Evaluation of molecular primers for sexing the magpie robin and green cucak via CHD1 gene amplification
DOI:
https://doi.org/10.46549/jipvet.v15i2.557Keywords:
Accuracy, Bird, PCR sexing, Primer, SelectionAbstract
Researchers and breeders are increasingly using the Polymerase Chain Reaction (PCR) method for bird sexing. However, the compatibility of the DNA template and the oligo primer used is the primary key to the success of PCR amplification. In the present study, we evaluated five types of popular PCR primers sexing to determine the sex of a pair of each the magpie robin (Copsychus malabaricus) and green cucak (Chloropsis sonnerati). We used DNA samples from each pair of males and females from the two species above, respectively, to test the five primers listed to amplify the target gene chromodomain helicase DNA binding 1 (CHD1). The results demonstrated that the primer sets of CHD1LF/CHD1LR, CHD1F/CHD1R, and P2/P8 could be used for Copsychus malabaricus, while only CHD1LF/CHD1LR was suitable for Chloropsis sonnerati. It is crucial to select the correct primer to improve bird sexing accuracy because several of the other primers examined in this study were unable to amplify the targeted CHD1 gene correctly.
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Adisti AA, Zulfikhar R. 2021. Mengenal berbagai macam burung di Indonesia. Repository Jurnal Polbangtan Yoma. 2(1):1–36.
Akrom AM, Indarjulianto S, Yanuartono Y, Susmiati T, Nururrozi A, Raharjo S, Permana RGS, Sitompul YY. 2020. Swab bukal sebagai bahan sexing piyikan burung kenari (Serinus canaria) dan burung merpati (Columba livia). Jurnal Sain Veteriner. 38(1):31–36. https://doi.org/10.22146/jsv.49032
Davitkov D, Vučićević M, Glavinić U, Skadric I, Nešić V, Stevanović J, Stanimirović Z. 2021. Potential of inter and intra species variability of CHD1 gene in birds as a forensic tool. Acta Veterinaria Beograd. 71(2):147–157. https://doi.org/10.2478/acve 2021 0013
Dobreva MP, Lynton Jenkins JG, Chaves JA, Tokita M, Bonneaud C, Abzhanov A. 2021. Sex identification in embryos and adults of Darwin’s finches. PLoS One. 16(3):e0237687. https://doi.org/10.1371/journal.pone.0237687
Elnomrosy SM, Hagag NM, AbdAllah MI, Kolenda R, Zacharski M. 2022. Application of Loop Mediated Isothermal Amplification (LAMP) in sex identification of parrots bred in Egypt. Biology. 11(4):565. https://doi.org/10.3390/biology11040565
Fitriana F, Resita R, Disastra Y, Setyorini DR, Haryanto A, Aziz F. 2023a. Evaluation of primers targeting chromo helicase DNA binding gene (CHD) for molecular sexing identification in four bird families. Livest Anim Res. 21(1):14–20. https://doi.org/10.20961/lar.v21i1.66998
Fitriana F, Setyorini DR, Artdita CA, Ummami R, Haryanto A, Aziz F. 2023b. Comparison of four types molecular sexing primers in different bird families. Journal of Tropical Animal and Veterinary Science. 13(1):51–57. https://doi.org/10.46549/jipvet.v13i1.359
Fridolfsson A K, Ellegren H. 1999. A simple and universal method for molecular sexing of non ratite birds. J Avian Biol. 30(1):116–121. https://doi.org/10.2307/3677252
Gebhardt KJ, Waits LP. 2008. High error rates for avian molecular sex identification primer sets applied to molted feathers. J Field Ornithol. 79(3):286–292. https://doi.org/10.1111/j.1557 9263.2008.00175.x
Green MR, Sambrook J. 2019. Polymerase Chain Reaction. CSH Protoc. 2019(6):pdb.top095109. https://doi.org/10.1101/pdb.top095109
Griffiths R, Double MC, Orr K, Dawson RJG. 1998. A DNA test to sex most birds. Molecular Ecology. 7(8):1071–1075. https://doi.org/10.1046/j.1365 294x.1998.00389.x
Iskandar J. 2014. Dilema antara hobi dan bisnis perdagangan burung serta konservasi burung. Chimica et Natura Acta. 2(3):180–185. https://doi.org/10.24198/cna.v2.n3.9165
