Ensaios com primers de ISSR e PCR para estudo da variabilidade genética de Tetragonisca angustula (Latreille, 1811) (Hymenoptera, Apidae, Meliponinae) na região de Campo Mourão - Paraná

Abstract

The bees belonging to Meliponini tribe live in tropical and sub-tropical areas in the planet, covering an average of 60 genders. Its importance role is pollination, regarding the key piece of ecosystem maintenance. Tetragonisca angustula is a Meliponini that spread in almost all Latin America, adjusts well to different substrates to nests, accepts been handled by man and produces pleasant sweet taste honey. Due to its importance, they must be conserved, in order to avoid inbreeding and a possible extinction, make it possible keep up with the biggest evolutionary potential. For such, genetic becomes allied with, in order to determine the differential degree of gene between them. This way, the present study aimed to analyze this variability in Tetragonisca angustula throughout molecular marker ISSR. The bees were collected in Iretama and Campo Mourão cities. Its was analyzed a worker bee by colony, in 12 different colonies. After collecting, the samples were put through laboratoty procedures of DNA extraction, PCR and agarose gel electrophoresis, next, there was no evidence DNA band migrate through agarose gel. Therefore, due to this result, 11 different primers ISSR were tested in order to find out ideal sequence and temperature of annealing to its reaction and amplification, and then see the DNA band, after that analyze differential degree of gene of them. To this end, no primers tested replied to what was expected, so it could not be seen any fragment of genetic material. This way, we can infer that ISSR primers functionality used in this work was not successful to the species and to temperatures related to PCR reaction. In this sense, this work was adapted to selection and optimization of methodology techniques adopted to the study of the differential degree of gene cited, keeping up the focus on importance of genetic study in natural population as bioindicators of forest fragmentation.