Average Effective Number Of Alleles Research Articles

Developmental and validation of a novel small and high-efficient panel of microhaplotypes for forensic genetics by the next generation sequencing

BackgroundIn the domain of forensic science, the application of kinship identification and mixture deconvolution techniques are of critical importance, providing robust scientific evidence for the resolution of complex cases. Microhaplotypes, as the emerging class of genetic markers, have been widely studied in forensics due to their high polymorphisms and excellent stability.Results and discussionIn this research, a novel and high-efficient panel integrating 33 microhaplotype loci along with a sex-determining locus was developed by the next generation sequencing technology. In addition, we also assessed its forensic utility and delved into its capacity for kinship analysis and mixture deconvolution. The average effective number of alleles (Ae) of the 33 microhaplotype loci in the Guizhou Han population was 6.06, and the Ae values of 30 loci were greater than 5. The cumulative power of discrimination and cumulative power of exclusion values of the novel panel in the Guizhou Han population were 1-5.6 × 10− 43 and 1-1.6 × 10− 15, respectively. In the simulated kinship analysis, the panel could effectively distinguish between parent-child, full-sibling, half-sibling, grandfather-grandson, aunt-nephew and unrelated individuals, but uncertainty rates clearly increased when distinguishing between first cousins and unrelated individuals. For the mixtures, the novel panel had demonstrated excellent performance in estimating the number of contributors of mixtures with 1 to 5 contributors in combination with the machine learning methods.ConclusionsIn summary, we have developed a small and high-efficient panel for forensic genetics, which could provide novel insights into forensic complex kinships testing and mixture deconvolution.

Genetic structure of cattle of the siberian branch by microsatellite loci

Molecular-genetic methods are essential tools for the utilization and conservation of animal genetic resources. These methods facilitate more efficient management and control of breeding programs within livestock production systems. For studying the genetic diversity of a population, the use of STR markers is relevant due to the high variability of repeats. This study presents a genetic characterization of a Holstein and Black Pied cattle population (n = 10233) in Western Siberia using 12 microsatellite loci (BM1818, BM1824, BM2113, ETH10, ETH225, ETH3, INRA023, SPS115, TGLA126, TGLA122, TGLA227, TGLA53). A total of 145 alleles were identified across all loci, with frequencies ranging from 0.00005 to 0.68961. The highest level of genetic diversity was observed at the TGLA122 locus (25 alleles) with an average number of effective alleles (Ne) of 4.5. The least polymorphic locus was BM1824 (7 alleles) with an average Ne of 3.27. The average observed (Ho) and expected (He) heterozygosity across all loci was 0.6. The highest variability was observed at the TGLA53 locus, with a Wright’s fixation index (Fis) of 0.161, indicating a heterozygote deficiency. A similar deficiency was observed at the BM1818 locus. All other loci exhibited a positive Fis, with the highest value observed at the ETH3 locus (-0.074), indicating an excess of heterozygotes. The average Fis across all loci was -0.02, suggesting a sufficient level of heterozygosity within the studied population. These findings provide valuable information for population studies and practical breeding programs aiming to manage genetic diversity and improve selection efficiency in this cattle population.

Exploring Sampling Strategies and Genetic Diversity Analysis of Red Beet Germplasm Resources Using SSR Markers

By using 14 SSR primer pairs, we here analyzed and compared the amplification results of 534 DNA samples from six red sugar beet germplasm resources under three treatments. These data were used to explore the sampling strategy for the aforementioned resources. Based on the sampling strategy results, 21 SSR primer pairs were used to evaluate the genetic diversity of 47 red sugar beet germplasm resources. The six population genetic parameters used for exploring the sampling strategy unveiled that individual plants within the population had a relatively large genetic distance. The genetic parameters Ne, I, and Nei’s of the randomly mixed sampling samples increased rapidly between 10 and 30 plants before decreasing. Therefore, when SSR technology was used to analyze the genetic diversity of the red sugar beet germplasm resources, the optimal sampling gradient for each population was the adoption of a random single-plant mixed sampling sample of no less than 10 plants and no more than 30 plants. The 21 SSR primer pairs were used to detect genetic diversity in 30 random mixed samples of 47 resources. The average polymorphic information content (PIC) was 0.5738, the average number of observed alleles (Na) was 4.1905, the average number of effective alleles (Ne) was 2.8962, the average Shannon’s information index (I) was 1.1299, the average expected heterozygosity (Nei’s) was 0.6127, and the average expected heterozygosity (He) was 0.6127. The genetic distance of the 47 germplasm resources ranged from 0.0225 to 0.551 (average: 0.316). According to the population structure analysis, the most suitable K value was six, which indicated the presence of six populations. Based on the clustering analysis results, the 47 germplasm resources were segregated into six groups, with obvious clustering and some germplasm resources noted for having groups with close genetic relationships. We here established a more accurate and scientific sampling strategy for analyzing the genetic diversity of red sugar beet germplasm resources by using SSR molecular markers. The findings provide a reference for collecting and preserving red sugar beet germplasms and protecting their genetic diversity.

