remains unclear how long each species has been present in the state, and while neither is currently recognized as causing serious economic or ecological damage in Hawai'i, the similarity of the newly-confirmed PSHB population to other damaging invasive PSHB populations around the world is discussed. Although the invasive PSHB populations in Hawai'i and California likely have different geographic origins within the beetle's native range, they share identical Fusarium and Graphium fungal symbionts, neither of which have been isolated from PSHB in that native range.
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Several species of egg parasitoids (Hymenoptera: Mymaridae and Trichogrammatidae) of the leafhopper pest of grapevines in Japan, Arboridia (Arboridia) apicalis (Nawa) (Hemiptera, Cicadellidae), were reared and identified for the first time. Using a combination of genetic and morphological evidence, Anagrus (Anagrus) arboridiae Triapitsyn & Adachi-Hagimori, sp. nov. (Mymaridae) is described and illustrated from Honshu Island (Shimane Prefecture) and Kyushu Island (Miyazaki Prefecture). It is shown to be different from Anagrus (Anagrus) japonicus Sahad and A. flaviapex Chiappini & Lin, to which it is most similar; the latter species was originally described from China and is newly recorded here from Okinawa Island, Japan. Mitochondrial and nuclear ribosomal DNA sequence data provide clear evidence for the separation of A. arboridiae from A. flaviapex, A. japonicus, and some other members of the Anagrus (Anagrus) atomus (L.) species group. Two other species of Anagrus Haliday, A. (Anagrus) avalae Soyka and A. atomus, are also identified in Japan from eggs of the leafhoppers Edwardsiana ishidae (Matsumura) and Eurhadina ? betularia Anufriev, respectively. An updated key to females of the Japanese species of Anagrus is given. Oligosita pallida Kryger (a new record for Japan), Oligosita sp., and an Aphelinoidea (Aphelinoidea) sp. (Trichogrammatidae) were the other, although much less abundant, apparent egg parasitoids of A. apicalis in Shimane Prefecture, mainly in non-organic vineyards.
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Carrillo, J.D., Rugman-Jones, PF., Husein, D., Stajich, J.E., Kasson, M.T., Carrillo, D., Stouthamer, R., and Eskalen, A. 2019. Members of the Euwallacea fornicatus species complex exhibit promiscuous mutualism with ambrosia fungi in Taiwan. A number of ambrosia beetles have come to prominence in recent years because of the damage they inflict on a variety of trees within invaded habitats across the globe. Ambrosia beetles rely on symbiotic microorganisms, mainly fungi, as a dedicated food source and carry those microorganisms around with them within specialized organs termed mycangia. Investigation of members of the Euwallacea fornicatus species complex and their fungal symbionts in Taiwan revealed promiscuous symbioses with ambrosial Fusaria clade (AFC) members, Graphium spp., and Paracremonium spp. based on co-phylogenetic analyses. For AFC members, a novel diagnostic PCR assay targeting mating type genes MAT1-1-1 and MAT1-2-1 was developed and validated by amplicon size and sequencing. Mating type screening of AFC members revealed the isolates screened are all heterothallic (self-sterile), with both MAT types represented and recovered from fungi vectored by E. fornicatus (tea shot hole borer), E. kuroshio (Kuroshio shot hole borer), and E. whitfordiodendrus (polyphagous shot hole borer) in Taiwan. Members of the Euwallacea fornicatus species complex and the variety of ambrosia fungi they utilize further confirms that their relationship with these fungi are more likely promiscuous in native areas, as opposed to strictly obligate to a specific combination of fungi as observed in invaded areas.
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Ascomicetos/fisiología , Simbiosis , Gorgojos/microbiología , Animales , FemeninoRESUMEN
BACKGROUND: Aphytis melinus DeBach (Hymenoptera: Aphelinidae) is a highly effective biocontrol agent of the California red scale Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae). It is commercially reared and used for augmentative releases within integrated pest management programs. However, mass rearing of biocontrol agents can result in population bottlenecks and high levels of inbreeding and/or adaptation to the factitious rearing conditions. Although these factors can all negatively impact field performance of biocontrol agents, few empirical studies have examined the genetic consequences of mass rearing. We used double-digest RAD sequencing (ddRADseq) to investigate the effect of traditional mass rearing on genetic variation among insectary colonies of A. melinus relative to wild populations in native (Pakistan) and introduced (California) ranges. RESULTS: Analyses of up to 9700 single nucleotide polymorphisms (SNPs) revealed that insectary populations had less genomic variation than introduced populations. This was evidenced by fewer private alleles, reduced heterozygosity, and greater missing data in the insectary populations. Further, California insectaries formed a distinct genomic cluster relative to the other samples, a surprising result given that the insectary colonies were putatively established at different times and from different source populations. These differences were evident across most data sets also after we filtered out contaminant DNA from the most common host species (Aspidiotus nerii Bouché and A. aurantii). CONCLUSION: We hypothesize that this pattern would only result if: (i) directional selection for 'captive' phenotypes produces convergent patterns of genomic variation across insectaries; or (ii) the California insectary colonies were all founded from a unifying source population and/or that the insectaries regularly exchange 'genetic' stocks. We show that RADseq is an effective method to investigate the effects of mass rearing on genetics of biocontrol agents. © 2019 Society of Chemical Industry.
