A Florida strain of sweet potato whitefly, Bemisia tabaci (Gennadius), was found to have an expanded range which includes several new food crops. To determine why, we examined how it processes plant nutrients. The amino acid and carbohydrate content of phloem sap from poinsettia and pumpkin and of honeydew produced by the Florida strain were analyzed Honeydews produced by a strain from Arizona feeding on both plants were also analyzed Poinsettia phloem sap contained 15 amino acids; 14 of these were in pumpkin phloem sap. Almost all the same amino acids were in the honeydews produced by the two strains on the two hosts. Carbohydrates in phloem sap and honeydew were common transport sugars, like sucrose. Both honeydews contained trahalulose, a disaccharide not previously associated with insects. Both strains processed phloem sap and honeydew from both plants in the same manner, but the Florida strain produced significantly larger quantities of honeydew; it is therefore assumed to process more phloem sap. Since this strain has access to more phloem sap it also has access to more of the amino acids, which are in short supply in the phloem sap of some plants, allowing it to broaden its range.

Although problems associated with the sweet potato whitefly, Bemisia tabaci (Gennadius), are not as dramatic as they were in 1992. they were still significant in 1993. Laboratory research in 1993 focused on defining the cues that result in migratory behavior, specifically host quality. In addition, field studies were conducted to learn more about timing, direction, and distance flown. Our goal is to develop a predictive model that can be used for forecasting whitefly movement. During our behavioral studies, B. tabaci was presented with two cues that lead to disparate behaviors. More than 70% of the whiteflies we tested ended their flights (within three presentations of the cue) when given a choice between settling on a 'host' (550 nm interference filter) and continued flight. Only 6% of the individuals we tested, demonstrated what would be considered to be true migration Both endogenoous and exogenous factors can play an important role in determining when insects will fly. Finally, although the oogenesis-flight syndrome is thought to be a strong component of insect migratory activity, whiteflies do not appear to postpone egg production until after they have engaged in flight. In the field marked whiteflies were also collected in the most distant of these traps. These field results support our hypothesis that most whitefly movement in the fall in the Yuma Valley is in a SW direction (prevailing winds are from the NE). Within a 3 hr time frame whiteflies can travel as far as 2.2 miles from the source field. We examined the effects of female flight distance and eggload. We found significant differences in the eggload of field collected whiteflies versus whiteflies collected in traps at all distances from the source field. There was no relationship between distance flown and eggload. These results may indicate that dispersing individuals are capable of delaying egg laying until a suitable host plant is located

ix, 152 leaves : photos, graphs, tables, map. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Entomology, 1980

Diss. (sammanfattning). Uppsala : Sveriges lantbruksuniv. / Härtill 4 uppsatser.

3 pp. / Growing degree days to reach various growth stages in small grains is presented in this publication, as well as the optimum timing of cultural practices relative to crop growth stage.

Culicoides molestus (Skuse) is the major species of biting midge that plagues human comfort in the estuarine Gold Coast region of southeast Queensland. Local authorities have initiated a search for an effective, non-chemical means of control, that would minimize human-midge interaction. The effectiveness of a program to control an organism, such as a biting midge, is dependent upon knowledge of the biology of the particular organism of interest. This project revolved examines the lifecycle of C. molestus in detail. It addresses questions regarding the location, seasonal distribution, and dispersal of its juvenile stages in the sand of infested beaches, and their response to chemical treatment, the monthly and annual cycles of the adult midge, and the possibilities of achieving laboratory oviposition, as a first step to laboratory colonisation. The distribution of eggs, larvae and pupae of C. molestus was found to be mostly concentrated around, but below, mean tide level. They also occurred well below the mean tide level. Eggs and larvae have been recovered from as deep as 10 cm in the sand. A seasonal study of the juveniles of this species indicated that they were more strongly influenced by tides than seasons. After a routine pest-control larviciding treatment, a beach recolonisation study revealed that beaches become suitable for oviposition approximately two months after treatment. Large larvae invaded the sprayed areas within days of treatment, which suggests the existence of a refuge outside of the reach of the insecticide. Larvae found in clean (egg- and larva-free), isolated sand containers, placed on the study beach, indicated that larvae could swim in or on the water as a way of moving around the beach. Extended bite-rate studies highlighted the existence of four peaks in adult midge biting activity during the course of a year, around the mid seasons. The strongest peaks of activity were found to be in autumn and spring, but the data suggest that the species undergoes four generations in a year. Through a series of trial-and-error experiments, oviposition under laboratory conditions was achieved. Although the time from blood-feeding to egg maturation is not yet well determined, it occurs within an eight day mean survival period. Blood quality appears critical for adult blood-fed midge survival. Midges fed on the blood of a volunteer who was frequently exposed to midge bites do not live long enough to mature its eggs. The partial ovarial development of one unfed adult female, reared in the laboratory, indicates that C. molestus is facultatively anautogenous.

