Selected Microsatellite Markers Research Articles

Detection of putative loci affecting milk yield in Turkish Awassi sheep using microsatellite markers.

On the basis of comparisons between bovine and ovine genome mapping information, the aim of the study was to analyze the genetic diversity of selected DNA microsatellites from the bovine genome and to investigate their correlation with the average daily milk yield in Awassi sheep. 18 informative microsatellite markers were selected from the significant QTL regions affecting milk yield identified in the bovine genome in previous studies. The selected microsatellite markers were then amplified by PCR as reciprocal amplifications on the genomic DNA of Awassi sheep, with standard daily milk yield records. Thus, in this study, 18 microsatellite markers associated with milk yield in the bovine genome were examined for both determination of genetic polymorphism within the flock and the effects of marker loci on average daily milk yield in Awassi sheep. Allele frequencies of markers were determined based on the results of fragment analysis. The analysis of variance showed that the 123bp allele at the marker locus BMS1341 on BTA2 significantly influenced the average daily milk yield of Ivesi sheep (P < 0.01). On the other hand, the BMS381 locus with a 115bp allele on BTA2, the MCM140 locus with a 185bp allele on BTA6, the BMS2721 locus with a 155bp allele, the BM1237 locus with 174 and 180bp alleles on BTA7, and finally, the BMS1967 locus with a 117bp allele, the BM4208 locus with 176 and 182bp alleles, and the INRA locus with a185 bp allele on BTA8 showed moderately significant effects on the average daily milk yield of Ivesi ewes (P < 0.05).

The use of microsatellite markers to analyze genetic diversity of ornamental chili peppers (Capsicum spp.) and their transferability to cross amplification

Ornamental chili peppers, as one of high economic value horticultural crops, present a high diversity which needs to be assessed. The aim of this study was to analyze the genetic diversity of ornamental chili pepper genotypes using microsatellite markers and to analyze the transferability of these markers to cross-amplification to ornamental chili pepper species other than Capsicum annuum L. Two generated main clusters revealed in this study, that the first cluster consisted entirely of genotypes from C. annuum species while the second cluster consisted of a mixture from C. annuum, C. chinense, and C. baccatum species. The highest genetic similarity was identified between Nazla IPB and Violeta IPB, and Viola IPB with Triwarsana IPB, with a similarity of 96% for both. Polymorphism analysis showed that all of the microsatellite markers used were classified as highly informative (PIC >0.5). Total markers were able to cross-species amplify all of the ornamental chili pepper genotypes, with an overall average amplification percentage of 96.25%, with most of them showing 100% cross-amplification. Selected microsatellite markers from this study could be used in species differentiation, molecular identification of interspecific hybrids, and assessing the genetic resource on broad genotypes of ornamental chili peppers.

GENETIC DIVERSITY AND PARENTAGE ANALYSIS FOR DNA MARKER-BASED FOREST REPRODUCTIVE MATERIAL TRACEABILITY SYSTEM IN LITHUANIA

Scots pine is economically and ecologically important forest tree species in Lithuania. According to the Regulation of Forest Planting and Restoration of Lithuania (2018) and the Regulations on Forest Reproductive Material (FRM) (2017) forests must be planted and replanted with the target tree species, using high quality seeds and seedlings from a given provenance. EU Council Directive (1999/105/EC) indicates that EU Member States are responsible for the regulation, requirements and implementation of FRM quality and traceability system. The existing FRM control system in Lithuania is based on the control actions of the supporting documents regarding the origin of FRM. However, this FRM traceability system is limited and, in some cases, cannot ensure that FRM used for reforestation/afforestation originates from the declared place of origin or belongs to a certain seed or plant lot. In this study we aimed to assess and compare genetic diversity of scots pine seed orchard clones with seeds, and to test DNA based parentage analysis methods with Jonava scots pine seed orchard clones and seeds. Up to now DNA markers based FRM traceability system in Lithuania has not been tested and implemented. Thus, our first results based on 12 nuclear microsatellite markers indicated that genetic diversity among Jonava seed orchard clones and collected seeds were moderately high and comparable with other studies on scots pine in Europe. The test of the maternity analysis software CERVUS has showed very positive results in maternity assignments. Therefore, selected microsatellite markers in combination with accurate sampling design and specific DNA analysis methods should be improved and can be used for FRM traceability system based on DNA “fingerprints”.

