تعیین جنسیت مولکولی ماهی آزاد دریای خزر (Salmo caspius) با استفاده از جایگاه تعیین جنسیت SRY

نوع مقاله : مقاله پژوهشی

نویسنده

موسسة تحقیقات بین‌المللی تاسماهیان دریای خزر، سازمان تحقیقات آموزش و ترویج کشاورزی، رشت، ایران.

10.22059/jfisheries.2025.390403.1450

چکیده

ماهی آزاد دریای خزر از ماهیان ارزشمند و بومی این دریاچه و رودخانه‌های جنوب و شمال آن است. اهمیت تحقیق روی این گونه از جهت حفظ ذخایر آن برای مباحث حفاظت گونه‌های بومی و با ارزش دریای خزر و همچنین تکثیر، پرورش و رشد این گونه به‌عنوان گونه ماهی سردآبی با خصوصیات بازارپسندی بالا از اهمیت ویژه‌ای برخوردار است. بدین‌جهت، شناسایی جنس نر و ماده و تعیین جنسیت ژنتیکی[1] ماهی آزاد دریای خزر قبل از بلوغ و تولید‌مثل و زمانی که از نظر ریختی[2] قابل تشخیص باشد؛ مطلوب بوده و اهمیت ویژه‌ای دارد. در این تحقیق، استخراج DNA با استفاده از روش فنل-کلروفرم روی 37 نمونه از مولدین آزاد ماهی نر و ماده دریای خزر شامل 21 عدد نر و 16 عدد ماده که از نظر ریختی جنسیت آنها مشخص بود  و براساس تخمک و اسپرم‌دهی یا باز شدن حفرة شکمی و رویت گنادها انجام شد. تکثیر باند جایگاه نر با استفاده از دو گروه متفاوت از آغازگرهای دو تحقیق روی جایگاه SRY با اندازة متفاوت، با استفاده از روش زنجیره‌ای پلی‌مراز (PCR) انجام شد. در دو گروه آغازگرهای مورد بررسی نشان داد که فنوتیپ نمونه‌های مولدین نر و ماده، مطابق با ژنوتیپ به‌دست آمده براساس دو گروه آغازگرهای مورد استفاده است و همین‌طور نتایج دو گروه آغازگر به‌کار برده شده از نظر تعیین جنسیت، کاملاً بر هم انطباق دارد. این تحقیق علاوه بر دستیابی به روش بهینه برای تعیین جنسیت ژنتیکی ماهی آزاد دریای خزر، تأیید کرد که جایگاه جنس نر و سیستم تعیین جنسیت هتروژنتیک[3] در آزاد ماهی دریای خزر همانند دیگر آزادماهیان است.
 
[1]Genotypic
[2]Phenotypic
[3]XY (Hetrogenic)

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Sex determination of Caspian trout (Salmo caspius) using the SRY sex-determining locus

نویسنده [English]

  • Shirin Jamshidi
International Sturgeon Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Rasht, Iran.
چکیده [English]

The Caspian trout is one of the valuable and native fish species of Caspian Sea and its surrounding river from the north to south. Researches regarding this species are particularly important for conserving its population, as a valuable and native species of the Caspian Sea. Additionally, due to its potential for reproduction, cultivation, and fish farming as a cold-water fish species with high marketability, it holds a special role in aquacuolture in Iran. Therefore, identifying male and female individuals and determining the genetic sex of Caspian Sea trout before they reach reproductive maturity when their sex can be morphologically determined is highly desirable and of great importance. In this study, DNA extraction was performed using the phenol-chloroform method on 37 broodstock samples of male and female Caspian Sea trout, including 21 males and 16 females. The morphological sex of these individuals was confirmed based on the presence of eggs and sperm or by opening the abdominal cavity and visually identifying the gonads. Amplification of the male-specific locus was conducted using two different groups of primers based on previous studies targeting the SRY locus, each with a different fragment size, using the Polymerase Chain Reaction (PCR) method. The analysis of the amplified bands on agarose gel and scoring of the bands from both primer groups showed that the phenotypic sex of the broodstock samples matched the genotypes identified using both sets of primers. Furthermore, the results from the two primer groups were fully consistent in terms of sex determination. This study not only established an optimized method for genetic sex determination in Caspian Sea trout, but also confirmed the presence of the male-specific locus and a heterogametic sex determination system in Caspian Sea salmon, similar to other salmonid species.