Diss. (sammanfattning) Alnarp : Sveriges lantbruksuniv. / Härtill 6 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2007. / Härtill 4 uppsatser.

Made available in DSpace on 2016-06-02T20:20:35Z (GMT). No. of bitstreams: 1 5217.pdf: 18042962 bytes, checksum: 6c55d259639b3b244326b14d21f608f1 (MD5) Previous issue date: 2012-02-29 / Universidade Federal de Minas Gerais / The sugarcane weevil, Sphenophorus levis, is an insect that feeds on the rhizome of sugarcane in its larval stage, boring channels that cause damage and death to the plant. Conventional methods of insect control have not been efficient. The aim of the present study was to generate knowledge on the biology of this insect on the molecular level using a transcriptomic approach to determine potential targets genes for the engineering of insect-resistant plants. After sequence processing and assembly using the dCAS program, 3804 sequences were grouped into 201 contigs and 1363 singlets, which were manually annotated. Several plant cell wall degrading enzymes were identified, including pectinases and cellulases. An invertasecontaining contig was identified for the first time in a coleopteran. Total probable digestive enzymes accounted for 19.3% and unknown genes accounted for 28.8% of the total number of expressed sequence tags. Considering the predominance of cathepsin L enzymes among the digestive enzymes found in the transcriptome (54.3%) and their importance to the breakdown of proteins in the insect digestive process, a cDNA clone encoding a cathepsin L enzyme, denominated Sl-CathL, was chosen for recombinant expression in Pichia pastoris cells, characterization and in vitro inhibition by the sugarcane cystatin CaneCPI-4 (Ki = 0.196 nM). Immunolocalization assays demonstrated the production of Sl-CathL in the midgut epithelium and secretion into the gut lumen from vesicles containing the enzyme. S. levis demonstrated sensitivity in triggering RNA interference machinery induced by dsRNA injections in the body cavity and gene-specific knockdown was confirmed by qRT-PCR. Larvae injected with V-ATPase E dsRNA died within three weeks after injection and serpin 1-silenced larvae either exhibited delayed development, arresting in the pupal stage, or died as pharate adults. In conclusion, transcriptome analyses, together with the inhibition of the main digestive enzyme by Cane-CPI-4 and gene silencing using RNAi, are promising procedures for the development of transgenic sugarcane plants to enhance resistance to Sphenophorus levis. / O bicudo da cana-de-açúcar, Sphenophorus levis, é um inseto que na fase larval se alimenta do rizoma da cana-de-açúcar cavando galerias que causam danos e levam à morte das plantas. Os métodos convencionais de controle disponíveis não tem sido eficientes contra esse inseto. O objetivo desse estudo foi gerar conhecimento sobre a biologia desse inseto em nível molecular utilizando uma abordagem transcriptômica para a identificação de possíveis genes alvo, visando futuros estudos para desenvolvimento de plantas transgênicas resistentes ao ataque deste inseto. Após o processamento e montagem das sequências utilizando o programa dCAS, 3804 sequências foram agrupadas em 201 contigs e 1363 singlets, os quais foram manualmente anotados. Foram identificados diversos genes de degradação de parede celular, incluindo pectinases e celulases. Pela primeira vez um contig contendo uma sequência que codifica uma invertase foi identificado em um coleóptero. As prováveis enzimas digestivas totalizam 19,3% e os genes desconhecidos representam 28,8% do total de sequências expressas. Considerando a predominância de catepsinas L dentre as enzimas digestivas identificadas no transcriptoma (54,3%) e a sua importância para a hidrólise de proteínas nos processos digestivos, um clone de cDNA codificando uma catepsina L, denominado Sl-CathL, foi escolhido para a expressão recombinante em células de Pichia pastoris, caracterização e inibição in vitro utilizando a cistatina da cana-de-açúcar CaneCPI-4 (Ki = 0,196 nM). Os ensaios de imunolocalização evidenciaram a produção da Sl-CathL no epitélio do intestino médio e a secreção de vesículas, contendo a enzima, no lúmen do intestino. Larvas do inseto S. levis também apresentaram sensibilidade para disparar a maquinaria de RNA de interferência induzida por injeções de dsRNA na cavidade corpórea e o silenciamento gênico específico foi confirmado por qRT-PCR. As larvas que receberam injeções com dsRNA da V-ATPase E morreram dentro de três semanas, enquanto as larvas que tiveram a serpina 1 silenciada exibiram atraso no desenvolvimento, aprisionamento na fase pupal, ou morreram como adultos farados. Desse modo, concluímos neste trabalho iniciado a partir da análise do transcriptoma, que a inibição da principal enzima digestiva pela Cane-CPI-4 e o silenciamento gênico via RNAi são alternativas promissoras para o estabelecimento de plantas resistentes ao inseto e podem ser aplicadas no desenvolvimento de plantas de cana-de-açúcar transgênicas com o objetivo de aumentar sua resistência ao Sphenophorus levis.