Development and Characterization of Novel Polymorphic Microsatellite Markers for Tapinoma indicum (Hymenoptera: Formicidae).

Tapinoma indicum (Forel) (Hymenoptera: Formicidae) is a nuisance pest in Asia countries. However, studies on T. indicum are limited, especially in the field of molecular biology, to investigate the species characteristic at the molecular level. This paper aims to provide valuable genetic markers as tools with which to study the T. indicum population. In this study, a total of 143,998 microsatellite markers were developed based on the 2.61 × 106 microsatellites isolated from T. indicum genomic DNA sequences. Fifty selected microsatellite markers were amplified with varying numbers of alleles ranging from 0 to 19. Seven out of fifty microsatellite markers were characterized for polymorphism with the Hardy–Weinberg equilibrium (HWE) and linkage disequilibrium (LD) analysis. All seven microsatellite markers demonstrated a high polymorphic information content (PIC) value ranging from 0.87 to 0.93, with a mean value of 0.90. There is no evidence of scoring errors caused by stutter peaks, no large allele dropout, and no linkage disequilibrium among the seven loci; although loci Ti-Tr04, Ti-Tr09, Ti-Te04, Ti-Te13, and Ti-Pe5 showed signs of null alleles and deviation from the HWE due to excessive homozygosity. In conclusion, a significant amount of microsatellite markers was developed from the data set of next-generation sequencing, and seven of microsatellite markers were validated as informative genetic markers that can be utilized to study the T. indicum population.

Polymorphism of Selected Microsatellite Markers against Breeding Performance Indexes of Polish Large White and Polish Landrace Sows

The study aimed to analyze 12 microsatellite markers located in the areas of quantitative trait loci related to litter size in 82 sows, including 45 Polish Large White and 37 Polish Landrace sows, kept on a farm in southwest Poland. Breeding documentation provided data on the total number of piglets born per litter and the numbers of live-born, stillborn and weaned piglets; the corresponding percentage values were also calculated. DNA isolation was performed from 15–20 hairs taken from live animals, and the markers used in the research were divided into four multiplexes. The influence of genotype at a given locus on the reproductive traits was analyzed only for genotypes represented by at least five animals. The results of the analyses for all the sows (treated as the maternal component and not as distinct breeds) showed statistically significant (p ≤ 0.05) differences between the genotypes and the values of the analyzed traits for markers S0008, SW160, SW245, SW714, SW1125 and SW2411. Among these markers, the breed-dependent analysis also showed significant differences in the Polish Large White breed for markers S0008, SW160 and SW1125, and in the Polish Landrace breed for SW245. Additionally, the breed-dependent analysis found significant differences in markers SW903 and SW1808 for the Polish Large White sows, and S0064, SW472 and SW903 in Polish Landrace sows. SW903 was the only marker for which differences in the analyzed reproductive traits differed between genotypes in both breeds (still analyzed separately), although in terms of different traits. The above results indicate the usefulness of microsatellite markers in researching the differentiation of litter size indicators. Although both breeds belong to the maternal component, they showed significant differences in terms of markers. This may suggest difficulties in finding universal (that is, working well for various breeds of the maternal component) markers, indicating the need to look for breed-specific markers, something that calls for further research into numerous animals.

