The Helicoverpa armigera stunt virus (HaSV) is a member of a family of small nonenveloped (+) ssRNA insect viruses currently known as the Tetraviridae. This family is unique in terms of the T=4 quasi-symmetry of its capsid particles and the unusually narrow host range and tissue tropism. Assembly of tetraviral particles has been well characterised and involves the combination of 240 copies of a single capsid precursor protein (VCap) into a procapsid followed by autoproteolytic cleavage to yield the major (β) and minor (γ) capsid subunits within the mature particle. HaSV has two genomic RNAs, RNA 1 encoding the replicase and RNA 2 encoding VCap and p17, the ORF of which lies upstream of and overlaping with the 5’ end of the VCap ORF. Prior to this study, Vlok (2009) used a plasmid expression system to study RNA packaging in HaSV VLPs assembled in Spodoptera frugiperda 9 (Sf9) cells co-expressing p17 and VCap. The study showed that the p17 ORF was required for the packaging of RNA 2 during capsid assembly but it was unclear whether p17 expression was required for packaging. In addition, expression from the transfected plasmids was sub-optimal affecting both the yield of VLPs and the detection of p17. The aim of this study was to use the plasmid system to test whether p17 expression was required for plasmid-derived VLP RNA packaging and then develop a baculovirus-mediated system to test this hypothesis. By using a plasmid in which the start codon of p17 was mutated, it was shown that p17 expression was required for RNA 2 packaging into plasmid-VLPs. For the baculovirus system, four recombinant baculoviruses based upon the pFastBac Dual expression system, were constructed. These included Bac20, expressing wild type RNA 2, Bac21, RNA 2 with p17 silenced, Bac23, RNA 2 and p17 expressed on a separate transcript and Bac24, RNA 2 with p17 silenced plus p17 expressed on a separate transcript. Assembly of VLPs was more efficient using the baculovirus expression system and p17 expression was observed in cells infected with Bac20, Bac23 and Bac24, but not Bac21. In contrast to the plasmid-VLPs, bac-VLPs did not require p17 for the encapsidation of RNA 2. In addition to RNA 2, Bac23 and Bac24 packaged the p17 mRNA transcribed separately from RNA 2. This insinuated that bac-VLPs may be packaging RNA non-selectively. It was proposed that p17 may play a role in packaging in an RNA-limiting environment (plasmid system) but functioned differently when viral RNA was in excess (baculovirus system). This data points to the importance of developing a replication system for the analysis of the packaging pathways of these viruses and this study has laid down the foundations for such a system in which RNA 1 and RNA 2 can be introduced into a single cell by means of a single recombinant virus.

Transgenic corn and cotton expressing crystalline (Cry) insecticidal proteins from Bacillus thuringiensis (Bt) were commercially introduced in 1996. This technology has greatly improved the control of several key lepidopteran insect pests tobacco budworm, Heliothis virescens (Farbricius), pink bollworm, Pectinophora gossypiella (Saunders), southwestern corn borer, Diatraea grandiosella (Dyar), and European corn borer, Ostrinia nubilalis (Hübner). The corn earworm, Helicoverpa zea, has been more difficult to control using Bt crops and supplemental insecticide applications are often needed to prevent economic losses. A major threat to the longevity of transgenic technology is the evolution of resistance, especially when an insect pest infests both Bt corn and cotton in the same growing season. Similar Cry proteins are currently expressed in both corn and cotton commercial production systems. At least one generation per year develops on Bt corn before infesting cotton. Given that H. zea infests both crop hosts at some point every year, the objectives of this study were to evaluate the contribution and influence of Genuity VT3 PRO corn (expressing Cry1A.105 and Cry2Ab) on H. zea density, fitness, and selection on BollGard II cotton (expressing Cry1Ac and Cry2Ab). Non-Bt and VT3 PRO corn fields were sampled for larval density and any observed larvae were collected. Non-Bt field corn supported 61% more larvae compared to VT3 PRO fields. Larvae infesting non-Bt corn developed faster than those infesting VT3 PRO corn. Larvae collected from VT3 PRO corn had significantly higher pupal weight two out of the three years of this study. Pupae from VT3 PRO corn also had longer pupal duration two out of three years compared to pupae of larvae collected from non-Bt corn. Offspring from larvae reared on VT3 PRO corn had a higher LC50 compared to offspring from larvae reared on non-Bt corn. H. zea susceptibility to Bt is highly variable but the results presented here indicate that dual-gene corn hybrids such as VT3 PRO can select for H. zea populations with a higher propensity for causing damage in Bt cotton.

