How does developmental plasticity differ between diverse types of nutrition regimes?

Tese de mestrado em Biologia Evolutiva e do Desenvolvimento, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2016

Antimicrobial activity in the tick Rhipicephalus (Boophilus) microplus eggs: Cellular localization and temporal expression of microplusin during oogenesis and embryogenesis

Arthropods display different mechanisms to protect themselves against infections, among which antimicrobial peptides (AMPs) play an important role, acting directly against invader pathogens. We have detected several factors with inhibitory activity against Candida albicans and Micrococcus luteus on the surface and in homogenate of eggs of the tick Rhipicephalus (Boophilus) microplus. One of the anti-M. luteus factors of the egg homogenate was isolated to homogeneity. Analysis by electrospray mass spectrometry (ESI-MS) revealed that it corresponds to microplusin, an AMP previously isolated from the cell-free hemolymph of X (B.) microplus. Reverse transcription (RT) quantitative polymerase chain reactions (qPCR) showed that the levels of microplusin mRNA gradually increase along ovary development, reaching an impressive highest value three days after the adult females have dropped from the calf and start oviposition. Interestingly, the level of microplusin mRNA is very low in recently laid eggs. An enhance of microplusin gene expression in eggs is observed only nine days after the onset of oviposition, achieving the highest level just before the larva hatching, when the level of expression decreases once again. Fluorescence microscopy analysis using an anti-microplusin serum revealed that microplusin is present among yolk granules of oocytes as well as in the connecting tube of ovaries. These results, together to our previous data. suggest that microplusin may be involved not only in protection of adult female hemocele, but also in protection of the female reproductive tract and embryos, what points this AMP as a considerable target for development of new methods to control R. (B.) microplus as well as the vector-borne pathogens. (c) 2009 Elsevier Ltd. All rights reserved.

Moscas-Das-Frutas em Cultivares de Cafeeiros de Presidente Prudente, SP

Coffee is considered the primary host of the Mediterranean fruit fly Ceratitis capitata (Wiedemann). The infestation of C. capitata in the State of São Paulo has reached economic importance in coffee plantations. Although little information about changes caused by the fly on the coffee beverage, it is known that the fruit fly infestation causes rapid change from cherry to raisin stage, causing qualitative damage on the parchment coffee production. The objective of this work was to study the population dynamics and diversity of Tephritidae and Lonchaeidae in coffee cultivars of Icatu Vermelho IAC 4045, Vermelho IAC 99, Novo Mundo 388-17-1, Obatã IAC 1669-20, Icatu Amarelo IAC 2944, grafted on Apoatã (IAC 2258) (Coffea canephora) and Icatu Vermelho IAC 4045 ungrafted and Apoatã (IAC 2258)-grafted with approximately 3 years. The experiment was conducted in Presidente Prudente, São Paulo State, Brazil, from June 2006 to July 2008. The experimental design was a randomized block with four replications in a factorial design (3 years X 6 cultivars). From each plot of 100 plants we randomly collected 250 mature fruits. The samples were taken at 15 day intervals. The population dynamics were evaluated by using one plastic McPhail trap per cultivar. After 26 months a total of 36,932 specimens of C. capitata were trapped in all cultivars, corresponding to 49.27% males and 50.73% females. Approximately 83.3% of the specimens were collected from January to December 2007. The population fluctuation showed population peaks in May, June and July, relative to fruit ripening period. We trapped 21 specimens of A. montei Lima and A. fraterculus (Wied.). The coffee fruits of Presidente Prudente, SP, are infested by the following species of Lonchaeidae: Neosilba pendula (Bezzi), N. zadolicha McAlpine & Steyskal, N. inesperata Strikis & Prado and Neosilba pradoi Strikis & Lerena. Neosilba pendula occurred in all evaluated cultivars and N. inesperata was recoverd only from Icatu Amarelo IAC 2944 and IAC Icatu Vermelho 4045/un-grafted. No Anastrepha specimen was recovered from the fruits. Cultivars did not differ due to tephritid and lonchaeid infestations, but in 2008 the highest infestation by C. capitata occurred in the field.