Development of microsatellite markers for Diaphanosoma dubium (Crustacea, Cladocera) and application to seasonal population dynamics

Diaphanosoma, the “tropical Daphnia”, is common and ubiquitous in South China. Like other ctenopods, Diaphanosoma has a reproductive mode similar to Daphnia’s, but its resting eggs are rarely observed and lack an ephippium. With limited dispersal and reduced buffer effect from resting egg banks, Diaphanosoma is expected to have a population genetic structure different from that of temperate Daphnia. To facilitate genetic comparison, we developed microsatellite markers using next-generation sequencing for the most common species in tropical and subtropical East Asia, Diaphanosoma dubium. Thirty-one polymorphic microsatellite markers were obtained, and 29 of them were efficient for the congeneric species D. excisum, D. orghidani, D. mongolianum and D. chankensis. The markers allowed intra- and interspecific genetic analysis, including population structure, hybridization and introgression. We used 11 selected microsatellite markers to analyze spatial and temporal heterogeneity of genetic diversity in four (sub)tropical D. dubium populations from two large reservoirs and two temporary ponds. In contrast to temperate Cladocera, higher genetic diversity in summer rather than in spring suggested weak contribution from resting eggs in spring. Clustering of DAPC and STRUCTURE analyses indicated a clear-cut genetic structure in the four populations. Variation partitioning revealed that water storage and depth were key factors in genetic differentiation. Within large reservoirs, we detected backward (reversing time) gene flow from resting egg banks. We conclude that resting eggs have an effective contribution to the genetic diversity in large water bodies during growing seasons and that large water bodies can host higher genetic diversity in summer due to environmental heterogeneity and high carrying capacity. Spatial and temporal heterogeneity of genetic diversity detected by our microsatellite markers showed the newly developed markers can be applied for further study of populations of D. dubium and other species of Diaphanosoma at a contemporary scale.

Altitude and hillside orientation shapes the population structure of the Leishmania infantum vector Phlebotomus ariasi

Despite their role in Leishmania transmission, little is known about the organization of sand fly populations in their environment. Here, we used 11 previously described microsatellite markers to investigate the population genetic structure of Phlebotomus ariasi, the main vector of Leishmania infantum in the region of Montpellier (South of France). From May to October 2011, we captured 1,253 Ph. ariasi specimens using sticky traps in 17 sites in the North of Montpellier along a 14-km transect, and recorded the relevant environmental data (e.g., altitude and hillside). Among the selected microsatellite markers, we removed five loci because of stutter artifacts, absence of polymorphism, or non-neutral evolution. Multiple regression analyses showed the influence of altitude and hillside (51% and 15%, respectively), and the absence of influence of geographic distance on the genetic data. The observed significant isolation by elevation suggested a population structure of Ph. ariasi organized in altitudinal ecotypes with substantial rates of migration and positive assortative mating. This organization has implications on sand fly ecology and pathogen transmission. Indeed, this structure might favor the global temporal and spatial stability of sand fly populations and the spread and increase of L. infantum cases in France. Our results highlight the necessity to consider sand fly populations at small scales to study their ecology and their impact on pathogens they transmit.

Case study of microsatellite polymorphism of European perch in selected commercially important lakes of Latvia

Polymorphism of selected microsatellite markers was described for European perch populations of inland Lake Kala and costal Lake Babites in Latvia. The data consisted of ten microsatellite loci analysed for 90 individuals from the Lake Babites (n = 45) and the Lake Kala (n = 45). Both lakes differ in water area, connection with the Gulf of Riga (Baltic Sea), lake type, and fish species. In the Lake Kala population, the average number of alleles per locus varied from 4 to 15 and in the population of Lake Babites from 4 to 12. Low but significant genetic differentiation was detected between populations (FST = 0.046). Observed and expected heterozygosity in both lakes was similar (Kala: Ho = 0.680 and He = 0.816; Babites: Ho = 0.693 and He = 0.815). The number of unique alleles per locus was 2.3 in Lake Kala and 0.6 in Lake Babites. The present study showed relatively high polymorphism of analysed microsatellite markers.

Genetic diversity and pathogenicity of Fusarium species associated with fruit rot disease in banana across Peninsular Malaysia.