کلیدواژه‌ها [English]

  • Caspian Sea
  • Male sex locus
  • Molecular
  • Sex determination
  • Salmonidae
Bertho, S., Herpin, A., Jouanno, E., Yano, A., Bobe, J., Parrinello, H., Journot, L., Guyomard, R., Muller, T., Swanson, P., McKinney, G., 2022. A nonfunctional copy of the salmonid sex-determining gene (sdy) is responsible for the “apparent” XY females in Chinook salmon, Oncorhynchus tshawytscha. G3 Genes/Genomes/Genetics, 12(2), 451. DOI: 10.1093/g3journal/jkab451
Bertho, S., Herpin, A., Schartl, M., Guiguen, Y., 2021. Lessons from an unusual vertebrate sex-determining gene. Philosophical Transactions of the Royal Society B, 376(1832), 20200092. DOI: 10.1098/rstb.2020.0092
Brown, M.S., Evans, B.S., Afonso, L.O., 2020. Discordance for genotypic sex in phenotypic female Atlantic salmon (Salmo salar) is related to a reduced sdY copy number. Scientific Reports, 10(1), 9651. DOI: 10.1038/s41598-020-66406-x
Cavileer, T.D., Hunter, S.S., Olsen, J., Wenburg, J., Nagler, J.J., 2015. A sex‐determining gene (sdY) assay shows discordance between phenotypic and genotypic sex in wild populations of Chinook Salmon. Transactions of the American Fisheries Society, 144(2), 423-430. DOI: 10.1080/00028487.2014.993479
Eisbrenner, W.D., Botwright, N., Cook, M., Davidson, E.A., Dominik, S., Elliott, N.G., Henshall, J., Jones, S.L., Kube, P.D., Lubieniecki, K.P. and Peng, S., 2014. Evidence for multiple sex-determining loci in Tasmanian Atlantic salmon (Salmo salar). Heredity, 113(1), pp.86-92.
Habibi, E., Kalbassi, M., Hosseini, S., Qasemi, S., 2013. Feasibility analysis of sex determination of caspian trout (Salmo trutta caspius, Kessler, 1877) using AFLP markers. Modern Genetics Journal 7(4). (In Persian)
Hashemzadeh Segherloo, I., Freyhof, J., Berrebi, P., Ferchaud, A.L., Geiger, M., Laroche, J., Levin, B.A., Normandeau, E., Bernatchez, L., 2021. A genomic perspective on an old question: Salmo trout or Salmo trutta (Teleostei: Salmonidae). Molecular Phylogenetics and Evolution, 162, 107204. DOI: 10.1016/j.ympev.2021.107204
Hashemzadeh Segherloo, I., Tabatabaei, S.N., Abdoli, A., Freyhof, J., Normandeau, E., Levin, B., Geiger, M.F., Laporte, M., Hallerman, E., Bernatchez, L., 2022. Biogeographic insights from a genomic survey of Salmo trout from the Aarlo-Caspian regions. Hydrobiologia, 849(19), 4325-4339. DOI: 10.1007/s10750-022-04993-8
Hattori, R.S., Murai, Y.U., Oura, M., Masuda, S., Majhi, S.K., Sakamoto, T., Sakamoto, J.I. Fernandino Somoza, G.M., Yokota, M., Strüssmann, C.A., 2012. A Y-linked anti-Mullerian hormone duplication takes over a critical role in sex-determination. Proceedings of the National Academy of Sciences of the United States of America 109(8), 2955-2959. DOI: 10.1073/pnas.1018392109
Hillis, D.M., Moritz, C., Mable, B.K., 1996. Molecular systematics (Vol. 23). Sinauer.
Ieda, R., Hosoya, S., Tajima, S., Atsumi, K., Kamiya, T., Nozawa, A., Aoki, Y., Tasumi, S., Koyama, T., Nakamura, O., Suzuki, Y., 2018. Identification of the sex-determining locus in grass puffer (Takifugu niphobles) provides evidence for sex-chromosome turnover in a subset of Takifugu species. PLoS One, 13(1), e0190635. DOI: 10.1371/journal.pone.0190635
Jouladeh-Roudbar, A., Ghanavi, H.R. and Doadrio, I., 2020. Ichthyofauna from Iranian freshwater: Annotated checklist, diagnosis, taxonomy, distribution and conservation assessment. Zoological Studies, 59. DOI: 10.6620/ZS.2020.59-21
Kamiya, T., Kai, W., Tasumi, S., Oka, A., Matsunaga, T., Mizuno, N., Fujita, M., Suetake, H., Suzuki, S., Hosoya, S., Tohari, S., 2012. A trans-species missense SNP in Amhr2 is associated with sex determination in the tiger pufferfish, Takifugu rubripes (fugu). PLoS Genetics, 8(7), e1002798. DOI: 10.1371/journal.pgen.1002798
Kijas, J., McWilliam, S., Naval Sanchez, M., Kube, P., King, H., Evans, B., Nome, T., Lien, S., Verbyla, K., 2018. Evolution of sex determination loci in Atlantic salmon. Scientific Reports, 8(1), 5664. DOI: 10.1038/s41598-018-23984-1
Kuhl, H., Guiguen, Y., Höhne, C., Kreuz, E., Du, K., Klopp, C., Lopez-Roques, C., Yebra-Pimentel, E.S., Ciorpac, M., Gessner, J., Holostenco, D., 2021. A 180 Myr-old female-specific genome region in sturgeon reveals the oldest known vertebrate sex determining system with undifferentiated sex chromosomes. Philosophical Transactions of the Royal Society B, 376(1832), 20200089. DOI: 10.1098/rstb.2020.0089