Hemoglobin polymorphism and its association with morphometric and egg production traits in Ethiopian indigenous and Sasso chicken breeds.

The present study investigated the biochemical polymorphism of hemoglobin (Hb) and its relationship with performance traits of Ethiopian indigenous and Sasso chicken breeds. A total of 284 chickens reared in three agro-ecologies were examined for genetic diversity and associations with productive traits at Hb locus using agarose gel electrophoresis. The results showed that the HbA allele was dominant in both breeds, and a higher proportion of male chickens were HbAA genotypes, while females were predominantly HbBB types. In the highland agro-ecology, chickens with the HbAA genotype were the most dominant, whereas in mid- and low-land agro-ecologies, chickens with HbBB and HbAB genotypes were found to be more frequent. A moderate level of expected heterozygosity was obtained with 0.47 and 0.445 for indigenous and Sasso chickens, respectively, with an average effective number of alleles per locus of 1.89 and 1.80. Moreover, chickens with HbAA genotypes showed significantly (p ≤ 0.05) higher body weight and linear body measurements than those of HbAB and HbBB genotypes. However, for appendage body structures (comb and wattle dimensions), chickens with the HbAB and HbBB genotypes had higher mean values. Additionally, clutch size (14.2 ± 0.4), clutch length (21.8 ± 0.7), and eight-month egg production (84.1 ± 1.2) were significantly (p ≤ 0.05) higher for hens with HbBB genotypes, followed by those with HbAB-types. Therefore, the considerable hemoglobin variability and significant associations of Hb variants with the performance traits can be sought as guiding information for further genetic improvement interventions in the chicken breeds under investigation. Further microsatellite marker-based genotyping is recommended to validate the higher morphometric values for HbAA genotypes and the better egg production for HbBB and HbAB genotypes.

Genetic and morphological diversity of introduced cultivars of almonds (Prunus amygdalus L.) in Bosnia and Herzegovina.

The main morphological and genetic characterization of seven introduced almond cultivars in Bosnia & Herzegovina was conducted. The almond cultivars included three from Italy (Tuono, Genco, Supernova), two from France (Ferragnes and Ferraduel), and two from the USA (Texas and Nonpareil). Genetic characterization was utilized by using 10 microsatellite markers, with nine markers from Prunus persicae and one from Prunus armeniaca. The results of genetic characterization revealed an average of 5.40 alleles per primer per locus. The average number of effective alleles for the 10 SSR loci of introduced cultivars was 3.92. The Shannon Information Index averaged 1.41. The observed heterozygosity (Ho) and expected heterozygosity (He) averaged 0.53 and 0.69, respectively. Morphological analyses of the fruit of introduced almond cultivars in Bosnia & Herzegovina indicated favorable agroecological conditions for their cultivation and spread. The results suggest that these introduced almond cultivars could be utilized in breeding programs to enhance the genetic diversity of the local almond population in Bosnia & Herzegovina.