Thesis (Ph. D.)--University of Missouri-Columbia, 2003. / Some text in Spanish. Typescript. Vita. Includes bibliographical references (leaves 103-114). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri-Columbia, 2003. / Some text in Spanish. Typescript. Vita. Includes bibliographical references (leaves 103-114). Also available on the Internet.

Thesis (Ph. D. )--Washington State University, August 2006. / Includes bibliographical references (p. 215-227).

H. armigera larvae are a key stage for pest management in conmercial irrigated cotton crops in South Africa. Effective survey methods for detecting larval populations in the field require an understanding of the biology of the larvae, particularly their feeding habits. Their feeding is central to the development of pest threshold levels for the implementation of integrated control programmes. This applies to routine surveys for the larvae as well as to the damage they cause. Biological characteristics of the larvae are described with the emphasis on the identification of the larval instars, which were consistently five in number in both field and laboratory populations. The distribution of H. armigera larvae on cotton plants in the field was examined, but was found to more or less random; had there been a clear preference for any height zones or compass direction this would have been an obvious avenue for improving the survey methods currently in use. Details of field and laboratory investigations of the selection of feeding sites by the larvae are given. The study confirmed a clear preference by the larvae for cotton buds, flowers and bolls (in the thesis collectively called "fruiting forms"), over leaves. There were indications that the larvae selected flowers more readily than buds or bolls. This "preference", however, is shown to be of no practical value for refining survey methods. Damage levels to cotton due to B. armigera are discussed. Both direct losses and indirect losses due to the abortion of fruiting forms are considered. These criteria are inadequate since they do not take into account the ability of cotton plants to compensate for these losses. It is concluded that this compensation by cotton plants should be taken into account in further studies of the pest status of B. armigera.

The Tachinid fly, Palexorista laxa (Curran) is an important parasitoid of Heliothis armigera (Hubner) in Africa and India. The biological characteristics of the parasitoid were studied with an emphasis on its host instar preference, host range, seasonality and percentage parasitism. The developmental stages were also studied and described. A high host mortality (48%-100%) due to parasite attack occurred when small larvae were parasitised (2nd & 3rd instars). A preference for the 4th and 5th host instars was shown by the flies. P. laxa was found to be a non-specific parasitoid and parasitised a number of lepidopterous larvae in the laboratory including Busseola fusca (Fuller), Chilo partellus (Swinhoe) and Eldana saccharina Walker, but preferred Ii. armigera. The percentage parasitism on H. armigera in the field was variable peaking at 48% in 1988/89. Of the various parasitoids recorded from H. armigera on sunflower P. laxa was by far the most abundant. Mating occurred immediately after emergence with the males emerging a day earlier than the females. Males were capable of multiple matings whereas females mated only once in a lifetime. P. laxa females produced incubated macrotype eggs which hatched almost immediately after oviposition. The duration of development of the incubated egg and larval stage was 6,7 days, the pupa 10,4 days for females and 9,4 days for males, and the adult lived up to 50 days at 25°C. A reduction in pupal weight, an increase in the duration of the pupal stage and higher pupal mortality accompanied an increase in density of parasitoids per host. A linear relationship was shown between adult mass and number of ovarioles and eggs in P. laxa females. The oviposition period was 26,9 days with a high variation in daily progeny production. The total fertility per female was 126,3 puparia. "Inexperienced" females deposited incubated eggs on all parts of the host body while "experienced" females confined oviposition to the head and thorax. P. laxa females did not discriminate between previously parasitised and non-parasitised hosts. The effect of temperature on larval and pupal survival and development time was studied at 20°, 22°, 25°, 28°and 30°C. As expected, duration of development decreased with an increase in temperature, and there was a reduction in mortality.