Li, J., Phillips, R.B., Harwood, A.S., Koop, B.F., Davidson, W.S., 2011. Identification of the sex chromosomes of brown trout (Salmo trutta) and their comparison with the corresponding chromosomes in Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss). Cytogenetic and Genome Research, 133(1), 25-33. DOI: 10.1159/000323410
Lubieniecki, K.P., Lin, S., Cabana, E.I., Li, J., Lai, Y.Y., Davidson, W.S., 2015. Genomic instability of the sex-determining locus in Atlantic salmon (Salmo salar). G3: Genes, Genomes, Genetics, 22, 2513-22. DOI: 10.1534/g3.115.020115
Makhrov, A.A., Artamonova, V.S., Kolmakova, O.V., Ponomareva, M.V., 2018, January. Sex determination model in pink salmon Oncorhynchus gorbuscha (Walbaum, 1792) (Salmonidae, Osteichthyes) controlled by multi-copy genes located in sex chromosomes. In Doklady Biochemistry and Biophysics (Vol. 478, No. 1, pp. 21-24). Moscow: Pleiades Publishing.
Martínez, P., Viñas, A.M., Sánchez, L., Díaz, N., Ribas, L., Piferrer, F., 2014. Genetic architecture of sex determination in fish: applications to sex ratio control in aquaculture. Frontiers in genetics, 5, p.340. DOI: 10.3389/fgene.2014.00340/full
Matsuda, M., Nagahama, Y., Shinomiya, A., Sato, T., Matsuda, C., Kobayashi, T., Morrey, C.E., Shibata, N., Asakawa, S., Shimizu, N., Hori, H., 2002. DMY is a Y-specific DM-domain gene required for male development in the medaka fish. Nature, 417(6888), 559-563. DOI: 10.1038/nature751
Matsuda, M., Shinomiya, A., Kinoshita, M., Suzuki, A., Kobayashi, T., Paul-Prasanth, B., Lau, E.L., Hamaguchi, S., Sakaizumi, M., Nagahama, Y., 2007. DMY gene induces male development in genetically female (XX) medaka fish. Proceedings of the National Academy of Sciences, 104(10), 3865-3870. DOI: 10.1073/pnas.0611707104
Myosho, T., Otake, H., Masuyama, H., Matsuda, M., Kuroki, Y., Fujiyama, A., Naruse, K., Hamaguchi, S. and Sakaizumi, M., 2012. Tracing the emergence of a novel sex-determining gene in medaka, Oryzias luzonensis. Genetics, 191(1), 163-170. DOI: 10.1534/genetics.111.137497
Ninua, L., Tarkhnishvili, D., Gvazava, E., 2018. Phylogeography and taxonomic status of trout and salmon from the Ponto‐Caspian drainages, with inferences on European Brown Trout evolution and taxonomy. Ecology and Evolution, 8(5), 2645-2658. DOI: 10.1002/ece3.3884
Quéméré, E., Perrier, C., Besnard, A.L., Evanno, G., Baglinière, J.L., Guiguen, Y., Launey, S., 2014. An improved PCR-based method for faster sex determination in brown trout (Salmo trutta) and Atlantic salmon (Salmo salar). Conservation Genetics Resources, 6, 825-827. DOI: 10.1007/s12686-014-0259-8
Salehi, M., Mousavi-Sabet, H., Eagderi, S., 2022. The Iranian native salmonids (Salmo caspius and S. trutta), distinct species or two forms of the same species; a geometric-morphometric based hypothesis. Aquaculture Sciences, 10(2), 25-34. (In Persian)
Schartl, M., 2004. A comparative view on sex determination in medaka. Mechanisms of Development, 121(7-8), 639-645. DOI: 10.1016/j.mod.2004.03.001
Shahhosseini, Gh., 2020. Meiotic gynogenesis inducing in Caspian Sea salmon (Salmo trutta caspius) using gamma irradiation followed by sex determination using SDY gene. Journal of Nuclear Science, Engineering and Technology (JONSAT), 41(3), 145-154. (In Persian)
Sutherland, B.J., Rico, C., Audet, C., Bernatchez, L., 2017. Sex chromosome evolution, heterochiasmy, and physiological QTL in the salmonid brook charr Salvelinus fontinalis. G3: Genes, Genomes, Genetics, 7(8), 2749-2762. DOI: 10.1534/g3.117.040915
Woram, R.A., Gharbi, K., Sakamoto, T., Hoyheim, B., Holm, L.E., Naish, K., McGowan, C., Ferguson, M.M., Phillips, R.B., Stein, J., Guyomard, R., 2003. Comparative genome analysis of the primary sex-determining locus in salmonid fishes. Genome Research, 13(2), 272-280.
Yano, A., Nicol, B., Jouanno, E., Quillet, E., Fostier, A., Guyomard, R., Guiguen, Y., 2013. The sexually dimorphic on the Y‐chromosome gene (sdY) is a conserved male‐specific Y‐chromosome sequence in many salmonids. Evolutionary Applications, 6(3), 486-496. DOI: 10.1111/eva.12032