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Estudio de Asociación del Genoma Completo , Hemípteros/parasitología , Control de Insectos , Control Biológico de Vectores , Polimorfismo de Nucleótido Simple , Avispas/genética , Animales , California , PakistánRESUMEN
BACKGROUND: Trichogrammatids are minute parasitoid wasps that develop within other insect eggs. They are less than half a millimeter long, smaller than some protozoans. The Trichogrammatidae are one of the earliest branching families of Chalcidoidea: a diverse superfamily of approximately half a million species of parasitoid wasps, proposed to have evolved from a miniaturized ancestor. Trichogramma are frequently used in agriculture, released as biological control agents against major moth and butterfly pests. Additionally, Trichogramma are well known for their symbiotic bacteria that induce asexual reproduction in infected females. Knowledge of the genome sequence of Trichogramma is a major step towards further understanding its biology and potential applications in pest control. RESULTS: We report the 195-Mb genome sequence of Trichogramma pretiosum and uncover signatures of miniaturization and adaptation in Trichogramma and related parasitoids. Comparative analyses reveal relatively rapid evolution of proteins involved in ribosome biogenesis and function, transcriptional regulation, and ploidy regulation. Chalcids also show loss or especially rapid evolution of 285 gene clusters conserved in other Hymenoptera, including many that are involved in signal transduction and embryonic development. Comparisons between sexual and asexual lineages of Trichogramma pretiosum reveal that there is no strong evidence for genome degradation (e.g., gene loss) in the asexual lineage, although it does contain a lower repeat content than the sexual lineage. Trichogramma shows particularly rapid genome evolution compared to other hymenopterans. We speculate these changes reflect adaptations to miniaturization, and to life as a specialized egg parasitoid. CONCLUSIONS: The genomes of Trichogramma and related parasitoids are a valuable resource for future studies of these diverse and economically important insects, including explorations of parasitoid biology, symbiosis, asexuality, biological control, and the evolution of miniaturization. Understanding the molecular determinants of parasitism can also inform mass rearing of Trichogramma and other parasitoids for biological control.
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Evolución Molecular , Control Biológico de Vectores , Avispas/clasificación , Avispas/genética , Animales , Genómica , Mariposas Nocturnas/parasitología , Filogenia , Avispas/patogenicidad , Secuenciación Completa del Genoma/métodosRESUMEN
Extracting and sequencing DNA from specimens can impose major time and monetary costs to studies requiring genotyping, or identification to species, of large numbers of individuals. As such, so-called direct PCR methods have been developed enabling significant savings at the DNA extraction step. Similarly, real-time quantitative PCR techniques (qPCR) offer very cost-effective alternatives to sequencing. High-resolution melt analysis (HRM) is a qPCR method that incorporates an intercalating dye into a double-stranded PCR amplicon. The dye fluoresces brightly, but only when it is bound. Thus, after PCR, raising the temperature of the amplicon while measuring the fluorescence of the reaction results in the generation of a sequence-specific melt curve, allowing discrimination of genotypes. Methods combining HRM (or other qPCR methods) and direct PCR have not previously been reported, most likely due to concerns that any tissue in the reaction tube would interfere with detection of the fluorescent signal. Here, we couple direct PCR with HRM and, by way of three examples, demonstrate a very quick and cost-effective method for genotyping large numbers of specimens, using Rotor-Gene HRM instruments (QIAGEN). In contrast to the heated-block design of most qPCR/HRM instruments, the Rotor-Gene's centrifugal rotor and air-based temperature-regulation system facilitate our method by depositing tissues away from the pathway of the machine's fluorescence detection optics.
