Abacá mosaic virus: a distinct strain of Sugarcane mosaic virus

Abacá mosaic virus (AbaMV) is related to members of the sugarcane mosaic virus subgroup of the genus Potyvirus. The ~2 kb 3′ terminal region of the viral genome was sequenced and, in all areas analysed, found to be most similar to Sugarcane mosaic virus (SCMV) and distinct from Johnsongrass mosaic virus (JGMV), Maize dwarf mosaic virus (MDMV) and Sorghum mosaic virus (SrMV). Cladograms of the 3′ terminal region of the NIb protein, the coat protein core and the 3′ untranslated region showed that AbaMV clustered with SCMV, which was a distinct clade and separate from JGMV, MDMV and SrMV. The N-terminal region of the AbaMV coat protein had a unique amino acid repeat motif different from those previously published for other strains of SCMV. The first experimental transmission of AbaMV from abacá (Musa textilis) to banana (Musa sp.), using the aphid vectors Rhopalosiphum maidis and Aphis gossypii, is reported. Polyclonal antisera for the detection of AbaMV in western blot assays and ELISA were prepared from recombinant coat protein expressed in E. coli. A reverse transcriptase PCR diagnostic assay, with microtitre plate colourimetric detection, was developed to discriminate between AbaMV and Banana bract mosaic virus, another Musa-infecting potyvirus. Sequence data, host reactions and serological relationships indicate that AbaMV should be considered a distinct strain of SCMV, and the strain designation SCMV-Ab is suggested.

Identification of a new Polerovirus (family Luteoviridae) associated with cotton bunchy top disease in Australia

Cotton bunchy top (CBT) disease has caused significant yield losses in Australia and is now managed by control of its vector, the cotton aphid (Aphis gossypii). Its mode of transmission and similarities in symptoms to cotton Blue Disease suggested it may also be caused by a luteovirus or related virus. Degenerate primers to conserved regions of the genomes of the family Luteoviridae were used to amplify viral cDNAs from CBT-affected cotton leaf tissue that were not present in healthy plants. Partial genome sequence of a new virus (Cotton bunchy top virus, CBTV) was obtained spanning part of the RNA-dependent-RNA-polymerase (RdRP), all of the coat protein and part of the aphid-transmission protein. CBTV sequences could be detected in viruliferous aphids able to transmit CBT, but not aphids from non-symptomatic plants, indicating that it is associated with the disease and may be the causal agent. All CBTV open-reading frames had their closest similarity to viruses of the genus Polerovirus. The partial RdRP had 90 % amino acid identity to the RdRP of Cotton leafroll dwarf virus (CLRDV) that causes cotton blue disease, while other parts of the genome were more similar to other poleroviruses. The sequence similarity and genome organization of CBTV suggest that it should be considered a new member of the genus Polerovirus. This partial genome sequence of CBTV opens up the possibility for developing diagnostic tests for detection of the virus in cotton plants, aphids and weeds as well as alternative strategies for engineering CBT resistance in cotton plants through biotechnology. © 2012 Australasian Plant Pathology Society Inc.

Honeydew as a source of nutrition for Lysiphlebus testaceipes (Cresson) (Hymenoptera: Braconidae): Effect of adult diet on lifespan and egg load

Parasitoid survival and fecundity is generally enhanced with access to carbohydrate food sources. In many agricultural ecosystems, there is often a scarcity of suitable carbohydrates for parasitoids. This study compared the suitability of aphid honeydew and buckwheat nectar as diet for the aphid parasitoid Lysiphlebus testaceipes. Wasp lifespan and egg load were both increased with access to carbohydrates, but no differences were detected between the various carbohydrates diets tested. As aphid honeydew is a sufficient source of nutrition and L.testaceipes is a short-lived species, adding additional sources of carbohydrates like floral nectar strips to the agricultural landscape is unlikely to significantly increase the biological control exerted by L.testaceipes. © 2012 Australian Entomological Society.

Modeling ecological and evolutionary processes in spatially structured populations

