Respondents of the 1986 Broccoli Growers Survey grew an average of 6.2 acres of broccoli in Fall 1986; popular varieties were Green Valiant (87.5% of respondents), Packman and Emperor (62.5%). Less than 5% of tobacco land was converted to broccoli production by 62.5% of the growers. Before heading, 2.4 insecticidal sprays were applied for worms at $20.51/acre. During heading, 2.4 insecticidal sprays were applied for worms at $19.40/acre. Under normal growing conditions, a nine-day mean of 8.4 and a 15-day mean of 12.1 cabbage looper larvae/plant did not significantly reduce yield in the Spring and Fall respectively. However, means of 7.4 and 11.1 cabbage looper larvae/plant reduced yield (head weight) under drought conditions. Harvest date was not affected by the above cabbage looper levels. In Fall 1986, a reduction in yield occurred when plants had 50% or more defoliation and harvest date was significantly delayed at 100% defoliation. In the drought stressed Spring 1987 planting, a stimulation in yield occurred at 25% defoliation and 75% defoliation was needed to significantly reduce yield, but there was no difference in harvest date. The imported cabbageworm was the most abundant pest of broccoli in Montgomery Co., VA. Based on counts of all immature stages of the insects for three seasons, Packman and Southern Comet varieties were less preferred compared to Green Defender. Laboratory studies indicated that imported cabbageworm larvae developed faster to the fifth instar on Packman broccoli than those reared on Green Defender, Southern Comet or Emperor. Difference in amount consumed between varieties was not detected. Pupae reared on Southern Comet weighed significantly more than those reared on Packman. / M.S.

The susceptibility of a population of diamondback moth, Plutella xvlostella L (Lepidoptera: Plutellidae), collected from Montgomery County, Virginia, and its ability to develop resistance to Bacillus thuringiensis was evaluated. The field population of diamondback moths was found to be susceptible to B. thuringiensis. Selection pressure at 153 I.U. of B. thuringiensis endotoxin per ml for nine generations did not cause any significant difference in mean mortality of third instar diamondback moths although there was a trend towards lower mortality, there was significant negative linear correlation between generation and mean mortality of diamondback moth larvae (P = 0.003, r²= 0.73). LC₅₀ increased from 264 I.U. of B. thuringiensis endotoxin per ml in generation I to 514 LU/ml in generation IX. The interrelationship of B. thuringiensis to diamondback moth and its primary parasitoid, Diadegma insulare Cress (Hymenoptera. Ichneumonidae), was studied by determining: the differential response of third instar diamondback moth, parasitized and unparasitized, to B. thuringiensis, and the ability of D. insulare to discriminate between B. thuringiensis-treated and untreated hosts. There was no significant difference (P > 0.05) between mean mortality of parasitized and unparasitized larvae at each of the three concentrations consisting of 154, 334, and 2237 I.U. of B. thuringiensis endotoxin per ml. The regressions of the response of parasitized and unparasitized larvae, however, were highly significant (P = 0.0001). The LC₅₀s of parasitized versus unparasitized larvae were 373 and 176 I.U./ml B. thuringiensis endotoxin, respectively. Female D. insulare did not discriminate between B. thuringiensis-treated and untreated hosts. The percentage of D. insulare females emerging from B. thuringiensis-treated larvae (41.4%) was not significantly different from that of untreated larvae (32.0 %). Mean mortality of third instar diamondback moth subjected to B. thuringiensis endotoxin at 153, 334, and 2237 I.U./ml were not significantly different at temperatures of 15 and 20 °C, but were significantly lower than that at 30 °C. The effects of B. thuringiensis endotoxin residues on leaves under room conditions [27 ± 1 °C, RH 27 ± 72%, and 8:16 (L:D)] were not significantly different at 2 and 192 hours after treatment. / Master of Science

The diamondback moth, Plutella xylostella (Linnaeus 1758.) (Lepidoptera: Plutellidae), is a major insect pest of Brassica crops in many parts of the world leading to economic losses amounting to an estimated US$ 4-5 billion. Although parasitoids (biological control agents) play a major role in suppressing the pest populations during November – May in South Africa, the pest reaches outbreak status during September and October due to low impact of parasitoids, which has necessitated regular application of insecticides. However, insecticide applications have often resulted in the pest developing resistance, and when coupled with the negative effects of several insecticides on parasitoids, integration of the two pest control