Moscas frugívoras, lepidópteros desfolhadores e seus parasitóides (Hymenoptera) associados a cultivo de café, em Cravinhos, SP

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Dinâmica populacional de moscas-das-frutas e de Triozoida limbata (Hemiptera: Psyllidae) e danos de Costalimaita ferruginea (Coleoptera: Chrysomelidae) e de T. limbata em pomar de goiaba submetido a sistema de racionalização de inseticidas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mass-rearing of Mediterranean fruit fly using low-cost yeast products produced in Brazil

Ceratitis capitata is one of the most important pests of fruits for exportation, and Sterile Insect Technique (SIT) has been the most efficient and environmental friendly technique used to control fruit fly populations around the world. A key goal in achieving a successful SIT program is a mass rearing system producing high quality insects at low cost. Providing adults with an artificial diet containing hydrolysed protein has been the major obstacle for bio-production facilities in Brazil, because it is expensive and has to be imported. Two other commercial products, autolysed yeast (AY) and yeast extract (YE), of domestic origin and low cost, were tested as substitutes of the imported hydrolyzed protein. To compare their efficiency we observed the female fecundity, adult survival and egg viability of flies raised on diets containing one of each of the different protein products. Flies reared on the domestic yeast products had equivalent or superior performance to the flies reared on imported protein. Both AY and YE can be a possible substitute for imported hydrolyzed protein for C. capitata mass-rearing, as they are cheaper and are readily available in the national market.

The host marking pheromone application on the management of fruit flies - a review

The aim of this work was to review the role of the host marking pheromone (HMP) and its application in integrated management programs for the fruit flies. Initially the oviposition behavior of tephritids has been analyzed with emphasis on Ceratitis capitata. The deposition of HMP, which consists in the last stage of the oviposition behavior has been characterized and discussed about evolutive aspects and the biological meaning of the tephritidae communication through the HMP. Finally, the perspectives on the use of HMP in the integrated management of fruit flies have been discussed.

The lepidopteran transposon vector, piggyBac, mediates germ-line transformation in the Mediterranean fruit fly

The piggyBac (IFP2) short inverted terminal repeat transposable element from the cabbage looper Trichoplusia ni was tested for gene transfer vector function as part of a bipartite vector–helper system in the Mediterranean fruit fly Ceratitis capitata. A piggyBac vector marked with the medfly white gene was tested with a normally regulated piggyBac transposase helper at two different concentrations in a white eye host strain. Both experiments yielded transformants at an approximate frequency of 3–5%, with a total of six lines isolated having pigmented eyes with various levels of coloration. G1 transformant siblings from each line shared at least one common integration, with several sublines having an additional second integration. For the first transformant line isolated, two integrations were determined to be stable for 15 generations. For five of the lines, a piggyBac-mediated transposition was verified by sequencing the insertion site junctions isolated by inverse PCR that identified a characteristic piggyBac TTAA target site duplication. The efficient and stable transformation of the medfly with a lepidopteran vector represents transposon function over a relatively large evolutionary distance and suggests that the piggyBac system will be functional in a broad range of insects.

Molecular clock or erratic evolution? A tale of two genes.

We have investigated the evolution of glycerol-3-phosphate dehydrogenase (Gpdh). The rate of amino acid replacements is 1 x 10(-10)/site/year when Drosophila species are compared. The rate is 2.7 times greater when Drosophila and Chymomyza species are compared; and about 5 times greater when any of those species are compared with the medfly Ceratitis capitata. This rate of 5 x 10(-10)/site/year is also the rate observed in comparisons between mammals, or between different animal phyla, or between the three multicellular kingdoms. We have also studied the evolution of Cu,Zn superoxide dismutase (Sod). The rate of amino acid replacements is about 17 x 10(-10)/site/year when comparisons are made between dipterans or between mammals, but only 5 x 10(-10) when animal phyla are compared, and only 3 x 10(-10) when the multicellular kingdoms are compared. The apparent decrease by about a factor of 5 in the rate of SOD evolution as the divergence between species increases can be consistent with the molecular clock hypothesis by assuming the covarion hypothesis (namely, that the number of amino acids that can change is constant, but the set of such amino acids changes from time to time and from lineage to lineage). However, we know of no model consistent with the molecular clock hypothesis that would account for the increase in the rate of GPDH evolution as the divergence between species increases.

