شناسایی مورفولوژیک و مولکولی گونه غالب خیار دریایی در منطقه اولی (خلیج فارس-استان بوشهر) و آنالیز برخی ترکیبات دیواره بدن آن

نوع مقاله : علوم جانوری

نویسندگان

1 گروه زیست شناسی دریا، دانشکده علوم دریایی و اقیانوسی، دانشگاه علوم و فنون دریایی خرمشهر، خرمشهر، ایران

2 پژوهشکده زیست فناوری دریا، مرکز تحقیقات زیست پزشکی شهدای خلیج فارس، دانشگاه علوم پزشکی بوشهر، بوشهر، ایران

3 مرکز تحقیقات آبزیان خلیج فارس، دانشگاه خلیج فارس بوشهر، بوشهر، ایران

چکیده

هدف از این پژوهش، شناسایی یکی از گونه­ های غالب خیار دریایی در منطقه اولی- بوشهر ، با استفاده از روش مورفولوژیک و مولکولی و بررسی برخی ترکیبات تشکیل ­دهنده دیواره بدن ازجمله پروتئین کلاژن بوده است. تا علاوه بر شناسایی دقیق گونه، با مطالعه ترکیبات دیواره بدن آن بتوان، از یافته­ های این تحقیق در جهت ارتقاء صنایع غذایی و دارویی در کشور استفاده نمود. در این راستا، ابتدا با شناسایی مورفولوژیک از طریق بررسی اوسیکل­ های دیواره بدن و سپس با شناسایی مولکولی از طریق تعیین توالی بخشی از ژنوم میتوکندریایی 16SrRNA، گونه مورد بررسی به ­عنوان گونه Holothuria leucospilota شناسایی گردید. در بررسی ترکیبات دیواره بدن، پروتئین و رطوبت با مقادیر بالای 70% بیش ­ترین ترکیبات تشکیل ­دهنده بدن گونه مورد نظر بود و چربی کم ­ترین مقدار را در بین سایر ترکیبات داشته است. از بین ترکیبات پروتئینی، استخراج پروتئین کلاژن به ­روش هضم آنزیمی با پپسین صورت گرفت و با استفاده از آزمون SDS-PAGE، نوع آن و زیر واحدهای تشکیل ­دهنده آن با توجه به پروفایل وزن مولکولی، در مقایسه با سایر تحقیقات، صورت گرفت. روش استخراج موثر تشخیص داده شد و کلاژن استخراجی از نوع I بوده که دارای سه زنجیره 1α با وزن مولکولی حدود 140 کیلو دالتون می ­باشد.

کلیدواژه‌ها

عنوان مقاله [English]

Morphological and molecular identification of sea cucumber dominant specie in Ouly marine area (Persian Gulf- Bushehr province) and analyzing of its certain body-wall compositions

نویسندگان [English]

  • Fatemeh Ghobadyan 1
  • Hossein Zolgharnein 1
  • Mohammad ali Salari 1
  • Iraj Nabipour 2
  • Amir Vazirizadeh 3
1 Department of Marine Biology, Faculty of marine sciences and oceanography, Khorramshahr University of Marine Science and Technology, Khorramshahr, Iran
2 Marine Biotechnology Research Institute, Persian Gulf Biomedical Research Center, Bushehr University of Medical Sciences, Bushehr, Iran
3 Persian Gulf Aquaculture Research Center, Persian Gulf University Bushehr, Bushehr, Iran
چکیده [English]

This research aimed to analyze some of body compositions also collagen protein, in one of dominant species of sea cucumbers in Ouly-Bushehr area, with species morphological and molecular identification. may be with this study, in addition of exact species Identification, our finding in case of its body wall compounants determination, help to more improvement of medical and nutritional industries in our country,Iran. the result of morphological identification according to body wall ossicles shape and then by molecular identification according to partial sequencing of 16SrRNA mitochondrial DNA indicated the species is “Holothuria leucospilota”. In case of body composions analyzing, protein and moisture were two aboundant compounants with more than 70% in amount and fat was the fewest one. Among protein compounants, collagen was extracted by pepsinal enzymatic hydrolyzing. The type of extracted collagen protein and its subunite composition was determined by SDS-PAGE method. As a result, the extraction method was effective and applicable and the collagen was type (I) with three α1 chains had about 140 kDa molecular weight.