Diante da busca por abordagens mais sustentáveis para o controle de insetos, os inibidores de peptidases de plantas surgem como uma ferramenta promissora para a proteção de culturas via obtenção de plantas transgênicas. Efeitos nocivos da ingestão de inibidores de peptidases de soja (IPS) sobre o desenvolvimento de Diatraea saccharalis já foram reportados em trabalhos anteriores, entretanto, esses efeitos não foram estudados em nível molecular para essa espécie. Esse trabalho teve como objetivos identificar e analisar a expressão gênica e a atividade de serino peptidases intestinais de D. saccharalis em resposta à ingestão de um concentrado ativo de inibidores de peptidases de soja, relacionando essas informações com a presença ou não de algum mecanismo adaptativo desse inseto em resposta à ingestão desses inibidores. Para isso, foi obtido o transcriptoma intestinal de lagartas de D. saccharalis no quinto ínstar do desenvolvimento, submetidas à ingestão crônica de IPS. Em seguida foram identificadas as sequências correspondentes a serino peptidases do tipo tripsinas e quimotripsinas nesse transcriptoma, as quais foram utilizadas para o estudo da expressão gênica relativa por RT-PCR quantitativo, em resposta à ingestão aguda (por 24 e 48 horas) e crônica e de 0,5% (m/v) de um concentrado ativo de IPS por lagartas de D. saccharalis no terceiro e no quinto ínstares do desenvolvimento larval. As mesmas sequências foram utilizadas para a proposição de filogenias, sendo comparadas inclusive com sequências de serino peptidases de Spodoptera frugiperda identificadas em outro estudo paralelo. Ensaios de atividade tríptica e quimotríptica foram realizados utilizando substratos específicos (BApNA e SApNA) para determinar o efeito da ingestão de IPS sobre a atividade enzimática e finalmente, a integridade e a atividade inibitória do IPS foram analisadas após sua passagem pelo trato digestório das lagartas. Os resultados obtidos permitiram concluir que, nas condições experimentais empregadas nesse estudo, as lagartas de D. saccharalis não foram capazes de modular a expressão gênica das serino peptidases em resposta à ingestão crônica ou aguda de IPS. Os resultados de atividade tríptica e quimotríptica em função da ingestão de IPS podem sugerir um sinergismo na ação dessas enzimas em lagartas do quinto instar. Não foram detectadas diferenças na atividade de tripsinas e quimotripsinas em lagartas do terceiro ínstar em função da ingestão de IPS. A separação eletroforética das proteínas das fezes de lagartas permitiu a visualização de uma banda com massa molecular em torno de 20 kDa possivelmente correspondente ao inibidor do tipo Kunitz presente em soja que, juntamente com os dados de atividade antitríptica das fezes, indicam que D. saccharalis não é capaz de hidrolisar esse inibidor como um mecanismo de adaptação à sua presença na dieta. / Given the importance of search for more sustainable approaches for pest control, plant peptidases inhibitors emerge as a promising tool for the protection of crops by obtaining transgenic plants. Harmful effects of soybean peptidases inhibitors (SPI) intake by Diatraea saccharalis on its development were already reported in previous studies, however, these effects have not been studied at the molecular level for this specie. This study aimed to identify and analyze the gene expression and the gut serine peptidases activity of D. saccharalis in response to the intake of an active extract of soybean peptidases inhibitors. These informations were related to the presence or absence of some adaptive mechanism of this insect in response to these inhibitors. The gut transcriptome of larvae of D. saccharalis in the fifth developmental instar was obtained, undergoing chronic intake of IPS. Then, sequences of serino peptidases like trypsin and chymotrypsin were identified and used for gene expression studies on a real-time PCR in response to acute (for 24 and 48 hours) and chronic intake of 0.5 % (w/v) of SPI active extract by larvae of D. saccharalis in the third and fifth instars of larval development. The same sequences were used to propose phylogenies, including comparisons with sequences of Spodoptera frugiperda serine peptidases identified in another parallel study. Tryptic and chymotryptic activity assays were performed using specific substrates (BApNA and SApNA) to determine the effect of the SPI intake on enzyme activity and finally, the integrity and the inhibitory activity of the SPI were analyzed after passing through the digestive tract of caterpillars. The results showed that under the experimental conditions employed in this study, the larvae of D. saccharalis were not able to modulate the gene expression of serine peptidases in response to acute or chronic SPI intake. The results of tryptic and chymotryptic activity due to the SPI intake can suggest a synergism in the action of these enzymes in larvae of the fifth developmental instar. SPI intake did not cause differences in enzymatic activity of third instar larvae. Electrophoretic separation of proteins from the feces of caterpillars enabled visualization of a band with a molecular mass around 20 kDa possibly corresponding to the Kunitz inhibitor present in soybean. The antitryptic activity of the feces indicates that D. saccharalis is not able to hydrolyze soybean peptidases inhibitor as a mechanism of adaptation to their presence in the diet.