strategies for effective management of P. xylostella population density has been difficult to achieve. One approach that has received little attention is integration of host plant resistance (bottom-up effect) and biological control (top-down effect) for effective management of P. xylostella. However, the interaction between host plants, the insect pest, and parasitoids is not simple and straight forward, as strong negative impact of host plants on fitness of the insect pest can be cascaded up the food chain and have a negative impact on a given parasitoid, which in turn may reduce the desired complementary effect between the two pest control strategies. To identify optimal interactions between cabbage (Brassica oleracea L. var. capitata, Brassicaceae), P. xylostella and its larval parasitoid Cotesia vestalis (Haliday 1834) (Hymenoptera: Braconidae), this study investigated (i) the effects of seven cabbage cultivars (Empowa, Hollywood F1, Megaton, Leano, Menzania, Beverley Hills and Karabo) on fitness parameters (survival, developmental time, pupal weights, longevity without food and oviposition rates) of P. xylostella; (ii) the influence of the same host plant cultivars on fitness parameters (developmental time, pupal weights, longevity xi without food, fecundity, emergence rate and sex ratio) of C. vestalis. Furthermore, net reproductive rates and the intrinsic rates of natural increase were calculated for C. vestalis that emerged from hosts fed on each of the cultivars. All experiments were conducted in climate-controlled laboratory rooms maintained at 22 ± 1 ºC (mean ± S.D.), 60 ± 5 % RH and 16L: 8D photoperiod. Under the no choice test, overall survival of P. xylostella immature stages was highest on Karabo (67.26%) and lowest on Megaton (44.92%). The larval and pupal developmental period, and generation time was prolonged on Empowa (18.48 days), Karabo (14.64 days) and Beverly Hills (17.48 days), while developmental period on Hollywood F1 (13.79 days) was shortest. Male and female P. xylostella pupal weights were lighter from larvae that fed on Megaton (4.13 and 4.65 mg), Menzania (4.53 and 4.91 mg), and Hollywood F1 (4.11 and 5.08 mg), whereas pupal weights from Karabo (6.0 and 6.82 mg) were the heaviest. Unfed female moths originally reared on Beverley Hills had the highest longevity (5.05 days), whereas those reared on Leano (3.54 days) and Megaton (3.89 days) had the shortest life span. Under the choice-test, P. xylostella moth laid significantly more eggs on Empowa (48.8%) and Hollywood F1 (45.6%) and least on Menzania (11.8%) and Leano (10.6%). Megaton was more resistant to P. xylostella due to lower survival rates of immature stages, lower pupal weights and moth longevity. The generation time of C. vestalis was shortest on Karabo (10.10 days) and Leano (10.38 days), and longest on Megaton (12.57 days) and Empowa (12.80 days). The highest pupal weight of C. vestalis was obtained from parasitoids reared from P. xylostella fed Menzania (5.4 mg), Megaton (5.25 mg) and Beverly Hills (4.85 mg) and the lightest on Karabo (3.8 mg). Parasitoids reared on larvae that fed on Hollywood F1 lived the longest (2.28 days) followed by Menzania (1.94 days) and Beverly Hills (1.8 days), whereas those whose hosts fed on Leano had shortest life span (0.83 days). Despite the parasitoids from Megaton hosts being heavier, their fecundity and number of female progeny per female (16.87 and 3.60, respectively) were lowest. Cotesia vestalis fecundity and daughters produced per female were highest on hosts fed on Menzania (38.00 and 9.13, respectively) and Beverly Hills (32.87 and 9.07, respectively). As a consequence, the net reproductive rate (R0) and intrinsic rate of increase (r) were higher on Menzania (7.87 and 0.58, respectively) and Beverly Hills (8.29 and 0.62, respectively). As survival and overall fitness of P. xylostella was lower on Megaton, this cultivar can play a major role in restricting population growth of this pest and thus generational number of eggs deposited on it during September and October. However, this strong bottom-up effect of Megaton on P. xylostella was cascaded up the food chain, as overall fitness of C. vestalis was lower on hosts developing on it. In contrast, the overall fitness of C. vestalis was higher on hosts that developed on Menzania and Beverly Hills. As these cultivars showed potential to sustain population density of C. vestalis at higher levels, it is also assumed that the period required for the parasitoid to reach the critical density to suppress the host population at a lower average density will be reached quicker than on other cultivars. Thus, their cultivation may improve biological control of P. xylostella during November–May in South Africa / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture)

