Estudo cinético na caracterização do status corporal de zinco na Síndrome de Berardinelli-Seip

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Análise citogenética de células-tronco derivadas do subendotélio da veia umbilical humana

Mesenchymal stem cells (MSCs) are known as a population of multi-potential cells able to proliferate and differentiate into multiple mesodermal tissues including bone, cartilage, muscle, ligament, tendon, fat and stroma. Several applications of the study of EC can be emphasized the therapeutic techniques such as guided bone regeneration by implantation of EC in the affected site, without the need for bone grafts, using titanium as a vehicle. The process of cryopreservation is essential for the maintenance of cell cultures, since the cell line is frozen, it can be maintained in liquid nitrogen for an indefinite period and then thawed for therapeutic or experimental purposes. The aim of this study was to isolate a population of MSCs derived from the subendothelium of the umbilical vein human (MSCs-SUVH) to assess cytogenetic analysis by the possibility of appearance of chromosomal changes in two different situations: MSCs-SUVH regarding the process of cryopreservation and MSCs-SUVH grown on the surface of titanium. Flow cytometry analysis revealed that, this cell population was positive for the markers CD29, CD73 and CD90, but there was no expression of hematopoietic lineage markers, such as CD14, CD34 and CD45 and demonstrated capacity for osteogenic differentiation. The chromosomes obtained from the primary culture of MSCs-SUVH were analyzed by GTW banding technique, and results are described as guidelines to ISCN 2005. There was not the emergence of clonal chromosomal changes in the MSCs-SUVH in different situations analyzed. However one of the strings presented a balanced paracentric inversion, probably a cytogenetic constitutional alterations, which was present before and after the experimental situations that the MSCs-SUVH was submitted

caracterização in situ e estrutura genética de populações de Gossypium mustelinum

Gossypium mustelinum Miers ex Watt is the only cotton species native from Brazil. It is endemic of the semi-arid region from North-east of the country, where it occur near from resilient water sources. The threats to the in situ conservation of the populations are caused by human interference in its habitat, mainly by excessive cattle graze and deforestation. Establish efficient strategies of in situ conservation depend on the accomplishment of a diagnosis of how the specie is found in its natural environment, and the knowledge about the genetic structure of the populations. The objectives of this work were i) to determine the in situ conditions of two populations present in rivers from basin of Rio Paraguaçu at the Bahia State, ii) to evaluate the structure and genetic variability presented in both populations, iii) to establish in situ and ex situ conservation strategies. It were realized collection in november 2007, when was realized in situ characterization of G. mustelinum. SSR markers were used for analyze 218 genotypes deriving from two populations of the G. mustelinum, localized at Tocó river and the Capivara river. The allelic frequencies, the heterozigosity and the F statics were estimated. All the plants were classified as wild and natives, and there was no evidence of the use the plants or its parts. The populations showed different conservation conditions in situ. Few plantlets were found in sites with excessive cattle feed, an indication that the damages in young plants should be high enough to compromise the renovation of the populations. On the other hand, populations were well preserved when the anthropic damages was low or inexistent. The 14 SSR primer pairs amplified 17 loci with a medium number of 5 alleles per locus (a total of 85 alleles). The high level of endogamy estimated (FIS=0,808) and the low observed heterozygosity (H0=0,093) were indicatives that the populations reproduce mainly by selfing, geitonogamy and crosses between related individuals. The genetic diversity was high (HE=0,482) and the differentiation between the populations was very high (FST=0,328). At least two sites from both populations of G. mustelinum must be preserved to achieve suitable in situ conservation. Actions that preserve the gallery forest and keep the cattle away should implemented, and could be as simple as erecting a fence. It is not possible anticipated if the in situ preservation will be possible. Therefore collections and ex situ preservation of representative specimens are essential to conserve the genetic diversity of native G. mustelinum

Novas propriedades do SKTI (Inibidor de tripsina de soja): inibição para elastase neutrofílica humana e efeitos no processo de injúria pulmonar aguda

Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 µg (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases

Avaliação demográfica, clinico-laboratorial e genética de indivíduos com lupus eritematoso sistêmico e artrite reumatóide residentes em região tropical

