Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
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2018-08-27
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Universidade Federal de Goiás
Resumo
Stryphnodendron adstringens presents a wide geographical distribution, being
predominant in regions of Cerrado sensu stricto and is popularly known as
"barbatimão". It is a species widely used in herbal medicine because of its anti-inflammatory,
antibacterial and antiulcerogenic potential. The Cerrado’s removal of vegetation cover
reduces significantly the number of several native species, among them S. adstringens.
Thus, it is important that efficient strategies for the use and conservation of this species are
drawn. In order to provide molecular tools for studies of genetic diversity and conservation of
S. adstringens, the present work had the objective of developing microsatellite markers
for the species. Genomic DNA was obtained from leaf tissue using the CTAB protocol. The
identification of the microsatellite regions and the design of the primers were performed using
the QDD program modules. From the identified microsatellite regions, 20 pairs of primers
were designed, 14 of which flank microsatellite regions composed of dinucleotides, four by
tetranucleotides and two by pentanucleotides. Initially, four individuals were used for the
standardization tests of the PCR protocol and annealing temperatures. Subsequently, 48
individuals were selected, distributed in three populations, to evaluate polymorphism via 6%
polyacrylamide gel. Of the 20 pairs of primers evaluated, 16 presented polymorphic
amplification products and four monomorphic amplification products. Considering the 16
polymorphic markers, the number of alleles varied between two (SadH19) and 13 (SadH13),
with a mean of seven alleles per locus. The observed heterozygosity (Ho) and expected (He)
and PIC values were 0.506, 0.543, 0.635, respectively. The mean Hmax value founded
(65,519) indicates values of genetic diversity that can be considered medians for this set of
loci evaluated in three populations of S. adstringens. On the other hand, although genetic
diversity is median, this set of 16 polymorphic markers exhibited a ombined probability of
paternity exclusion high (0.9999983) and combined probability of genetic identity low
(3,49x10-15). The analysis of variance of allelic frequencies presented significant values for
two of the three estimated statistics with f not significant 0.050, significant θ equal to 0,329
and F in the overall value also significant 0.360. Thus, it can be concluded that the panel of
polymorphic markers developed for S. adstringens is highly informative and indicated for
population genetic studies for the species. Another important factor is that these markers can
be tested in other evolutionarily close species for the availability of microsatellite markers,
without the need to develop new primers.
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BARATELI, L. O. Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae). 2018. 59 f. Dissertação (Mestrado em Genética e Melhoramento de Plantas) - Universidade Federal de Goiás, Goiânia, 2018.