Heterologous Microsatellite Research Articles

Genetic characterization and diversity assessment in ‘Bhangor’ indigenous swamp buffalo population using heterologous microsatellite markers

‘Bhangor’ newly identified swamp buffalo population from North East Indian, was characterized using microsatellite markers. Genomic DNA was isolated from blood samples of 76 unrelated animals, 15 microsatellite markers (CSSM33, BM1818, CSRM60, HEL13, ILSTS019, ILSTS025, ILSTS028, ILSTS029, ILSTS033, ILSTS036, ILSTS056, ILSTS058, ILSTS061, ILSTS089 and ETH003) were found to be highly polymorphic in the population of the selected markers. A total of 114 alleles were observed, which ranged from 3 in CSRM60 and ILSTS025 locus to 12 in ILSTS056 and ILSTS061. The mean effective number of alleles across all polymorphic loci was found to be 3.76. The overall mean expected heterozygosity and unbiased expected heterozygosity values were 0.67 and 0.68, ranging from 0.067 (ILSTS025) to 0.85 (ILSTS058) and 0.068 (ILSTS025) to 0.86 (ILSTS058), respectively. Within the population, the inbreeding estimates (FIS) ranged between −0.4352 and 0.804, with an average FIS of 0.114 ± 0.033. The outcome for infinite allele model (IAM), two-phase model (TPM) and test for mode shift revealed the absence of any recent bottleneck in the investigated buffalo population. The population was found to be in optimum diversity based on polymorphic microsatellite markers. With fast changing agro-climatic conditions; there is an urgent need to characterize the nondescript livestock populations.

Genetic characterization of donkeys of Braj region of India

The grey type donkeys of Braj region of Uttar Pradesh in India were genetically characterized using ten heterologous microsatellite loci. At these loci the PCR product size ranged from 75-95 bp at locus HTG6 to 251-277 bp at locus COR18. The observed number of alleles varied from 4 (HTG15) to 10 (HTG7 and AHT4) with a mean of 7.50 ± 1.96. The observed heterozygosity ranged from 0.46 (HTG6 and NVHEQ54) to 0.79 (COR18) with a mean of 0.66 ± 0.12. The mean genetic diversity estimate (FIS) was 0.157. When these donkeys were compared on the basis of allelic frequency data at these loci to the brown type donkeys of Ladakh, Spiti and Rayalseema regions in the union territories/states of Ladakh, Himachal Pradesh and Andhra Pradesh, respectively, they clustered independently from these three donkey populations in a dendrogram based on Goldstein’s average square distances indicating their genetic distinctness.

Obtaining microsatellite markers for Pseudoplatystoma reticulatum using heterologous primers

Studies on genetic composition in fish populations contribute to conservationist practices and inbreeding control in fish stocks. To this end, molecular tools such as microsatellite markers (SSRs) are often used, but they are expensive and time-consuming to develop. A species-specific heterologous marker emerges as an alternative, which can be used in taxonomically related species in a fast way. Our goal was to test SSRs markers of Brachyplatystoma rousseauxi and Pseudoplatystoma punctifer in P. reticulatum in an unprecedented way. For this purpose, DNA was extracted from fragments of the caudal fin of 222 P. reticulatum adults, using a NaCl-based method. Then, DNA samples were amplified by Polymerase Chain Reaction (PCR) using six markers, four from B. rousseauxi (BR38, 47, 51, and 61) and two from P. punctifer (PPU13 and PPU15). Two primers showed non-specific amplification and were disregarded (BR38 and PPU13). In the remaining four primers, the number of alleles per locus varied between two (BR47) to sixteen (BR51), and the average size of alleles was between 142 and 400 bp. Mean effective number of alleles per locus ranged from 10,650 (BR51) to 1,784 (BR47), with null or low-frequency alleles in all studied loci. Observed heterozygosity ranged from 0.299 (BR47) to 0.640 (BR51) and was always lower than the expected heterozygosity. Hardy-Weinberg balance was significant (p < 0.05) in all loci, and inbreeding coefficient (FIS) was always positive. Polymorphic Information Content (PIC) confirmed the efficiency of the markers since they had moderate (BR47) to high levels of information (BR51, BR61, and PPU15). Transferability test showed that the heterologous microsatellite molecular markers, originally for B. rousseauxi and P. punctifer, were efficient in P. reticulatum, producing three primers with high information content. Therefore, these markers can be safely used in future population studies of this species.