The aetiology of autoimmunes disease is multifactorial and involves interactions among environmental, hormonal and genetic factors. Many different genes may contribute to autoimmunes disease susceptibility. The major histocompatibility complex (MHC) genes have been extensively studied, however many non-polymorphic MHC genes have also been reported to contribute to autoimmune diseases susceptibility. The aim of the present study was to evaluate the influence of SLC11A1 gene in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Ninety-six patients with SLE, 37 with RA and 202 controls enrolled in this case-control study, were evaluated with regard to demographic, genetic, laboratorial and clinical data. SLE mainly affects females in the ratio of 18 women for each man, 88,3% of the patients aged from 15 to 45 years old and it occurs with similar frequency in whites and mulattos. The rate of RA between women and men was 11:1, with 77,1% of the cases occurring from 31 to 60 years. The genetic analysis of the point mutation -236 of the SLC11A1 gene by SSCP did not show significant differences between alleles/genotypes in patients with SLE or RA when compared to controls. The most frequent clinical manifestations in patients with SLE were cutaneous (87%) and joint (84.9%). In patients with RA, the most frequent out-joint clinical manifestation were rheumatoid nodules (13,5%). Antinuclear antibodies were present in 100% of the patients with SLE. There was no significant relation between activity of disease and presence of rheumatoid factor in patients with RA, however 55,6% of patients with active disease presented positive rheumatoid factor. Significant association between alleles/genotypes of point mutation -236 and clinical manifestations was not found

Estrutura genética populacional de Heliconius erato e Heliconius melpomene (Lepidoptera: Nymphalidae) em fragmentos de Mata Atlântica do Rio Grande do Norte

Extensive studies using molecular markers on butterflies have shown how a highly fragmented landscape may result in the reduction of gene flow among patches of habitat and, consequently, increase genetic differentiation among populations. However, little is known about Heliconius geographical structure and the effects of fragmentation on the connectivity of populations. Furthermore, findings on the effects of the population structure on the dynamics of mimicry evolution in Heliconius butterflies need to be tested in H. erato and H. melpomene specimens found in other locations other than Central and northern South Americas. For the present study, we had two motivations: (1) compare the population structure of H. erato and H. melpomene given the highly fragmented Brazil s Atlantic Forest habitat; and (2) studying population structure of co-mimics could give us insights into the dynamics of mimicry evolution. For this, we analysed the spatial structure and connectivity of eight populations of Heliconius butterflies, in a total of 137 H. erato specimens and 145 H. melpomene specimens, using nine microsatellites loci, 1144 AFLPs markers and 282 mitochondrial DNA sequences. In general, both species exhibited evidence of population subdivision but no isolation by distance indicating some extent of genetic differentiation among populations. Contrary to Kronforst & Gilbert s (2008) Costa Rican Heliconius, H. melpomene exhibited more genetic differentiation than H. erato based on nuclear markers. However, for mitochondrial DNA, H. erato populations showed more genetic differentiation than H. melpomene. Our results corroborate to other studies on Heliconius butterflies concerning the pronounced population subdivision and local genetic drift found in this genus. Nevertheless, the pattern of this differentiation varies significantly from the pattern found in studies conducted in Central America, where H. erato is generally more differentiated and structured than H. melpomene, based on nuclear markers. This different pattern may reflect different evolutionary histories of Heliconius species in Northeastern Brazil s Atlantic Forest

Estrutura genética de Chromis multilineata (Guichenot, 1853) (Perciformes Pomacentridae) na costa NE do Brasil e ilhas oceânicas: evidências de um passado evolutivo instável.

