scholarly journals Targeted genome-wide SNP genotyping in feral horses using non-invasive fecal swabs

2021 ◽  
Author(s):  
Stefan Gavriliuc ◽  
Salman Reza ◽  
Chanwoori Jeong ◽  
Fitsum Getachew ◽  
Philip Dunstan McLoughlin ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Snp Genotyping ◽  
Nucleotide Polymorphisms ◽  
Feral Horses ◽  
Non Invasive ◽  
Genetics Research ◽  
Snp Data ◽  
Genome Wide ◽  
Snp Microarrays ◽  
Template Dna

Abstract The development of high-throughput sequencing has prompted a transition in wildlife genetics from using microsatellites toward sets of Single Nucleotide Polymorphisms (SNPs). However, genotyping large numbers of targeted SNPs using non-invasive samples remains challenging due to relatively large DNA input requirements. Recently, target enrichment has emerged as a promising approach requiring little template DNA. We assessed the efficacy of Tecan Genomics’ Allegro Targeted Genotyping (ATG) for generating genome-wide SNP data in feral horses using DNA isolated from fecal swabs. Total and host-specific DNA were quantified for 989 samples collected as part of a long-term individual-based study of feral horses on Sable Island, Nova Scotia, Canada, using dsDNA fluorescence and a host-specific qPCR assay, respectively. Forty-eight samples representing 44 individuals containing at least 10ng of host DNA (ATG’s recommended minimum input) were genotyped using a custom multiplex panel targeting 279 SNPs. Genotyping accuracy and consistency were assessed by contrasting ATG genotypes with those obtained from the same individuals with SNP microarrays, and from multiple samples from the same horse, respectively. 62% of swabs yielded the minimum recommended amount of host DNA for ATG. Ignoring samples that failed to amplify, ATG recovered an average of 86.7% targeted sites per sample, while genotype concordance between ATG and SNP microarrays was 98.5%. The repeatability of genotypes from the same individual approached unity with an average of 99.9%. This study demonstrates the suitability of ATG for genome-wide, non-invasive targeted SNP genotyping, and will facilitate further ecological and conservation genetics research in equids and related species.

scholarly journals Genome-wide SNPs redefines species boundaries and conservation units in the freshwater mussel genus Cyprogenia of North America

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kyung Seok Kim ◽  
Kevin J. Roe
Keyword(s):  
Phylogenetic Analyses ◽  
Freshwater Mussel ◽  
Conservation Strategies ◽  
Nucleotide Polymorphisms ◽  
Conservation Units ◽  
Genetic Structuring ◽  
The North ◽  
Snp Data ◽  
Genome Wide ◽  
Significant Difference

AbstractDetailed information on species delineation and population genetic structure is a prerequisite for designing effective restoration and conservation strategies for imperiled organisms. Phylogenomic and population genomic analyses based on genome-wide double digest restriction-site associated DNA sequencing (ddRAD-Seq) data has identified three allopatric lineages in the North American freshwater mussel genus Cyprogenia. Cyprogenia stegaria is restricted to the Eastern Highlands and displays little genetic structuring within this region. However, two allopatric lineages of C. aberti in the Ozark and Ouachita highlands exhibit substantial levels (mean uncorrected FST = 0.368) of genetic differentiation and each warrants recognition as a distinct evolutionary lineage. Lineages of Cyprogenia in the Ouachita and Ozark highlands are further subdivided reflecting structuring at the level of river systems. Species tree inference and species delimitation in a Bayesian framework using single nucleotide polymorphisms (SNP) data supported results from phylogenetic analyses, and supports three species of Cyprogenia over the currently recognized two species. A comparison of SNPs generated from both destructively and non-destructively collected samples revealed no significant difference in the SNP error rate, quality and amount of ddRAD sequence reads, indicating that nondestructive or trace samples can be effectively utilized to generate SNP data for organisms for which destructive sampling is not permitted.

scholarly journals Identification of genome-wide single-nucleotide polymorphisms among geographically diverse radish accessions