Evidence of facultative parthenogenesis in three Neotropical pitviper species of the Bothrops atrox group.

We examined four suspected cases of facultative parthenogenesis in three species of a neotropical lineage of pitvipers of the Bothrops atrox group. Reproduction without mating was observed in captive females of B. atrox, B. moojeni and B. leucurus housed alone for seven years (the two former species) and nine years (the latter one). In addition to the observation of captivity data, we investigated molecularly this phenomenon using heterologous microsatellites. DNA was extracted from the mothers’ scales or liver, from embryo and newborn fragments, and yolked ova. Four of the microsatellites showed good amplification using Polymerase Chain Reaction and informative band segregation patterns among each mother and respective offspring. Captivity information, litter characteristics (comparison of the number of newborns, embryos and yolked ova) and molecular data altogether agreed with facultative parthenogenesis predictions in at least three out of the four mothers studied: B. atrox (ID#933) was heterozygous for three out of the four markers, and the sons S1 and S2 were homozygous; B. moojeni (BUT86) was heterozygous for two out of four markers, offspring S1, S3, E2, and E4, and O1 to O6 were homozygous; and B. leucurus (MJJS503) was heterozygous for three out of four markers, and son E1 and O1 were homozygous. B. moojeni (BUT44) was homozygous for all loci analyzed in the mother and offspring, which although not informative is also consistent with parthenogenesis. This study represents the first molecular confirmation of different pitviper species undergoing facultative parthenogenesis among Neotropical endemic snakes.

Genetic variability, management, and conservation implications of the critically endangered Brazilian pitviper Bothrops insularis.

Information on demographic, genetic, and environmental parameters of wild and captive animal populations has proven to be crucial to conservation programs and strategies. Genetic approaches in conservation programs of Brazilian snakes remain scarce despite their importance for critically endangered species, such as Bothrops insularis, the golden lancehead, which is endemic to Ilha da Queimada Grande, coast of São Paulo State, Brazil. This study aims to (a) characterize the genetic diversity of ex situ and in situ populations of B. insularis using heterologous microsatellites; (b) investigate genetic structure among and within these populations; and (c) provide data for the conservation program of the species. Twelve informative microsatellites obtained from three species of the B. neuwiedi group were used to access genetic diversity indexes of ex situ and in situ populations. Low‐to‐medium genetic diversity parameters were found. Both populations showed low—albeit significant—values of system of mating inbreeding coefficient, whereas only the in situ population showed a significant value of pedigree inbreeding coefficient. Significant values of genetic differentiation indexes suggest a small differentiation between the two populations. Discriminant analysis of principal components (DAPC) recovered five clusters. No geographic relationship was found in the island, suggesting the occurrence of gene flow. Also, our data allowed the establishment of six preferential breeding couples, aiming to minimize inbreeding and elucidate uncertain parental relationships in the captive population. In a conservation perspective, continuous monitoring of both populations is demanded: it involves the incorporation of new individuals from the island into the captive population to avoid inbreeding and to achieve the recommended allelic similarity between the two populations. At last, we recommend that the genetic data support researches as a base to maintain a viable and healthy captive population, highly genetically similar to the in situ one, which is crucial for considering a reintroduction process into the island.

Transferability of heterologous primers in Brycon falcatus

ABSTRACT The genusBryconcomprises fish species of significant socioeconomic and biological importance in Brazil. Despite that, the genetic knowledge about these species is scarce, especially regardingBrycon falcatus. Thus, the objective of this study was to evaluate the transferability of heterologous microsatellite primers inB. falcatus for the first time. Heterologous primers obtained from B. opalinus, B. hilarii, B. insignis, B. orbignyanus, B. amazonicus, Prochilodus argenteus, Prochilodus lineatus, Piaractus mesopotamicus, and Colossoma macropomum were evaluated. The primers that showed the best amplification patterns were applied to a sample of 22 individuals and the genetic parameters were calculated. Nine primers displayed satisfactory cross-amplification withB. falcatus: BoM5 (Brycon opalinus); Bh8, Bh13 and Bh16 (B. hilarii); Borg59 (B. orbignyanus); Bag22 (B. amazonicus); Par12 and Par80 (P. argenteus), and Cm1A8 (C. macropomum). The genetic parameters (number of alleles, effective alleles, allele richness, and expected and observed heterozygosity) and the polymorphic information content (PIC) confirmed the viability of these primers for population genetics analyses. Our study demonstrates the potential of transferability of microsatellite markers from related species and even different genera to B. falcatus, providing usefull tools for future population genetic studies in this species.