The fauna of Brazilian reef fishes comprises approximately 320 species distributed along the coast of the mainland and islands ocean. Little is known about the levels of connectivity between their populations, but has been given the interest in the relations between the offshore and the islands of the Brazil, in a biogeographical perspective. The oceanic islands Brazilian hosting a considerable number of endemic species, which are locally abundant, and divide a substantial portion of its reef fish fauna with the Western Atlantic. Among the richest families of reef fish in species are Pomacentridae. This study analyzed through analysis of sequences of the mitochondrial DNA control region (D-loop), the standards-breeding population of C. Multilineata in different areas of the NE coast of Brazil, involving both oceanic islands (Fernando de Noronha Archipelago and of St. Peter and St. Paul) and continental shelf (RN and BA). To this aim, partial sequences were used in the region HVR1 of mtDNA (312pb). The population structure and parameters for the estimates of genetic variability, molecular variance (AMOVA), estimation of the index for fixing (FST) and number of migrants were determined. The phylogenetic relationships between the populations were estimated using neighbor-joining (NJ) method. A group of Bayesian analysis was used to verify population structure, according to haplotype frequency of each individual. The genetic variability of populations was extremely high. The populations sampled show moderate genetic structure, with a higher degree of genetic divergence being observed for the sample of the Archipelago of St. Peter and St. Paul. At smaller geographical scale, the sample of Rio Grande do Norte and the Archipelago of Fernando de Noronha do not have genetic differentiation. Three moderately differentiated population groups were identified: a population group (I), formed by the Rio Grande do Norte (I') and the archipelago of Fernando de Noronha (I''), and two other different groups formed by the island population of the archipelago of Saint Peter and St. Paul (II) and Bahia (III). The genetic patterns found suggest that the species has suffered a relatively recent radiation favoring the absence of shared haplotypes. C. multilineata seems to constitute a relatively homogenous population along the West Atlantic coast, with evidence of a moderate population genetic structure in relation to the Archipelago of St. Peter and St. Paul. These data supports the importance of the dispersal larvae by marine current and the interpopulation similarity this species.

Caracterização in situ e diversidade genética de algodoeiros mocós (Gossypium hissutum raça marie galante) da Região Nordeste do Brasil

Brazil is one of the major centers of diversity for polyploid cotton plants; these plants belong to the genus Gossypium, which has three known species: G. hirsutum, G. barbadense and G. mustelinum. The Northeast is the only region where the three species occur, the last group being endemic. Northeast s cotton plants can be important sources of variability for genetic breeding. It is believed that great part of local diversity is being lost, due to economic, political, cultural and agricultural problems. In an attempt to mitigate this loss and delineate conservation strategies it is necessary to know how the species are found where they occur. The objective was to characterize and determine how plants are maintained in situ in the states of Maranhão, Piauí, Ceará, Rio Grande do Norte and Paraíba at the beginning of the XXI century. The in situ characterization of G. hirsutum and G. barbadense was conducted through structured interviews with the cotton plants owners and through the analysis of the environment. The data were collected during expeditions undertaken between the years 2004 to 2005. Twenty-two plants were collected in the state of Paraíba, forty-four in the state of Rio Grande do Norte, one hundred and forty-six in the state of Ceará, forty in the state of Maranhão and ninety-one plants in the state of Piauí. All plants collected in the states of Paraíba and Rio Grande do Norte belonged to moco type. Moco cotton plants also predominated in the other states, representing 92%, 62% and 78% of plants collected in Ceará, Piauí and Maranhão, respectively. The other cotton plants collected belong to the species G. barbadense. The cotton plants were found in situ as dooryard plants, roads side, feral populations, cultivation or local varieties. Great part were dooryard plants (45.2%), being major in Piauí and Maranhão. Cultivation predominated in Ceará; in Rio Grande do Norte feral populations were the most frequent and, in Paraíba, local varieties. The maintenance of moco plants is related, mainly, to the phytotherapic domestic use (20.9%) and to confection of lamp wicks (29.7%). Few inhabitants in Paraíba, Rio Grande do Norte, Piauí and none in Maranhão used harvest the plants, storage the seeds or gin; however, in Ceará, 40.5% of owners affirmed that they harvested and commercialized the fiber. It was found that the maintenance of species is dependent of the fragile cultural habits of local inhabitants, therefore the maintenance in situ is not a suitable way to conservation of genetic resources. The efforts must be directed to the continuity of collections, maintenance and characterization ex situ

Caracterização genética e in situ de Gossypium barbadense na região norte do Brasil