In field experiments, thresholds, spray timings, and bollworm oviposition were conducted to evaluate the current recommended action thresholds for bollworm (Lepidoptera: Noctuidae) in cotton. Based on studies conducted evaluating thresholds, insecticide applications varied across Bt technologies. Bollgard 3 required less sprays when compared to Bollgard II and non-Bt. A second experiment highlighted the importance of timely insecticide applications for managing bollworm populations in Bollgard II cotton based on insect damage. The highest yields were associated with insecticide applications made during the timeframe that cotton is the most susceptible to bollworm damage. The last experiment was conducted to determine if oviposition varied throughout the plant canopy. Oviposition occurred throughout the whole plant, however, the greatest relationship of egg lay was observed in the top three nodes of the plant. From this research, studies would suggest that the current recommended bollworm thresholds in Mississippi cotton production systems, at this time, do not need to be adjusted.

Desde a década de 50, os cientistas tem utilizado radiação ionizante para esterilizar insetos, que são liberados na natureza para acasalar, mas sem nenhuma descendência. Conhecida como a técnica do inseto estéril (TIE), este método de controle de inseto vem tradicionalmente utilizando radiações ionizantes para esterilizar os insetos, sendo uma técnica que não gera resíduo, e pode agir em sinergia com as demais técnicas dentro do manejo integrado de pragas. O Brasil por vários anos vem lutando contra o aumento de pragas, introduzindo novas táticas e técnicas dentro dos programas de MIP, para driblar a resistência dos produtos químicos, como: diminuir os resíduos dos agrotóxicos; para algumas culturas importantes do o nosso país, temos um largo espectro de pragas ocorrendo do inicio ao fim da colheita, uma delas é a cultura do algodoeiro e dentre as pragas chave dessa cultura, temos algumas lagartas extremamente importantes, entre elas Heliothis virescens e Helicoverpa armigera. Essas espécies são parecidas morfologicamente, sendo a segunda identificada a poucos anos aqui no Brasil. Ainda não há trabalhos no Brasil utilizando TIE como ferramenta adicional para lepidópteros, portanto objetivou-se com esse estudo avaliar o efeito de doses de radiação gama nas diferentes fases do ciclo evolutivo de Heliothis virescens e Helicoverpa armigera, bem como avaliar a esterilidade na geração P e a capacidade de competição dos insetos irradiados com os não irradiados. A fase pupa foi a que apresentou um melhor resultado, pois com 75 Gy atingiu a esterilidade em Heliothis virecens e 100 Gy esterilizou Helicoverpa armigera, portanto contemplou a fase e a dose escolhida para avaliar a competição entre os insetos irradiados e os insetos normais de ambas as espécies. E tanto Heliothis virecens como Helicoverpa armigera apresentaram um resultado satisfatório, pois os insetos irradiados conseguiram reduzir significativamente a viabilidade dos ovos na proporção de 9: 1: 1 / Since the 1950s, scientists have used ionizing radiation to sterilize insects, which are released in nature to mate, but without any progeny. Known as the sterile insect technique (TIE), this method of insect control has traditionally used ionizing radiation to sterilize insects, a technique that does not generate residues, and can act in synergy with the other techniques within integrated pest management. For several years, Brazil has been fighting against the increase of pests, introducing new tactics and techniques within the IPM programs, to overcome the resistance of chemical products, such as: reducing the residues of agrochemicals; For some important crops of our country, we have a wide spectrum of pests occurring from the beginning to the end of the harvest, one of them is the cotton crop and among the key pests of this crop, we have some extremely important caterpillars, among them Heliothis virescens and Helicoverpa armigera These species are morphologically similar, the second being identified a few years ago in Brazil. There are still no studies in Brazil using TIE as an additional tool for Lepidoptera, therefore the purpose of this study was to evaluate the effect of doses of gamma radiation in the different phases of the evolutionary cycle of Heliothis virescens and Helicoverpa armigera, as well as to evaluate the sterility in generation P And the ability of insects to irradiate with non-irradiated insects. The pupal phase presented the best result because 75 Gy achieved sterility in Heliothis virecens and 100 Gy sterilized Helicoverpa armigera, therefore it contemplated the phase and dose chosen to evaluate the competition between the irradiated insects and the normal insects of both species. Both Heliothis virecens and Helicoverpa armigera presented a satisfactory result, as the irradiated insects managed to significantly reduce the viability of the eggs in a ratio of 9: 1: 1