An economic method for the identification of catfish hybrids using microsatellites: a case study of Pseudoplatystoma metaense × Leiarius marmoratus hybrids

The hybrids of pimelodid catfish provide products of excellent genetic value for aquaculture due to their greater growth in size, resistance to diseases and adaptation to feed concentrate. However, this practice involves risks such as genetic contamination, the commercialization of hybrid products as pure species, the introduction of exotic species and escapes of fish farming products. Thus, the aim of this study was to evaluate microsatellites for the identification of hybrids between Pseudoplatystoma metaense and Leiarius marmoratus. Genomic DNA extraction was achieved from fin tissue and nine heterologous microsatellites previously reported in the literature were amplified: Pcor01, Pcor02, Pcor05, Pcor10, Phrac02, Phrac07, Phrac09, Prt03, and Prt30. The DNA extraction resulted in high concentrations and a good DNA quality with absorbance values at 260/280 nm between 1.96 and 2.07. The microsatellites Pcor10, Phrac02, Phrac07 and Prt03 indicated heterozygosity patterns in the agarose gel for hybrids, with the presence of two diagnostic bands. Our results showed that these microsatellites are a powerful and economic tool for identifying interspecific hybridization, without requiring genotyping or sequencing. Here, we made the first report of amplification of these microsatellites for the endemic species of the Orinoco basin Pseudoplatystoma metaense, useful information for population and phylogeny studies. We also highlight the importance of using molecular identification techniques for hybrids and we invite the aquaculture industry to conserve good management practices in the fish farming areas. Thereby, hybrids do not compromise the sustainable development of aquaculture or pose serious risks for native and wild species.

Transferability of heterologous microsatellite primers in Leiarius marmoratus

The Amazonian Jundiá (Leiarius marmoratus) (Siluliformes: Pimelodidae) is a species of catfish with social and economic importance in some South American countries such as Brazil and Colombia. Genetic evaluation of this species is limited due to the lack of specific molecular markers, hindering studies on genetic diversity and structure in animals under captive conditions or in natural populations. The objective of the present study was to evaluate the transferability of heterologous microsatellite markers in Leiarius marmoratus. Thirty-two heterologous primers were tested in L. marmoratus. The primers that presented the best standards were applied to 20 specimens, and the number of alleles (Na), number of effective alleles (Ne), gene diversity per Locus (GdL) and percentage of amplification failure (Md) were calculated. Eleven primers demonstrated satisfactory transferability patterns, all from the fish of the Pimelodidae family, of which, seven were monomorphic and four polymorphic. The eleven markers presented low percentage of Md (mean was 5.9% samples per locus). Na varied from one to two alleles per locus, revealing low polymorphism in the evaluated samples. The mean Ne and GdL numbers were 1.77 and 0.32, respectively. The transferability of the heterologous microsatellite loci in L. marmoratus was shown to be possible. However, further tests are needed to apply these markers in population genetic studies.

Genetic Diversity Analysis of Five Egyptian Buffalo Populations Using Microsatellite Markers

For assessing the genetic diversity and genetic characterization of five Egyptian buffalo populations a total of 12 microsatellite markers were used. The total number of buffaloes sampled was 80, collected at random from five farms in five different governorates; Cairo, Kafr El-Sheikh, Shebeen El-Kom, Menoufia, and Beni Suef. The genetic parameters (allelic diversity, allelic frequencies, observed heterozygosity, unbiased expected heterozygosity, and polymorphic information content) were calculated using three different programs. All used microsatellites were polymorphic and ranged from four alleles (Loci; CSSM029, CSSM036, CSSM038, CSSM043, CSSM046, and ILSTS005) to nine alleles (Loci; BM1818 and CSSM047) with a total of 64 alleles in the whole population. Allelic richness for the whole population ranged between 3.297 (in locus CSSM029) and 6.806 (in locus CSSM047) with overall mean 4.574. Within populations, Kafr El-Sheikh population had the highest average of allelic richness (4.384). This indicates the potential of this population to adapt with environmental changes in future compared with other populations. BMC1013, BM1818, CSSM019, and CSSM047 showed the highest allelic richness. PIC estimates were high and ranged between 0.65 (in locus CSSM029) and 0.92 (in locus CSSM022) with an average of 0.82. Values of Ho were lower than values of HNb for all populations, which denoting depression of heterozygotes in these populations and may be attributable to existence of null alleles and inbreeding. This study as well proves the usefulness of heterologous bovine microsatellite markers in evaluation of the genetic variability in Egyptian buffalo populations due to high polymorphism, informativeness of these markers which can be used to develop future breeding strategies and conservation decisions on our indigenous breed.