DNA Research ◽  
2020 ◽  
Vol 27 (1) ◽  
Author(s):  
Hiroto Kobayashi ◽  
Kenta Shirasawa ◽  
Nobuko Fukino ◽  
Hideki Hirakawa ◽  
Takashi Akanuma ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphisms ◽  
Southeast Asia ◽  
East Asia ◽  
Near East ◽  
Dna Polymorphisms ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
South And Southeast Asia ◽  
Snp Data ◽  
Genome Wide

Abstract Radish (Raphanus sativus L.) is cultivated around the world as a vegetable crop and exhibits diverse morphological and physiological features. DNA polymorphisms are responsible for differences in traits among cultivars. In this study, we determined genome-wide single-nucleotide polymorphisms (SNPs) among geographically diverse radish accessions using the double-digest restriction site-associated DNA sequencing (ddRAD-Seq) method. A total of 52,559 SNPs was identified in a collection of over 500 radish accessions (cultivated and wild) from East Asia, South and Southeast Asia, and the Occident and Near East. In addition, 2,624 SNP sites without missing data (referred to as common SNP sites) were identified among 510 accessions. Genetic diversity analyses, based on the common SNP sites, divided the cultivated radish accessions into four main groups, each derived from four geographical areas (Japan, East Asia, South and Southeast Asia, and the Occident and Near East). Furthermore, we discuss the origin of cultivated radish and its migration from the West to East Asia. SNP data generated in this work will facilitate further genetic studies on the radish breeding and production of DNA markers.

scholarly journals Phased Genotyping-by-Sequencing Enhances Analysis of Genetic Diversity and Reveals Divergent Copy Number Variants in Maize

2017 ◽  
Vol 7 (7) ◽  
pp. 2161-2170 ◽  
Author(s):  
Heather Manching ◽  
Subhajit Sengupta ◽  
Keith R Hopper ◽  
Shawn W Polson ◽  
Yuan Ji ◽  
...  
Keyword(s):  
Copy Number ◽  
High Throughput Sequencing ◽  
Copy Number Variants ◽  
Genotyping By Sequencing ◽  
Genomic Libraries ◽  
Reduced Representation ◽  
Large Samples ◽  
Snp Data ◽  
Genome Wide ◽  
Missing Data Problem

Abstract High-throughput sequencing (HTS) of reduced representation genomic libraries has ushered in an era of genotyping-by-sequencing (GBS), where genome-wide genotype data can be obtained for nearly any species. However, there remains a need for imputation-free GBS methods for genotyping large samples taken from heterogeneous populations of heterozygous individuals. This requires that a number of issues encountered with GBS be considered, including the sequencing of nonoverlapping sets of loci across multiple GBS libraries, a common missing data problem that results in low call rates for markers per individual, and a tendency for applicability only in inbred line samples with sufficient linkage disequilibrium for accurate imputation. We addressed these issues while developing and validating a new, comprehensive platform for GBS. This study supports the notion that GBS can be tailored to particular aims, and using Zea mays our results indicate that large samples of unknown pedigree can be genotyped to obtain complete and accurate GBS data. Optimizing size selection to sequence a high proportion of shared loci among individuals in different libraries and using simple in silico filters, a GBS procedure was established that produces high call rates per marker (>85%) with accuracy exceeding 99.4%. Furthermore, by capitalizing on the sequence-read structure of GBS data (stacks of reads), a new tool for resolving local haplotypes and scoring phased genotypes was developed, a feature that is not available in many GBS pipelines. Using local haplotypes reduces the marker dimensionality of the genotype matrix while increasing the informativeness of the data. Phased GBS in maize also revealed the existence of reproducibly inaccurate (apparent accuracy) genotypes that were due to divergent copy number variants (CNVs) unobservable in the underlying single nucleotide polymorphism (SNP) data.

scholarly journals Comparison of single nucleotide polymorphisms and microsatellites in non-invasive genetic monitoring of a wolf population

2012 ◽  
Vol 64 (1) ◽  
pp. 321-335 ◽  
Author(s):  
Elena Fabbri ◽  
R. Caniglia ◽  
Nadia Mucci ◽  
H.P. Thomsen ◽  
K. Krag ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphisms ◽  
Error Rates ◽  
Snp Genotyping ◽  
Genetic Monitoring ◽  
Taqman Probe ◽  
Degraded Dna ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Non Invasive ◽  
Probe Assay

