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Master's Dissertation
DOI
https://doi.org/10.11606/D.46.2019.tde-24102019-113712
Document
Author
Full name
Bárbara Bacelar Nascimento
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2019
Supervisor
Committee
Terra, Walter Ribeiro (President)
Tanaka, Aparecida Sadae
Winck, Flavia Vischi
Winter, Carlos Eduardo
Title in Portuguese
Análise do transcriptoma intestinal do Hemiptera Dysdercus peruvianus orientada fisiologicamente
Keywords in Portuguese
Absorção de nutrientes
Análise de transcriptoma
Dysdercus peruvianus
Abstract in Portuguese
As espécies de insetos do gênero Dysdercus são conhecidas por causar prejuízos à agricultura. O estudo do funcionamento da fisiologia do intestino de insetos desse gênero pode levar à identificação de possíveis alvos moleculares para proposta de novos métodos de controle de insetos. Neste trabalho, foram identificados e localizados sítios de expressão das sequências de enzimas digestivas principais e também de transportadores potencialmente envolvidos na absorção de nutrientes, íons e água ao longo do intestino, a partir de dados transcriptômicos. Para isso foi feito uma identificação e anotação de proteínas por bioinformática a partir do transcriptoma de D. peruvianus e foi analisada a expressão gênica dessas proteínas ao longo do intestino médio. Foram identificados transportadores e enzimas envolvidos no processo de digestão e a expressão diferencial desses foi determinada e analisada. O ensaio com as enzimas α-galactosidase e α-N-acetil-D-galactosaminidase demonstrou que ambas enzimas estão presentes no inseto, sendo que a primeira é mais expressa no intestino e a segunda é típica de carcaça, permitindo a identificação de suas sequências. A análise das enzimas envolvidas na formação de diacilglicerol mostrou que provavelmente a via mais utilizada consiste na acilação do monoacilglicerol a diacilglicerol, para ser levado a lipoforina circulante na hemolinfa. Os dados resultaram na elaboração de modelo de organização de processo digestivo e absortivo no intestino de D. peruvianus.
Title in English
Physiologically-oriented Hemiptera Dysdercus peruvianus midgut transcriptoma analysis
Keywords in English
Dysdercus peruvianus
Nutrient absorption
Transcriptome analysis
Abstract in English
Insect species of the genus Dysdercus are known to cause damage to agriculture. The study of the functioning of insect midgut physiology of this genus can lead to the identification of possible molecular targets for the proposal of new methods of insect control. In this work, expression sites of the sequences of the main digestive enzymes and of transporters potentially involved in the absorption of nutrients, ions and water along the intestine were identified and located from transcriptomic data. For this, a bioinformatics identification and annotation of proteins was made from the transcriptome of D. peruvianus and the gene expression of these proteins were analyzed along the middle gut. Transporters and enzymes involved in the digestion process were identified and the differential expression of these was determined and analyzed. The enzimatic assay of α-galactosidase and α-N-acetyl-Dgalactosaminidase demonstrated that both enzymes are present in the insect, the former being more expressed in the midgut and the second is typical of carcass, allowing the identification of their sequences. Analysis of the enzymes involved in the formation of diacylglycerol showed that the most commonly used pathway is probably the acylation of monoacylglycerol to diacylglycerol, to be taken to circulating lipoforin in hemolymph. The data resulted in the elaboration of a model of the organization of digestive and absorptive process in the midgut of D. peruvianus.
 
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Publishing Date
2019-11-14
 
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