Assessment of genetic diversity and bottleneck in Purnathadi buffaloes using short tandem repeat markers

This study is the maiden attempt for genetic characterization of the Purnathadi, a phenotypically distinct buffalo population of the western Vidarbha region of Maharashtra and to explore genetic diversity using STR markers. A total of 48 unrelated Purnathadi buffaloes from the entire native tract were genotyped using a battery of 25 heterologous microsatellite markers. 5′ end of forward primer of each microsatellite marker was labeled with one of the fluorescent dyes, viz., FAM (Blue), VIC (Green), NED (Yellow) or PET (Red) to assess the fragment length of genotyped PCR product with automated DNA sequencer (ABI 3100). 23 microsatellite loci (except ETH003 and ILSTS030) amplified successfully and adequately high allelic diversity (observed: 0.615 ± 0.043 and expected: 0.655 ± 0.037) was reported with 162 distinct microsatellite alleles. Sufficiently high Shannon index and PIC indicated the suitability of markers to evaluate genetic diversity in Purnathadi buffaloes. Within-population inbreeding estimates (FIS ) for Purnathadi buffalo ranged between −0.171 and 0.495 with global FIS average of 5.9%. The outcome for IAM, TPM and test for mode shift revealed the absence of any recent bottleneck in Purnathadi buffalo. Pairwise FST (genetic differentiation) and gene flow between Purnathadi, Nagpuri and Marathwadi buffaloes were estimated using genotype data of 19 microsatellite markers. Lowest FST (0.031) was observed between Nagpuri and Purnathadi buffaloes with highest gene flow of 7.91% and highest FST (0.094) was between Purnathadi and Marathwadi populations. Present findings will definitely support in designing breeding plan for genetic improvement, as well as for developing conservation strategies of Purnathadi buffalo population. The comparative molecular study with other breeds of the regions is needed.

Genetic Diversity and Population Structure of Genypterus chilensis, a Commercial Benthic Marine Species of the South Pacific

The red cusk-eel (Genypterus chilensis, Guichenot 1848) is an iconic species in Chilean gastronomy, with a cultural significance that extends beyond its economic value, worthy of conservation. Despite the decline in fishery productivity across most of the distribution range for this species, little effort has been devoted to assessing stock management. In the present study, seven heterologous microsatellite loci (cmrGb3.8.1, cmrGb5.2B, cmrGb4.2A, cmrGb4.2B, cmrGb5.9, cmrGb4.11 and cmrGb2.6.1) were genotyped in a total of 153 individuals from four locations in central and southern Chile (30.5oS–37.6oS), comprising ~1000 km of coastline. Five microsatellite loci were consistently amplified with a low frequency of null alleles (<2.5%). These markers showed high average heterozygosity (HO=0.886 and HE=0.884) and PIC (0.869). Significant genetic differentiation among locations (global FST=0.043, global DJost=0.252) suggested the presence of at least three different genetic groups along the Chilean coast, with moderate levels of admixture between the two central populations (0.168 <Qmean<0.822). The southern population showed no admixture with the central populations (Qmean=0.985), less allelic richness and a small effective population size, indicating that urgent management measures must be developed for this fishery. The present study provides baseline information to assist fishery and aquaculture management, contribute to recovery of declining populations, promote sustainable fishing and aquaculture and avoid the collapse of red cusk-eel production.