Khan NW, St John J, Quinn TW. 1998. Chromosome specific intron size differences in the avian CHD gene provide an efficient method for sex identification in birds. The Auk. 115(4):1074–1078. https://doi.org/10.2307/4089527
Kroczak A, Wierzbicki H, Urantówka AD. 2022. The length polymorphism of the 9th intron in the avian CHD1 gene allows sex determination in some species of Palaeognathae. Genes. 13(3):507. https://doi.org/10.3390/genes13030507
Kroczak A, Wołoszyńska M, Wierzbicki H, Kurkowski M, Grabowski KA, Piasecki T, Galosi L, Urantówka AD. 2021. New bird sexing strategy developed in the order Psittaciformes involves multiple markers to avoid sex misidentification: debunked myth of the universal DNA marker. Genes. 12(6):878. https://doi.org/10.3390/genes12060878
Lee JCI, Tsai LCT, Hwa PY, Chan CL, Huang A, Chin SC, Wang LC, Lin JT, Linacre A, Hsieh HM. 2010. A novel strategy for avian species and gender identification using the CHD gene. Mol Cell Probes. 24(1):27–31. https://doi.org/10.1016/j.mcp.2009.08.003
Liang S, Chen M, Gao C, Yan H, Zhang G, Wang X. 2019. Sex identification of pigeons using polymerase chain reaction analysis with simple DNA extraction. Avian Biology Research. 12(2):45–48. https://doi.org/10.1177/1758155919832141
Molle MNV, Maker UP, Kilmaskossu A. 2019. Jenis jenis burung kicau dan aspek konservasinya pada komunitas kicau mania Manokwari. Vogelkop: Jurnal Biologi. 2(1):21–27. https://doi.org/10.30862/vogelkopjbio.v2i1.43
Pamulang YV, Haryanto A. 2021. Short communication: molecular bird sexing on kutilang (Pycnonotus sp.) based on amplification of CHD Z and CHD W genes by using PCR method. Biodiversitas. 22(1):449–452. https://doi.org/10.13057/biodiv/d220155
Purwaningrum M, Nugroho HA, Asvan M, Karyanti K, Alviyanto B, Kusuma R, Haryanto A. 2019. Molecular techniques for sex identification of captive birds. Vet World. 12(9):1506–1513. https://doi.org/10.14202/vetworld.2019.1506 1513
Putranto HD, Okvianto D, Prakoso H. 2018. Studi reproduksi burung murai batu (Copsychus malabaricus) pada penangkaran lokal di Kota Bengkulu. Jurnal Sain Peternakan Indonesia. 13(2):130–139.
Ramos CN, Barros TB, de Antonio ES, Mirabal B, da Silva MB, Fraga RE, Tomazi L. 2021. Sexing as a tool for reinsertion of Amazona aestiva parrots to nature: use of less invasive technique. Research Society and Development. 10(12):e380101220601. https://doi.org/10.33448/rsd v10i12.20601
Savitri D, Putri I, Nugrahani WP, Purwaningrum M, Haryanto A. 2021. Molecular bird sexing of sulphur crested cockatoo (Cacatua galerita) by PCR method. Indonesian Journal of Biotechnology. 26(1):1–6. https://doi.org/10.22146/ijbiotech.54611
Setiadi A. 2023. Konservasi keanekaragaman hayati endemik melalui ecology, socio economic, dan socio cultural approach (studi pada Taman Kehati Kokolomboi, Sulawesi Tengah). Learning Society: Jurnal CSR, Pendidikan dan Pemberdayaan Masyarakat. 4(1):244–254. https://doi.org/10.30872/ls.v4i1.2313
Turcu MC, Bel LV, Collarile T, Pusta DL. 2020. Comparative evaluation of two techniques of sex determination in lovebirds (Agapornis spp.). Bull Univ Agric Sci Vet Med Cluj Napoca. 77(2):106–114. https://doi.org/10.15835/buasvmcn vm:2020.0030
van der Velde M, Haddrath O, Verkuil YI, Baker AJ, Piersma T. 2017. New primers for molecular sex identification of waders. Wader Study. 124(2):147–151. https://doi.org/10.18194/ws.00069
Vučićević M, Stevanov Pavlovic M, Stevanovic J, Bosnjak J, Gajic B, Aleksic N, Stanimirovic Z. 2013. Sex determination in 58 bird species and evaluation of CHD gene as a universal molecular marker in bird sexing. Zoo Biol. 32(3):269–276. https://doi.org/10.1002/zoo.21010
Yuda P, Saputra AW. 2021. Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo). F1000Res. 9:599. https://doi.org/10.12688/f1000research.23712.4
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