Genetic Diversity and Construction of Core Collection of Sterculia Germplasm

Amplification of 107 Sterculia accessions with 12 ISSR primers resulted in 235 bands, the percentage of polymorphic sites was 100%, with the average number of effective alleles (Ne) of 1.2464, the average Nei’s gene diversity index (H) of 0.1556, the average Shannon's information index (I) of 0.2563, and the average polymorphism information content (PIC) of 0.8594, indicating that the collected 107 accessions had high genetic diversity. The genetic diversity was unevenly distributed among Sterculia samples collected from different provinces, and the highest value was observed in samples from Guangdong Province, followed by those from Yunnan Province, and the lowest was from Guangxi Province. The core collection construction of the 107 Sterculia accessions was carried out by using the SCR and LDSS methods, respectively, and the results showed that the LDSS method was more suitable for Sterculia germplasm. Different core collections were constructed based on sampling proportions of 35, 30, 25, 20 and 15%, respectively, the t-test and principal coordinates analysis results showed that the genetic diversity of the original germplasm can be fully represented at a sampling proportion of 30%. Bangladesh J. Bot. 53(2): 319-325, 2024 (June)

Microsatellite analysis of genetic diversity and population differentiation of Acanthopagrus schlegelii in China

The study utilized 9 self-selected polymorphic microsatellite loci to amplify and investigate the genetic diversity and differentiation of Acanthopagrus schlegelii populations in 10 wild populations along the Chinese coast. The values of Polymorphic Information Content (PIC) ranged from 0.4433 to 0.5804, with an average value of 0.5143. The average number of alleles per locus (A) ranged from 3.0000 to 3.4444, with a mean value of 3.2333 and an overall value of 3.6667. The average number of effective alleles per locus (Ae) ranged from 2.0130 to 2.6014, with a mean value of 2.2994 and an overall value of 2.8013. The observed heterozygosity (Ho) ranged from 0.4607 to 0.5789, with an average value of 0.5278 and an overall value of 0.5261. The expected heterozygosity (He) ranged from 0.4657 to 0.5967, with an average value of 0.5183 and an overall value of 0.6224. The results of genetic differentiation showed the genetic differentiation coefficient (Fst) was 0.1806, and the gene flow (Nm) was 1.1342. The UPGMA cluster analysis indicated significant genetic differentiation among A. schlegelii populations along the Chinese coast, with the populations being divided into two groups, namely the northern and southern populations. The genetic differentiation structure of the southern population was inconsistent with its geographical distribution. This study utilized microsatellite molecular markers to investigate the genetic diversity and genetic differentiation of 10 wild A. schlegelii populations along the coast of China. The research results indicate that the genetic diversity of the 10 wild A. schlegelii populations is at a moderate level, with evidence of north-south differentiation. This provides a scientific basis for the conservation, germplasm improvement, and aquaculture of A. schlegelii resources in China. It also offers background information for recent stock enhancement and release projects and their impact on the genetic resources of A. schlegelii.

Microsatellite marker-based analysis of the genetic diversity and population structure of three Arnebiae Radix in western China

Arnebiae Radix is an important medicinal and perennial herb found in Western China, particularly in the Xinjiang region. However, the assessment, utilization and conservation of Arnebiae Radix resources are still unexplored. In this study, we evaluated the genetic diversity of three Arnebiae Radix populations across 47 regions (Ae = 16, Ag = 16, Ad = 15) in Xinjiang, China, using inter-simple sequence repeat (ISSR) molecular markers. In total, 48 alleles were amplified by six pairs of primers screened with ISSR markers. The average number of effective alleles (Ne) was 1.5770. The percentage of interspecific genetic polymorphisms in A. guttata (Ag = 89.58 %) was greater than that in A. euchroma. and A. decumbens (Ae = Ad = 87.50 %). Intraspecific genetic polymorphisms, Bo Le (BL) population of A. euchroma exhibited the highest percentage of polymorphic bands (PPB% =58.33 %, Na = 1.313, Ne = 1.467, I = 0.0.366, H = 0.255), which indicated high genetic diversity. In contrast, the Tuo Li (TL) population of A. guttata had the lowest values for these parameters (PPB% =0.00 %, Na = 0.313, Ne = 1,000, I = 0.000, H = 0.000). The Arnebiae Radix germplasms were classified into two major groups (I and II) based on UPGMA cluster analysis (Fig. 8a) and principal coordinate analysis (PCOA). In addition, A. decumbens is placed in a separate category due to its high differentiation coefficient. The AMOVA and genetic differentiation coefficient results indicated that the genetic variation in Arnebiae Radix was predominantly due to intrapopulation differences (78 %). Additionally, the gene flow index (Nm) between populations was 2.4128, which further indicated that the genetic diversity of Arnebiae Radix was greater at the intrapopulation level. The destruction of the ecological environment leads to the continuous reduction and degradation of the genetic diversity of Arnebiae Radix germplasm resources. In this study, we used ISSR molecular markers to analyze the genetic diversity and relatedness of Arnebiae Radix, which revealed the genetic relationship of Arnebiae Radix germplasm resources at the molecular level and provided a scientific basis for future research on selecting and breeding good varieties, evaluating the quality of Arnebiae Radix, and conserving and utilizing its resources.