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Escarabajos/clasificación , Genotipo , Himenópteros/clasificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Animales , ADN , Fluorescencia , Especificidad de la EspecieRESUMEN
The Red Palm Weevil (RPW) Rhynchophorus ferrugineus (Olivier) is a voracious pest of palm species. In recent decades its range has expanded greatly, particularly impacting the date palm industry in the Middle East. This has led to conjecture regarding the origins of invasive RPW populations. For example, in parts of the Middle East, RPW is commonly referred to as the "Pakistani weevil" in the belief that it originated there. We sought evidence to support or refute this belief. First reports of RPW in Pakistan were from the Punjab region in 1918, but it is unknown whether it is native or invasive there. We estimated genetic variation across five populations of RPW from two provinces of Pakistan, using sequences of the mitochondrial cytochrome oxidase subunit I gene. Four haplotypes were detected; two (H1 and H5) were abundant, accounting for 88% of specimens across the sampled populations, and were previously known from the Middle East. The remaining haplotypes (H51 and H52) were newly detected (in global terms) and there was no geographic overlap in their distribution within Pakistan. Levels of haplotype diversity were much lower than those previously recorded in accepted parts of the native range of RPW, suggesting that the weevil may be invasive in Pakistan. The affinity of Pakistani haplotypes to those reported from India (and the geographical proximity of the two countries), make the latter a likely "native" source. With regards the validity of the name "Pakistani weevil", we found little genetic evidence to justify it.
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ADN Mitocondrial/genética , Variación Genética , Proteínas de Insectos/genética , Gorgojos/genética , Distribución Animal , Animales , Especies Introducidas , Pakistán , Gorgojos/enzimologíaRESUMEN
In this study we assessed the relationship between the laboratory and field performance of different isofemale lines of Trichogramma pretiosum Riley. In comparative assays, we used three rare mitochondrial haplotypes as genetic markers of the isofemale lines, and by introgressing these mitochondrial haplotypes into each of 15 genetically different nuclear lines, also tested the assumption that mitochondria are neutral markers. In a laboratory trial, 45 isofemale lines (15 nuclear genotypes x three mitochondrial haplotypes) were ranked in three categories (best, intermediate and worst) according to the mean offspring production and the proportion of female offspring. Subsequently, lines from each of the three categories were selected for field releases to quantify field parasitism on Ephestia kuehniella. Temporally separate releases were done in a transgenic Bt cornfield, with four plots, each with 50 points of recapture. The points of recapture consisted of trap cards with eggs of E. kuehniella collected daily. The trap cards were maintained in the laboratory at 25°C until the adult wasps emerged, and the maternal identity of the wasps was determined using qPCR and high-resolution melt curve analysis to determine the mitochondrial haplotype. The results showed that these measures of laboratory performance (fecundity and offspring sex ratio) were good predictors of field success in T. pretiosum. We also report strong evidence discrediting the assumption that mitochondria are neutral, in view of the correlation between performance and mitochondrial haplotype.
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Laboratorios , Mariposas Nocturnas/parasitología , Óvulo/parasitología , Avispas/fisiología , Animales , Distribución de Chi-Cuadrado , ADN Mitocondrial/química , ADN Mitocondrial/genética , Ecosistema , Femenino , Haplotipos , Interacciones Huésped-Parásitos , Endogamia , Modelos Lineales , Masculino , Desnaturalización de Ácido Nucleico , Reproducción/genética , Reproducción/fisiología , Avispas/genéticaRESUMEN
Thousand cankers disease (TCD) of walnut trees (Juglans spp.) results from aggressive feeding in the phloem by the walnut twig beetle (WTB), Pityophthorus juglandis, accompanied by inoculation of its galleries with a pathogenic fungus, Geosmithia morbida. In 1960, WTB was only known from four U.S. counties (in Arizona, California, and New Mexico), but the species has now (2014) invaded over 115 counties, representing much of the western USA, and at least six states in the eastern USA. The eastern expansion places TCD in direct proximity to highly valuable (> $500 billion) native timber stands of eastern black walnut, Juglans nigra. Using mitochondrial DNA sequences, from nearly 1100 individuals, we examined variation among 77 samples of WTB populations across its extended range in the USA, revealing high levels of polymorphism and evidence of two divergent lineages. The highest level of genetic diversity for the different lineages was found in the neighboring Madrean Sky Island and Western New Mexico regions, respectively. Despite their proximity, there was little evidence of mixing between these regions, with only a single migrant detected among 179 beetles tested. Indeed, geographic overlap of the two lineages was only common in parts of Colorado and Utah. Just two haplotypes, from the same lineage, predominated over the vast majority of the recently expanded range. Tests for Wolbachia proved negative suggesting it plays no role in "driving" the spread of particular haplotypes, or in maintaining deep levels of intraspecific divergence in WTB. Genotyping of ribosomal RNA corroborated the mitochondrial lineages, but also revealed evidence of hybridization between them. Hybridization was particularly prevalent in the sympatric areas, also apparent in all invaded areas, but absent from the most haplotype-rich area of each mitochondrial lineage. Hypotheses about the specific status of WTB, its recent expansion, and potential evolutionary origins of TCD are discussed.