The aims of this study were to identify the Fusarium isolates based on translation elongation factor (tef) 1α sequence, to determine the genetic diversity among isolates and species using selected microsatellite markers and to examine the pathogenicity of Fusarium isolates causing fruit rot disease of banana. One-hundred and thirteen microfungi isolates were obtained from fruit rot infected banana in Peninsular Malaysia. However, this study was focused on the dominant number of the discovered microfungi that belongs to the genus Fusarium; 48 isolates of the microfungi have been identified belonging to 11 species of Fusarium, namely Fusarium incarnatum, Fusarium equiseti, Fusarium camptoceras, Fusarium solani, Fusarium concolor, Fusarium oxysporum, Fusarium proliferatum, Fusarium verticillioides, Fusarium sacchari, Fusarium concentricum and Fusarium fujikuroi. All Fusarium isolates were grouped into their respective clades indicating their similarities and differences in genetic diversity among isolates. Out of 48 Fusarium isolates tested, 42 isolates caused the fruit rot symptom at different levels of severity based on Disease Severity Index (DSI). The most virulent isolate was F. proliferatum B2433B with DSI of 100%. All the isolated Fusarium species were successfully identified and some of them were confirmed as the causal agents of pre- and postharvest fruit rot in banana across Peninsular Malaysia. Our results will provide additional information regarding new report of Fusarium species in causing banana fruit rot and in the search of potential biocontrol agent of the disease.

Survey of genetic structure of geese using novel microsatellite markers.

ObjectiveThe aim of this study was to create a set of microsatellite markers with high polymorphism for the genetic monitoring and genetic structure analysis of local goose populations.MethodsNovel microsatellite markers were isolated from the genomic DNA of white Roman geese using short tandem repeated probes. The DNA segments, including short tandem repeats, were tested for their variability among four populations of geese from the Changhua Animal Propagation Station (CAPS). The selected microsatellite markers could then be used to monitor genetic variability and study the genetic structures of geese from local geese farms.Results14 novel microsatellite loci were isolated. In addition to seven known loci, two multiplex sets were constructed for the detection of genetic variations in geese populations. The average of allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.09, 5.145, 0.499, 0.745, and 0.705, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting white Roman cluster and a spreading Chinese cluster. In white Roman populations, the CAPS populations were depleted to roughly two clusters when K was set equal to 6 in the Bayesian cluster analysis. The founders of private farm populations had a similar genetic structure. Among the Chinese geese populations, the CAPS populations and private populations represented different clads of the phylogenetic tree and individuals from the private populations had uneven genetic characteristics according to various analyses.ConclusionBased on this study’s analyses, we suggest that the CAPS should institute a proper breeding strategy for white Roman geese to avoid further clustering. In addition, for preservation and stable quality, the Chinese geese in the CAPS and the aforementioned proper breeding scheme should be introduced to geese breeders.