Genetic background of lily germplasm resources based on SSR markers

To study the genetic background of lily (Lilium spp.) germplasm resources, and accurately evaluate and select excellent germplasm for genetic improvement of lily, we analyzed the genetic background of 62 lily germplasm accessions from 11 provinces of China by using simple sequence repeat (SSR) molecular markers. The results showed that 15 out of 83 pairs of lily SSR primers were polymorphic. A total of 157 allelic loci were amplified, with the number of alleles per locus ranging from 5 to 19 and the average number of effective alleles per locus being 4.162 8. The average observed heterozygosity and expected heterozygosity were 0.228 2 and 0.694 1, respectively. The average polymorphic information content was 0.678 8. The average Nei's diversity index and Shannon's information index were 0.694 1 and 1.594 9, respectively, indicating that the tested lily germplasm had high genetic diversity. The 62 germplasm accessions were classified into 5 groups by the unweighted pair group method with arithmetic mean (UPGMA) and into 3 groups by the principal component analysis. The two analyses revealed a geographic correlation among different groups. The majority of lily germplasm accessions from the same source tended to cluster together. The population structure analysis classified the lily accessions into 4 populations and 1 mixed population. The above results provide a theoretical basis and genetic resources for the precise identification and breeding of lily germplasm resources.

Особенности генетической структуры популяции Pinus sylvestris L. в степной зоне Европейской России

The article presents the results of studу of the genetic structure of Scots pine (Pinus sylvestris L.) growing in the ecologically favorable territory of the steppe zone of European Russia. 35–year-old pine forest plantations (the Voronezh Region, Kantemirovskiy District, the 3rd bonitet class, forest site type – A1, a random sample, 60 trees) have been chosen as the object of the study. Genetic diversity has been examined on the basis of the microsatellite analysis of 18 SSR-loci (14 EST-SSR-loci and 4 nSSR-loci) and two isozymic loci (shikimate dehydrogenase and glutamate dehydrogenase – or Skdh and Gdh, respectively). It has been revealed that in the studied population, all the used microsatellite loci, with the exception of lw_isotig02842, are polymorphic. The average values of genetic variability calculated from the microsatellite loci have been obtained: the proportion of the polymorphic loci – 94.44 %, the average number of alleles per locus – 3.500, the average effective number of alleles – 2.466, the observed and expected heterozygosity – 0.209 and 0.493 respectively; the Wright fixation index – 0.577. In terms of the level of allelic diversity, the plantation is characterized by relatively lower values of genetic and statistical population parameters (the average observed and expected heterozygosity) compared to the natural forest-steppe populations of the Central Chernozem Region. A high positive Wright fixation index indicates the presence of an increased proportion of inbreeding in the steppe population trees. The reasons and possible nature of the revealed phenomenon are discussed. Based on the isoenzyme analysis, the genetic structure of the Skdh and Gdh loci is examined. The results of the analysis indicate a high frequency of the rapid allele Skdh-11 (82 %) and the embryonic semi-lethal allele Gdh-11 (22 %), which, apparently, can be considered as one of the mechanisms of the population adaptation to the conditions of a warmer and drier climate of the steppe region. A deficiency of glutamate dehydrogenase heterozygotes has been established. It has also been noted that the groups of EST-SSR-loci and nSSR-loci differ in the level of genetic variability and population structure. It has been shown that the combined use of different types of genetic markers makes it possible to obtain more complete and objective information on the distinctive features of the genetic structure of pine forests in the optimal and pessimal zones of the range. The studied 18 microsatellite loci can be used to assess the genetic diversity of Scots pine populations and trees in the steppe region of European Russia.