Single nucleotide polymorphisms (SNPs) which represent the most widespread source of sequence variation in genomes, are becoming a routine application in several fields such as forensics, ecology and conservation genetics. Their use, requiring short amplifications, may allow a more efficient genotyping of degraded DNA. We provide the first application of SNP genotyping in an Italian non-invasive genetic monitoring project of the wolf. We compared three different techniques for genotyping SNPs: pyrosequencing, SNaPshot? and TaqMan? Probe Assay in Real-Time PCR. We successively genotyped nine SNPs using the TaqMan Probe Assay in 51 Italian wolves, 57 domestic dogs, 15 wolf x dog hybrids and 313 wolf scats collected in the northern Apennines. The obtained results were used to estimate genetic variability and PCR error rates in SNP genotyping protocols compared to standard microsatellite analysis. We evaluated the cost, laboratory effort and reliability of these different markers and discuss the possible future use of VeraCode, SNPlex and Fluidigm EP1 system in wild population monitoring.

scholarly journals Sample identification and pedigree reconstruction in Wolverine (Gulo gulo) using SNP genotyping of non-invasive samples

2021 ◽  
Author(s):  
Robert Ekblom ◽  
Malin Aronsson ◽  
Franziska Elsner-Gearing ◽  
Malin Johansson ◽  
Toby Fountain ◽  
...  
Keyword(s):  
Snp Array ◽  
Snp Genotyping ◽  
Individual Identification ◽  
Nucleotide Polymorphisms ◽  
Genetic Management ◽  
Gulo Gulo ◽  
Non Invasive ◽  
Species Of Conservation Concern ◽  
Different Types ◽  
Improved Performance

AbstractFor conservation genetic studies using non-invasively collected samples, genome-wide data may be hard to acquire. Until now, such studies have instead mostly relied on analyses of traditional genetic markers such as microsatellites (SSRs). Recently, high throughput genotyping of single nucleotide polymorphisms (SNPs) has become available, expanding the use of genomic methods to include non-model species of conservation concern. We have developed a 96-marker SNP array for use in applied conservation monitoring of the Scandinavian wolverine (Gulo gulo) population. By genotyping more than a thousand non-invasively collected samples, we were able to obtain precise estimates of different types of genotyping errors and sample dropout rates. The SNP panel significantly outperforms the SSR markers (and DBY intron markers for sexing) both in terms of precision in genotyping, sex assignment and individual identification, as well as in the proportion of samples successfully genotyped. Furthermore, SNP genotyping offers a simplified laboratory and analysis pipeline with fewer samples needed to be repeatedly genotyped in order to obtain reliable consensus data. In addition, we utilised a unique opportunity to successfully demonstrate the application of SNP genotype data for reconstructing pedigrees in wild populations, by validating the method with samples from wild individuals with known relatedness. By offering a simplified workflow with improved performance, we anticipate this methodology will facilitate the use of non-invasive samples to improve genetic management of many different types of populations that have previously been challenging to survey.

scholarly journals Genome-wide SNP Data Reveal an Overestimation of Species Diversity in a Group of Hawkmoths

2019 ◽  
Vol 11 (8) ◽  
pp. 2136-2150 ◽  
Author(s):  
Anna K Hundsdoerfer ◽  
Kyung Min Lee ◽  
Ian J Kitching ◽  
Marko Mutanen
Keyword(s):  
Valid Species ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Contemporary Debate ◽  
Snp Data ◽  
Genome Wide ◽  
Genome Wide Data ◽  
Research Outcome ◽  
Biological Entities ◽  
Hyles Euphorbiae