کلیدواژه‌ها [English]

  • Sea cucumber
  • Collagen
  • body composition
  • Molecular
  • Morphological
  • Identification

مراجع

  1. عطاران فریمان، گ.؛ طاهری، ع. و برزکار، ن.، 1395. ارزیابی کلاژن دیواره بدن خیار دریایی گونه Stichopus horrens خلیج چابهار و ژلاتین حاصل از آن. مجله فصلنامه علوم و صنایع غذایی. دوره 52، شماره 13، صفحات 79 تا 89.
  2. Abedin, M.Z.; Karim, A.A.; Ahmed, F.; Latiff, A.A.; Gan, C.Y. and Ghazali, F. C., 2013. Isolation and characterization of pepsin-solubilized collagen from the integument of sea cucumber (Stichopus vastus). J Sci Food Agric. Vol. 93, pp: 1083-1088.
  3. Adibzadeh, N.; Aminzadeh, S.; Jamili, S.; Karkhane, A.A. and Farrokhi, N., 2014. Purification and Characterization of Pepsin-Solubilized Collagen from Skin of Sea Cucumber Holothuria parva. Appl Biochem Biotechnol. Vol. 173, No. 6, pp: 143-154.
  4. Altschul, S.F.; Gish, W.; Miller, W.; Myers, E.W. and Limpman, D.J., 1990. Basic local alighnment search tools. Mol Biol.Vol.215, No. 3, pp: 403-410.
  5. Amos, B. and Hoelzel, A.R., 1992. Application of Molecular Genetic Techniques to The Conservation of small populations. Biol Conserv. Vol. 61, pp: 133-144.
  6. AOAC. 1990. OfficialMethods of Analyses of Association of Analytical Chemists. AOAC, Washington, DC (15th edn.)
  7. Aydin, M.; Sevgili, H.; Tufan, B.; Emre, Y. and Kose, S., 2011. Proximate composition and fatty acid profile of three different fresh and dried commercial sea cucumbers from Turkey. Int J Food Sci Technol. Vol. 46, pp: 500-508.
  8. Aydın, M.; Sevgili, H.; Tufan, B.; Emre, Y. and Köse, S., 2011. Proximate composition and fatty acid profile of three different fresh and dried commercial sea cucumbers from Turkey. Int J Food Sci Technol. Vol. 46, No. 3, pp: 500-508.
  9. Bilgin, Ş. and İzci, L., 2016. The effects of drying & boiling process on nutritional components of Holothuria forskali (Delle Chiaje, 1823). J Food Health Sci. Vol. 2, No. 1, pp: 1-8.
  10. Bilgin, S. and Tanrikulu, O.H., 2018. The changes in chemical composition of Holothuria tubulosa (Gmelin, 1788) with ambient-drying and oven-drying methods. Food Sci Nutr. Vol. 2, pp: 1-6.
  11. Billett, D.S.M., 1991. Deep Sea Holothurians. Oceang Mar Biol. Vol. 29, pp: 259-317.
  12. Birkeland, Ch., 1989. The influence of echinoderms on coral reef communities: pp: 1-79. In: Jangoux, M. and Lawrence, J.M., (eds.). 1989. Echinoderm Studies. Balkema, Rotterdam, Brookfield. 383 P.
  13. Çaklı, Ş.; Cadun, A.; Kışla, D. and Dinçer, T., 2004. Determination of quality characteristics of Holothuria tubulosa (Gmelin, 1788) in Turkish Sea (Aegean Region) depending on sun drying process step used in Turkey. J Aquat Food Product Techol. Vol. 13, No.3, pp: 69-78.
  14. Chang-Lee, M.V.; Price, R.J. and Lampila, L.E., 1989. Effect of processing on proximate composition and mineral content of sea cucumbers (Parastichopus spp.). J Food Sci. Vol. 54, pp: 567-572.
  15. Chukwu, O., 2009. Influences of drying methods on nutritional properties of tilapia fish (Oreochromis nilotieus). World J Agric Sci. Vol. 5, pp: 256-258.
  16. Ciarlo, A.S.; Paredi, M.E. and Fraga, A.N., 1997. Isolation of soluble collagen from hake skin (Merluccius hubbsi). J Aquat Food Product Techol. Vol. 6, No.1, pp: 65-77.
  17. Clouse, R.; Janies, D. and Kerr, A.M., 2005. Resurrection of Bohadschia bivittata from B. marmorata (Holothuroidea: Holothuriidae) based on behavioral, morphological, and mitochondrial DNAevidence. Zoology. Vol. 108, No. 1, pp: 27-39.
  18. Conand, C., 1998. Holothurians (Sea Cucumber, Class Holothuridea). FAO, Rome, FAO species identification guid for fishery parpose. pp: 1158-1190.