Doctor of Philosophy / Department of Entomology / Raymond A. Cloyd / James R. Nechols / Raymond A. Cloyd / James R. Nechols / Greenhouse studies were conducted to examine direct effects of light intensity and nitrogen source on primary and secondary metabolism of pac choi (Brassica rapa L. var. chinensis cv. ‘Mei Qing Choi’) and indirect effects on diamondback moth (Plutella xylostella L.)(DBM). In the first study, plants were exposed to high and low light intensities during different times of the year, resulting in a range of light intensities. From four experiments, plants exhibited higher phenolic content, greater shoot biomass, and higher C:N ratios under high light intensity, whereas plants under low light intensity contained higher protein. Ferulic acid increased under high light intensity, and this increase was negatively correlated with male DBM body weights. However, DBM developed faster on plants in the August experiment (high light), compared to the July experiment (lower light). This implies that light intensity may not be affecting DBM through plant-mediated changes unless reduced male weights confer a reduction in larval consumption. In the nitrogen source study, application of an organic source of nitrogen (fish hydrolysate fertilizer) was compared to a conventional fertilizer to determine whether nitrogen source directly impacts pac choi chemistry and biomass, thus indirectly impacting DBM fitness. In two experiments, there was no significant effect of fertility treatment on pac choi nutrients or biomass, with the exception of percent leaf phosphorus, which was significantly higher in the conventional fertility treatment. For DBM, percent survival and cohort development were significantly reduced on pac choi receiving the organic fertilizer. Calcium and magnesium were significantly higher in pac choi infested with DBM larvae than plants without DBM. In addition, calcium was negatively correlated with female DBM body weights in one experiment for the organic treatment. Overall, this study demonstrated that pac choi plants that received the organic fertilizer were similar to pac choi plants that received a conventional fertilizer with the exception of phosphorus. Furthermore, female DBM body weights were negatively impacted by calcium in the organic treatment. As multiple fitness traits for DBM were negatively affected in the organic treatment, pac choi crops grown with fish hydrolysate fertilizer may experience less feeding from DBM.

Insect pests represent major threats to food production, biodiversity conservation, and human and animal health. Currently, the most widespread strategy to control their populations is through the spraying of synthetic chemical insecticides. However, the overuse of these compounds has had significant negative environmental consequences. Additionally, our reliance on insecticides has resulted in major reductions in their efficacy through pest-evolved resistance. To successfully manage insect pests, while avoiding environmental degradation, thus requires the development of novel, more sustainable, pest management strategies. Recent advances in our understanding of recombinant DNA methods and molecular biology have allowed the application of transgenic tools to pest management. Here, synthetic genes can be engineered, transformed into the genomes of pest species, and transported into wild target populations through the natural mating behaviour of the insect. A strategy in which these transgenes are lethal to those insects inheriting them in the field is known as RIDL – Release of Insects carrying a Dominant Lethal. A variant of RIDL limits this lethality to females – female specific RIDL (fsRIDL) – which explicitly targets the reproductive capacity of a target population. The aim of this thesis is to investigate the application of such an fsRIDL strategy to the diamondback moth (Plutella xylostella L.). This economically important pest of brassica crops is highly adept at developing resistance to insecticides and is considered extremely difficult to manage effectively. I present findings which demonstrate the power of diamondback moth lines transformed with fsRIDL transgenes to eliminate target pest populations, and combine synergistically with other transgenic control strategies such as Bt crops in counteracting the evolution of pesticide resistance. Additionally, an exploration into an alternative gene expression system to that used in current RIDL strategies – the Q system – suggests that not all expression systems will be suitable for transgene control within this highly specific framework. It is hoped that this work will contribute towards the effective control of the diamondback moth, and form a model for the sustainable control of other lepidopteran species through genetic pest management.

