57º Congresso Brasileiro de Genética
Resumos do 57º Congresso Brasileiro de Genética • 30 de agosto a 2 de setembro de 2011
Centro de Convenções do Hotel Monte Real Resort • Águas de Lindóia • SP • Brasil
www.sbg.org.br - ISBN 978-85-89109-06-2
125
Transcriptome analysis – Comparing
primitively social and social bees
Marco Antonio, DS1*; Cristino, AS2; Hartfelder, K3
1
Departamento de Genética, Faculdade de Medicina de Ribeirão Preto – Universidade de São Paulo, Ribeirão Preto
email: [email protected] 2 The Queensland Brain Institute, The University of Queensland, St Lucia, Brisbane,
Australia, 3 Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de
Ribeirão Preto – Universidade de São Paulo, Ribeirão Preto
Keywords: Transcriptome, Bee, Social bees, Inseticide sensitivity, Gene family
Expressed Sequence Tags (ESTs) are fragments of cDNA between 200-800 bp. They are generated from mRNA
through reverse transcription and are very usefull for gene identification and prediction. Futhermore, through BLAST
analysis it’s possible to associate predicted functions to each EST. The clusterization of ESTs based in annotated
genes improves prediction, and unclusterized ESTs can represent specific genes of the species. Adult workers of
Melipona quadrifasciata anthidioides and Frieseomelitta varia were collected when returning from flights and insight nest.
Adult females of Euglossa cordata and Eulaema nigrita were collected in the field when visiting flowers. The bees were
immediately frozen in liquid nitrogen. The brains of 15-30 bees were dissected and total RNA extracted in TRIzol.
Sequencing was done on a Roche 454 platform. After validation of ESTs quality they were submited to CAP3 analysis
generating contigs and singlets. These sequences were used to assemble the database of each specie. BLAST algorithm
were used for alignment analysis with cutoff E-value<10-5 and gap/penalities default. Data mining algorithms were
especially designed for the transcriptomes using Shell script and Python (v.2.4 and above). The Apis mellifera genome
was chosen as anchor due to phylogenetic proximity and a completely sequenced annotated genome. The dbEST of
NCBI encompasses 79,179 Apis mellifera ESTs, of a normalized brain library.For each transcriptome a “best hit” list
was created using blastn analysis against Apis mellifera (Official Gene Set 2). A second list was created through blastx
searches using the first list against Drosophila melanogaster genome. The resulting data were used to retrieve ontologies
from Babelomics3. Every functional attribute here described is based on bioinformatically inferred evidence according with functionality based on Drosophila melanogaster which has extensive annotated genome. The Euglossa cordata
transcriptome has 49,830 ESTs of which 18,568 related with Apis mellifera genes. Gene Ontology (GO) information
was obtained for 5.862 ESTs. The Eulaema nigrita transcriptome has 56,689 ESTs of which 19,354 matched with Apis
gene. 5,904 ESTs have GO information with 43% being related with biological processes at level 3. Frieseomellita varia
has 50,808 ESTs with 13,795 positively matched against Apis genes. We found 6,070 ESTs being related with biological processes at level 3 and 56% related with molecular function. Melipona quadrifasciata has 54,525 ESTs with 17,521
being related with Apis genes. Interestingly we found GO information for 13,140 ESTs. All species have proportionally the same quantity of ESTs with cellular metabolism, primary metabolism and macromolecules metabolism. They
share 4,681 genes based on NCBI non-redundant database.
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Transcriptome analysis – Comparing primitively social and social bees