Les virus de l’enroulement de la vigne (Grapevine leafroll-associated virus, GLRaV) sont répandus mondialement et transmis à la vigne uniquement par cochenilles (Coccoidea). En France, l’enroulement viral affecte particulièrement les vignobles des régions septentrionales.L’approche biologique de la vection a montré la capacité de Phenacoccus aceris à transmettre à la vigne les GLRaV-1, -3, -4, -5, -6, -9 et ceux du bois strié Grapevine virus A et B. Cette étude est la première démonstration de la transmission du GLRaV-6 et confirme l’absence de spécificité des cochenilles dans la transmission des Ampelovirus. Les larves néonates de P. aceris et de Neopulvinaria innumerabilis représentent un stade de développement efficace pour la transmission de ces virus. En conséquence, leurs capacités vectrices, associées à leur fort potentiel de dissémination anémophile, impliquent un risque important de dispersion naturelle de ces virus dans un vignoble infesté. Les relevés sur quatre parcelles distinctes montrent que Parthenolecanium corni, Pulvinaria vitis, Heliococcus bohemicus et P. aceris sont communes, chaque vignoble différant par la diversité spécifique, le taux de ceps infestés et l’abondance des cochenilles. L'étude épidémiologique prouve le rôle des cochenilles dans la dispersion de l’enroulement viral dans les vignobles septentrionaux. A Bonzon, la responsabilité de P. aceris dans la diffusion rapide du GLRaV-1 est mise en évidence. Cette découverte représente la première preuve en Europe d’une dispersion naturelle du GLRaV-1. A Marsannayla-Côte, l’incidence du GLRaV-1 reste faible, la colonie de P. aceris ne semblant avoir qu’un rôle très limité dans la diffusion de la maladie. L'épidémiologie moléculaire à Bonzon révèle une diversité génétique importante du GLRaV-1 à l’échelle parcellaire et fournit pour la première fois des données sur le polymorphisme génétique d'une population de GLRaV-1 ayant été dispersée par des cochenilles. / Grapevine leafroll viruses (Grapevine leafroll-associated virus, GLRaV) are present worldwide and transmitted to grapevine only by scale insect vectors (Coccoidea). In France, leafroll disease is present in all vine-growing areas, particularly in north-eastern regions. The biological approach of transmission allowed us to show the capacity of the mealybug Phenacoccus aceris to transmit the viruses GLRaV-1, -3, -4, -5, -6, -9 and the rugose wood viruses Grapevine virus A and B. This study represents the first evidence of the transmission of GLRaV-6 and confirms the absence of mealybug specificity in the transmission of Ampelovirus. First instar nymphs of P. aceris and of Neopulvinaria innumerabilis represent a very efficient development stage in the transmission of leafroll and rugose wood viruses. As a consequence, their vector capacities associated with the high potential of dispersal of these nymphs imply an important risk of natural spread of viruses in an infested vineyard. The entomological monitoring on 4 plots shows that Parthenolecanium corni, Pulvinaria vitis, Heliococcus bohemicus and P. aceris are common in vineyards, each site differing by the specific diversity, the level of infested stocks and the abundance of scale insects on stocks. The epidemiological study proves the role of scales insects in the dispersal of leafroll disease in the vineyards of north-eastern France. In Bonzon, the major role of P. aceris in the rapid spread of the GLRaV-1 is demonstrated. This finding represents the first report in Europe of a natural spread of GLRaV-1. In Marsannay-la-Côte, the incidence of the GLRaV-1 remains low and the colony of P. aceris, not associated to grapevine, seems to have only a very limited role in the disease spread. The molecular epidemiology study in Bonzon reveals an important genetic diversity of GLRaV-1 within a signle plotand supplies for the first time information on the genetic polymorphism of a GLRaV-1population being spread by scale insects.