Genetic characterization and assessment of diversity in Pandharpuri buffalo breed of India using heterologous microsatellite markers

The present study was aimed at genetic characterization and diversity analysis of Pandharpuri buffalo population using the Food and Agriculture Organization (FAO) recommended bovine microsatellite markers. Genomic DNA was isolated from blood samples of 50 unrelated animals and a total of 23 microsatellite loci were amplified using polymerase chain reaction. Among 23 recommended microsatellite markers, 14 markers (BM2113, BM1818, CSSM66, HEL13, INRA037, ILSTS05, HAUT27, INRA023, INRA035, HEL5, ETH3, NRA063, MM12 and ETH10) were found to be highly polymorphic in Pandharpuri population. The amplified products were run on polyacrylamide gel electrophoresis (PAGE) apparatus along with a ladder; subsequently, the allele typing was done based on silver staining of the gel. The effective and observed number of alleles, heterozygosity (expected and unbiased) estimates and polymorphic information content (PIC) levels were estimated for each locus. A total of 87 alleles were secured for 14 polymorphic loci studied giving an overall average of 6.21 alleles per locus. The number of alleles ranged from 2 (INRA063) to 9 (BM1818 and HEL13). The mean effective number of alleles across all polymorphic loci was found to be 4.28. The overall mean expected heterozygosity and unbiased expected heterozygosity values were 0.77 and 0.76, ranging from 0.50 (INRA063) to 0.88 (BM1818) and 0.50 (INRA063) to 0.88 (BM1818), respectively. The average PIC estimate across all polymorphic loci was 0.73 ranging from 0.373 (INRA063) to 0.864 (BM1818). In the present study, the characterization and diversity estimates are reported for Pandharpuri population. It was found to maintain optimum diversity based on 14 polymorphic microsatellite markers.

Microsatellite-based Genetic Diversity and Mutation-Drift Equilibrium in Dharwadi Buffalo (Bubalus bubalis) of India

Genetic diversity was assessed in a buffalo population (Dharwadi) of Karnataka by utilizing 25 heterologous bovine microsatellite markers. Genotypic status of individuals at each locus was identified by automated DNA sequencer, and allelic data were analyzed for genetic diversity parameters. Observed number of alleles varied between 5 and 19 with 13.12 ± 0.80 alleles per locus. High level of heterozygosity (0.63) indicated plenty of existing genetic diversity in this buffalo population. Within-population inbreeding estimate (FIS) was significantly positive (0.186) and 88% of the investigated loci contributed toward the homozygote excess. Accordingly, significant deviation from Hardy–Weinberg equilibrium was observed at 24 of 25 analyzed loci. The population did not suffer reduction in effective population size in the last few generations. Assessment of Dharwadi buffalo population for mutation-drift equilibrium did not reveal significant heterozygosity excess under all the models of microsatellite evolution as per Sign test. No shift was recorded in the frequency distribution of alleles, and a normal L-shaped curve indicated absence of any recent genetic bottleneck. The study highlighted sufficient genetic variability within Dharwadi buffalo population that can be utilized in combination with phenotype and production information for registration as a distinct breed of Indian buffalo.

Transferability of heterologous microsatellite primers in Brycon gouldingi

ABSTRACT: Brycon gouldingi is a species of neotropical fish of socioeconomic and environmental importance in the Tocantins-Araguaia Basin. Genetic studies on this species are still limited, making it difficult to evaluate the population structure and genetic diversity in natural and captive stocks. Here, we aimed to evaluate the transferability of heterologous microsatellite primers in B. gouldingi. A total of 30 primers for eight species were evaluated: Brycon hilarii, Brycon opalinus, Brycon cephalus, Brycon orbignyanus, Prochilodus lineatus, Prochilodus argenteus, Piaractus mesopotamicus, and Colossoma macropomum. The primers that showed the best amplification patterns were applied to 20 specimens of B. gouldingi, and their genetic parameters were assessed. Among the 30 primers, seven showed satisfactory transferability, six of which belonged to the genus Brycon: Bh13 (B. hilarii), BoM5, BoM13 (B. opalinus), Borg9, Borg13, and Borg59 (B. orbignyanus), and one belonged to P. argenteus (Par80). The primers for the other species tested showed non-specificity or monomorphism; and were therefore, excluded from the analyses. The number of alleles ranged between two (Borg13 and Borg59) and three (Bh13, BoM5, BoM13, Borg9 and Par80), with sizes varying between 103 bp (BoM5) and 430 bp (Borg9). Four primers showed evidence of null alleles (BoM13, Borg9, Borg13, and Par80), which could probably be attributed to the respective Hardy-Weinberg deviation. Thus, seven primers were validated for cross-amplification in B. gouldingi, which may be used in future studies involving this species.