Caracterização da rebentação da tangerineira ‘Setubalense’ em anos de safra e de contrassafra

A produção de citrinos encontra-se amplamente disseminada por todos os continentes, representando uma atividade económica geradora de milhões de euros. As crescentes exigências dos consumidores conduzem ao aparecimento de novas variedades, em detrimento das cultivares tradicionais. A produção de tangerineira ‘Setubalense’ tem vindo a sofrer um acentuado declínio, mas os consumidores mais exigentes em aromas cítricos e fragrâncias intensas continuam a apreciar esta cultivar. Por isso, esta cultivar tem algum espaço comercial para mercados diferenciados em que se valorizam as cultivares tradicionais. Estas características e a sua época de maturação, que implica ser pouco afetada pela Ceratitis capitata, fazem da ‘Setubalense’ uma cultivar recomendável para o modo de produção biológico. Com o objetivo de caracterizar o padrão produtivo alternante e tendo por objeto de estudo um pomar de 5 anos de tangerineira ‘Setubalense’ enxertado sobre citranjeira ‘Troyer’ [Citrus sinensis (L.) Osbeck x Poncirus trifoliata L.], foi avaliada a composição da rebentação e da floração nos anos de safra e de contrassafra. A intensidade e as características da rebentação dependem do quadrante da árvore e do ano (safra ou contrassafra). Nos anos de safra a percentagem de nós rebentados oscila entre 67,6 a 79,5 %, que contrasta com os 25,4 a 28,2 % dos anos de contrassafra, função do quadrante analisado. Nos anos produtivos os rebentos formados são essencialmente florais e nos improdutivos quase exclusivamente vegetativos. A formação de elevado número de flores nos anos de safra conduz a uma intensa abcisão de órgãos generativos. O número de folhas formada por cada 100 nós é similar nos anos de safra e de contrassafra. Os resultados obtidos refletem o padrão alternante desta cultivar, evidenciando o cuidado particular que esta cultura tem de ter na sua condução.

Bioecologia, monitoramento e controle de Drosophila suzukii na cultura do morangueiro.

O Brasil ocupa a terceira posição no ranking mundial de produção de frutas. A produção é diversificada devido às condições climáticas do País, que permitem produzir frutas tropicais, subtropicais e temperadas. A maior parte da produção tem como destino o mercado interno e apenas 2,5% destinam-se à exportação. Com a globalização da economia e as dificuldades de monitoramento das fronteiras brasileiras, a produção agrícola está vulnerável a diversos problemas fitossanitários. A fruticultura tem sido um dos setores mais afetados pelo registro de novas pragas, que aumentam os custos de produção e comprometem a qualidade das frutas devido às injurias causadas, e pelo maior risco da presença de resíduos de inseticidas utilizados para o seu controle, além de dificultar as exportações devido às barreiras quarentenárias. Dentre essas pragas, as moscas-das-frutas têm sido uma ameaça constante. Historicamente, a primeira espécie registrada no País foi a mosca do mediterrâneo, Ceratitis capitata, em 1901. Quase 100 anos depois, em 1996, foi registrada a ocorrência da mosca da carambola, Bactrocera carambolae, no Estado do Amapá, e em 1999 foi detectado Zaprionus indianus, em São Paulo. Diferentemente das duas primeiras espécies, que são tefritídeos, Z. indianus é um drosofilídeo, mas causa os mesmos danos nos frutos. Em 2013, no Rio Grande do Sul, foi confirmada a presença da drosófila da asa manchada Drosophila suzukii, considerada uma das principais pragas das chamadas ?pequenas frutas?, que incluem morango, mirtilo, amora-preta, framboesa e cereja, mas a drosófila da asa manchada também ataca outras frutíferas, como videira, nectarineira, pessegueiro, entre outros. No Brasil, já foram constatadas perdas em morango e teme-se que o inseto possa se adaptar às nossas condições e causar perdas econômicas significativas em outros cultivos. Dada a tradição da Embrapa Clima Temperado e da Embrapa Uva e Vinho em pesquisas sobre morangueiro, este documento tem o objetivo de disponibilizar informações sobre a identificação, a bioecologia e as estratégias para o monitoramento e controle da drosófila da asa manchada, baseadas nos trabalhos realizados em outros países e nas pesquisas conduzidas pelos autores do texto.