L’enroulement de la vigne cause des pertes de rendement et de la qualité des vins au niveau mondial. Il est causé par quatre espèces de Grapevine leafroll-associated virus(GLRaVs) ; GLRaV-1, -2, -3, -4-like. Si le GLRaV-2 (genre Closterovirus) ne possède pas de vecteur connu, les trois autres espèces (genre Ampelovirus) sont transmises par cochenilles(Coccoidea) qui contribuent à leur dispersion dans et entre les vignobles. Les vignobles de la France septentrionale (Alsace, Bourgogne, Champagne) sont les plus impactés par l’enroulement viral. Ce travail a porté sur le rôle vecteur de Phenacoccus aceris, espèce connue pour son efficacité de transmission et de dispersion des ampélovirus, ainsi que de vitivirus souvent associés. Les interactions cellulaires et moléculaires entre virus et cochenille restent peu connues. Une approche pluridisciplinaire combinant entomologie,virologie, biologie cellulaire et moléculaire a été développée pour étudier la biologie de la vection du GLRaV-1 et du Grapevine virus A (GVA) par P. aceris. Des expériences de transmission ont montré que ces virus sont transmis selon le mode semi-persistant non circulant. L’étude préliminaire du comportement alimentaire de P. aceris sur vigne par électropénétrographie a révélé une activité similaire à celle d'autres espèces de cochenilles déjà décrites et suggère un effet de l’infection sur le comportement alimentaire. L’anatomie des pièces buccales de P. aceris, organes directement impliqués dans la transmission et la rétention de virus non-circulants a été décrite et une méthode basée sur l’acquisition de virus purifié sur membrane a été développée pour rechercher les sites de rétention virale dans le vecteur. / Grapevine leafroll disease affects grape yield and wine quality worldwide. It is caused by four species of Grapevine leafroll-associated virus (GLRaVs) (GLRaV-1, -2, -3, -4-like).While GLRaV-2 (genus Closterovirus) has no known vector, the other three (genus Ampelovirus) are transmitted by mealybugs (Coccoidea) and thus prone to be dispersed within and between vineyards. In north-eastern France (Alsace, Bourgogne et Champagne),vineyards are more impacted by Grapevine leafroll disease. This thesis focusses on the vector role of the species Phenacoccus aceris, known for its efficiency in transmission and dissemination of ampeloviruses, as well as often associated vitiviruses. Molecular and cellular interactions between viruses and mealybugs remain poorly known. A multidisciplinary approach, combining entomology, virology, molecular and cellular biology, was developed to analyse the vector biology of GLRaV-1 and Grapevine virus A (GVA) by P. aceris.Transmission experiments showed that GLRaV-1 and GVA transmission follows the semipersistent non-circulative mode. A preliminary study of P. aceris feeding behavior on grape using electropenetrography revealed an activity similar to that of other mealybug species already described and suggested a potential effect of infection on Ph. aceris feeding behavior. The anatomy of mouth parts, directely implied in transmission and retention of non circulative viruses was described and a method for membrane acquisition of purified virus was developed to search for virus retention sites within the vector.

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2017-03-20T12:23:05Z No. of bitstreams: 1 PDF - Vanessa da Silva Guedes.pdf: 12640326 bytes, checksum: 799248fdb9844a9994517fcd5c5aebe4 (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2017-03-22T15:19:41Z (GMT) No. of bitstreams: 1 PDF - Vanessa da Silva Guedes.pdf: 12640326 bytes, checksum: 799248fdb9844a9994517fcd5c5aebe4 (MD5) / Made available in DSpace on 2017-03-22T15:19:41Z (GMT). No. of bitstreams: 1 PDF - Vanessa da Silva Guedes.pdf: 12640326 bytes, checksum: 799248fdb9844a9994517fcd5c5aebe4 (MD5) Previous issue date: 2017-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Cotton crop has been well known to man since ancient times, but one of the main problems of this culture is the attack of pests. Phenacoccus solenopsis Tinsley (Hemiptera:Pseudococcidae) has a high morphological diversity, biological adaptations and ecological adjustments that give this species a high capacity to feed host plants. Chemical control is the main method used to control population outbreaks of this mealybug. However, it is difficult to fully control this mealybug with conventional insecticides because of the waxy material that covers the bodies of adult females. The objective of this research was to study the biological aspects of P. solenopsis in cotton leaf discs with and without kaolin. Kaolin is a white clay mineral with various agricultural applications. The film of kaolin particles creates a physical or mechanical barrier of protection against arthropods and pathogens. The work was carried out in the Embrapa Algodão Entomology Laboratory, Campina Grande, State of Paraíba, Brazil, in