Somaclones of mandacaru (Cactaceae) with high morphological divergence may generate new varieties of ornamental cacti

Somaclones of mandacaru cactus ( Cereus peruvianus Mill.) with typical and atypical shoots were regenerated from a callus. Since the genetic relationship among regenerated somaclones has not been investigated, current study employed heterologous microsatellite primers to examine the molecular diversity within and among somaclones, showing typical and atypical phenotypes with high morphological divergence. Nei identity value calculated between the somaclones was high (I = 0.929) and AMOVA showed higher genetic variation within (96%) than among (4%) the samples of somaclones. The polymorphism in the microsatellite loci indicated high levels of mean observed and expected heterozygosity in atypical somaclones, presumably with high adaptive potential and as source of genotypes for generation of new varieties of ornamental cacti. On the other hand, low molecular divergence between typical and atypical morphologies of somaclones is a promising perspective for use of the atypical somaclones as source of chemical compounds of commercial and industrial interest. The somaclonal variations occurring in vitro callus culture has generated phenotypically differentiated subpopulations with low molecular divergence, however with high genetic variability, enough to be recommended as a source of genotypes to generate new varieties of ornamental cacti and of plants with new traits, necessary for breeding programs.

Genetic characterization of brown type donkeys of Andhra Pradesh using microsatellite markers

The brown type donkeys of Andhra Pradesh, which are mainly concentrated in Kurnool and Anathapur districts, were evaluated for within breed genetic diversity and bottlenecks using heterologous microsatellite markers. The genomic DNA, isolated from 28 blood samples collected from Kurnool district, were amplified by PCR using FAM and HEX labeled primers and resolved for alleles on automatic DNA sequencer. In all, twenty loci of the horse origin were tested and only 12 loci gave scorable results. Rest of the loci either did not amplify (HMS3 and HMS7) or did not resolve properly (VHL20) or showed less than 4 alleles (HMS5, HMS6, HTG4, ASB17 and COR22) in the studied population. At the 12 loci included in the final analysis, the PCR product size range varied from 76-92 bp at locus HTG6 to 257-273 bp at locus COR18. The observed number of alleles varied from 4 (VHL209) to 10 (AHT5 and HTG7) with a mean of 6.92±1.83. The effective number of alleles ranged from 1.62 (VHL209) to 7.91 (AHT5) with a mean of 4.21±2.06. The observed heterozygosity ranged from 0.32 (HMS2) to 0.92 (AHT5) with a mean of 0.57±0.2. The expected heterozygosity ranged between 0.39 (VHL209) to 0.89 (AHT5 and HTG7) with a mean of 0.72±0.14. The mean genetic diversity estimate (FIS) was 0.21 indicating moderately high levels of inbreeding. The cumulative exclusion probability (PE) of these loci was 0.999892 indicating their suitability for parentage testing in these donkeys. The sign test, standardized differences test, the Wilcoxon test using the allelic frequency data at the studied loci as well as normal 'L' shaped distribution of the allelic frequency indicated the absence of any recent genetic bottleneck in Brown type donkeys of Andhra Pradesh. When these donkeys were compared to Spiti donkeys of Himachal Pradesh on the basis of allelic frequency data at these loci, they showed Nei's standard and unbiased genetic distances of 0.32 and 0.29, respectively.

Phenotypic and genotypic characterization and compatibility among genotypes to select elite clones of cupuassu

ABSTRACT The expansion of the genetic base of cultivated materials is an ongoing activity of the cupuassu (Theobroma grandiflorum) breeding program. However, the parents involved need to be genotypically and phenotypically characterized to ensure compatibility of crossings, as well as to assist in the selection of more promising individuals for hybridization. This study aimed to identify and select T. grandiflorum clones that are compatible and genetically divergent using tools such as the estimates of genotypic, phenotypic, and combined distances, as well as the compatibility rates among clones. The genetic distance analysis of the clones was performed with 14 heterologous microsatellite primers of cocoa (Theobroma cacao) that amplify the DNA of cupuassu. Phenotypic characterization was based on 14 variables related to fruit production. The joint dissimilarity matrix was obtained by means of the sum of the phenotypic and molecular dissimilarity matrices. The intra- and inter-clonal compatibility was estimated through controlled crossings. A low correlation was noted between the dissimilarity matrices based on the molecular and agronomic data. As for compatibility, all clones were self-incompatible, with different compatibility rates when crossed. The compatibility index was strongly influenced by the degree of relationship of the clones. It was possible to identify and select the most promising sets of cupuassu clones to be used in breeding programs, despite their genetic relationship.