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Escarabajos/genética , Polimorfismo de Longitud del Fragmento de Restricción , Animales , Secuencia de Bases , ADN Mitocondrial/genética , Genes de Insecto , Datos de Secuencia Molecular , Filogeografía , ARN Ribosómico/genéticaRESUMEN
The red palm weevil (RPW) is a major pest of palms. It is native to southeast Asia and Melanesia, but in recent decades has vastly expanded its range as the result of multiple accidental anthropogenic introductions into the Middle East, Mediterranean Basin, Caribbean, and U.S.A. Currently regarded as a single species, Rhynchophorus ferrugineus (Olivier), RPW displays remarkable color variation across its range, and consequently has a taxonomic history littered with new species descriptions and synonymization. We compared DNA sequences of the mitochondrial cytochrome oxidase subunit I (COI) gene from RPW populations throughout the native and invaded ranges, to investigate the specific status and invasion history of this serious economic pest, and to identify possible common routes of entry. Analyses of COI haplotype data provide conclusive support, corroborated by sequences of additional nuclear gene regions, for the existence of at least two predominantly allopatric species. The true R. ferrugineus is native only to the northern and western parts of continental southeast Asia, Sri Lanka and the Philippines, and is responsible for almost all invasive populations worldwide. In contrast, the second species, which is currently synonymized under R. ferrugineus and should be resurrected under the name R. vulneratus (Panzer), has a more southern distribution across Indonesia, and is responsible for only one invasive population; that in California, U.S.A. The distribution of COI haplotypes is used to discuss the possible existence of further cryptic species, sources and routes of entry of different invasive populations, and the implications of our findings for current control methods.
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Biología Molecular , Gorgojos/genética , Animales , Especies Introducidas , Análisis de Secuencia de ADN , Especificidad de la Especie , Gorgojos/anatomía & histologíaRESUMEN
The polyphagous shot hole borer (PSHB) is an invasive ambrosia beetle that forms a symbiosis with a new, as-yet-undescribed Fusarium sp., together causing Fusarium dieback on avocado and other host plants in California and Israel. In California, PSHB was first reported on black locust in 2003 but there were no records of fungal damage until 2012, when a Fusarium sp. was recovered from the tissues of several backyard avocado trees infested with PSHB in Los Angeles County. The aim of this study was to determine the plant host range of the beetle-fungus complex in two heavily infested botanical gardens in Los Angeles County. Of the 335 tree species observed, 207 (62%), representing 58 plant families, showed signs and symptoms consistent with attack by PSHB. The Fusarium sp. was recovered from 54% of the plant species attacked by PSHB, indicated by the presence of the Fusarium sp. at least at the site of the entry hole. Trees attacked by PSHB included 11 species of California natives, 13 agriculturally important species, and many common street trees. Survey results also revealed 19 tree species that function as reproductive hosts for PSHB. Additionally, approximately a quarter of all tree individuals planted along the streets of southern California belong to a species classified as a reproductive host. These data suggest the beetle-disease complex potentially may establish in a variety of plant communities locally and worldwide.
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Over the past three decades, Western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), has become a major worldwide pest of many agricultural and horticultural crops. In response, much time, money, and effort have been put into pure and applied research focusing on the biology and control of this pest. Western flower thrips is native to Western North America and widespread in California. High levels of variation in basic biology, pest status, and resistance to insecticides bring into question the specific status of Western flower thrips. We used nuclear-mitochondrial barcoding to compare DNA sequences of nuclear and mitochondrial genes between Western flower thrips populations across California, looking for association between these unlinked loci. Sequences of D2 domain of 28S and cytochrome c oxidase I gene revealed the existence of two distinct but sympatric genetic entities, and we describe a simple polymerase chain reaction-based method for diagnosing these entities. The complete association of these nuclear and mitochondrial loci in areas of sympatry is indicative of reproductive isolation and the existence of two cryptic species, both of which key out to Western flower thrips by using morphological characters. The finding that Western flower thrips is a complex of two species has important implications for past, current, and most importantly future research on these pests.