Genetic variability and population divergence of Rhododendron platypodum Diels in China in the context of conservation

Genetic diversity in endangered species is of special significance in the face of escalating global climate change and alarming biodiversity declines. Rhododendron platypodum Diels, an endangered species endemic to China, is distinguished by its restricted geographical range. This study aimed to explore genetic diversity and differentiation among its populations, gathering samples from all four distribution sites: Jinfo Mountain (JFM), Zhaoyun Mountain (ZYM), Baima Mountain (BMM), and Mao’er Mountain (MEM). We employed 18 pairs of Simple Sequence Repeat (SSR) primers to ascertain the genetic diversity and structural characteristics of these samples and further utilized 19 phenotypic data points to corroborate the differentiation observed among the populations. These primers detected 52 alleles, with the average number of observed alleles (Na) being 2.89, the average number of effective alleles (Ne) being 2.12, the average observed heterozygosity (Ho) being 0.57, and the expected heterozygosity (He) being 0.50. This array of data demonstrates the efficacy of the primers in reflecting R. platypodum’s genetic diversity. SSR-based genetic analysis of the populations yielded Ho, He, and Shannon index (I) values ranging from 0.47 to 0.65, 0.36 to 0.46, and 0.53 to 0.69, respectively. Notably, the ZYM population emerged as the most genetically diverse. Further analysis, incorporating molecular variance, principal component analysis, UPGMA cluster analysis, and structure analysis, highlighted significant genetic differentiation between the Chongqing (BMM, JFM, ZYM) and Guangxi (MEM) populations. Morphological data analysis corroborated these findings. Additionally, marked genetic and morphological distinctions were evident among the three Chongqing populations (BMM, JFM, and ZYM). This suggests that, despite the observed regional differentiation, R. platypodum’s overall genetic diversity is relatively constrained compared to other species within the Rhododendron genus. Consequently, R. platypodum conservation hinges critically on preserving its genetic diversity and protecting its distinct populations.

Performance of a 74-Microhaplotype Assay in Kinship Analyses.

Microhaplotypes (MHs) consisting of multiple SNPs and indels on short stretches of DNA are new and interesting loci for forensic genetic investigations. In this study, we analysed 74 previously defined MHs in two of the populations that our laboratory provides with forensic genetic services, Danes and Greenlanders. In addition to the 229 SNPs that originally made up the 74 MHs, 66 SNPs and 3 indels were identified in the two populations, and 45 of these variants were included in new definitions of the MHs, whereas 24 SNPs were considered rare and of little value for case work. The average effective number of alleles (Ae) was 3.2, 3.0, and 2.6 in Danes, West Greenlanders, and East Greenlanders, respectively. High levels of linkage disequilibrium were observed in East Greenlanders, which reflects the characteristics of this population that has a small size, and signs of admixture and substructure. Pairwise kinship simulations of full siblings, half-siblings, first cousins, and unrelated individuals were performed using allele frequencies from MHs, STRs and SNPs from Danish and Greenlandic populations. The MH panel outperformed the currently used STR and SNP marker sets and was able to differentiate siblings from unrelated individuals with a 0% false positive rate and a 1.1% false negative rate using an LR threshold of 10,000 in the Danish population. However, the panel was not able to differentiate half-siblings or first cousins from unrelated individuals. The results generated in this study will be used to implement MHs as investigative markers for relationship testing in our laboratory.

