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Research Article

volume 53 issue 12 (december 2019) : 1577-1582,   Doi: 10.18805/ijar.B-951

Molecular identification of sex in the monomorphic breed of pigeons

J
J. Gruszczyńska
A
A. Alama
M
M. Miąsko
P
P. Florczuk - Kołomyja
B
B. Grzegrzółka
1Department of Genetics and Animal Breeding, Faculty of Animal Sciences, Warsaw University of Life Sciences-SGGW, ul. Ciszewskiego 8, 02-786 Warszawa, Poland.
Cite article:- Gruszczyńska J., Alama A., Miąsko M., Kołomyja - Florczuk P., Grzegrzółka B. (2019). Molecular identification of sex in the monomorphic breed of pigeons. Indian Journal of Animal Research. 53(12): 1577-1582. doi: 10.18805/ijar.B-951.

ABSTRACT

In many avian species, especially in monomorphic species and breeds, sex identification creates a serious problem, as they do not show any phenotypic differences. One of such breeds is the Wroclaw Meat Pigeon. In this study, molecular identification of sex with P2 and P8 primers used for the CHD1 (chromo-helicase-DNA-binding-protein) gene amplification was performed. Peripheral blood samples were analyzed from 46 birds, and their DNA was isolated with the phenol-chloroform method. The fragments (370 bp CHD1-Z; 350 bp CHD1-W) obtained from the PCR were cut with the BsuRI. Only the sequence in the Z chromosome was cut into fragments of 305 and 65 bp by the restriction enzyme. The difference between CHD1-Z and CHD1-W was visualized in 3% agarose gel. A single band was identified as male, whereas two bands (plus 1 invisible) were identified as female. Consequently, 23 specimens in each sex were identified. 