GENETIC ANALYSIS GERMAN SHEPHERD BREED DOGS USING MICROSATELLITE DNA MARKERS

Modern methods of breeding dogs are based on getting of stable phenotypic uniformity by using close inbreeding or breeding by one line, but such strategy leads to a loss of genetic diversity, and as a result there are genetic defects in breeds which have no external manifestations or manifest in adulthood of dogs and are transmitted from generation to generation. Therefore, to prevent use of dogs with genetic abnormalities in breeding, and to develop standards for a breed and make an accurate pedigree, must carry out the genetic evaluation of animals. One of the modern tools for dogs’ genetic evaluation is DNA-testing using microsatellite loci permitting to match the parental couple effectively, identify (to certify) animals, undertake a comprehensive assessment for heterozygous and homozygous genotypes in populations, permitted for use in the selection process, and illustrate clearly the impact of artificial selection on the genetic characteristics of breeds. The study was conducted in Ukrainian Laboratory of Quality and Safety of Agricultural Products in Department of Molecular Biology Research. For the genetic analysis 42 German Shepherd dogs, used for breeding in kennels of Ukrainian Kennel Union (UKU), were selected. The material for the research was DNA isolated from dogs’ buccal epithelium cells and blood. Genomic DNA was extracted using a standard set of reagents for DNA isolation. Level of theoretically expected heterozygosity (Hexp) varied between 0.385 (PEZ1) to 0.835 (PEZ8). On average theoretically expected heterozygosity with coefficient of 0.657 had not significant advantage over value of actual heterozygosity (0.629), it also shows that the status of the sample of dogs is close to balance. The same is observed in actual and expected heterozygosity for PEZ 6 (0.629) and PEZ 8 (0.657) loci, which also shows the balance. For FHC2010 loci actual heterozygosity is higher than expected, indicating increasing the number of heterozygous individuals. For FHC2054 locus, by contrast, theoretically expected heterozygosity (0.670) dominates the actual (0.429), indicating the lack of heterozygous genotypes in this micropopulation. The value of PIC (polymorphism information content) of the analysed loci ranged from 0.325 to 0.740 with average value 0.574. PEZ6, PEZ8, FHC 2010 and FHC 2054 loci optimally meets their suitability for genetic certification of genotypes because their frequency varies from 0.587 to 0.740. The reduced average index of polymorphism for PEZ1 locus with coefficient of 0.325 confirmed the insufficient level of its polymorphism for full genetic evaluation of the micropopulation of German Shepherd dogs (PIC &lt; 0.500), as confirmed by Chinese researcher J.-H. Ye, according to his data PIC value for PEZ1 locus was 0,320, which correlates with our results. And PIC value for PEZ8 locus was 0.740 in our studies, whereas according to J.-H. Ye – 0,720, which, by contrast, indicates high polymorphism and confirms the effectiveness of its use in genotyping of dogs. Probability of exclusion of accidental allele coincidence (PE), which is 0.675 on average, indicates a lack of the number and informativeness of the selected microsatellite markers for German Shepherd as in this case a combined probability (CPE) of accidental allele coincidence is 0.933886 or 93.3%. The chosen microsatellite loci to study the genetic structure of the German Shepherd dog population, show a sufficiently high informativeness of chosen system of molecular genetic DNA markers. However, there is the need for using additional microsatellite markers which will increase the combined probability of accidental allele coincidence (CPE) from 93.3% to 99.9%. The analysis of heterozygosity is important in studying the dynamics of genetic processes in populations, because heterozygosity has an effect on many factors, including mutations, selection, non-random mating, genetic drift, etc., so continuous monitoring of genetic diversity is required for their timely identification and development of measures to improve breeding work on biodiversity in different dog breeds.

MICROSATELLITE MARKERS POLYMORPHISM BETWEEN TWO EGYPTIAN GOAT POPULATIONS (Capra hircus)

Two native goat populations present in El-Farafra and Siwa oases, located in the Western Desert of Egypt were geno-typed using ten microsatellite molecular markers (SSR). Blood samples taken from a total of 34 individual goats, 14 from Farafra and 20 from Siwa, were subjected to DNA extraction and subsequently to SSR-PCR amplification. The number of alleles ranged from two for MAF70 marker to seven for OarFCB48 and SRCRSP8 loci, the average number per population for Farafra goats was 3.0 and 3.4 for Siwa goats with a total number of a 42 alleles for both populations. The mean observed heterozygosity (Ho) and expected heterozygosity (HE) for both populations varied from 0.21 to 0.24 and 0.56 to 0.59, respectively. Fixation indices revealed a 0.7063 variation referring to differences among individuals versus total variance (Fit). While, among populations differences versus total variance had a lower fixation index (Fst = 0.2371) indicating low level of genetic differentiation between Farafra and Siwa populations. A pair wise difference between Farafra and Siwa goat populations was (0.6151) based on among breeds F index (Fis).The highest PIC was observed for SRCRSP8 microsatellite marker (0.791) and the lowest PIC was 0.375 for SRCRSP23. The average PIC of the ten markers was 0.530 and 0.570 for Farafra and Siwa goat populations, respectively, which indicated that the ten microsatellite markers contained highly polymorphic loci in both Egyptian goat populations. In the genetic diversity analysis, microsatellite markers with PIC>0.7 were taken as the most ideal selected markers. From the selected microsatellite markers in the present study, the PIC of OarFCB48 and SRCRSP8 (Siwa) exceeded 0.7 which indicated that these loci could be used as genetic markers for genetic diversity analysis of Siwa goat population. The markers generated by OarFCB48 and SRCRSP8 loci could be utilized in marker assisted selection (MAS) to improve the performance of Egyptian goat populations.