Abstract The interface between populations and evolving young species continues to generate much contemporary debate in systematics depending on the species concept(s) applied but which ultimately reduces to the fundamental question of “when do nondiscrete entities become distinct, mutually exclusive evolutionary units”? Species are perceived as critical biological entities, and the discovery and naming of new species is perceived by many authors as a major research aim for assessing current biodiversity before much of it becomes extinct. However, less attention is given to determining whether these names represent valid biological entities because this is perceived as both a laborious chore and an undesirable research outcome. The charismatic spurge hawkmoths (Hyles euphorbiae complex, HEC) offer an opportunity to study this less fashionable aspect of systematics. To elucidate this intriguing systematic challenge, we analyzed over 10,000 ddRAD single nucleotide polymorphisms from 62 individuals using coalescent-based and population genomic methodology. These genome-wide data reveal a clear overestimation of (sub)species-level diversity and demonstrate that the HEC taxonomy has been seriously oversplit. We conclude that only one valid species name should be retained for the entire HEC, namely Hyles euphorbiae, and we do not recognize any formal subspecies or other taxonomic subdivisions within it. Although the adoption of genetic tools has frequently revealed morphologically cryptic diversity, the converse, taxonomic oversplitting of species, is generally (and wrongly in our opinion) accepted as rare. Furthermore, taxonomic oversplitting is most likely to have taken place in intensively studied popular and charismatic organisms such as the HEC.

scholarly journals Characterization of genome-wide genetic variation between two varieties of tea plant (Camellia sinensis) and development of InDel markers for genetics research

2019 ◽  
Author(s):  
Shengrui Liu ◽  
Yanlin An ◽  
Wei Tong ◽  
Xiuju Qin ◽  
Lidia Samarina ◽  
...  
Keyword(s):  
Genetic Variation ◽  
Camellia Sinensis ◽  
Distribution Patterns ◽  
Tea Plant ◽  
Whole Genome ◽  
Nucleotide Polymorphisms ◽  
Genetics Research ◽  
Indel Markers ◽  
Genome Wide ◽  
Caffeine Biosynthesis

Abstract Single nucleotide polymorphisms (SNPs) and Insertions/Deletions (InDels) are the major genetic variations and distributed extensively across the plant whole genome. Few investigations of these variations, however, have been performed in the long-lived perennial tea plant. Here, we have investigated the genome-wide genetic variation between Camellia sinensis var. sinensis ‘Shuchazao’ and Camellia sinensis var. assamica ‘Yunkang 10’, generating 7,511,731 SNPs and 255,218 InDels based on their whole genome sequences, and subsequently analyzed their distinct types and distribution patterns. A total of 48 InDel markers that yielded polymorphic and unambiguous fragments were developed when screening six tea cultivars. These markers were further deployed on forty-six tea cultivars for transferability and genetic diversity analysis, exhibiting informative with an average 4.02 of the number of alleles ( Na ) and 0.457 of polymorphism information content (PIC). The dendrogram showed that the phylogenetic relationships among these tea cultivars are highly consistent with their genetic backgrounds or original places. Interestingly, we observed that the content of catechin/caffeine between ‘Shuchazao’ and ‘Yunkang 10’ were significantly different, and a large number of SNPs/InDels were identified within catechin/caffeine biosynthesis-related genes. The identified genome-wide genetic variation and newly-developed InDel markers will provide a valuable resource for tea plant genetics and genomics studies, especially those SNPs/InDels within catechin/caffeine biosynthesis-related genes can be served as pivotal candidates for elucidating the molecular mechanism of catechin/caffeine biosynthesis.

scholarly journals High-throughput sequencing reveals inbreeding depression in a natural population

2014 ◽  
Vol 111 (10) ◽  
pp. 3775-3780 ◽  
Author(s):  
Joseph I. Hoffman ◽  
Fraser Simpson ◽  
Patrice David ◽  
Jolianne M. Rijks ◽  
Thijs Kuiken ◽  
...  
Keyword(s):  
Natural Population ◽  
Inbreeding Depression ◽  
High Throughput Sequencing ◽  
Natural Populations ◽  
Harbor Seals ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Peromyscus Polionotus ◽  
Genome Wide ◽  
Underlying Mechanisms

Proxy measures of genome-wide heterozygosity based on approximately 10 microsatellites have been used to uncover heterozygosity fitness correlations (HFCs) for a wealth of important fitness traits in natural populations. However, effect sizes are typically very small and the underlying mechanisms remain contentious, as a handful of markers usually provides little power to detect inbreeding. We therefore used restriction site associated DNA (RAD) sequencing to accurately estimate genome-wide heterozygosity, an approach transferrable to any organism. As a proof of concept, we first RAD sequenced oldfield mice (Peromyscus polionotus) from a known pedigree, finding strong concordance between the inbreeding coefficient and heterozygosity measured at 13,198 single-nucleotide polymorphisms (SNPs). When applied to a natural population of harbor seals (Phoca vitulina), a weak HFC for parasite infection based on 27 microsatellites strengthened considerably with 14,585 SNPs, the deviance explained by heterozygosity increasing almost fivefold to a remarkable 49%. These findings arguably provide the strongest evidence to date of an HFC being due to inbreeding depression in a natural population lacking a pedigree. They also suggest that under some circumstances heterozygosity may explain far more variation in fitness than previously envisaged.