REFERENCES

  1. Bermúdez-Humarán, L.G., Chávez-Zamarripa, P., Guzmán-Velasco, A., Leal-Garza, C.H., Montes de Oca-Luna, R. (2002). Loss of restriction site Ddei, used for avian molecular sexing in oreophasis derbianus. Reprod. Domest. Anim., 37: 321-323.
  2. Cerit, H., Avanus, K. (2007). Sex determination by CHDW and CHDZ genes of Avian Sex Chromosomes In Nymphicus hollandicus. Turkish. J. Vet. Anim. Sci., 31: 371-374.
  3. Dash, S., Kumarasamy, P., Malik, H., Thiagarajan, V. (2013). Phylogenetic analysis of ratite bird emu (Dromaius novaehollandiae) using sex specific sequences. Indian J. Anim. Res., 47: 247-250.
  4. Dubiec, A., Zagalska-Neubauer, M. (2006). Molecular techniques for sex identification in birds. Biological Lett., 43(1): 3-12.
  5. Fridolfsson, A.K., Ellegren, H. (1999). A Simple and Universal method for molecular sexing of non-ratite birds. J. Avian Biol., 30: 116-121.
  6. Griffiths, R., Daan, S., Dijkstra, C. (1996). Sex identification in birds using two CHD genes. Proc. R. Soc. Lond., B, Biol. Sci., 263: 1249-1254.
  7. Griffiths, R., Double, M., Orr, K., Dawson, R. (1998). A DNA test to sex most birds. Mol. Ecol., 7: 1071-1075. 
  8. Griffiths, R., Tiwari, B. (1993). The isolation of molecular genetic markers for the identification of sex. Proc. Natl. Acad. Sci. U.S.A., 90: 8324-8326.
  9. Griffiths, R., Tiwari, B. (1995). Sex of the last Wild Spix’s macaw. Nature, 375: 454.
  10. Grzegrzó³ka, B., Gruszczyñska, J., Domaga³a, E., S³onecki, R., Bartyzel, B.J., Czerniawska-Pi¹tkowska, E. (2016). Morphological assessment of sexual dimorphism abnormality cases in Japanese quail (Coturnix japonica). Reprod. Domest. Anim., 15 Suppl. 2: 96.
  11. Jaiswal, N.R., Gottigalla, B.Y. (2016). Sex identification in pigeons using CHD based molecular markers. PARIPEX-Indian J. Res., 5: 209-210.
  12. Jensen, T., Pernasetti, F.M., Durrant, B. (2003). Conditions for rapid sex determination in 47 avian species by PCR of Genomic DNA From Blood, Shell-Membrane Blood Vessels, and Feathers. Zoo Biol., 22: 561-571.
  13. Journal of Laws, (2015). Ustawa z dnia 15 stycznia 2015 r. o ochronie zwierz¹t wykorzystywanych do celów naukowych lub edukacyjnych [In Polish] Dz. U. 2015, item 266, 1-25.
  14. Kahn, N.W., John, J.S., Quinn, T.W. (1998). Chromosome-specific intron size differences In the avian CHD gene pro vide an efficient metod for sex identification in birds. Auk, 115(4): 1074-1078.
  15. Khaerunnisa, I., Sari, E., Ulfah, M., Sumantri, J.C. (2013). Avian sex determination based on Chromo Helicase DNA-binding (CHD) genes using polymerase chain reaction (PCR). Media Peternakan, 36(2): 85-90.
  16. Longmire, J.L., Maltbie, M., Pavelka, R.W., Smith, L.M., Witte, S.M., Ryder, O.A., Ellsworth, D.L., Baker, R.J. (1993). Gender identification in birds using microsatellite DNA fingerprint analysis. Auk, 110(2): 378-381.
  17. Maron, J.L., Myers, J.P. (1984). A description and evaluation of two techniques for sexing wintering sanderlings. J. Field Ornithol., 55(3): 336-342.
  18. Morinha, F., Cabral, J.A., Bastos, E. (2012). Molecular sexing of birds: A comparative rewiev of polymerase chain re action (PCR)-based methods. Theriogenology, 78: 703-714. 
  19. Pawlina, E. (2011). The Wroclaw Meat Pigeon bred at the Wroc³aw University of Life Siences. Przegl¹d Hodowlany, 10: 28-30. 
  20. Richner, H. (1989). Avian laparoscopy as a field technique for sexing birds and an assessment of its effects on wild birds. J. Field Ornithol., 60(2): 137-142.
  21. Sacchi, P., Soglia, D., Maione, S., Meneguz, G., Campora, M., Rasero, R. (2004). A non-invasive test for sex identification in short-    toed eagle (Circaetus gallicus). Mol. Cell. Probes., 18: 193-196.
  22. Singh, J., Mukhopadhyay, C.S., Brah, G.S., Saini, S. (2014). Z-chromosome linked dinucleotide STRs: association with reproduction traits in chicken. Indian J. Anim. Res., 48(6): 521-526. 
  23. Smith, C.A., Roeszler, K.N., Hudson, Q.J., Sinclair, A.H. (2007). Avian sex determination: what, when and where? Cytogenet. Genome. Res., 117: 165-173. 
  24. Thanou, E., Giokas, S., Goutner, V., Liordos, V., Fraguedakis-Tsolis, S. (2013). Efficiency and accuracy of PCR-based sex determination methods in the European Phalacrocoracidae. Ann. Zool. Fennici, 50(1-2): 52-63. 
  25. Väli, Ü., Elts, J. (2002). Molecular sexing of Eurasian Woodcock Scolopax rusticola. WSGB, 98: 48.
  26. Volodin, I., Kaiser, M., Matrosova, E.V., Klenova, A., Filatova, O., Kholodova, M. (2009). The technique of noninvasive distant sexing for four monomorphic Dendrocygna whistlings duck species by their loud Whistler. Bioacoustics, 18: 277-290.
  27. Vucicevic, 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: 269-276.
  28. Wu, Ch., Horng, Y., Wang, R., Yang, K., Huang, M. (2007). A novel sex-specific DNA marker in Columbidae birds. Theriogenology, 67: 328-333.
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 12 (december 2019) : 1577-1582,   Doi: 10.18805/ijar.B-951