Genome-wide analysis of loss of heterozygosity in breast infiltrating ductal carcinoma distant normal tissue highlights arm specific enrichment and expansion across tumor stages.

Studies have shown concurrent loss of heterozygosity (LOH) in breast infiltrating ductal carcinoma (IDC) and adjacent or distant normal tissue. However, the overall extent of LOH in normal tissue and their significance to tumorigenesis remain unknown, as existing studies are largely based on selected microsatellite markers. Here we present the first autosome-wide study of LOH in IDC and distant normal tissue using informative loci deduced from SNP array-based and sequencing-based techniques. We show a consistently high LOH concurrence rate in IDC (mean = 24%) and distant normal tissue (m = 54%), suggesting for most patients (31/33) histologically normal tissue contains genomic instability that can be a potential marker of increased IDC risk. Concurrent LOH is more frequent in fragile site related genes like WWOX (9/31), NTRK2 (10/31), and FHIT (7/31) than traditional genetic markers like BRCA1 (0/23), BRCA2 (2/29) and TP53 (1/13). Analysis at arm level shows distant normal tissue has low level but non-random enrichment of LOH (topped by 8p and 16q) significantly correlated with matched IDC (Pearson r = 0.66, p = 3.5E-6) (topped by 8p, 11q, 13q, 16q, 17p, and 17q). The arm-specific LOH enrichment was independently observed in tumor samples from 548 IDC patients when stratified by tumor size based T stages. Fine LOH structure from sequencing data indicates LOH in low order tissues non-randomly overlap (∼67%) with LOH that usually has longer tract length (the length of genomic region affected by LOH) in high order tissues. The consistent observations from multiple datasets suggest progressive LOH in the development of IDC potentially through arm-specific pile up effect with discernible signature in normal tissue. Our finding also suggests that LOH detected in IDC by comparing to paired adjacent or distant normal tissue are more likely underestimated.

Genetic diversity of DNA microsatellite for Tibetan Yak

To assess the genetic diversity and relationship of the Tibetan yak breeds. The genetic diversity and phylogenies of a total of 480 individual from 11 Tibetan yak groups were analyzed using PCR and multiplex gel electrophoresis of silver staining technology with eight pairs of microsatellite markers.The result showed that these markers were highly polymorphic loci with rich genetic diversity in the Tibetan yak populations.The average polymorphic information content (PIC) in 11 groups of yak were higher than 0.5. The highest HEL13 was 0.8496, and the lowest TGLA57 was 0.7349. Among them, the PICof Dingqing yak was minimum (0.7505), indicating that the group is relatively pure.Sangri Yak had the highest PIC value (0.7949) indicating greater genetic variationwithinthe groups. Among the 11 groups examined, the order of heterozygosity size wasSangri(0.8193)>Jiangda(0.8190)>Sangsang(0.8157)>Baqing(0.8150)>Kangbu(0.8123)> Jiali(0.8087)>Gongbujiangda(0.8054)>Sibu(0.8041)>Leiwuqi(0.8033)>Pali(0.8031)>Dingqing(0.7831). The groups from eastern Tibet had grater genetic diversity than those from Western Tibet, which indicate that Tibet may be one of the cradles of the yak.According to the genetic distance, the cluster relationship constructed with UPGMA and NJ methods showed that 11 yak groups in Tibet could be divided into three forms. In summary,Tibet yak has abundant genetic diversity and the selected microsatellite markers can be used to evaluategenetic diversity of Tibetan yak.