Morphological and molecular diversity of eggplant accessions (Solanum melongena L) using Simple Sequence Repeats (SSR) markers

The evaluation of various desirable characteristics in eggplant genotypes has facilitated the efficient process of selecting and improving their traits. Morphological parameters have proven to be valuable in assessing the similarities or differences among different accessions, while molecular data have been used to support the conclusions drawn from the morphological analysis. This study was conducted to evaluate the performance of 42 eggplant genotypes collected from Malaysia, China, and Thailand. Data for growth, yield, and related traits were collected and subjected to analysis of variance (ANOVA) using SAS 9.4. The field trial experiment was designed as a completely randomized block design with three replications. Analysis of variance (ANOVA) showed that the traits under study were highly significant (p ≤ 0.01). MV18 (5.97 kg) and TV17 (5.59 kg) were observed to have high yields per plant. The SSR markers used exhibited moderate average values for the number of alleles (2.53). The major allele frequency displayed a high average value (0.53) and a moderate average number of effective alleles (2.31). Additionally, the observed Shannon's information index, expected heterozygosity, and PIC were high (0.84, 0.54, and 0.45, respectively). 42 accessions were grouped into five major clusters based on similarities using the unweighted pair-group method with arithmetic averages based on similarity matrices UPGMA Dendrogram and the EIGEN module. The findings of this study indicate that the use of simple sequence repeat (SSR) markers can effectively estimate genetic diversity and analyze phylogenetic relationships. Moreover, these markers can assist eggplant breeders in selecting desirable quantitative traits within their breeding program.

ИЗУЧЕНИЕ ПОЛИМОРФИЗМА МИКРОСАТЕЛЛИТНЫХ ЛОКУСОВ ОБРАЗЦОВ ВИНОГРАДА С НЕИДЕНТИФИЦИРОВАННОЙ СОРТОВОЙ ПРИНАДЛЕЖНОСТЬЮ АМПЕЛОГРАФИЧЕСКОЙ КОЛЛЕКЦИИ «МАГАРАЧ» С ИСПОЛЬЗОВАНИЕМ МОЛЕКУЛЯРНЫХ МАРКЕРОВ

The paper presents the results of DNA profiling of 28 grape samples with unidentified varietal affiliation, selected from the Magarach Ampelographic Collection, and provides polymorphism assessment of their microsatellite loci. Genotyping was performed using 9 nuclear (VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VrZAG79, VVS2) and 3 chloroplast (ccmp3, ccmp5, ccmp10) SSR markers. For nuclear loci, 105 alleles were identified, the average number of alleles per locus (na) was 11.7; the average effective number of alleles (ne) is 7.23; Shannon information index (I) for the studied batch of samples was 2.13. The highest allele frequency was observed in samples with VVS2133 and VVS2135 (p = 0.250), VVMD5238 (p = 0.232), VVMD7247 (p = 0.232), VVMD25239 (p = 0.250), VVMD27180 (p = 0.232), VVMD28236 (p = 0.143), VVMD32272 (p = 0.304), VrZAG62188 (p = 0.179), VrZAG79251 (p = 0.232) alleles. The average value of actual heterozygosity Heto was 0.8333, expected heterozygosity Hetе was 0.8712. For chloroplast loci, 7 alleles were identified: ccmp3106, ccmp3107, ccmp5104, ccmp5105, ccmp10114, ccmp10115, ccmp10116. Also 4 chlorotypes were identified: A (1 sample), B (9), C (13), D (5). A dendrogram of genetic similarity was constructed using the UPGMA method, which showed the presence of three clusters containing, respectively, 11, 11 and 6 samples. There was a uniform distribution of chlorotypes B and D over three clusters, and chlorotype C – over two clusters. It is concluded that the results can be used to assess the genetic diversity of Magarach Ampelographic Collection.

Analysis of the genetic diversity and population structures of black locust (Robinia pseudoacacia L.) stands in Poland based on simple sequence repeat markers

Abstract Black locust (Robinia pseudoacacia L.) was introduced in Poland over 200 years ago, and its distribution area now covers the whole country, with the highest concentration of occurrence in the western part. Breeding of this species has been started in Poland for about 20 years, albeit on a limited scale, and two selected seed stands, 34 plus trees and two seed orchards have been registered. So far, selection efforts have not been coupled with the recognition of variation in the genetic structure of local populations. To fill the knowledge gap on genetic diversity, we selected the seven qualitatively best populations of R. pseudoacacia for our study. To clarify the origin of the Polish population of R. pseudoacacia, a sample from a seed stand in eastern Germany and from a seed orchard with Hungarian clones from the Oborniki Śląskie Forest District was added. In this study, three microsatellites were used to evaluate genotypes: Rops15, Rops16 and Rops18, with the Rops15 locus (motif AG) showing hypermutability. Moderate or high variability was observed in all microsatellite loci examined, with a total of 27 alleles identified. In all stands, the average observed number of alleles per locus was higher than the average effective number of alleles per locus. Three main clusters have been identified in the genetic structure of the population, one of which is represented by the population from the Regional Directorate of State Forests in Zielona Góra. The second group comprised the populations from Mieszkowice, the populations Pińczów and Wołów, and the population from Germany. The last one included population from Strzelce and Oborniki Śląskie, where the population from Hungary is represented. This may be of great importance for practice. Perhaps it is worth postulating the separation of two seed regions. Under most scenarios, climate change is projected to change the distribution of forest types and tree species in all biomes. It can be assumed that the importance of black locust in times of climate change is likely to increase.