Many species inhabit fragmented landscapes, resulting either from anthropogenic or from natural processes. The ecological and evolutionary dynamics of spatially structured populations are affected by a complex interplay between endogenous and exogenous factors. The metapopulation approach, simplifying the landscape to a discrete set of patches of breeding habitat surrounded by unsuitable matrix, has become a widely applied paradigm for the study of species inhabiting highly fragmented landscapes. In this thesis, I focus on the construction of biologically realistic models and their parameterization with empirical data, with the general objective of understanding how the interactions between individuals and their spatially structured environment affect ecological and evolutionary processes in fragmented landscapes. I study two hierarchically structured model systems, which are the Glanville fritillary butterfly in the Åland Islands, and a system of two interacting aphid species in the Tvärminne archipelago, both being located in South-Western Finland. The interesting and challenging feature of both study systems is that the population dynamics occur over multiple spatial scales that are linked by various processes. My main emphasis is in the development of mathematical and statistical methodologies. For the Glanville fritillary case study, I first build a Bayesian framework for the estimation of death rates and capture probabilities from mark-recapture data, with the novelty of accounting for variation among individuals in capture probabilities and survival. I then characterize the dispersal phase of the butterflies by deriving a mathematical approximation of a diffusion-based movement model applied to a network of patches. I use the movement model as a building block to construct an individual-based evolutionary model for the Glanville fritillary butterfly metapopulation. I parameterize the evolutionary model using a pattern-oriented approach, and use it to study how the landscape structure affects the evolution of dispersal. For the aphid case study, I develop a Bayesian model of hierarchical multi-scale metapopulation dynamics, where the observed extinction and colonization rates are decomposed into intrinsic rates operating specifically at each spatial scale. In summary, I show how analytical approaches, hierarchical Bayesian methods and individual-based simulations can be used individually or in combination to tackle complex problems from many different viewpoints. In particular, hierarchical Bayesian methods provide a useful tool for decomposing ecological complexity into more tractable components.

The X Chromosome of Hemipteran Insects: Conservation, Dosage Compensation and Sex-Biased Expression

Insects of the order Hemiptera (true bugs) use a wide range of mechanisms of sex determination, including genetic sex determination, paternal genome elimination, and haplodiploidy. Genetic sex determination, the prevalent mode, is generally controlled by a pair of XY sex chromosomes or by an XX/XO system, but different configurations that include additional sex chromosomes are also present. Although this diversity of sex determining systems has been extensively studied at the cytogenetic level, only the X chromosome of the model pea aphid Acyrthosiphon pisum has been analyzed at the genomic level, and little is known about X chromosome biology in the rest of the order. In this study, we take advantage of published DNA- and RNA-seq data from three additional Hemiptera species to perform a comparative analysis of the gene content and expression of the X chromosome throughout this clade. We find that, despite showing evidence of dosage compensation, the X chromosomes of these species show female-biased expression, and a deficit of male-biased genes, in direct contrast to the pea aphid X. We further detect an excess of shared gene content between these very distant species, suggesting that despite the diversity of sex determining systems, the same chromosomal element is used as the X throughout a large portion of the order.

烟田烟蚜茧蜂的活动规律及其对烟蚜的防治效果

田间释放烟蚜茧蜂能很好地抑制烟蚜种群数量的增长。以常规施药烟田、不施药烟田作对照 ,在前期其对烟蚜的相对防效分别为 8.4%和 5 2 .8% ,中期为 6 4.0 %～ 79.0 %和 6 8.6 %～ 82 .3% ,后期为 93.0 %和 93.5 %。放蜂田烟蚜茧蜂成虫喜欢在烟株中下部叶片活动 ,1 3∶0 0～ 1 4∶0 0是其在烟株中下部活动的高峰期。烟蚜茧蜂对烟株下部叶片上烟蚜的较强选择性与烟蚜密度无关 ,下部叶片上的僵蚜数量均显著高于中、上和顶部。 1 3∶0 0～ 1 4∶0 0利用生物农药对烟株上部叶片上的烟蚜进行防治 ,既是烟蚜茧蜂与生物农药集成组装的切合点 ,又是保护利用田间烟蚜茧蜂的有效措施。

Mass rearing of Aphidius gifuensis (Hymenoptera : Aphidiidae) for biological control of Myzus persicae (Homoptera : Aphididae)

The green peach aphid, Myzus persicae, is a major pest of tobacco, Nicotiana tabacum, in Yunnan province, China, where its control still depends on the use of insecticides. In recent years, the local government and farmers have sought to improve the biological control of this tobacco pest. In this paper, we present methods for mass rearing Aphidius gifuensis, a dominant endoparasitoid of M. persicae on tobacco plants in this region. The tobacco cultivar K326 (N. tabacum) was used as the host plant and M. persicae as the host insect. In the greenhouse, we collected tobacco seedlings for about 35 days (i.e., until the six-true-leaf stage), transferred them to 7.5-cm diameter pots, and kept these plants in the greenhouse for another 18 days. These pots were then transferred to an insectary-greenhouse, where the tobacco seedlings were inoculated with five to seven wingless adult M. persicae per pot. After 3 days, the infested seedlings were moved to a second greenhouse to allow the aphid population to increase, and after an additional 4 +/- 1 days when 182 +/- 4.25 aphid adults and nymphs were produced per pot, they were inoculated with A. gifuensis. With this rearing system, we were able to produce 256 +/- 8.8 aphid mummies per pot, with an emergence rate of 95.6 +/- 2.45%; 69% were females. The daily cost of parasite production (recurring costs only) was US$ 0.06 per 1000 aphid mummies. With this technique, we released 109 800 parasitoids in 1998, 196 000 in 1999, 780 000 in 2000, and 5 600 000 in 2001 during a 2-month period each year This production method is discussed with respect to countrywide usage in biological control and integrated control of M. persicae.