Discrimination of korean native chicken lines using fifteen selected microsatellite markers.

In order to evaluate the genetic diversity and discrimination among five Korean native chicken lines, a total of 86 individuals were genotyped using 150 microsatellite (MS) markers, and 15 highly polymorphic MS markers were selected. Based on the highest value of the number of alleles, the expected heterozygosity (He) and polymorphic information content (PIC) for the selected markers ranged from 6 to 12, 0.466 to 0.852, 0.709 to 0.882 and 0.648 to 0.865, respectively. Using these markers, the calculated genetic distance (Fst), the heterozygote deficit among chicken lines (Fit) and the heterozygote deficit within chicken line (Fis) values ranged from 0.0309 to 0.2473, 0.0013 to 0.4513 and −0.1002 to 0.271, respectively. The expected probability of identity values in random individuals (PI), random half-sib (PIhalf-sibs) and random sibs (PIsibs) were estimated at 7.98×10−29, 2.88×10−20 and 1.25×10−08, respectively, indicating that these markers can be used for traceability systems in Korean native chickens. The unrooted phylogenetic neighbor-joining (NJ) tree was constructed using 15 MS markers that clearly differentiated among the five native chicken lines. Also, the structure was estimated by the individual clustering with the K value of 5. The selected 15 MS markers were found to be useful for the conservation, breeding plan, and traceability system in Korean native chickens.

Genetic variability of microsatellites in autochthonous Podolian cattle breeds in Croatia

The aim of the study was to analyse the status of the genetic structure of two autochthonous Croatian Podolian breeds (Istrian cattle and Slavonian Syrmian Podolians). The study comprised 20 randomly selected blood samples of each cattle breed. The genotyping was performed on 11 microsatellites (TGLA227, BM2113, TGLA53, ETH10, SPS115, TGLA126, TGLA122, INRA23, ETH3, ETH225, BM1824) as recommended by International Society of Animal Genetics and Food and Agriculture Organization. The procedure enabled the identification of 423 (96.14%) genotypes, indicating the validity of DNA analyses in blood samples and adequacy of the selected microsatellite markers. Likewise, there was the total number of 203 alleles with minimal differences among the studied groups. The differences (p&gt; 0.05)were observed within the genotypes of the studied groups regarding the microsatellite loci. The distribution of the genetic relationship was presented by Wright F coefficients. The FIShad negative values for the TGLA 227 locus in both studied groups and for ETH3, ETH225 and BM1834 loci in the Slavonian Syrmian Podolians. This obtained data with relative high values of heterozygosity also shows a genetical influence of other breeds. Almost all FITvalues were appropriate (0.31905) as a result of excessive selection effects. The mean value of FST(0.07336) indicates that the average degree of genetic variability of the studied breeds was not significantly different despite certain differences in individual loci. The observed specific polymorphism of loci shows that the Istrian cattle and the Slavonian Syrmian Podolians have similar genetic characteristics, but also that there are certain differences between the breeds and even inter-breed differences. The findings indicate a relationship between the breeds and their suitable purity for interbreeding but they also indicate certain danger to certain parts of genome within the studied species. The results can be applied in the protection and breeding programs for the preservation of autochtonous Croatian Podolian breeds.

Wstepne badania zr�znicowania genetycznego wybranych polskich lokalnych odmian kur

The study was aimed at evaluation of the applicability of selected microsatellite markers in examining genetic diversity in five Polish local chicken varieties (two groups of native crested chickens, crestless native chickens from Subcarpathian region and two groups of native miniature chickens) and reveal their genetic relationships to five purebreds (Leghorn, Rhode Island Red, Sussex, Greenleg Partridge, Minorca). About 22.8% (overall FST = 0.228) of the genetic variation was explained by differences between breeds and varieties. The DA genetic distances between five Polish native varieties and five purebreds varied between 0.154 and 0.440. The results obtained in this study indicate the great value of the collection and suggest the need for formal protection of these local Polish chicken varieties.