  19. Cui, F.X.; Xue, C.H.; Li, Z.J.; Zhang, Y.Q.; Dong, P.; Fu, X.Y. and Gao, X., 2007. Characterization and subunit composition of collagen from the body wall of sea cucumber Stichopus japonicus. Food Chem. Vol. 100, No. 3, pp: 1120-1125.
  20. FAO. 2008. Sea cucumbers: A global review of fisheries and trade. FAO FAO Fish Aquac Tech Pap. Vol. 516, pp: 1-317.
  21. Foegeding, E.A.; Lanier, T.C. and Hultin, H.O., 1996. Collagen. In O. R. Fennema (Ed.), Food chem (3rd ed): New York: Marcel Dekker, Inc. pp: 902-906.
  22. Friess, W., 1998. Collagen biomaterial for drug delivery. Eur J Pharm Biopharm. Vol. 45, pp: 113-136.
  23. Gao, Y.; Li, Z.; Qi, Y.; Guo, Z.; Lin, Y.; Li, W.; Hu, Y. and Zhao, Q., 2016. Proximate composition and nutritional quality of deep sea growth sea cucumbers (Stichopus japonicus) fromdifferent origins. J Sci Food Agric. Vol. 96, pp: 2378-2383.
  24. Granda, V.T., 2006. The biological and trade status of sea cucumber in the families Holothuridae and Stichopodidae. In: Convention on International trade in Endangered Species of Wild Fauna and Flora, Twenty- Second meeting of the animals Committee. pp: 7-13.
  25. Hartl, D.L. and Clark, A.G., 1989. Principles of Population Genetics. 2nd ed. Massachusetts, USA: SinauerAssociates, Inc. 682 p.
  26. Kamarudin, K.R.; Hashim, R. and Usup, G., 2010. Phylogeny of Sea Cucumber (Echinodermata: Holothuroidea) as Inferred from 16S Mitochondrial rRNA Gene Sequences. Journal of Sains Malaysiana. Vol. 39, No. 2, pp: 209-218.
  27. Kamarudin, K.R.; Rehan, A.M.; Hashim, R.; Usup, G.; Fauzanahmad, H.; Anua, H.A. and Yamanidris, M., 2011. Molecular Phylogeny of Holothuria (Mertensiothuria) leucospilota (Brandt 1835) as Inferred from Cytochrome C Oxidase I Mitochondrial DNA Gene Sequences. Journal of Sains Malaysiana. Vol. 40. No. 2, pp: 125-133.
  28. Kamarul R.K.; Aisyah, M.R.; Ahmad Lutfi, L.; Hajar Fauzan, A.; Mohd Hanafi, A.; Noor Faizul Hadry, N.; Ridzwan, H.; Rosnah, H. and Gires, U., 2009. Coral Reef Sea Cucumbers in Malaysia. Malay J Sci. Vol. 28, No. 2, pp: 171-186.
  29. Kamarul, R.K. and Ridzwan, B.H., 2005. Distribution and Taxonomic Revision of Sea Cucumbers (Echinodermata: Holothuroidea) in Several Populations of Malaysia. International Conference on Biogeography and Biodiversity: Wallace in Sarawak- 150 Years Later, 13-15 July, Kuching, Sarawak, Malaysia. 225 p.
  30. Kerr, A.M.; Janies, D.A.; Clouse, R.M.; Samyn, Y.; Kuszak, J. and Kim, J., 2005. Molecular Phylogeny of Coral-Reef Sea Cucumbers (Holothuriidae: Aspidochirotida) Based on 16S Mitochondrial Ribosomal DNASequence. Mar Biotechnol. Vol. 7, pp: 53-60.
  31. Kimura, S. and Tanaka, H., 1983. Characterization of top shell muscle collagen comprising three identical α1chains. Bull Jpn Soc Sci Fisheries. Vol. 49, pp: 229-232.
  32. Kittiphattanabawon, P.; Benjakul, S.; Visessanguan, W.; Nagai, T. and Tanaka, M., 2005. Characterisation of acid soluble collagen from skin and bone of bigeye snapper (Priacanthus tayenus). J Food Chem. Vol. 89, pp: 363-372.
  33. Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature. Vol. 227, pp: 680-685.
  34. Larkin, M.A.; Blackshields, G.; Brown, N.P.; Chenna, R.; McGettigan, P.A.; McWilliam, H.; Valentin, F.; Wallace, I.M.; Wilm, A.; Lopez, R.; Thompson, J.D.; Gibson, T.J. and Higgins, D.G., 2007. Clustal W and Clustal X version 2.0. Bioinformatic. Vol. 23, No. 21, pp: 2947-2948.
  35. Lavery, S.; Chan, T.K.; Tam, Y.K. and Chu, K.H., 2004. “Phylogenetic relationship and evolutionary of the shrimp genus Penaeus s.l. drived from mitochondrial DNA. Mol Phyl and Evol. Vol. 31, pp: 39-49.
  36. Lewis, M.S. and Piez, K.A., 1964. The characterization of collagen from the skin of the dogfish shark, Squalus acanthias. J Biol Chem. Vol. 239, No. 10, pp: 3336-3340.