climatic chamber of type B.O.D. (Biochemical Oxygen Demand), adjusted to a temperature of 25 ± 1 °C, relative humidity of 60 ± 10% and photophase of 12 hours. The experimental design was completely randomized with two treatments and fifty replicates. The treatments consisted of spraying the immature and newly emerged adult stages of P. solenopsis and immersing cotton leaf discs in kaolin suspension (with kaolin) in distilled -1 water at 60 gL the immature stages of P. solenopsis were determined and, in the adult phase, the periods of or in distilled water (control, without kaolin). The duration and survival of pre-oviposition, oviposition, post-oviposition, longevity, fecundity, number of eggs/day, number of eggs/female/day. The parameters of the fertility and life expectancy tables of P. solenopsis were also estimated to determine the population growth potential of this mealybug in both treatments. The results obtained in this research demonstrate that the survival of the immature stages of P. solenopsis in cotton leaves disks was not affected by kaolin. However, it was observed a shorter duration of development of the second and third instars and of the egg period to the adults of P. solenopsis females in the treatment with kaolin suspension and a longer period of oviposition, fecundity and number of eggs per female in that same treatment. In addition, the shortest time required for the P. solenopsis population to double time (TD) associated with the highest values of gross (TBR) and net reproductive rate (Ro), intrinsic (rm) and finite rates of increase (λ), survival, specific fertility (mx), reproductive value (VRx) and life expectancy (ex) in treatment with kaolin suspension indicate that spraying of this product on cotton plants infested by mealybug should be avoided because they favor its growth populational. / A cultura do algodão é bem conhecida do homem desde antiguidade, porém um dos principais problemas dessa a cultura é o ataque de pragas. A cochonilha Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) apresenta uma grande diversidade morfológica, adaptações biológicas e ajustes ecológicos que conferem a essa espécie uma alta capacidade de se alimentar de plantas hospedeiras. O controle químico é o principal método empregado para controlar os surtos populacionais dessa cochonilha. No entanto, é difícil controlar plenamente essa cochonilha com inseticidas convencionais, devido ao material ceroso que recobre os corpos das fêmeas adultas. O objetivo deste trabalho foi estudar os aspectos biológicos de P. solenopsis em discos de folhas de algodão com e sem caulim. O caulim é um mineral de argila branca com diversas aplicações agrícolas. O filme de partículas de caulim cria uma barreira física ou mecânica de proteção contra os artrópodes e patógenos. O trabalho foi conduzido no Laboratório de Entomologia da Embrapa Algodão, município de Campina Grande, Estado da Paraíba, Brasil, em câmaras climatizadas do tipo B.O.D. (Biochemical Oxygen Demand), ajustadas para à temperatura de 25 ± 1 ºC, umidade relativa de 60 ± 10% e fotofase de 12 horas. O delineamento experimental foi inteiramente casualizado com dois tratamentos e cinquenta repetições. Os tratamentos consistiram da pulverização dos estágios imaturos e adultos recém-emergidos de P. solenopsis e da imersão dos discos de folhas de -1 algodoeiro em suspensão de caulim (com caulim) em água destilada a 60 gL destilada, testemunha (sem caulim). sobrevivência dos estágios imaturos de P. solenopsis e, na fase adulta, os períodos de pré- ou em água Foram determinados o tempo de duração e a oviposição, oviposição, pós-oviposição, longevidade, fecundidade, número de ovos/dia, número de ovos/fêmea/dia. Foram também estimados os parâmetros das tabelas de fertilidade e de esperança de vida de P. solenopsis para se determinar o potencial de crescimento populacional dessa cochonilha em ambos os tratamentos. Os resultados obtidos nesta pesquisa demonstram que a sobrevivência dos estágios imaturos de P. solenopsis em discos de folhas de algodoeiro não foi prejudicada pelo caulim. No entanto, observou-se menor duração do desenvolvimento dos segundo e terceiro instares e do período de ovo à adultos de fêmeas de P. solenopsis no tratamento com suspensão de caulim e um maior período de oviposição, fecundidade e número de ovos por fêmea nesse mesmo tratamento. Além disso, o menor tempo necessário para a população de P. solenopsis duplicar em número de indivíduos (TD) associado aos maiores valores das taxas bruta (TBR) e líquida de reprodução (Ro), razões intrínseca (rm) e finita de crescimento (λ), sobrevivência, fertilidade específica (mx), valor reprodutivo (VRx) e esperança de vida (ex) no tratamento com suspensão de caulim, indicam que as pulverizações desse produto nas lavouras de algodoeiro infestadas pela cochonilha devem ser evitadas porque favorecem seu crescimento populacional.