The use of microsatellite markers in Neotropical studies of wild birds: a literature review.

Despite extensive habitat fragmentation, the Neotropical region possesses 30% of the world´s bird species. Microsatellites have remained one of the most popular genetic markers and have been used in ecological and conservation studies since the 1990's. We conducted a literature review comparing the number of papers published from January 1990 to July 2015 that used microsatellite markers for studies of wild birds in the Neotropical region, USA and some European countries. We assigned the articles to three categories of studies: population genetics, animal behavior/kinship analysis and the development of species-specific bird microsatellite markers. We also compared the studies in the Neotropics that used heterologous versus species-specific markers and provide a list of heterologous markers of utility in multiple birds. Despite the rich bird fauna in the Neotropics, the number of articles published represents only 5.6% of that published by the USA and selected European countries. Within the Neotropical region, Brazil possessed 60.5% of the total papers published, with the remaining 39.5% shared between five countries. We conclude that the lack of specialized laboratories and resources still represents a limit to microsatellite-based genetic studies of birds within the Neotropical region. To overcome these limitations, we suggest the use of heterologous microsatellite markers as a cost-effective and time-effective tool to assist ecological studies of wild birds.

SSR-enriched genetic linkage maps of bermudagrass (Cynodon dactylon×transvaalensis), and their comparison with allied plant genomes.

We report SSR-enriched genetic maps of bermudagrass that: (1) reveal partial residual polysomic inheritance in the tetraploid species, and (2) provide insights into the evolution of chloridoid genomes. This study describes genetic linkage maps of two bermudagrass species, Cynodon dactylon (T89) and Cynodon transvaalensis (T574), that integrate heterologous microsatellite markers from sugarcane into frameworks built with single-dose restriction fragments (SDRFs). A maximum likelihood approach was used to construct two separate parental maps from a population of 110 F1 progeny of a cross between the two parents. The T89 map is based on 291 loci on 34 cosegregating groups (CGs), with an average marker spacing of 12.5cM. The T574 map is based on 125 loci on 14 CGs, with an average marker spacing of 10.7cM. Six T89 and one T574 CG(s) deviated from disomic inheritance. Furthermore, marker segregation data and linkage phase analysis revealed partial residual polysomic inheritance in T89, suggesting that common bermudagrass is undergoing diploidization following whole genome duplication (WGD). Twenty-six T89 CGs were coalesced into 9 homo(eo)logous linkage groups (LGs), while 12 T574 CGs were assembled into 9 LGs, both putatively representing the basic chromosome complement (x = 9) of the species. Eight T89 and two T574 CGs remain unassigned. The marker composition of bermudagrass ancestral chromosomes was inferred by aligning T89 and T574 homologs, and used in comparisons to sorghum and rice genome sequences based on 108 and 91 significant blast hits, respectively. Two nested chromosome fusions (NCFs) shared by two other chloridoids (i.e., zoysiagrass and finger millet) and at least three independent translocation events were evident during chromosome number reduction from 14 in the polyploid common ancestor of Poaceae to 9 in Cynodon.

Genetic diversity of the pampas deer (Ozotoceros bezoarticus) population in the Brazilian Pantanal assessed by combining fresh fecal DNA analysis and a set of heterologous microsatellite loci.

The pampas deer (Ozotoceros bezoarticus) is close to being classified as ‘globally threatened’, with the largest population occurring in the Brazilian Pantanal. Since capture is stressful to these animals, non-invasive sampling methods such as the use of feces can provide reliable sources of DNA. The aim of this study was to use fecal samples to evaluate the genetic variability of the Brazilian Pantanal population of pampas deer. Six heterologous microsatellite markers were used to screen 142 stool specimens. Seventy-four deer were identified, of which 50 adults were used to determine the genetic characteristics of the population. The Pantanal population showed high genetic diversity (mean number of alleles per locus = 11.5, expected heterozygosity = 0.75). This is the first investigation to characterize a South American deer species using fecal DNA and demonstrates the usefulness and efficiency of this approach, as well as the feasibility of obtaining information that could not have been easily obtained by traditional DNA sampling. Our findings suggest that management strategies for this species may be much more effective if applied now when the population still shows high genetic variability.