Development and use of parasitoids (Hymenoptera : Aphidiidae & Aphelinidae) for biological control of aphids in China

A review of aphid parasitoids in China with special emphasis on their production, utilization, and conservation is presented with a brief history of Chinese biological control. Twenty genera, 99 species of Aphidiidae and two genera, 11 species of Aphelinidae were recorded in China. Each parasitoid is listed with a brief description of aphids, host plants, areas of study such as taxonomy, biology, bionomics, geographic distribution, rearing, and literature citations. Achievements, status, and problems in aphid parasitoid production, utilization, conservation, and future prospects are detailed for dominant aphid parasitoids such as Aphidius gifuensis Ashmead, A. ervi Haliday and Aphelinus mali Haldeman. Finally, opportunities and challenges of commercialization commercialization of natural enemies, especially aphid parasitoids, in China, are analyzed and discussed.

Studies on Formica lugubris Zetterstedt in Ireland (Hymenoptera, Formicidae)

This thesis is based on studies of Formica lugubris from 1972-1975. While this species' range is diminishing in Ireland, the nests are quite common in the State plantations of South Tipperary. It is not certain that the species is indigenous. Above-ground activity occurs from late-February to the end of October; foraging begins in April. Two territorial "spring-battles" between neighbouring nests are described. Most active nests produced alatae of both sexes and flights were observed on successive June mornings above l7.5°C air temperature. Both polygyny and polycaly seem to be rare. Where the nests occur commonly, the recorded densities are similar to those reported from the continent. Most nests persisted at the same site since 1973. The nest-sites are described by recording an array of nest, soil, tree, vegetation and location variables at each site. Pinus sylvestris is the most important overhead tree. Nests seem to be the same age as their surrounding plantation and reach a maximum of c. 30 years. Nearest-neighbour analysis suggests the sites are overdispersed. Forager route-fidelity was studied and long-term absence from the route, anaesthetization and "removal" of an aphid tree had little effect on this fidelity. There were no identifiable groups of workers specifically honeydew or prey-carriers. Size-duty relationships of workers participating in adult transport are described. Foraging rhythms were studied on representative days: the numbers foraging were linearly related to temperature. Route-traffic passed randomly and an average foraging trip lasted c. four hours. Annual food intake to a nest with 25 000 foragers was estimated at approximately 75 kg honeydew and 2 million prey-items. Forager-numbers and colony-size were estimated using the capture-mark - recapture method: paint marking was used for the forager estimate and an interval radiophosphorus mark, detected by autoradiography, was used for the colony-size estimate. The aphids attended by lugubris and the nest myrmecophiles are recorded.

Identification and analysis of mechanisms involved in a broad-spectrum disease resistant mutant of the winter wheat cv. Guardian

M66 an X-ray induced mutant of winter wheat (Triticum aestivum) cv. Guardian exhibits broad-spectrum resistance to powdery mildew (Blumeria graminis f. sp. tritici), yellow rust (Puccinia striiformis f. sp. tritici), and leaf rust (Puccinia recondita f. sp. tritici), along with partial resistance to stagnonospora nodorum blotch (caused by the necrotroph Stagonosporum nodorum) and septoria tritici blotch (caused by the hemibiotroph Mycosphaerella graminicola) compared to the parent plant ‘Guardian’. Analysis revealed that M66 exhibited no symptoms of infection following artificial inoculation with Bgt in the glasshouse after adult growth stage (GS 45). Resistance in M66 was associated with widespread leaf flecking which developed during tillering. Flecking also occurred in M66 leaves without Bgt challenge; as a result grain yields were reduced by approximately 17% compared to ‘Guardian’ in the absence of disease. At the seedling stage, M66 exhibited partial resistance. M66, along with Tht mutants (Tht 12, Tht13), also exhibit increased tolerance to environmental stresses (abiotic), such as drought and heat stress at seedling and adult growth stages, However, adult M66 exhibited increased susceptibility to the aphid Schizaphis graminum compared to ‘Guardian’. Resistance to Bgt in M66 was characterized with increased and earlier H2O2 accumulation at the site of infection which resulted in increased papilla formation in epidermal cells, compared to ‘Guardian’. Papilla formation was associated with reduced pathogen ingress and haustorium formation, indicating that the primary cause of resistance in M66 was prevention of pathogen penetration. Heat treatment at 46º C prior to challenge with Bgt also induced partial disease resistance to Blumeria graminis f. sp. tritici in ‘Guardian’ and M66 seedlings. This was characterized by a delay in primary infection, due to increased production of ROS species, such as hydrogen peroxide, ROS-scavenging enzymes and Hsp70, resulting in cross-linking of cell wall components prior to inoculation. This actively prevented the fungus from penetrating the epidermal cell wall. Proteomics analysis using 2-D gel electrophoresis identified primary and secondary disease resistance effects in M66 including detection of ROS scavenging enzymes (4, 24 hai), such as ascorbate peroxidase and a superoxidase dismutase isoform (CuZnSOD) in M66 which were absent from ‘Guardian’. Chitinase (PR protein) was also upregulated (24 hai) in M66 compared to ‘Guardian’.Monosomic and ditelosomic analysis of M66 revealed that the mutation in M66 is located on the long arm of chromosome 2B (2BL). Chromosome 2BL is known to have key genes involved in resistance to pathogens such as those causing stripe rust and powdery mildew. The TaMloB1 gene, an orthologue of the barley Mlo gene, is also located on chromosome 2BL. Sanger sequencing of part of the coding sequence revealed no deletions in the TaMloB1 gene between ‘Guardian’ and M66.