The diamondback moth, Plutella xylostella (Linnaeus 1758.) (Lepidoptera: Plutellidae), is a major insect pest of Brassica crops in many parts of the world leading to economic losses amounting to an estimated US$ 4-5 billion. Although parasitoids (biological control agents) play a major role in suppressing the pest populations during November – May in South Africa, the pest reaches outbreak status during September and October due to low impact of parasitoids, which has necessitated regular application of insecticides. However, insecticide applications have often resulted in the pest developing resistance, and when coupled with the negative effects of several insecticides on parasitoids, integration of the two pest control strategies for effective management of P. xylostella population density has been difficult to achieve. One approach that has received little attention is integration of host plant resistance (bottom-up effect) and biological control (top-down effect) for effective management of P. xylostella. However, the interaction between host plants, the insect pest, and parasitoids is not simple and straight forward, as strong negative impact of host plants on fitness of the insect pest can be cascaded up the food chain and have a negative impact on a given parasitoid, which in turn may reduce the desired complementary effect between the two pest control strategies. To identify optimal interactions between cabbage (Brassica oleracea L. var. capitata, Brassicaceae), P. xylostella and its larval parasitoid Cotesia vestalis (Haliday 1834) (Hymenoptera: Braconidae), this study investigated (i) the effects of seven cabbage cultivars (Empowa, Hollywood F1, Megaton, Leano, Menzania, Beverley Hills and Karabo) on fitness parameters (survival, developmental time, pupal weights, longevity without food and oviposition rates) of P. xylostella; (ii) the influence of the same host plant cultivars on fitness parameters (developmental time, pupal weights, longevity xi without food, fecundity, emergence rate and sex ratio) of C. vestalis. Furthermore, net reproductive rates and the intrinsic rates of natural increase were calculated for C. vestalis that emerged from hosts fed on each of the cultivars. All experiments were conducted in climate-controlled laboratory rooms maintained at 22 ± 1 ºC (mean ± S.D.), 60 ± 5 % RH and 16L: 8D photoperiod. Under the no choice test, overall survival of P. xylostella immature stages was highest on Karabo (67.26%) and lowest on Megaton (44.92%). The larval and pupal developmental period, and generation time was prolonged on Empowa (18.48 days), Karabo (14.64 days) and Beverly Hills (17.48 days), while developmental period on Hollywood F1 (13.79 days) was shortest. Male and female P. xylostella pupal weights were lighter from larvae that fed on Megaton (4.13 and 4.65 mg), Menzania (4.53 and 4.91 mg), and Hollywood F1 (4.11 and 5.08 mg), whereas pupal weights from Karabo (6.0 and 6.82 mg) were the heaviest. Unfed female moths originally reared on Beverley Hills had the highest longevity (5.05 days), whereas those reared on Leano (3.54 days) and Megaton (3.89 days) had the shortest life span. Under the choice-test, P. xylostella moth laid significantly more eggs on Empowa (48.8%) and Hollywood F1 (45.6%) and least on Menzania (11.8%) and Leano (10.6%). Megaton was more resistant to P. xylostella due to lower survival rates of immature stages, lower pupal weights and moth longevity. The generation time of C. vestalis was shortest on Karabo (10.10 days) and Leano (10.38 days), and longest on Megaton (12.57 days) and Empowa (12.80 days). The highest pupal weight of C. vestalis was obtained from parasitoids reared from P. xylostella fed Menzania (5.4 mg), Megaton (5.25 mg) and Beverly Hills (4.85 mg) and the lightest on Karabo (3.8 mg). Parasitoids reared on larvae that fed on Hollywood F1 lived the longest (2.28 days) followed by Menzania (1.94 days) and Beverly Hills (1.8 days), whereas those whose hosts fed on Leano had shortest life span (0.83 days). Despite the parasitoids from Megaton hosts being heavier, their fecundity and number of female progeny per female (16.87 and 3.60, respectively) were lowest. Cotesia vestalis fecundity and daughters produced per female were highest on hosts fed on Menzania (38.00 and 9.13, respectively) and Beverly Hills (32.87 and 9.07, respectively). As a consequence, the net reproductive rate (R0) and intrinsic rate of increase (r) were higher on Menzania (7.87 and 0.58, respectively) and Beverly Hills (8.29 and 0.62, respectively). As survival and overall fitness of P. xylostella was lower on Megaton, this cultivar can play a major role in restricting population growth of this pest and thus generational number of eggs deposited on it during September and October. However, this strong bottom-up effect of Megaton on P. xylostella was cascaded up the food chain, as overall fitness of C. vestalis was lower on hosts developing on it. In contrast, the overall fitness of C. vestalis was higher on hosts that developed on Menzania and Beverly Hills. As these cultivars showed potential to sustain population density of C. vestalis at higher levels, it is also assumed that the period required for the parasitoid to reach the critical density to suppress the host population at a lower average density will be reached quicker than on other cultivars. Thus, their cultivation may improve biological control of P. xylostella during November–May in South Africa / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture)