Les virus de l'enroulement de la vigne (Grapevine leafroll-associated virus, GLRaV) sont répandus mondialement et transmis à la vigne uniquement par cochenilles (Coccoidea). En France, l'enroulement viral affecte particulièrement les vignobles des régions septentrionales.L'approche biologique de la vection a montré la capacité de Phenacoccus aceris à transmettre à la vigne les GLRaV-1, -3, -4, -5, -6, -9 et ceux du bois strié Grapevine virus A et B. Cette étude est la première démonstration de la transmission du GLRaV-6 et confirme l'absence de spécificité des cochenilles dans la transmission des Ampelovirus. Les larves néonates de P. aceris et de Neopulvinaria innumerabilis représentent un stade de développement efficace pour la transmission de ces virus. En conséquence, leurs capacités vectrices, associées à leur fort potentiel de dissémination anémophile, impliquent un risque important de dispersion naturelle de ces virus dans un vignoble infesté. Les relevés sur quatre parcelles distinctes montrent que Parthenolecanium corni, Pulvinaria vitis, Heliococcus bohemicus et P. aceris sont communes, chaque vignoble différant par la diversité spécifique, le taux de ceps infestés et l'abondance des cochenilles. L'étude épidémiologique prouve le rôle des cochenilles dans la dispersion de l'enroulement viral dans les vignobles septentrionaux. A Bonzon, la responsabilité de P. aceris dans la diffusion rapide du GLRaV-1 est mise en évidence. Cette découverte représente la première preuve en Europe d'une dispersion naturelle du GLRaV-1. A Marsannayla-Côte, l'incidence du GLRaV-1 reste faible, la colonie de P. aceris ne semblant avoir qu'un rôle très limité dans la diffusion de la maladie. L'épidémiologie moléculaire à Bonzon révèle une diversité génétique importante du GLRaV-1 à l'échelle parcellaire et fournit pour la première fois des données sur le polymorphisme génétique d'une population de GLRaV-1 ayant été dispersée par des cochenilles.

Phenacoccus peruvianus (Hemiptera: Pseudococcidae) is an invasive mealybug of Neotropical origin, first reported in the Mediterranean Basin in Almeria (Spain) in 1999. In the following years the mealybug spread into other Mediterranean regions and has also been recorded in Portugal and France, as well as in Sicily, Corsica and the Balearic Islands. Phenacoccus peruvianus is a polyphagous species and damages economically important ornamental plants. Since this was a relatively unknown species, during the first years of invasion, the mealybug was managed by the application of chemical treatments with wide-spectrum pesticides. However, the latest European directive on pesticide use reduces or even forbids pesticide applications in a wide range of urban green areas, giving significant priority to biological control (European Parliament and Council 2009). This thesis sets the basis for introducing biological control into a P. peruvianus management program in urban landscapes, focusing on its characterization, sampling, biology and control. In order to facilitate the identification of this and other mealybug species, we characterised 33 mealybug populations infesting crops and ornamental plants in Eastern Spain, using a combination of molecular and morphological techniques. This characterisation led to the identification of ten mealybug species and made routine identification possible through DNA sequencing or the use of derived species-specific molecular tools. The sequences obtained also add to the phylogenetic knowledge of the Pseudococcidae family and provide insight into the invasion history of some species. Phenacoccus peruvianus populations were high in bougainvillea plants during spring and summer, declining to almost undetectable levels in autumn and winter. The mealybug was mainly found in bracts and there were no significant migrations between plant strata. Phenacoccus peruvianus showed a high aggregated distribution on bracts, leaves and twigs. We recommend a binomial sampling of 200 leaves and an action threshold of 55% infested leaves for IPM purposes in urban landscapes. Its most abundant natural enemies were found to be the primary parasitoids Acerophagus n. sp. near coccois and Leptomastix epona Walker (Hymenoptera: Encyrtidae). We also identified several predator species from the Anthocoridae, Coccinellidae, Chamaemyiidae, and Chrysopidae families. Phenacoccus peruvianus populations were lower during the second and third year of the survey, coinciding with an increase in the parasitoid Acerophagus sp. populations, which displaced the native L. epona. Differential female offspring and resource preemption are discussed as the main reasons for this displacement. To obtain further information on the biology of the new parasitoid Acerophagus sp. we determined some traits of its reproductive and feeding strategies. Acerophagus sp. egg load reached its maximum when it was 5 days old with almost 30 mature eggs. Phenacoccus peruvianus second and third nymphal instars and adults were suitable for parasitism and efficient encapsulation was low (10.76 ± 0.31 %). The parasitoid always preferred older instars when different host instars were available. Acerophagus sp. developed as a solitary parasitoid in the second instar and as a gregarious parasitoid in older instars (2¿4 parasitoids per host). Moreover, it reproduced parthenogenetically and all the emerged offspring were females. Immature development lasted between 20 and 22 days at 25°C and 65% HR. Under these conditions, adults lived for longer than 20 days when fed on honey, but fewer than 3 days when fed on naturally occurring sugar sources (host honeydew and Bougainvillea glabra flowers). / Beltrà Ivars, A. (2014). Biology and management of the invasive mealybug Phenacoccus peruvianus (Hemiptera: Pseudococcidae) in urban landscapes [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/37233