Brazil has been considered one of the diversity centers of Gossypium barbadense species. It is believed that a relatively big erosion genetic process occurs with the species, due to economic, cultural and agricultural problems. A local diagnostic about species situation is the first step for reducing the diversity loss and establishing conservation strategies in situ. This research aimed the identification of the presence of Gossypium populations, characterization, determination of the main risks and collection of the accesses to store in germoplam banks, in Para and Amapa States. Expeditions were conducted in November 2004. An interview was carried out with the plant proprietor for characterizing in situ of G. barbadense species and of the environment where the plants were inserted. On hundred seventy nine plants in 22 municipal districts were collected in Para State and 117 plants in nine municipal districts in Amapa State. The majority of plants belong to G. barbadense species (98% in Amapa and 94% in Para). Plants occur in back yards, beside roads and spontaneously. That ones from back yards were more abundant (97% in Amapa and 95% in Para) and maintained as medicinal plants as the principal reason. Plants in natural environments in both states evaluated were not found, therefore, the creation of reserves and the application of others conventional methods of maintenance in situ are not applicable. The plant proprietors do not use to store or process seeds. Seed storage was reported as a practice by only 1% of the plant proprietors from Para and 11% from Amapa. The most plants collected were from two to three years of age (58% in Amapa and 93% in Para). As conclusions G. barbadense is the species most spread in the two studied states and are found in back yards. In Amapa State the botanical variety barbadense or Quebradinho is predominant, whereas in Para State the predominant variety is brasiliense or Rim-de-boi. Adequate conservation of thestudied species must be carried out in germoplasm collections maintained ex situ

Estrutura genética de populações de Amphobotrys ricini, agente causal do mofo cinzento da mamoneira

The gray mold, causal organism Amphobotrys ricini, is one of the major diseases of castor bean. Difficulties in managing plant disease arises form the limited understanding of the genetic structure of A. ricini, their complexity and variability make it difficult to control. Genetic structure can be used to infer the relative impact of different forces that influence the evolution of pathogen populations, that allow to predict the potencial for pathogen populations to envolve in agricultural ecosystems. Growers protect their crop by applying fungicides, but there aren t fungicides to provide significant control of gray mold of castor bean. The objectives of this work were use RAPD to determine the genetic structure of A. ricini subpopulations in Paraíba and assay the sensitivity of A. ricini isolates to azoxystrobin and carbendazim. To determine the genetic structure of A. ricini subpopulations in Paraíba, 23 isolates were colleted from two different geographic location (subpopulation). These isolates were analysed by RAPD using 22 random decamer primers, purchased from OPERON, produced a total of 80 markers polimorphics. The resulting matrixes were analysed using PopGene version 1.32. Sensitivity to azoxystrobin and carbendazim of 30 isolates, colleted form Paraíba and Alagoas, was estimated based on spore germination and colony growth inhibition. The stock solutions were added toV8 medium after sterilization to produce final concentrations of 0, 0.01, 0.1, 1, 10, and 100 µg/ml of carbendazim and 0, 0.001, 0.01, 0.1, 1, and 10 µg/ml of azoxystrobin. All statistical analyses were performed using SAS to estimate the dose that inhibited fungal growth by 50% (ED50 values). The genetic diversity within subpopulations (Hs=0,271) accounted for 92% of the total genetic diversity (Ht=0,293), while genetic diversity between subpopulations (Gst = 0,075) represented only 7,5%. The estimated number of migrants per generation (NM ) was 6,15. Nei s average gene identity across 80 RAPD loci was 0,9468. Individual ED50 values, for the 30 isolates screened for their sensitivity to azoxystrobin, ranged From a maximum of 0,168 µg/ml to a minimum of 0,0036 µg/ml. The ED50 values for carbendazim varied within the range of 0,026 to 0,316 µg/ml

Caracterização da resistência do algodoeiro a Ramularia areola e variabilidade molecular do patógeno