Made available in DSpace on 2014-06-11T19:25:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-02-22Bitstream added on 2014-06-13T18:26:16Z : No. of bitstreams: 1 carvalho_js_me_jabo.pdf: 532415 bytes, checksum: cfa12555262ca20f28efd7aad4d084ec (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O trabalho foi desenvolvido no Laboratório de Biologia e Criação de Insetos (LBCI) da Faculdade de Ciências Agrárias e Veterinárias, Unesp – Jaboticabal, SP, para estudar os efeitos de diferentes concentrações de sinigrina aplicada na superfície foliar de couve e brócolis, em alguns parâmetros biológicos de Plutella xylostella. Para realização dos testes, sinigrina foi aplicada à parte ventral/dorsal das folhas de brássicas das cultivares ‘Da Geórgia’ (couve-manteiga) e ‘Ramoso Piracicaba Precoce’ (brócolis), em solução 5% de Tween 20® em diferentes concentrações (0,0; 0,2; 0,4; 0,8; 1,6 e 3,2 mg/mL). Assim como, avaliou-se o consumo, pelas lagartas, através de medidas de peso fresco, peso seco, área foliar e escala visual de notas para os danos. Observou-se que concentração baixa de sinigrina, em couve e em brócolis, não prejudicou o desenvolvimento de P. xylostella, porém altas concentrações afetaram o parâmetro viabilidade. A análise de agrupamentos, pelo método de “cluster”, mostrou que a dose 0,2mg/mL de sinigrina em ambas variedades apresentou os maiores contrastes, não pertencendo a nenhum grupo. O índice Potencial Reprodutivo Corrigido foi determinante apenas para couve, indicando boa capacidade reprodutiva das lagartas alimentadas com a menor concentração de sinigrina. Nas doses extremas, 0,2 e 3,2mg/mL, observou-se aumento no consumo foliar em couve e diminuição em brócolis. O consumo foi maior pelas lagartas alimentadas com folhas de brócolis. Em geral, o comportamento da traça foi diferente nas cultivares testadas, mas essas diferenças podem estar relacionadas com a quantidade de sinigrina presente naturalmente nas folhas de cada espécie. / The work was carried out in the Laboratório de Biologia e Criação de Insetos (LBCI), at the Faculdade de Ciências Agrárias e Veterinárias, Unesp, Jaboticabal, SP, with the objective to evaluate the effect of sinigrin applied on foliar surface of cabbage and broccoli, on the biological aspects of Plutella xylostella. It was evaluated: the biology aspects of diamondback moth feeding on cabbage ‘Da Georgia’ and broccoli ‘Ramoso Piracicaba Precoce’ treated with different sinigrin concentrations (0.0, 0.2, 0.4, 0.8, 1.6 e 3.2mg/mL in 5% of Tween 20® solution, as well as, the consumption, measuring fresh weight, dry weight, foliar area and visual grade scale to the damage. It was observed that low sinigrin concentration, in cabbage and broccoli, does not affect P. xylostella development, however, high concentrations affect the viability. The group analysis by “cluster method”, showed that 0.2mg/mL of sinigrin of both varieties presented the biggest contrast, not belonging any group. The Corrected Reproductive Potential index was determinate just for cabbage, indicating good reproductive capacity with low sinigrin concentration. The extreme sinigrin doses, 0.2, 0.4 and 3.2mg/mL, increase the cabbage leafs consumption and decrease in broccoli. The consumption was bigger by the caterpillars fed on broccoli leafs. In general, the insect behavior was different in the tried cultivars, but these differences can be related to sinigrin quantity present naturally in the leaves of each specie.

Submitted by CAROLINE PLACIDI DE BORTOLI null (carubortoli@yahoo.com.br) on 2018-02-22T10:06:56Z No. of bitstreams: 1 Tese_Caroline_Placidi_De_Bortoli.pdf: 20556112 bytes, checksum: 3c463875d34d1f0b87961046250a6dab (MD5) / Approved for entry into archive by Alexandra Maria Donadon Lusser Segali null (alexmar@fcav.unesp.br) on 2018-02-22T12:45:28Z (GMT) No. of bitstreams: 1 debortoli_cp_dr_jabo.pdf: 20556112 bytes, checksum: 3c463875d34d1f0b87961046250a6dab (MD5) / Made available in DSpace on 2018-02-22T12:45:28Z (GMT). No. of bitstreams: 1 debortoli_cp_dr_jabo.pdf: 20556112 bytes, checksum: 3c463875d34d1f0b87961046250a6dab (MD5) Previous issue date: 2018-02-16 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Plutella xylostella é uma praga de grande importância para as crucíferas em todo o mundo. Embora seja controlada com inseticidas sintéticos e biológicos, ela pode desenvolver resistência rapidamente a uma variedade de inseticidas. Os biopesticidas mais comuns utilizados para controlar P. xylostella são baseados na bactéria entomopatogênica Bacillus thuringiensis. Embora muitos estudos tenham