Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo)

Yuda, Pramana; Saputra, Andie Wijaya

doi:10.12688/f1000research.23712.4

Cited by 6 publications

(5 citation statements)

References 26 publications

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“…New primer set was designed manually based on the published CHD1-Z and CHD1-W sequences (Acc. No: MT074328.1 and Acc.No: MT074330.1) of Macrocephalon maleo (Yuda and Saputra 2020). Those sequences were a result of the amplification of CHD1 using primer set 2550F/2718R developed by Fridolfsson and Ellegren (1999).…”

Section: Primer Designmentioning

confidence: 99%

“…In addition, P2/P8 primer set needs post-PCR digestion with a restriction enzyme such as BamHI or HaeIII to obtain distinguishing fragments CHD1-Z and CHD1-W (Romanoev et al 2019). Using a primer set 2550F/2817R, Yuda and Saputra (2020) identified the sexes of Maleo Senkawor (Macrocephalon maleo) chicks. However, the primer set was only successfully identified the sexes of 63.7% of 162 chicks.…”

Section: Introductionmentioning

confidence: 99%

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Short Communication: A new primer set in CHD1 gene for bird sex identification

et al. 2021

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Abstract. Vera F, Wajjwalku W, Yuda P, Daryono BS. 2021. Short Communication: A new primer set in CHD1 gene for bird sex identification. Biodiversitas 22: 4977-4982. Determine sex is difficult for many bird species that are sexually monomorphic or only dimorphic in the adult stage. Many molecular markers have been developed for DNA sexing, which were mostly based on identifying Z and W chromosomes of avians. The sex determination of birds was mostly applied universal primer set such as P2/P8 or 2550F/2718R. However, those universal primers that were designed sometimes could not good result consistency in non-ratite birds or ratite birds. Therefore, we improved a specific primer design with deletion site in W chromosome to amplify the female-specific segments on Chromodomain-helicase DNA binding protein-1 (CHD1) gene from alignment sequences CHD1-Z and CHD1-W of Macrocephalon maleo S. Müller, 1846. This study aims to design a new pair of primer targeting a section of the CHD1 gene that can be amplified in non-ratite birds, the name of a new primer is In-Sex F/In-Sex R. A new primer set amplified DNA fragments in around 650 or 550 base pairs of CHD1-Z and also, 350 base pairs of CHD1-W. The result has successfully amplified the sex of multiple species on orders Galliformes, Passeriformes, Accipitriformes, and Strigiformes. This analysis can be helpful to the effort of in-situ or ex-situ management and conservation programs e.g the mating system in birds. And also, the analysis can be helpful for sexing data in the case of captive birds before releasing in natural habitat or reintroduction programs.

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Section: Primer Designmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Short Communication: A new primer set in CHD1 gene for bird sex identification

et al. 2021

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“…Birds sexing using the Polymerase Chain Reaction (PCR) technique is increasingly being used by researchers and breeders for breeding and sustainable conservation efforts. The PCR technique has high sensitivity, can be performed at all ages from juvenile to adults, and even could identify the sexes using the eggshell membrane as a DNA sample (Begović et al, 2017;Akrom et al, 2020;Yuda and Saputra, 2021;Turcu et al, 2023). PCR technique has advantages over conventional methods such as external morphological observation, vent sexing, laparoscopy, karyotyping, faecal sexing steroids, and ultrasound for oviduct identification (Khaerunnisa et al, 2013;Casana et al, 2019;Turcu et al, 2020;Hidayat et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Appropriate Primer Selection Improves Molecular Bird Sexing Accuracy

Fitriana,

Al Qodry,

De Lucas

et al. 2023

BuletinPeternak

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Birds sexing utilize the Polymerase Chain Reaction (PCR) technique is increasingly being used by researchers and breeders. The PCR technique has high sensitivity, but its success is influenced by the specificity of the DNA template with the oligo primer used. This study aimed to evaluate 5 types of PCR primers P2/P8, 2550F/2718R, CHD1F/CHD1R, 1237L/1272H, and CHD1LF/CHD1LR to determine the sex of Phasianidae, Anatidae, Muscicapidae, and Psittacidae families. This research was conducted by tested primers mentioned above to amplify the target gene chromodomain helicase DNA binding 1 (CHD1) on DNA samples of each pair of males and females from four bird families, respectively. The results indicated that CHD1LF/CHD1LR PCR primer gave the best results and was recommended to determine the sex of four families tested. Some of other primers tested in this study failed to amplify targeted gene correctly, it is important to use appropriate primer to increase bird sexing accuracy.

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“…Molecular tests based on DNA characteristics unique for each sex, especially those using PCR (Polymerase Chain Reaction) technology, have proven their reliability and simplicity. Additionally, they are inexpensive, fast, and have the advantage that DNA can be extracted from readily available samples such as feathers, buccal cells, fecal matter, and post-hatched egg-shell membrane [ 14 , 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

The Length Polymorphism of the 9th Intron in the Avian CHD1 Gene Allows Sex Determination in Some Species of Palaeognathae

Kroczak

Wierzbicki

Urantówka

2022

Genes

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In palaeognathous birds, several PCR-based methods and a range of genes and unknown genomic regions have been studied for the determination of sex. Many of these methods have proven to be unreliable, complex, expensive, and time-consuming. Even the most widely used PCR markers for sex typing in birds, the selected introns of the highly conserved CHD1 gene (primers P2/P8, 1237L/1272H, and 2550F/2718R), have rarely been effective in palaeognathous birds. In this study we used eight species of Palaeognathae to test three PCR markers: CHD1i9 (CHD1 gene intron 9) and NIPBLi16 (NIPBL gene intron 16) that performed properly as Psittaciformes sex differentiation markers, but have not yet been tested in Palaeognathae, as well as the CHD1iA intron (CHD1 gene intron 16), which so far has not been used effectively to sex palaeognathous birds. The results of our research indicate that the CHD1i9 marker effectively differentiates sex in four of the eight species we studied. In Rhea americana, Eudromia elegans, and Tinamus solitarius, the electrophoretic patterns of the amplicons obtained clearly indicate the sex of tested individuals, whereas in Crypturellus tataupa, sexing is possible based on poorly visible female specific bands. Additionally, we present and discuss the results of our in silico investigation on the applicability of CHD1i9 to sex other Palaeognathae that were not tested in this study.

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Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo)

Cited by 6 publications

References 26 publications

Short Communication: A new primer set in CHD1 gene for bird sex identification

Short Communication: A new primer set in CHD1 gene for bird sex identification

Appropriate Primer Selection Improves Molecular Bird Sexing Accuracy

The Length Polymorphism of the 9th Intron in the Avian CHD1 Gene Allows Sex Determination in Some Species of Palaeognathae

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