    Molecular identification of sex in the monomorphic breed of pigeons

    J
    J. Gruszczyńska
    A
    A. Alama
    M
    M. Miąsko
    P
    P. Florczuk - Kołomyja
    B
    B. Grzegrzółka
    1Department of Genetics and Animal Breeding, Faculty of Animal Sciences, Warsaw University of Life Sciences-SGGW, ul. Ciszewskiego 8, 02-786 Warszawa, Poland.
    Cite article:- Gruszczyńska J., Alama A., Miąsko M., Kołomyja - Florczuk P., Grzegrzółka B. (2019). Molecular identification of sex in the monomorphic breed of pigeons. Indian Journal of Animal Research. 53(12): 1577-1582. doi: 10.18805/ijar.B-951.

    ABSTRACT

    In many avian species, especially in monomorphic species and breeds, sex identification creates a serious problem, as they do not show any phenotypic differences. One of such breeds is the Wroclaw Meat Pigeon. In this study, molecular identification of sex with P2 and P8 primers used for the CHD1 (chromo-helicase-DNA-binding-protein) gene amplification was performed. Peripheral blood samples were analyzed from 46 birds, and their DNA was isolated with the phenol-chloroform method. The fragments (370 bp CHD1-Z; 350 bp CHD1-W) obtained from the PCR were cut with the BsuRI. Only the sequence in the Z chromosome was cut into fragments of 305 and 65 bp by the restriction enzyme. The difference between CHD1-Z and CHD1-W was visualized in 3% agarose gel. A single band was identified as male, whereas two bands (plus 1 invisible) were identified as female. Consequently, 23 specimens in each sex were identified. 