Improving DNA mixtures analysis using compound markers composed of InDels and SNPs screened from the whole genome with next-generation sequencing.

Next-generation sequencing (NGS) allows for better identification of insertion and deletion polymorphisms (InDels) and their combination with adjacent single nucleotide polymorphisms (SNPs) to form compound markers. These markers can improve the polymorphism of microhaplotypes (MHs) within the same length range, and thus, boost the efficiency of DNA mixture analysis. In this study, we screened InDels and SNPs across the whole genome and selected highly polymorphic markers composed of InDels and/or SNPs within 300bp. Further, we successfully developed and evaluated an NGS-based panel comprising 55 loci, of which 24 were composed of both SNPs and InDels. Analysis of 124 unrelated Southern Han Chinese revealed an average effective number of alleles (Ae ) of 7.52 for this panel. The cumulative power of discrimination and cumulative probability of exclusion values of the 55 loci were 1-2.37×10-73 and 1-1.19×10-28 , respectively. Additionally, this panel exhibited high allele detection rates of over 97% in each of the 21 artificial mixtures involving from two to six contributors at different mixing ratios. We used EuroForMix to calculate the likelihood ratio (LR) and evaluate the evidence strength provided by this panel, and it could assess evidence strength with LR, distinguishing real and noncontributors. In conclusion, our panel holds great potential for detecting and analyzing DNA mixtures in forensic applications, with the capability to enhance routine mixture analysis.

Genetic diversity, population structure, and DNA fingerprinting of Ailanthus altissima var. erythrocarpa based on EST-SSR markers

Ailanthus altissima var. erythrocarpa is an A. altissima variety with high economic, ecological and ornamental value, but there have been no reports on the development of SSR primers for it. According to the SSR primer information provided by the transcriptome of A. altissima var. erythrocarpa, 120 individuals with different redness levels were used to screen polymorphic primers. Transcriptomic analysis revealed 10,681 SSR loci, of which mononucleotide repeats were dominant (58.3%), followed by dinucleotide and trinucleotide repeats (16.6%, 15.1%) and pentanucleotide repeats (0.2%). Among 140 pairs of randomly selected primers, nineteen pairs of core primers with high polymorphism were obtained. The average number of alleles (Na), average number of effective alleles (Ne), average Shannon’s diversity index (I), average observed heterozygosity (Ho), average expected heterozygosity (He), fixation index (F) and polymorphic information content (PIC) were 11.623, 4.098, 1.626, 0.516, 0.696, 0.232 and 0.671, respectively. Nineteen EST-SSR markers were used to study the genetic diversity and population structure of A. altissima var. erythrocarpa. The phylogenetic tree, PCoA, and structure analysis all divided the tested resources into two categories, clearly showing the genetic variation between individuals. The population showed high genetic diversity, mainly derived from intraspecific variation. Among nineteen pairs of primers, 4 pairs (p33, p15, p46, p92) could effectively distinguish and be used for fingerprinting of the tested materials. This study is of great significance for genetic diversity analysis and molecular-assisted breeding of A. altissima var. erythrocarpa.