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Complejo IV de Transporte de Electrones/genética , Insectos/genética , ARN Ribosómico 28S/genética , Animales , California , Núcleo Celular/genética , ADN Mitocondrial/genética , Plantas , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
'Hass' avocado, Persea americana Mill., fruit imported into California from Mexico are infested with high levels of armored scale insects (Hemiptera: Diaspididae), constituting several species. The paucity and delicate nature of morphological characters traditionally used to diagnose armored scales often require careful preparation of slide-mounted specimens and expert knowledge of the group, for their accurate identification. Here, we present a simple, quick, and accurate means to identify armored scales on Mexican avocados, based on amplification of the internal transcribed spacer two of ribosomal DNA, by using the polymerase chain reaction (PCR). This region seems to show a high level of intraspecific conformity among scale specimens originating from different localities. A suite of species-specific reverse PCR primers are combined in a single reaction, with a universal forward primer, and produce a PCR product of a unique size, that after standard gel electrophoresis, allows the direct diagnosis of six diaspidid species: Abgrallaspis aguacatae Evans, Watson & Miller; Hemiberlesia lataniae (Signoret); Hemiberlesia sp. near latania; Hemiberlesia rapax (Comstock); Acutaspis albopicta (Cockerell); and Pinnaspis strachani (Cooley). Two additional species, Diaspis miranda (Cockerell) and Diaspis sp. near miranda, also are separated from the others by using this method and are subsequently diagnosed by secondary digestion of the PCR product with the restriction endonuclease smaI.
Asunto(s)
Hemípteros/genética , Persea/parasitología , Animales , Secuencia Conservada , ADN/genética , Cartilla de ADN , ADN Ribosómico/genética , Hemípteros/clasificación , México , Reacción en Cadena de la Polimerasa/métodos , Especificidad de la Especie , Transcripción GenéticaRESUMEN
Conspecific sperm precedence (CSP) has been identified as an important post-copulatory, pre-zygotic mechanism that can act to reduce gene flow between populations. The evolution of CSP is thought to have arisen as a by-product of male and female coevolution in response to intraspecific post-copulatory sexual selection. However, little is known about the mechanisms that generate CSP. When Callosobruchus subinnotatus females copulate with both C. subinnotatus and Callosobruchus maculatus males, regardless of mating order, the majority of eggs are fertilized by conspecific sperm. The low number of heterospecific fertilizations does not result from general differences in the viability of sperm in the female reproductive tract, as heterospecific sperm fertilized equivalent numbers of eggs as conspecific sperm in the absence of sperm competition. Instead, CSP results from disadvantages to heterospecific sperm that are manifest only when in competition with conspecific sperm. CSP in C. subinnotatus appears to result from two, not mutually exclusive, mechanisms. First, conspecific sperm are better able to displace heterospecific sperm from female storage. Second, conspecific sperm achieve disproportionately higher numbers of fertilizations relative to their proportional representation in the fertilization set. Thus, we provide evidence of differential sperm use from the female spermatheca.
Asunto(s)
Escarabajos/fisiología , Conducta Sexual Animal/fisiología , Espermatozoides/fisiología , Animales , Masculino , Reproducción/fisiología , Especificidad de la EspecieRESUMEN
Effective plant quarantine and biological control initiatives require rapid and accurate identification of exotic and potentially invasive taxa that may cause high economic losses or environmental damage. The genus Scirtothrips Shull includes several species that are serious agricultural pests, and, because of their minute size and cryptic behavior, prone to undetected transport through international trade of plant material. Although assigning specimens to the genus Scirtothrips is straightforward using traditional taxonomic methods, identification of species is much more difficult and requires expert knowledge of the genus. Furthermore, the validity of some Scirtothrips species is questionable. Therefore, an easy, accurate, and highly reliable technique is desirable for Scirtothrips identification. Here, we provide a simple molecular key based on the internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) of nuclear ribosomal DNA. Individual specimens can be identified by amplification of their ITS1 and ITS2 regions with general primers and determining the size of the products by using standard agarose gel electrophoresis, followed in some instances by DNA digestion with the restriction enzymes SacII or PspOM I. The advantage of this identification system is that nonspecialists are able to quickly and cheaply identify individual specimens. Material analyzed for this work was collected in the United States (California), India, South Africa, Kenya, Mexico, Guatemala, Honduras, Nicaragua, Costa Rica, Panama, Australia, New Zealand and Raiatea (Society Islands French Polynesia). We have identified seven pest species with the molecular-based methods described here. It is hoped that this system can be extended to other members of the genus as their ITS1 and ITS2 sequences become available. We also provide molecular confirmation for two new Scirtothrips species, one species from Honduras and one species from New Zealand.