Genetic structure of Patagonian toothfish populations from otolith DNA

2016 ◽  
Vol 28 (5) ◽  
pp. 347-360 ◽  
Author(s):  
Lola Toomey ◽  
Dirk Welsford ◽  
Sharon A. Appleyard ◽  
Andrea Polanowski ◽  
Cassandra Faux ◽  
...  
Keyword(s):  
Population Genetics ◽  
Genetic Differentiation ◽  
High Throughput Sequencing ◽  
Nucleotide Polymorphisms ◽  
Nuclear Markers ◽  
Genetic Stock ◽  
Mitochondrial Markers ◽  
Genetics Research ◽  
Spatial Coverage ◽  
Patagonian Toothfish

AbstractThe Patagonian toothfish, Dissostichus eleginoides, is a valuable fishery species and has a discontinuous distribution across the Southern Ocean. Identification of the genetic stock structure of toothfish would allow evaluation of the suitability of the spatial scale at which fisheries management operates. Genetic subdivision seems likely given the species distribution. Population genetics studies of this species have been performed; however, they have been limited by sample size, spatial coverage and/or the type of markers investigated. As a potential solution, we developed methods for extracting toothfish DNA from otoliths that are available in large numbers from collections held at several research institutes. Genetic differentiation between the three oceanic sectors was investigated. Four mitochondrial and four nuclear markers with multiple single nucleotide polymorphisms were sequenced by high throughput sequencing for samples from six locations. Genetic differentiation was found between three sectors with nuclear markers. However, only the Pacific sector was differentiated from other sectors with mitochondrial markers. This study demonstrates the usefulness of otolith DNA as a means of increasing sample sizes for population genetics research of fish. Additionally, the combination of nuclear and mitochondrial markers may allow insight into how the observed differences in movements between male and female toothfish impact population structure.

scholarly journals Performance comparison of two reduced-representation based genome-wide marker-discovery strategies in a multi-taxon phylogeographic framework

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Philipp Kirschner ◽  
◽  
Wolfgang Arthofer ◽  
Stefanie Pfeifenberger ◽  
Eliška Záveská ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Performance Comparison ◽  
Nucleotide Polymorphisms ◽  
Aflp Data ◽  
Single Nucleotide ◽  
Reduced Representation ◽  
Genome Wide ◽  
Taxonomic Groups ◽  
Marker Discovery ◽  
Study Species

AbstractMulti-locus genetic data are pivotal in phylogenetics. Today, high-throughput sequencing (HTS) allows scientists to generate an unprecedented amount of such data from any organism. However, HTS is resource intense and may not be accessible to wide parts of the scientific community. In phylogeography, the use of HTS has concentrated on a few taxonomic groups, and the amount of data used to resolve a phylogeographic pattern often seems arbitrary. We explore the performance of two genetic marker sampling strategies and the effect of marker quantity in a comparative phylogeographic framework focusing on six species (arthropods and plants). The same analyses were applied to data inferred from amplified fragment length polymorphism fingerprinting (AFLP), a cheap, non-HTS based technique that is able to straightforwardly produce several hundred markers, and from restriction site associated DNA sequencing (RADseq), a more expensive, HTS-based technique that produces thousands of single nucleotide polymorphisms. We show that in four of six study species, AFLP leads to results comparable with those of RADseq. While we do not aim to contest the advantages of HTS techniques, we also show that AFLP is a robust technique to delimit evolutionary entities in both plants and animals. The demonstrated similarity of results from the two techniques also strengthens biological conclusions that were based on AFLP data in the past, an important finding given the wide utilization of AFLP over the last decades. We emphasize that whenever the delimitation of evolutionary entities is the central goal, as it is in many fields of biodiversity research, AFLP is still an adequate technique.

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