sido realizados com Bt, o modo de ação ainda não é totalmente compreendido. Uma grande diversidade de genes é diferencialmente expressa no intestino médio de insetos resistentes, o que sugere que vários processos celulares podem estar envolvidos na resistência. Descobertas recentes mostraram que as mutações no gene que codifica o transportador ABCC2 são responsáveis pela resistência às toxinas Bt em diferentes espécies de insetos. O objetivo desta pesquisa foi testar a hipótese de que a susceptibilidade de P. xylostella a Bt se correlaciona com o nível de expressão dos componentes desse regulador de estresse. Os níveis de expressão dos genes ALP, APN, CDKAL 1, MAP4K4 e ABCC2 foi comparado utilizando qPCR, entre populações suscetíveis e resistentes de P. xylostella. A investigação da sequência de DNA do cDNA do ABCC2 foi realizada, por PCR e sequenciamento, para testar a hipótese de que a susceptibilidade de P. xylostella a Bt se correlaciona com mutações no gene ABCC2. Foram realizados retrocruzamentos entre populações suscetíveis e resistentes e cruzamentos de complementação entre populações resistentes. Nossa pesquisa demonstrou que não há padrões na expressão dos genes testados demonstrando nenhuma associação entre expressão e resistência/susceptibilidade. No entanto, ao investigar a sequência do gene ABCC2, encontrou-se uma mutação no gene da população brasileira resistente, que poderia ser responsável pela causa da resistência da população estudada neste trabalho. Os ensaios de retrocruzamentos não confirmaram que a resistência foi devida à supressão de 1 pb encontrada, no entanto, os ensaios complementares indicaram que a população brasileira compartilha um alelo de resistência com a população havaiana resistente. / Plutella xylostella is a major insect pest of cruciferous crops worldwide. Although controlled with both synthetic and biological insecticides it can rapidly evolve resistance to a variety of insecticides. The most common biopesticides used to control P. xylostella are based on the entomopathogenic bacterium Bacillus thuringiensis. Although many studies have been performed on Bt, the mode of action is still not fully understood. A wide diversity of genes are differentially expressed in the midgut of resistant insects, this suggests that a variety of cell processes may be involved in the preservation of resistance. Recent discoveries have shown that mutations in the gene encoding an ABCC2 transporter are responsible for resistance to Bt toxins in various different insect species. The aim of this research was to test the hypothesis that the susceptibility of P. xylostella to Bt correlates with the level of expression of components of this putative stress-response regulon. The level of expression of ALP, APN, CDKAL 1, MAP4K4 and ABCC2 genes were compared using qPCR, between susceptible and resistant P. xylostella populations. The investigation of the DNA sequence of ABCC2 cDNA was performed, through PCR and sequencing, to test the hypothesis that the susceptibility of P. xylostella to Bt correlates with mutations on the ABCC2 gene. Backcrosses between susceptible and resistant populations and complementation cross between resistant populations were performed. Our research demonstrated that there were no patterns in the expression of the genes tested demonstrating no association between expression and resistance/susceptibility. However, when investigating the ABCC2 gene sequence, a mutation in the gene of the Brazilian resistant population was found, which could be responsible for the resistance of the Brazilian population studied in this research. Backcrossing assays didn’t confirm that the resistance was due to the 1bp deletion found, however complementation assays indicated that the resistant Brazilian population shares a resistance allele with the resistant Hawaiian population. / FAPESP: 2015/05891-6 / FAPESP: 2016/04868-3 / CNPq: 140916/2014-8 / CNPq: 166510/2017-3

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No. of bitstreams: 1 magalhaes_go_dr_jabo.pdf: 1456860 bytes, checksum: 9272f5a2364e18df572c352231781eec (MD5) Previous issue date: 2016-06-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo deste trabalho foi avaliar a biologia de linhagens de P. xylostella em criações para fins de pesquisa e verificar o efeito de B. thuringiensis sobre P. nigrispinus em associação de métodos para o manejo da praga em ambiente protegido. As larvas de P. xylostella foram coletadas em Brassica oleracea var. italica - brócolis Piracicaba, e mantidas em Brassica oleracea var. acephala - couve Manteiga híbrida HS-20. A cada geração aspectos biológicos dos insetos foram avaliados em ensaios de laboratório. As características biológicas analisadas foram: período larval, viabilidade de larvas, período pupal, viabilidade de pupas, peso pupal, razão sexual, número de ovos por fêmea, fertilidade de ovos, longevidade de machos e longevidade de fêmeas. Também, com os dados obtidos foi construída a tabela de vida de fertilidade. Já com predador, P. nigrispinus, foi avaliada a