Isolamento, caracterização e transferibilidade de marcadores microssatélites de Byrsonima cydoniifolia A. Juss. (Malpighiaceae)
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2014-03-13
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Universidade Federal de Goiás
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Molecular markers are identifiable DNA sequences found at specific sites in the genome and transmitted by the standard laws of inheritance from one generation to the next. Markers known as microsatellite are indicated for genetic population analysis; due they are widely distributed in the genome and their high polymorphic information content per locus. To support different management practices, genebanks, conservation studies is important to perform the analysis of genetic variability in natural populations of the species of interest. In the biome Cerrado there are inumerous medicinal and fruit trees that are used in a extractive way, among them are those of the gender Byrsonima sp. popularly known as murici, belonging to the family Malpighiaceae. In Brazil presents about 70 species with wide distribution in the Cerrado. The murici is a multi- potential plant and although not be domesticated, its economic exploitation can be viable. In this context, the objective was to develop a set of microsatellite markers for species Byrsonima cydoniifolia A. JUSS. and test the potential transferability of this set of markers for species Byrsonima crassifolia L. Kunth. The microsatellite regions were isolated using genomic libraries enriched for repetitive regions and then they were used for primer design. Using the primers designed temociclagem conditions were tested and the products were characterized using the DNA of 90 individuals of B. cydoniifolia and 24 indivuduals of B. crassifolia. Of these, 14 loci were polymorphic and 3 monomorphic in three populations of B. cydoniifolia. The number of alleles ranged from 3 to 17, with an average of 10.45 alleles per locus. The observed heterozygosity was similar to the expected heterozygosity, the values ranged from 0.20 to 0.92 and 0.43 to 0.92; respectively. 39 private alleles were identified. This set of markers showed high combined probability of paternity exclusion (Q) equal to 0.999 and combined probability of identity (I) equal to I=7.84x10-1. The global values for the fixation index (FIS) were not significant (P<0.05), whereas the values of FST=0,082 and FIT=0,143 were significant. For Byrsonima crassifolia species, nine pairs of primers produced good patterns of cross amplification in the study population, with a mean of 7.56 alleles per locus. The observed heterozygosity ranged from 0.250 to 0.958, which was higher than the expected heterozygosity ranging from 0.223 to 0.905. The combined probability of paternity exclusion (Q) was equal to 0.99847 and high combined probability of identity equal to 6.42x10-9. The strategy of development of microsatellite markers from the genomic library enriched methodology was efficient to generate a panel with eleven polymorphic microsatellite markers for B. cydoniifolia and nine polymorphic microsatellite markers successfully transferred to B. crassifolia, providing a useful tool for population genetic studies not only for the species B. cydoniifolia and B. crassifolia, but possibly for other related species evolutionarily.
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Bernardes, V. Isolamento, caracterização e transferibilidade de marcadores microssatélites de Byrsonima cydoniifolia A. Juss. (Malpighiaceae). 2014. 95 f. Dissertação (Mestrado em Genetica e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2014.