Geography and Genetics of Ecological Speciation in Pea Aphids

During ecological speciation, divergent natural selection drives evolution of ecological specialization and genetic differentiation of populations on alternate environments. Populations diverging onto the same alternate environments may be geographically widespread, so that divergence may occur at an array of locations simultaneously. Spatial variation in the process of divergence may produce a pattern of differences in divergence among locations called the Geographic Mosaic of Divergence. Diverging populations may vary in their degree of genetic differentiation and ecological specialization among locations. My dissertation examines the pattern and evolutionary processes of divergence in pea aphids (Acyrthosiphon pisum) on alfalfa (Medicago sativa) and clover (Trifolium pretense). In Chapter One, I examined differences among North American aphid populations in genetic differentiation at nuclear, sequence-based markers and in ecological specialization, measured as aphid fecundity on each host plant. In the East, aphids showed high host-plant associated ecological specialization and high genetic differentiation. In the West, aphids from clover were genetically indistinguishable from aphids on alfalfa, and aphids from clover were less specialized. Thus, the pattern of divergence differed among locations, suggesting a Geographic Mosaic of Divergence. In Chapter Two, I examined genomic heterogeneity in divergence in aphids on alfalfa and clover across North America using amplified fragment length polymorphisms (AFLPs). The degree of genetic differentiation varied greatly among markers, suggesting that divergent natural selection drives aphid divergence in all geographic locations. Three of the same genetic markers were identified as evolving under divergent selection in the eastern and western regions, and additional divergent markers were identified in the East. In Chapter Three, I investigated population structure of aphids in North America, France, and Sweden using AFLPs. Aphids on the same host plant were genetically similar across many parts of their range, so the evolution of host plant specialization does not appear to have occurred independently in every location. While aphids on alfalfa and clover were genetically differentiated in most locations, aphids from alfalfa and clover were genetically similar in both western North America and Sweden. High gene flow from alfalfa onto clover may constrain divergence in these locations.

Efecto de la temperatura sobre el pulgón del repollo (Brevicoryne brassicae L.) (Homóptera : Aphidoidea)

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Funcionalidad biodemográfica del pulgón Nasonovia Ribisnigri (Mosley) (Hemiptera : Aphididae) en cultivares de lechuga Lactuca sativa (l.) tipo mantecoso en condiciones controladas

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The relationship between agricultural intensification and biological control: experimental tests across Europe

Agricultural intensification can affect biodiversity and related ecosystem services such as biological control, but large-scale experimental evidence is missing. We examined aphid pest populations in cereal fields under experimentally reduced densities of (1) ground-dwelling predators (-G), (2) vegetation-dwelling predators and parasitoids (-V), (3) a combination of (1) and (2) (-G-V),compared with open-fields (control), in contrasting landscapes with low vs. high levels of agricultural intensification (AI), and in five European regions. Aphid populations were 28%, 97%, and 199% higher in -G, -V, and -G -V treatments, respectively, compared to the open fields, indicating synergistic effects of both natural-enemy groups. Enhanced parasitoid : host and predator : prey ratios were related to reduced aphid population density and population growth. The relative importance of parasitoids and vegetation-dwelling predators greatly differed among European regions, and agricultural intensification affected biological control and aphid density only in some regions. This shows a changing role of species group identity in diverse enemy communities and a need to consider region-specific landscape management.

Effect of temperature and prey in the biology of Scymnus subvillosus (Goeze) (Coleoptera: Coccinellidae)

Dissertação de Mestrado, Biotecnologia em Controlo Biológico, 18 de Dezembro de 2013, Universidade dos Açores.