história de vida, o consumo de lagartas de P. xylostella em plantas de couve tratadas com bioinseticidas à base de Bacillus thuringiensis e também sua atividade de fitofagia nestas plantas, em casa de vegetação. Nas avaliações de P. xylostella, indivíduos coletados no campo apresentaram duração larval (8,2 dias) e pupal (3,8 dias) maiores, diminuindo ao longo de gerações. A viabilidade pupal (86,9%) e a fertilidade (93,0%) também foram maiores para essa população, com menor peso de pupa (4,7 mg). Além disso, apresentaram menor taxa líquida de aumento populacional (Ro), com 39 fêmeas/fêmea/dia, enquanto que, na população de P. xylostella proveniente do laboratório foi de 47,8 fêmeas/fêmea/dia. As características biológicas dos predadores, de modo geral, não sofreram alterações quando consumiram presas diretamente em plantas de couve tratadas com os bioinseticidas. Além disso, a fitofagia foi semelhante indicando que os bioinseticidas utilizados, mesmo o produto formulado, não apresentam substâncias que são repelentes para os predadores. Em conclusão, os parâmetros biológicos de P. xylostella demonstraram que o baixo desempenho em algumas características nas primeiras gerações pode ser melhorado ao longo das gerações e que o isolado HD1 ou o produto comercial Agree® à base de B. thuringiensis podem ser utilizados em associação com o predador P. nigrispinus em ambiente protegido visando o controle de P. xylostella. / This study aimed to evaluate the biology of P. xylostella strains in mass rearing for research and verify the effects of B. thuringiensis on P. nigrispinus in association methods for the management of this pest in protected environment. Larvae collected from Brassica oleracea var. italica - broccoli Piracicaba, were kept in Brassica oleracea var. acephala - kale Manteiga hibrida HS-20. The biological aspects of insects generations were evaluated in laboratory bioassays. The biological parameters evaluated were: larval period, larval viability, pupal period, pupal viability, pupal weight, sex ratio, number of eggs per female, eggs fertility, longevity of males and females. The data were used to construc of a life table fertility. The predator consuming P. xylostella larvae in kale plants treated with bionsecticides based on Bacillus thuringiensis and also their phytophagy activity in these plants, in green house was evaluated. The individuals collected in the field presented larval period (8.2 days) and pupal (3.8 days) longer, decreasing over generations. 86.9% of pupal viability and 93.0% of fertility were also high in this population, however prepal weight was significanthy low (4.7 mg). In addition, the lower net rate of population increase (Ro) with 39 females / female / day, whereas in P. xylostella population from the laboratory was 47.8 females / female / day. In general, the biological characteristics of predators did not change when the predators consumed prey directly on kale plants treated with biopesticides. In addition, phytophagy was similar indicating that biopesticides used, even the formulated product did not have substances that were repellent to predators. In conclusion, the biological parameters of P. xylostella demonstrated that the poor performance in some biological in the first generation can be improved over the generations, and HD1 strain or commercial product Agree® based of B. thuringiensis can be used in combination with the predator P. nigrispinus in a protected environment to control P. xylostella.

Submitted by AMANDA APARECIDA FERNANDES LEMES null (amanda.aflemes@gmail.com) on 2017-11-09T11:32:59Z No. of bitstreams: 1 Dissertação_Amanda_Aparecida_Fernandes_Lemes.pdf: 1634439 bytes, checksum: 3da6c1a6b018e117ef6f8d99907d3496 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-11-21T14:03:34Z (GMT) No. of bitstreams: 1 lemes_aaf_me_jabo.pdf: 1634439 bytes, checksum: 3da6c1a6b018e117ef6f8d99907d3496 (MD5) / Made available in DSpace on 2017-11-21T14:03:34Z (GMT). No. of bitstreams: 1 lemes_aaf_me_jabo.pdf: 1634439 bytes, checksum: 3da6c1a6b018e117ef6f8d99907d3496 (MD5) Previous issue date: 2017-10-09 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Um dos principais problemas para o cultivo de brássicas é o dano causado pela traça-das-crucíferas, Plutella xylostella (Linnaeus, 1758) (Lepidoptera: Plutellidae), o que implica na aplicação de grandes volumes de agrotóxicos. Diversos inseticidas são empregados no controle de P. xylostella. Indoxacarbe foi o primeiro inseticida bloqueador de canais de sódio comercializado, com mecanismo de ação diferente do DDT e dos piretroides, apresentando várias vantagens, como baixa toxicidade aos organismos benéficos. Porém, aplicações frequentes de inseticidas para controle de P. xylostella, combinado com fatores biológicos da praga, podem favorecer a seleção de indivíduos resistentes. Até o momento, P. xylostella desenvolveu níveis de resistência a 95 ingredientes ativos, existindo na literatura mais de 55 relatos de resistência para o indoxacarbe, em sete países de diferentes continentes, inclusive no Brasil. Nesse