Submitted by Caroline Periotto (carol@ufscar.br) on 2016-10-10T17:24:48Z No. of bitstreams: 1 TeseACMR.pdf: 2424700 bytes, checksum: 8f6c5b8a916ee974192846236c7e1dc9 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-10-20T18:33:03Z (GMT) No. of bitstreams: 1 TeseACMR.pdf: 2424700 bytes, checksum: 8f6c5b8a916ee974192846236c7e1dc9 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-10-20T18:33:09Z (GMT) No. of bitstreams: 1 TeseACMR.pdf: 2424700 bytes, checksum: 8f6c5b8a916ee974192846236c7e1dc9 (MD5) / Made available in DSpace on 2016-10-20T18:33:16Z (GMT). No. of bitstreams: 1 TeseACMR.pdf: 2424700 bytes, checksum: 8f6c5b8a916ee974192846236c7e1dc9 (MD5) Previous issue date: 2016-06-03 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / This study objective to identify the diversity of predators through the use of serology producing specific antiserums for the main pests found in maize, Spodoptera frugiperda, Helicoverpa armigera and H. zea, Rhopalosiphum maidis and uses it to determine their predators. Pest samples were macerated in 0.85% saline solution. The macerated were centrifuged and the supernatants used as immunizing antigens for obtaining antiserum. For this purpose, a rabbit was immunized with 3.0 ml of the immunizing antigen on the lymph node region. Homologous serological tests were performed in double diffusion in agar. Homologous serological reactions were positive after seven days of antigen inoculation. The technique has sensitivity to detect predation of the four pests studied in this paper. The tests were positive for a prey in the digestive tract of the predator to 96 hours of ingestion. Regarding the number of preys, there was no difference in the lines, they only grew stronger and sharper. In laboratory, D. luteipes, O. vnigrum and C. externa presented preference for R. maidis and E. annulipes for S. frugiperda. In tests where predators were collected in the field, serological tests showed that there was a certain food preference of D. luteipes and O. v-nigrum by S. frugiperda. E. annulipes, H. axyridis, E. connexa and C. externa, by the aphid. After a single injection of antigen in rabbit’s lymph node region, it was possible to get an antiserum specific for pests of corn. / O presente trabalho teve como objetivo identificar a diversidade alimentar dos predadores através do uso da serologia produzindo os antissoros específicos para as principais pragas encontradas na cultura do milho Spodoptera frugiperda, Helicoverpa armigera, Helicoverpa zea, Rhopalosiphum maidis. Amostras das pragas foram maceradas em solução salina NaCl (0,85%). Os macerados foram centrifugados, e os sobrenadantes, utilizados como antígenos imunizantes para obtenção do antissoro. Para esse propósito um coelho foi imunizado com 3,0 ml do antígeno imunizante na região do linfonódulo. Testes serológicos homólogos foram realizados em dupla difusão em ágar. Reações serológicas homólogas foram positivas após sete dias após da imunização do antígeno. A técnica tem sensibilidade para detectar predação de das quatro pragas estudadas nesse trabalho. Os testes foram positivos para uma presa no trato digestivo do predador até 96hs de sua ingestão. Com relação ao número de presas ingeridas não houve diferença nas linhas de precipitação, apenas ficaram mais fortes e nítidas. Em laboratório a preferência alimentar de Doru luteipes, Olla v-nigrum e Chrysoperla externa apresentaram preferência alimentar por R. maidis e Euborellia annulipes por S. frugiperda. Nos testes onde os predadores foram coletados no campo os testes serológicos mostraram que houve uma preferência alimentar de D. luteipes e O. v-nigrum por S. frugiperda. E. annulipes, H. axyridis, E. connexa, e C. externa por pulgão. Concluiu-se que após uma única injeção do antígeno na região do linfonódulo do coelho, foi possível obter-se antissoro específico para as pragas do milho.