This research was conducted with the aim to study the genetic and pathogenic structure of Ramularia areola isolates collected in Brazil and to characterize the resistance response in cotton plants to ramularia spot. The genetic variability of 28 isolates of R. areola was studied using RAPD markers. The pathogenicity evaluation was realized by the inoculation of 6 isolates on cotton varieties Guazuncho-2 (Gossypium hirsutum) and VH8-4602 (Gossypium barbadense). The inheritance of disease resistance was studied using an artificially inoculated population of F2 individuals derived from the intercross of Guazuncho-2 (susceptible variety) end VH8-4602 (resistant variety), and also the parents and F1 individuals. Molecular polymorphism between the G. hisutum varieties DeltaOpal (suscetible) and CNPA CO-11612 (resistant) was estimated by 118 SSR and 24 AFLP markers. The parental genotypes Guazuncho-2 and VH8-4602 were selected for mapping, and then Recombinant Inbred Lines (RIL´s) derived from this crossing were evaluated with SSR 12 markers. The analysis of population structure of R. areola revealed that the three subpopulations were genetically simillar (Gst=0.18), and the isolates from Goiás and Minas Gerais were more similar to each other (0,92). This probability can be related to the relatively high gene flow among the three subpopulations (Nm=2.20). The isolates R. areola 9.1, from Minas Gerais State and 8.1 and 8.3 from Goiás State were the most aggressive ones to the susceptible variety Guazuncho-2. The variety VH8-4602 presented high level of resistance to ramularia spot. No differential interaction was observed between the pathogens and the analyzed varieties, and the resistance was classified as horizontal. The quantification of disease by number of necrotic lesions and number of spores in individual plants of F1 and F2 generations from the crossing between the varieties Guazuncho-2 and VH8-4602 presented continuous distribution, suggesting polygenic resistance. The resistance is probabilly recessive, since necrotic lesions and sporulation were observed on F1 plants. The molecular polymorphism between DeltaOpal e CNPA CO-11612 lineages was low (6%), then would be difficult to accomplish molecular mapping of disease resistance using this intercross. With the genotyping of the RIL s it was verified that 25% of the markers segregated in the proportions proposed by Mendel s Law and 75% of the studied markers presented segregation distortion in favor to the parental G. hirsutum. Both the low genetic variability of the pathogen and the number of resistance genes suggest that durable genetic resitance may be achieved

Análise molecular dos éxons 8 a 11 do gene da p53 em amostras de câncer de colo de útero no Rio Grande do Norte

Mutations on TP53 gene are common in human cancer but not in cervical cancer where they are rarely found and the inactivation and degradation of p53 protein are attributed to the action of E6 viral oncogene from high risk human papillomavirus (HPV). Analysis of cervical cancer cell lines suggests that HPV negative samples shows mutation on TP53, but clinical approaches didn t confirmed this hypothesis. However, in most TP53 mutations studies on cervical cancer, only the exons 5 to 8 were analyzed. Approximately 90% of mutations described are on this region. Recent studies on several cancer suggests that mutation frequency in the other exons must be considered. The aim of this work was to verify whether mutations on coding and non-coding regions occur in cancer tissue from cervical cancer in patients from Rio Grande do Norte using Denaturing Gradient Gel Electrophoresis (DGGE) as screening tool. Exons 8 to 11 were analyzed including some introns from 80 tumor samples and 8 peripheral blood samples from healthy women. DNA were submitted to PCR using primers with GC clamp on the end of one of them. The results were observed for each region after DGGE and silver staining. It was observed no amplified fragment with different migration profile from those obtained from DNA of peripheral blood. These results agree with those from literature where TP53 mutations in cervical cancer have been described in a very low frequency

Caracterização genética e evolução dos vírus dengue no Estado do Rio Grande do Norte

Dengue is considered as the most important arthropod-borne viral disease throughout the world due to the high number of people at risk to be infected, mainly in tropical and subtropical regions of the planet. The etiologic agent is Dengue Virus (DENV), it is a single positive-stranded RNA virus of the family Flavivirus, genus Flaviviridae. Four serotypes are known, DENV-1, DENV-2, DENV-3 and DENV-4. One of the most important characteristic of these viruses is the genetic variability, which demands phylogenetic and evolutionary studies to understand key aspects like: epidemiology, virulence, migration patterns and antigenic characteristics. The objective of this study is the genetic characterization of dengue viruses circulating in the state of Rio Grande does Norte from January 2010 to December 2012. The complete E gene (1485 pb) of DENV1, 2 e 4 from Brazilian (Rio Grande do Norte) patients was sequenced. Phylogenetic analysis was performed using MEGA 5.2 software, Tamura-Nei model and Neighbor-Joining trees were inferred for the datasets. In Brazil, there is just one DENV-1 genotype (genotype V), one DENV-2 genotype (Asian/American) and two DENV-4 genotypes (genotypes I and II). Brazilian strains of DENV-1 are subdivided in two different lineages (BR-I and BR-II), the Brazilian strains of DENV-2 are subdivided in four lineages (BRI-IV) and genotype II of DENV-4 is subdivided in three Brazilian lineages (BRI-III). The viruses isolated in RN belong to lineage BR-II (DENV-1), BR-IV (DENV-2) and BR-III (DENV-4).The Caribbean and near Latin American countries are the main source of these viruses to Brazil. Amino acids substitutions were detected in three domains of E protein, this makes clear the necessity of studies that associate epidemiological and molecular data to better understand the effects of these mutations. This is the first study about genetic characterization and evolution of Dengue viruses in Rio Grande do Norte, Brazil