Cross Species Amplification of Pennisetum glaucum Microsatellite Markers in Pennisetum purpureum and Genetic Diversity of Napier Grass Accessions

ABSTRACTNapier grass (Pennisetum purpureum Schum.) is an important forage crop in tropical areas although little is known about its genome information, and few molecular markers have been developed for this species. This work aimed to check the viability of cross‐species amplification of microsatellite markers between pearl millet (Pennisetum glaucum) and Napier grass and to evaluate the genetic diversity among Napier grass germplasm accessions. Fifty‐four microsatellite markers previously described in pearl millet were tested against Napier grass, and 30 markers (55.5%) showed successful cross‐amplification. From them, 18 microsatellite markers were selected to study the genetic diversity in the Embrapa Active Germplasm Bank of Napier Grass (Embrapa‐BAGCE). A total of 180 alleles were identified by these selected microsatellite markers in 107 Napier grass accessions and four pearl millet samples. The average similarity coefficient (Dice) calculated among the Embrapa‐BAGCE accessions was 0.651, ranging from 0.254 to 1.0. Some accessions showed similarity coefficients equal to one, indicating that they have common progenitors or that they might be the same accessions with different denomination. To our knowledge, this work is the first to describe microsatellite markers in Napier grass and represents a significant advance regarding the use of molecular markers in this species.

Analysis of molecular genetic diversity of cardoon (Cynara cardunculus L.) in Tunisia

The objective of this study is to investigate the genetic diversity, the relationships among six Tunisian wild cardoon populations (Cynara cardunculus var. sylvestris) and a Tunisian's population of cultivated cardoon (Cynara cardunculus var. altilis DC) in seven different geographical locations (Tiurif, Bahra, Zriba, Bouficha, Enfidha, Beja and Wad mliz) from semi-arid and wet regions of Tunisia. Twenty-three selected microsatellite markers are used for a sample of 98 cardoon genotypes. The total of 243 alleles is detected in the studied populations and the number of alleles per locus ranged from six to 23. The dendrogram based on Nei's (1972) UPGMA method divides the seven studied populations to five clusters. These preliminary results show that microsatellites are effective tools for plant species characterization and the analysed populations have a high genetic variability and will be suitable as genetic stocks for conservation and sustainable utilization programs of Cynara cardunculus L. in Tunisia.

Development of microsatellite markers for the small yellow croaker Larimichthys polyactis (Sciaenidae) by cross-species amplification

The small yellow croaker (Larimichthys polyactis) is a highly valued fish for human consumption found in the Western Pacific that was considered endangered until recently because of overfishing. We selected microsatellite markers for this species from markers developed for Miichthys miiuy, also of the family Sciaenidae. Among 43 markers polymorphic for M. miiuy, 11 were found to be polymorphic for L. polyactis. Characterization of these 11 loci was made based on 30 L. polyactis individuals collected by trawling in the Zhoushan Fishing Ground, Zhejiang Province, China. Total genomic DNA was isolated from fin clips. The number of alleles per locus ranged from 4 to 10, with a mean of 5.82, while the effective number of alleles ranged from 1.64 to 10.00, with a mean of 3.22. Observed and expected heterozygosities ranged from 0.17 to 0.72 and from 0.39 to 0.81, respectively. Significant deviation from Hardy-Weinberg equilibrium was found at four loci, after applying Bonferroni's correction. There was no significant association between any of the pairs of microsatellite loci, hence allelic variation at these loci was considered independent. These 11 polymorphic microsatellite loci will be useful for genetic diversity analysis and molecular-assisted breeding for L. polyactis.