Pragas da mangueira e seus inimigos naturais.

A cultura da mangueira esta em expansão no território nacional e o país já e o segundo maior produtor de frutos dessa planta no mundo. A produção de manga em seus aspectos quantitativos e qualitativos acha-se prejudicada pela infestação de pragas em todos os estágios fonológicos da cultura. Este trabalho relata o resultado de levantamentos de principais pragas e seus inimigos naturais em quatro épocas do ano, em Campinas - SP (variedade Haden), Cafelândia - SP (variedade Tommy Atkins) e Lins - SP (variedade Keitt). As cochonilhas Aulacaspis tubercularis e Lecanium sp. ocorrem em baixas infestações nas tres variedades. O himenóptero parasitóide da familia Braconidae ataca 12 a 60% das cochonilhas, mantendo-as em baixos níveis populacionais. A mal-formação de ponteiros atribuída a ácaros eriofiideos ocorreu em 0,3 a 17% dos ramos. Na época de frutificação 5 a 12% dos frutos estavam infestados pelas moscas-das-frutas Ceratitis capitata e Anastrepha sp.

Verificacao da atividade inseticida de alguns vegetais brasileiros.

Neste trabalho foram estudadas 30 espécies vegetais, quanto a atividade inseticida. Dessas espécies foram preparados 240 extratos a partir das diferentes partes dos vegetais, utilizando hexano, etanol, acetona e solução hidroalcoolica como líquidos extratores. Os testes foram efetuados contra os insetos: Mosca domestica, Ceratilis capitata, Zambrotes subfasciatus, Spodoptera frugiperda e Anthonomus grandis. Dos vegetais testados, apenas os extratos das sementes de Annona cacan, A. crassiflora, A. squamosa e raízes de Potomorphe umbellata apresentaram alguma atividade.

Bioecologia do parasitoide de ovos Fopius arisanus (Sonan, 1932) (Hymenoptera: Braconidae) em moscas-das-frutas.

2016

The gene transformer-2 of Anastrepha fruit flies (Diptera, Tephritidae) and its evolution in insects

Abstract Background In the tephritids Ceratitis, Bactrocera and Anastrepha, the gene transformer provides the memory device for sex determination via its auto-regulation; only in females is functional Tra protein produced. To date, the isolation and characterisation of the gene transformer-2 in the tephritids has only been undertaken in Ceratitis, and it has been shown that its function is required for the female-specific splicing of doublesex and transformer pre-mRNA. It therefore participates in transformer auto-regulatory function. In this work, the characterisation of this gene in eleven tephritid species belonging to the less extensively analysed genus Anastrepha was undertaken in order to throw light on the evolution of transformer-2. Results The gene transformer-2 produces a protein of 249 amino acids in both sexes, which shows the features of the SR protein family. No significant partially spliced mRNA isoform specific to the male germ line was detected, unlike in Drosophila. It is transcribed in both sexes during development and in adult life, in both the soma and germ line. The injection of Anastrepha transformer-2 dsRNA into Anastrepha embryos caused a change in the splicing pattern of the endogenous transformer and doublesex pre-mRNA of XX females from the female to the male mode. Consequently, these XX females were transformed into pseudomales. The comparison of the eleven Anastrepha Transformer-2 proteins among themselves, and with the Transformer-2 proteins of other insects, suggests the existence of negative selection acting at the protein level to maintain Transformer-2 structural features. Conclusions These results indicate that transformer-2 is required for sex determination in Anastrepha through its participation in the female-specific splicing of transformer and doublesex pre-mRNAs. It is therefore needed for the auto-regulation of the gene transformer. Thus, the transformer/transfomer-2 > doublesex elements at the bottom of the cascade, and their relationships, probably represent the ancestral state (which still exists in the Tephritidae, Calliphoridae and Muscidae lineages) of the extant cascade found in the Drosophilidae lineage (in which tra is just another component of the sex determination gene cascade regulated by Sex-lethal). In the phylogenetic lineage that gave rise to the drosophilids, evolution co-opted for Sex-lethal, modified it, and converted it into the key gene controlling sex determination.