Petiolate Oak (Quercus robur) polymorphism evaluation by SSR-analyzing

The results of molecular-genetic studies of populations of oak petiolate from 11 regions of Russia are presented. Using 10 microsatellite primers, 1049 alleles were identified. The studied samples differ insignificantly in the observed and effective number of alleles. To assess the genetic variability of populations, an indicator of expected and observed heterozygosity was calculated, which indicates a deficiency of heterozygous genotypes. It was found that each individual tree in the studied populations shows 87 % deficiency of heterozygotes relative to the population and 85,7 % relative to the species. It was noted that among all the studied alleles, 81 % were unique, and they occur only in one locus. The dendrogram based on the genetic distance shows clustering of the populations of the oak petiolate into several separate groups.

Molecular characterization of oats (Avena sativa L.) germplasm with microsatellite markers

Assessment of genetic diversity between selected genotypes serves as an essential tool to exploit the genetic variability. In the present study, Ninety nine accessions of oat acquired from different geographical regions were used to study the molecular genetic variability by the use of 91 microsatellite primers. A total of 248 alleles from 91 primer pairs in the 99 oat genotypes were detected. The average alleles per primer pair were recorded as 2.69. The primer pairs having the highest PIC values viz., AM 6 (0.50), AM 55 (0.49) and M 83381 (0.49) can be further used in the association mapping studies. Clustering of the oat genotypes into two main clusters A and B was done by using the symmetric matrix of DICE coefficient based on UPGMA method in Software – DARwin 6.0. In this study, the moderate amount of dissimilarity was observed between the whole set of accessions, the values ranging from 0.33 to 0.79. The individuals belonging to the different clusters may be useful in hybridization studies of oat improvement programmes.

Development of additional microsatellite primers for the mangrove tree species Avicennia germinans

Objective: The objective of this study was to develop additional microsatellite primers for the mangrove tree species Avicennia germinans that work in multiplex PCR panels to enable cost effective population analyses of this species at a finer scale. Results: Primer sets were identified from whole genome sequencing data and combined into multiplex PCR panels. Five multiplex reactions containing 20 novel primer sets with trinucleotide repeats were successfully developed. Fifteen of the microsatellite loci were polymorphic in the samples tested, with 1-4 alleles per locus.

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.

Are there differences in heterozygosity of strains obtained from intercrossed and self-fertilized onion plants?

ABSTRACT The commercial use of onion hybrids is preferred by producers. In contrast, the production of hybrid onion seeds is extremely inefficient. This is due to the use of lineages obtained by successive self-fertilizations assuming the effect of inbreeding depression per se. Therefore, it is necessary to understand new alternatives to reduce the effect of inbreeding depression in the strains. The objective of this study was to evaluate possible differences in polymorphism and levels of heterozygosity of strains obtained from intercrossed and self-fertilized plants. Twelve onion populations belonging to Bayer’s breeding program (Granex, IPA-1, IPA-2 and IPA-3) were used, obtained by self-fertilization of one plant, intercrossing of two plants or intercrossing of three plants. Three individuals from each strain were used in the analyzes. The amplifications were performed using 8 microsatellite primers with greater polymorphism, according to germplasm characterization studies carried out by CITA. Heterozygosity generally decreases with self-fertilization and increases as more plants are used in the cross. The SSR markers used in the present study were efficient in detecting variability in different genetic backgrounds. With the results obtained, it is suggested to carry out the obtaining of hybrids between the different combinations and to analyze the performance per se of the different modalities of obtaining strains proposed in the present research.

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.

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.

Development and characterization of polymorphic microsatellite markers for Spathoglottis plicata (Orchidaceae)

We developed novel and polymorphic microsatellite primers for Spathoglottis plicata, a tropical and subtropical terrestrial orchid, to investigate the genetic patterns and population structure among wild populations, and also to identify the varieties and hybrids of S. plicata in horticultural industry. The 12 novel microsatellites from S. plicata were developed by using polymerase chain reaction (PCR)-based isolation of microsatellite arrays. These markers that were successfully PCR amplified exhibited polymorphisms in S. plicata. The number of alleles, observed heterozygosity, expected heterozygosity and polymorphism information content values across loci ranged from 2.000 to 8.000, 0.000 to 0.756, 0.208 to 0.813 and 0.405 to 0.805 in total populations, respectively. The newly developed microsatellite markers exhibited variation in S. plicata. These markers can be used as a tool to further investigate the genetic diversity, conservation genetics and variety/hybrid identification of S. plicata.