Tamanho genômico e marcadores microssatélites para Pterodon pubescens e Pterodon emarginatus (Leguminosae)

• Autor(a) principal: Melo, Priscila Zei
• Data de Publicação: 2018
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UFG
• dARK ID: ark:/38995/0013000008hks
• Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/9359
• topic: Illumina MiSeq; Marcadores moleculares; Sucupira-branca; Molecular markers; GENETICA::GENETICA VEGETAL

PterodonpubescensBenth (Leguminosae) is popularly known as sucupira-branca or faveiro, widely distributed in the Cerrado, widely used as a medicinal plant and in civil construction. Microsatellite markers are valuable tools for accessing the genetic variability of plants and their population genetic structure, due to characteristics such as the polymorphic nature, co- dominant inheritance and abundance in the genome.Thus, the objective of this work was to estimate the genomic size, to assemble the partial genome, to develop microsatellite markers for the P. pubescens species and to test the transferability of these markers to P. emarginatus. The size of the P. pubescensgenome obtained by floxo cytometry to verify how much of the genome was sequenced. The DNA of an individual of the P. pubescens species was sequenced according to the Nextera DNA (Illumina) protocol and the sequences assembled with the Dipspades software. The identification of perfect microsatellite regions and primer design were performed using the QDD software. Pairs of primers designed were tested for the amplification efficiency and identification of polymorphic locos in P. pubescens and P. emarginatusspecies. Pairs of primers were chosen to be fluorescently labeled for the evaluation of polymorphism and genetic variability of the locos, using 24 individuals of P. pubescens and 24 of P. emarginatus. The estimated haploid size of the genome of P. pubescens was 0,725 pg, which corresponds to 117 Mpb. Mounting the genome of P. pubescens resulted in a total of 711 Mb, so the assembled sequences represent 16,4% of the genome of the species. In these sequences 6,090 microsatellite regions were detected, with the number of tandem repeats ranging from 5 to 30 times. Thus, it was possible to design 17.546 pairs of primers for microsatellite regions. Of these, 30 primer pairs were synthesized for the amplification tests. Of the 30 pairs of primers tested in P.pubescens27 presented a good pattern of amplification product and 26 locos were polymorphic.In P.emarginatus 26 pairs of primers were successfully transferred and all locos were polymorphic. Thus it was possible to evaluate a population of P. pubescens and another of P. emarginatus with two systems of multipliex composed by 10 locos. The mean number of alleles per locus for P. pubescens and P. emarginatus was 7,1 and 7,7 respectively. The expected mean heterozygosity was 0,684 and 0,646, and the mean heterozygosity observed was 0,662 and 0,578, respectively for P. pubescens and P. emarginatus. There was a significant deviation of the HWE (p <0.05) locos Pem 21 and Pem 22 in P. pubescens and Pem 22 and Pem 26 and Pem 24.It was foundevidence of linkage imbalance in the locos (Pem 21 / Pem 22, Pem 15 / Pem 21, Pem 23 / Pem 22, Pem 21 / Pem23 and Pem 10/22) for P. pubescens, while in P. emarginatus in the locos (Pem 21 / Pem 22, Pem 21 / Pem23, Pem 15/21, Pem 23 / Pem 22, Pem 10 / Pem 22). The combined probability of identity of P. pubescens and P. emarginatus was (I) 7,9 x 10-10 and 1.4 x 10-6 respectively and paternity exclusion probability was 0,999 for the two species. The available microsatellite markers are a promising tool for future population genetic studies for the species, besides the possibility of being transferred to species of the same family.