Análise da estrutura genética populacional da piraúna (cephalopholis fulva: serranidae) ao longo da costa e ilhas oceânicas brasileiras

Brazil has about 8,500 km of coastline and on this scale, fishing is a historically important source of animal protein for human consumption. The national fishing background shows a growth of marine fishery production until 1985 and within this period it was recorded a steady decline. From the year 2003 fishing statistics aim to some "recovery" of the total fisheries production, which probably is related to a change in industry practice. The target of commercial fishing became smaller species with low commercial value, but very abundants. The coney, Cephalopholis fulva (Serranidae), is one of these species that have been suffering a greater fishing pressure in recent years. In order to provide data about the current situation of the genetic diversity of these populations, several molecular markers have been being used for this purpose. The prior knowledge of genetic variability is crucial for management and biodiversity conservation. To this end, the control region sequences (dloop) of mtDNA from Cephalopholis fulva (Serranidae) from five geographical points of the coast of Brazil (Ceará, Rio Grande do Norte, Bahia and Espírito Santo) and the Archipelago of Fernando de Noronha (FN) were sequenced and their genetic diversity analyzed. The FST values were very low (0.0246 to 0.000), indicating high gene flow between the sampled spots. The indices h and indicate a secondary contact between previously allopatric lineages differentiated or large and stable populations with long evolutionary history. Tests of Tajima and Fu showed expansion for all populations. In contrast, the mismatch distribution and SSD indicated expansion just for coastal populations. Unlike other species of the Atlantic which have been deeply affected by events on later Pleistocene, the population-genetic patterns of C. fulva may be related to recent events occurred approximately 130,000 years ago. Moreover, the data presented by geographical samples of the specie C. fulva showed high genetic diversity, also indicating the absence of deleterious effects of over-exploitation on this specie, as well as evidence of complete panmixia between all sampled populations

Desenvolvimento de marcadores microssatélites e estrutura genética espacial da Copernicia prunifera (Miller) H. E. Moore (Arecaceae)

The present study aimed to develop microsatellite markers (SSR) for Copernicia prunifera; and characterize the demographic pattern and the spatial genetic structure (SGS) in different development stages of C. prunifera in a natural population of Rio Grande do Norte (RN) by using ISSR molecular markers. 17 SSR primers pairs were developed, which were tested by using DNA from samples of different populations. The demographic and genetic spatial structure was assessed in a plot with an area of 0.55 ha, where all individuals were georeferenced. The molecular analyses with the use of microsatellite markers pointed out that all built primers pairs, when submitted to PCR, had amplification. They showed sizes of base pairs ranging between 113 and 250 bp. The demographic analyses showed a clustered standard of spatial distribution in the first distance classes, random between 40 and 50 m and segregated in higher distances. Eight ISSR primers were used, thereby producing a total of 102 loci, with 100 of them being polymorphic. Among the three stages, the young showed the highest Nei’s genetic diversity index (He = 0.37); whilst the lowest index was found in the reproductive adults (He = 0.34). The AMOVA results showed a greater genetic differentiation within the development stages (98.61%) in comparison to the interval among the stages (1.39%). The total population (n = 161) showed a positive and significant relationship of kinship in the first distance class (12.3 m). The young showed a significant kinship up to 10.5 m and negative in the fifth distance class (37.6 m). The non-reproductive adults had a positive relationship of kinship in the first distance class (11.0 m) and random distribution of genotypes in the remaining classes. The reproductive adults showed genotypes spatially distributed in a random way. The values for the genetic bottleneck tests proved that the number of loci with excess observed heterozygosity was greater than expected. The SGS results reflect the restricted dispersion of the species, and the bottleneck tests reflect the reduction genotypes provoked by the anthropization of natural environments of C. prunifera.