Rapid and Low-Cost Molecular Sexing of a Corvid Songbird Using a Single Protocol with Two Universal Primer Sets

Lois‐Milevicich, Jimena; Gómez, Raúl O.; Ursino, Cynthia A.; Lois, Nicolás A.; Colina, Alicia de la

doi:10.13157/arla.68.2.2021.sc1

Cited by 6 publications

(5 citation statements)

References 34 publications

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“…The findings in owls mentioned above contrast with most other bird groups, where CHD1W is generally shorter than CHD1Z, and the differences between fragments are comparatively more minor 28 , 37 . For example, matching the results of 2550F/2718R, the plush-crested jays ( Cyanocorax chrysops ), Passeriformes, were characterised by Z-fragment ∼ 650 bp and W-fragment ∼ 450 bp in length 46 and the sex of brown eared pheasants ( Crossoptilon mantchuricum ), Galliformes, were revealed by Z-fragment ∼ 600 bp and W-fragment ∼ 450 bp long 19 . Primers CHD1F/CHD1R produced 443 bp in Z chromosome amplicon and 327 bp in W one in mallard duck ( Anas platyrhynchos ), Anseriformes 29 .…”

Section: Discussionsupporting

confidence: 53%

Comparison of three primer pairs for molecular sex determination in Eurasian pygmy owls (Glaucidium passerinum)

Stehlíková Sovadinová,

Mekadim,

Korpimäki

et al. 2024

Sci Rep

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Bird sex determination is fundamental in various ecological and biological studies, although many avian species cannot be sexed visually due to their monomorphic and/or monochromatic appearance. Thus, reliable laboratory methods for sexing are a prerequisite. Most avian nestlings lack sex-related signs, including the Eurasian pygmy owl (Glaucidium passerinum). We performed laboratory sex determination analysis of this species using blood samples of 242 juveniles and nine adults. It relied on the qPCR of the specific intron from the chromo-helicase DNA-binding protein 1 gene. We tested three primer sets, the P2/P8, 2550F/2718R, and CHD1F/CHD1R, commonly used for bird laboratory sexing. The outcomes were displayed on an agarose gel electrophoresis and a plot from melt curve analysis, which had not been previously conducted in Eurasian pygmy owls. We found that only primer set CHD1F/CHD1R proved reliable, as the only one determined sex with one and two band/s and peak/s on the electrophoresis and the melt curve plot for males and females, respectively. The other two primer pairs failed and depicted one band/peak in all specimens regardless of their sex. Therefore, we recommend performing Eurasian pygmy owls’ laboratory sexing by qPCR with CHD1F/CHD1R primers only.

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Section: Discussionsupporting

confidence: 53%

Comparison of three primer pairs for molecular sex determination in Eurasian pygmy owls (Glaucidium passerinum)

Stehlíková Sovadinová,

Mekadim,

Korpimäki

et al. 2024

Sci Rep

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“…Although there are other powerful approaches for sexing individuals (e.g. Griffiths et al 1998;Lois-Milevicich et al 2021), the use of sex-specific SNP markers located in unique fragments of Z-and W-chromosomes, as we propose here, will contribute to reliably sex individuals while genotyping samples for other purposes. While the approach requires reference sequences to identify and annotate sex chromosomes, this information is becoming increasingly available in non-model species through fast and affordable next generation (NGS) and whole-genome sequencing (WGS).…”

Section: Discussionmentioning

confidence: 99%

Identification of sex-linked SNP markers in wild populations of monomorphic birds

Garcia‐Raventós

Muñoz‐Mérida

Lapiedra

et al. 2023

Preprint

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Single-nucleotide polymorphism (SNP) analyses are a powerful tool for population genetics, pedigree reconstruction and phenotypic trait mapping. SNPs could also be useful for sexing individuals in species with reduced sexual dimorphism, yet this possibility remains poorly explored. Here, we develop a novel protocol for molecular sexing of birds based on the detection of unique Z- and W-linked SNP markers. Our method is based on the identification of two unique loci, one in each sexual chromosome. Individuals are considered males when they are heterozygotic for the Z-linked SNP and females when they are homozygote for the Z-linked SNP and have the W-linked SNP. We validated the method in the Jackdaw (Corvus monedula), a species whose reduced sexual dimorphism makes it difficult to sex individuals in the wild. We assessed the reliability of the method with 36 individuals of known sex, and found that their sex was correctly assigned in 100% of cases. The sex-linked markers also proved to be widely applicable to discriminate males and females from a sample of 927 genotyped individuals of different maturity stages with an accuracy of 99.5%. Given that SNP markers are increasingly used in quantitative genetic analyses of wild populations, the approach we propose has a great potential to be integrated into broader genetic research programmes without the need of additional sexing techniques.

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“…While there are other powerful approaches for sexing individuals (e.g. Griffiths et al, 1998;Lois-Milevicich et al, 2021), the use of sex-specific SNP markers located in unique fragments of Z-and W-chromosomes, as we propose here, contributes to reliably sex individuals while genotyping samples for other purposes. Although the approach requires reference sequences to identify and annotate sex chromosomes, this information is becoming increasingly available in nonmodel species through fast and affordable next-generation (NGS) and whole-genome sequencing (WGS).…”

Section: Discussionmentioning

confidence: 99%

Identification of sex‐linked SNP markers in wild populations of monomorphic birds

Garcia‐Raventós,

Muñoz‐Mérida,

Lapiedra

et al. 2023

Molecular Ecology Resources

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Single‐nucleotide polymorphism (SNP) analysis is a powerful tool for population genetics, pedigree reconstruction and phenotypic trait mapping. However, the untapped potential of SNP markers to discriminate the sex of individuals in species with reduced sexual dimorphism or of individuals during immature stages remains a largely unexplored avenue. Here, we developed a novel protocol for molecular sexing of birds based on the detection of unique Z‐ and W‐linked SNP markers. Our method is based on the identification of two unique loci, one in each sexual chromosome. Individuals are considered males when they show no calls for the W‐linked SNP and are heterozygous or homozygous for the Z‐linked SNP, while females exhibit both Z‐ and W‐linked SNP calls. We validated the method in the Jackdaw (Corvus monedula). The reduced sexual dimorphism in this species makes it difficult to identify the sex of individuals in the wild. We assessed the reliability of the method using 36 individuals of known sex and found that their sex was correctly assigned in 100% of cases. The sex‐linked markers also proved to be widely applicable for discriminating males and females from a sample of 927 genotyped individuals at different maturity stages, with an accuracy of 99.5%. Since SNP markers are increasingly used in quantitative genetic analyses of wild populations, the approach we propose has great potential to be integrated into broader genetic research programmes without the need for additional sexing techniques.

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Rapid and Low-Cost Molecular Sexing of a Corvid Songbird Using a Single Protocol with Two Universal Primer Sets

Cited by 6 publications

References 34 publications

Comparison of three primer pairs for molecular sex determination in Eurasian pygmy owls (Glaucidium passerinum)

Comparison of three primer pairs for molecular sex determination in Eurasian pygmy owls (Glaucidium passerinum)

Identification of sex-linked SNP markers in wild populations of monomorphic birds

Identification of sex‐linked SNP markers in wild populations of monomorphic birds

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