    REFERENCES

    1. Bermúdez-Humarán, L.G., Chávez-Zamarripa, P., Guzmán-Velasco, A., Leal-Garza, C.H., Montes de Oca-Luna, R. (2002). Loss of restriction site Ddei, used for avian molecular sexing in oreophasis derbianus. Reprod. Domest. Anim., 37: 321-323.
    2. Cerit, H., Avanus, K. (2007). Sex determination by CHDW and CHDZ genes of Avian Sex Chromosomes In Nymphicus hollandicus. Turkish. J. Vet. Anim. Sci., 31: 371-374.
    3. Dash, S., Kumarasamy, P., Malik, H., Thiagarajan, V. (2013). Phylogenetic analysis of ratite bird emu (Dromaius novaehollandiae) using sex specific sequences. Indian J. Anim. Res., 47: 247-250.
    4. Dubiec, A., Zagalska-Neubauer, M. (2006). Molecular techniques for sex identification in birds. Biological Lett., 43(1): 3-12.
    5. Fridolfsson, A.K., Ellegren, H. (1999). A Simple and Universal method for molecular sexing of non-ratite birds. J. Avian Biol., 30: 116-121.
    6. Griffiths, R., Daan, S., Dijkstra, C. (1996). Sex identification in birds using two CHD genes. Proc. R. Soc. Lond., B, Biol. Sci., 263: 1249-1254.
    7. Griffiths, R., Double, M., Orr, K., Dawson, R. (1998). A DNA test to sex most birds. Mol. Ecol., 7: 1071-1075. 
    8. Griffiths, R., Tiwari, B. (1993). The isolation of molecular genetic markers for the identification of sex. Proc. Natl. Acad. Sci. U.S.A., 90: 8324-8326.
    9. Griffiths, R., Tiwari, B. (1995). Sex of the last Wild Spix’s macaw. Nature, 375: 454.
    10. Grzegrzó³ka, B., Gruszczyñska, J., Domaga³a, E., S³onecki, R., Bartyzel, B.J., Czerniawska-Pi¹tkowska, E. (2016). Morphological assessment of sexual dimorphism abnormality cases in Japanese quail (Coturnix japonica). Reprod. Domest. Anim., 15 Suppl. 2: 96.
    11. Jaiswal, N.R., Gottigalla, B.Y. (2016). Sex identification in pigeons using CHD based molecular markers. PARIPEX-Indian J. Res., 5: 209-210.
    12. Jensen, T., Pernasetti, F.M., Durrant, B. (2003). Conditions for rapid sex determination in 47 avian species by PCR of Genomic DNA From Blood, Shell-Membrane Blood Vessels, and Feathers. Zoo Biol., 22: 561-571.
    13. Journal of Laws, (2015). Ustawa z dnia 15 stycznia 2015 r. o ochronie zwierz¹t wykorzystywanych do celów naukowych lub edukacyjnych [In Polish] Dz. U. 2015, item 266, 1-25.
    14. Kahn, N.W., John, J.S., Quinn, T.W. (1998). Chromosome-specific intron size differences In the avian CHD gene pro vide an efficient metod for sex identification in birds. Auk, 115(4): 1074-1078.
    15. Khaerunnisa, I., Sari, E., Ulfah, M., Sumantri, J.C. (2013). Avian sex determination based on Chromo Helicase DNA-binding (CHD) genes using polymerase chain reaction (PCR). Media Peternakan, 36(2): 85-90.
    16. Longmire, J.L., Maltbie, M., Pavelka, R.W., Smith, L.M., Witte, S.M., Ryder, O.A., Ellsworth, D.L., Baker, R.J. (1993). Gender identification in birds using microsatellite DNA fingerprint analysis. Auk, 110(2): 378-381.
    17. Maron, J.L., Myers, J.P. (1984). A description and evaluation of two techniques for sexing wintering sanderlings. J. Field Ornithol., 55(3): 336-342.
    18. Morinha, F., Cabral, J.A., Bastos, E. (2012). Molecular sexing of birds: A comparative rewiev of polymerase chain re action (PCR)-based methods. Theriogenology, 78: 703-714. 
    19. Pawlina, E. (2011). The Wroclaw Meat Pigeon bred at the Wroc³aw University of Life Siences. Przegl¹d Hodowlany, 10: 28-30. 
    20. Richner, H. (1989). Avian laparoscopy as a field technique for sexing birds and an assessment of its effects on wild birds. J. Field Ornithol., 60(2): 137-142.
    21. Sacchi, P., Soglia, D., Maione, S., Meneguz, G., Campora, M., Rasero, R. (2004). A non-invasive test for sex identification in short-    toed eagle (Circaetus gallicus). Mol. Cell. Probes., 18: 193-196.
    22. Singh, J., Mukhopadhyay, C.S., Brah, G.S., Saini, S. (2014). Z-chromosome linked dinucleotide STRs: association with reproduction traits in chicken. Indian J. Anim. Res., 48(6): 521-526. 
    23. Smith, C.A., Roeszler, K.N., Hudson, Q.J., Sinclair, A.H. (2007). Avian sex determination: what, when and where? Cytogenet. Genome. Res., 117: 165-173. 
    24. Thanou, E., Giokas, S., Goutner, V., Liordos, V., Fraguedakis-Tsolis, S. (2013). Efficiency and accuracy of PCR-based sex determination methods in the European Phalacrocoracidae. Ann. Zool. Fennici, 50(1-2): 52-63. 
    25. Väli, Ü., Elts, J. (2002). Molecular sexing of Eurasian Woodcock Scolopax rusticola. WSGB, 98: 48.
    26. Volodin, I., Kaiser, M., Matrosova, E.V., Klenova, A., Filatova, O., Kholodova, M. (2009). The technique of noninvasive distant sexing for four monomorphic Dendrocygna whistlings duck species by their loud Whistler. Bioacoustics, 18: 277-290.
    27. Vucicevic, 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: 269-276.
    28. Wu, Ch., Horng, Y., Wang, R., Yang, K., Huang, M. (2007). A novel sex-specific DNA marker in Columbidae birds. Theriogenology, 67: 328-333.
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