sentido, este estudo objetivou determinar os efeitos subletais e residuais do indoxacarbe em duas populações de P. xylostella, sendo uma coletada no campo e outra de laboratório (suscetível), avaliando e comparando a toxicidade desse inseticida em diferentes concentrações e analisando o comportamento destas duas populações e assim formar uma base teórica para melhorar o manejo desta praga. Foram realizados bioensaios com duas populações (PC- de campo e PL- de laboratório-suscetível) de P. xylostella com o inseticida indoxacarbe na dose comercial (100mg.L-1), avaliando os efeitos residuais do produto 1, 3, 7, 14 e 21 dias após a aplicação. Também foi estimada a CL50 e avaliados os efeitos subletais desse produto aplicado e avaliado após 48 h utilizando-se as concentrações subletais de CL5 (0,37 e 1,01 mg.L-1), CL15 (0,87 e 2,06 mg.L-1) e CL25 (1,44 e 3,16 mg.L-1) para as populações PL e PC, analisando-se algumas características biológicas das populações. A dose comercial de indoxacarbe pulverizada em plantas de couve matou todas as larvas com 1 dia e 3 dias após a aplicação. Os bioensaios de imersão foliar mostraram que o indoxacarbe apresentou alto nível de toxicidade para as lagartas de P. xylostella e os valores de CL50 foram de 3,7 e 6,9 mg.L-1 para a população suscetível e de campo, respectivamente. Os efeitos subletais foram indicados por menor sobrevivência larval e pupal e diminuição das taxas de sobrevivência da prole, devido ao efeito crônico do inseticida para ambas as populações. Houve também um aumento no consumo foliar e diminuição na sobrevivência de adultos na população suscetível e diminuição da fecundidade na população de campo. Os valores médios das taxas reprodutiva líquida (R0), intrínseca de aumento (rm) e de crescimento finito (Λ) foram significativamente menores nos tratamentos do que no grupo controle para ambas as populações. Houve diferença significativa entre as respostas das duas populações. O indoxacarbe mostrou-se eficaz contra P. xylostella, sendo que suas concentrações subletais influem negativamente nas características biológicas do inseto. / The main problem for brassica crops is the damage caused by the diamondback moth, Plutella xylostella, which has led to the application of large volumes of pesticides. Several insecticides are employed in the control of P. xylostella. Indoxacarbe was the first commercially available sodium channel blocker insecticide, with a different mechanism of action than DDTs and pyrethroids, has several advantages, such as low toxicity to beneficial organisms. However, frequent applications of these insecticides to control P. xylostella, combined with biological factors of the pest are favoring the selection of resistant individuals. So far P. xylostella has developed levels of resistance to 95 active ingredients and there are in the literature more than 55 reports of resistance to indoxacarb in seven countries on different continents, including Brazil. In this sense, the objective was to determine the residual and sublethal effects of indoxacarb in two populations of P. xylostella, one collected in the field and the other in the laboratory (susceptible), evaluating and comparing the toxicity of this insecticide in different concentrations and analyzing the behavior of these two populations and thus form a theoretical basis to improve the management of this pest. Bioassays were performed with two populations (PC- field and PL- laboratory) of P. xylostella with the insecticide in commercial dose indoxacarb (100 mg.L-1), evaluating the effects of residual products 1, 3, 7, 14 and 21 days after spray. LC50 was also estimated and the sublethal effects of this product applied and evaluated after 48 h were evaluated using the sublethal concentrations of LC5 (0.37 and 1.01 mg.L-1), LC15 (0.87 and 2.06 mg. L-1) and LC25 (1.44 and 3.16 mg.L-1) for PL and PC populations by analyzing some biological characteristics of the individuals. The commercial dose of indoxacarb sprayed on cabbage plants killed all larvae up to 1 day and 3 days after spray. Leaf-dip bioassays showed that indoxacarb showed a high level of toxicity against P. xylostella larvae and the 48 h LC50 values were 3.7 and 6.9 mg.L-1 for the susceptible and field population, respectively. The sublethal effects were indicated by lower larval and pupal survival and decrease in offspring survival rates, due to the chronic effect of the insecticide for both populations. There was also an increase in foliar consumption and a decrease in adult survival in the susceptible population and decrease in fecundity in the field population. The mean values of net reproductive rate (R0), intrinsic rate of increase (rm), finite rate of increase (Λ) were significantly lower in the treatments than in the control group for both populations. There were significant differences between the responses of the two populations. The indoxacarb was effective against P. xylostella and its sublethal concentrations negatively influence the biological characteristics of the insect. / CNPq: 134536/2015-0