Exploring the Genetic Diversity and Population Structure of Daphnia cucullata Sars, 1862 in Boreal Lakes (Latvian Lakeland) Based on Microsatellites

We have used Daphnia cucullata (Crustacea: Cladocera) as a model organism for the first time in the four deepest Latvian lakes from the Boreal biogeographical region in order to find the genetic diversity of these populations. During the research, we detected the most appropriate microsatellite markers for future genetic studies of Daphnia cucullata populations of lakes Svente, Riča, Dridzis and Geraņimovas-Ilzas in the Boreal biogeographical region. Based on these microsatellite markers, we determined the genetic diversity of these populations. The loci Dgm105 and Dgm101 had the maximum number of alleles and the maximum number of private alleles. The specific locus Dgm105 had five private alleles (62% of all detected alleles), and locus Dgm101 had four private alleles (57% of all detected alleles) in these loci. We determined the observed heterozygosity (Hobs) and the expected heterozygosity (Hexp) level (via Hardy–Weinberg equilibrium), the number of polymorphic loci, the number of detected alleles in each analyzed microsatellite locus, the average number of alleles at the locus (Na), the average effective number of alleles at the locus (Ne), the FST of the population’s genetic differentiation, the genetic distance (D) (following Nei) and the significance (χ2-test) of differences between the levels of observed and expected heterozygosity. It was shown that Daphnia cucullata populations from lakes with a low number of zooplankton taxa (Riča and Geraņimovas-Ilzas) have a higher genetic diversity compared to lakes with a high number of zooplankton taxa (Dridzis and Svente). It was found that Daphnia cucullata populations from lakes Dridzis and Svente have the least genetic distance, and these populations form a single genetic group, as confirmed via clustering.

SSR marker based profiling and population structure analysis in peach (Prunus persica) germplasm

For breeding programmes to be successful and for germplasm conservation, it is essential to characterize and analyze the genetic diversity of available germplasm. The present experiment was conducted at Molecular Biology and Genetic Engineering Laboratory of Uttarakhand Council for Biotechnology, Haldi, Uttarakhand during 2022 to study the molecular profile of 41 peach [Prunus persica (L.) Stokes] accessions using 23 polymorphic SSR markers. The number of alleles detected ranged from 3 to 8 with an average of 4.65 alleles per locus (Na) and a total of 107 alleles were amplified. The average effective number of alleles (Ne) were 2.89 per marker. The SSR marker MA015a produced maximum number of 8 alleles followed by BPPCT 015 and CPPCT14 which produced 7 alleles each. The polymorphic information content (PIC) varied between 0.317–0.836 with a mean value of 0.563. The observed heterozygosity examined was lower (Ho = 0.02) and the expected heterozygosity (He = 0.61) ranged between 0.34 to 0.85. The presence of a higher Shannon’s information index (I) of 1.17 indicates higher diversity in the given set of peach genotypes. Jaccard’s similarity coefficient ranging from 0.533 to 1, indicated a pair-wise relationship among the peach accessions. The cluster dendrogram partitioned the accessions into two main clusters. However, the total accessions were stratified into 3 groups (K=3) based on population structure analysis which was further confirmed by Principal Coordinate Analysis (PCoA). The information generated in the study may have great implications in molecular characterization, fingerprinting and documentation of accessions in the peach improvement programme.

김해 야생차 군락지들의 차나무에 대한 유전적 다양성 분석

This study examined the genetic diversity of 13 tea tree samples to establish a variety discrimination system for the three Gimhae wild tea colonies using ISSR markers and to provide basic data for genetic diversity analysis. Seven primers that showed polymorphic and reproducible results among 100 ISSR universal primers were selected and used to amplify the genomic DNA of the 13 tea trees from three Gimhae wild tea colonies. The results demonstrated that all seven primers generated 117 bands with a 100% polymorphic band ratio. The average observed number of alleles was 2.0. The average effective number of alleles was 1.5323, while the average Nei's gene diversity was 0.3271. The average Shannon's Information index was 0.5001. The genetic similarity among the 13 tea tree samples ranged from 0.4444 to 0.8718, while the genetic distance ranged from 0.1372 to 0.8109. The 13 tea trees in three Gimhae wild colonies displayed a high level of genetic diversity, and the Dongsangdong and Sangdong samples in the three Gimhae wild colonies had a genetic similarity greater than 0.8 with Chinese tea species, indicating a close relationship with Chinese tea. Overall, these findings provide a theoretical basis for preserving and cultivating Gimhae wild tea trees.