  37. Liu, Z.; Oliveira, A.C. and Su, Y.C., 2010. Purification and characterization of pepsin-solubilized collagen from skin and connective tissue of giant red sea cucumber (Parastichopus californicus). J Agric Food Chem. Vol. 58, No. 2, pp: 1270-1274.
  38. Love, R.M.; Yamaguchi, Y.; Creach, Y. and Lavety, J., 1976. The connective tissues and collagens of cod during starvation. Comp. Biochem and Physio. Part B. Vol. 55, pp: 487-492.
  39. Michonneau, F.; McPherson1, S.; Mark O’Loughlin, P. and Paulay, G., 2015. More than meets the eye: diversity and geographic patterns in sea cucumbers. https://www. biorxiv.org/content/biorxiv/early/2015/01/23/014282.
  40. Mizuta, S.; Yoshinaka, R.; Sato, M.; Itoh, Y. and Sakaguchi, M., 1992. Subunit composition of distinct types of collagens in the muscle of the kuruma prawn Penaeus japonicus. Comp Biochem and Physio. Vol. 102, pp: 803-811.
  41. Montero, P.; Borderias, J.; Turnay, J. and Leyzarbe, M.A., 1990. Characterization of hake (Merluccius merluccius L.) and trout (Salmo irideus Gibb) collagen. J Agric Food Chem. Vol. 38, No. 3, pp: 604-609.
  42. Muyonga, J.H.; Cole, C.G.B. and Duodu, K.G., 2004. Characterization of acid soluble collagen from skins of young and adult Nile perch (Lates niloticus). J Food Chem. Vol. 85, No. 1, pp:81-89.
  43. Nagai, T.; Worawattanamateekul, W.; Suzuki, N.; Nakamura, T.; Ito, T. and Fujiki, K., 2000. Isolation and characterization of collagen from rhizostomous jelly fish (Rhopilema asamushi). J Food Chem. Vol. 70, pp: 205-208.
  44. Nalinanon, S.; Benjakul, S.; Visessanguan, W. and Kishimura, H., 2007. Use of pepsin for collagen extraction from the skin of bigeye snapper (Priacanthus tayenus). Food Chem. Vol. 104, pp: 593-601.
  45. O’Loughlin, P.M.; Paulay, G.; Vandenspiegel, D. and Samyn, Y., 2007. New Holothuria species from Australia (Echinodermata: Holothuroidea: Holothuriidae), with comments on the origin of deep and cool holothuriids. Mem Mus Vic. Vol. 64, pp: 35-52.
  46. Oedjoe, M.D.R., 2017. Composition of Nutritional Content of Sea Cucumbers (Holothuroidea) in Mania Waters, Sabu Raijua Regency, East Nusa Tenggara. J Aquac Res Dev. Vol. 8, No. 7, pp: 1-3.
  47. Ogawa, M.; Portier, R.J.; Moody, M.W.; Bell, J.; Schexnayder, M.A. and Losso, J.N., 2004. Biochemical properties of bone and scale collagens isolated from the subtropical fish black drum (Pogonia cromis) and sheepshead seabream (Archosargus probatocephalus).  J Food Chem. Vol. 88, pp: 495-501.
  48. Ozer, N.P.; Mol, S. and Varlik, C., 2004. Effect of the handling procedures on the chemical composition of sea cucumber. Turk J Fish Aquat Sci. Vol. 4, pp: 71-74.
  49. Palumbi, S.; Martin, A.; Romano, S.; McMillan, W.O.; Stice, L. and Grabowski, G., 1991. The Simple Fool’s Guide to PCR. Honolulu: Department of Zoology and Kewalo Marine Laboratory, Universiti of Hawaii. 47 p.
  50. Park, S.Y.; Lim, H.K.; Lee, S.; Hwang, H.C.; Cho, S.K. and Cho, M., 2012. Pepsin-solubilised collagen (PSC) from Red Sea cucumber (Stichopus japonicus) regulates cell cycle and the fibronectin synthesis in HaCaT cell migration. J Food Chem. Vol. 132, pp: 487-492.
  51. Pérez-Ruzafa, A.; Entrambasaguas, L. and Bacallado, J.J., 1999. Fauna de equinodermos (Echinodermata) de los fondos rocosos infralitorales del archipiélago de Cabo Verde. Rev Acad Canar. Cienc. Vol. 11, pp: 43-62.
  52. Price, A.R.G., 1983. Echinoderms of Saudi Arabia. Echinoderms of the Arabian Gulf (Persian Gulf) coast of Saudi Arabia. Fauna of Saudi Arabia. Vol. 5, pp: 28-108.
  53. Sadowska, M.; Kolodziejska, I. and Niecikowska, C., 2003. Isolation of collagen from the skins of Baltic cod (Gadus morhua). J Food Chem. Vol. 81, pp: 257-262.
  54. Saiki, R.; Gelfand, D.H.; Stoffel, S.; Scharf, S.J.; Higuchi, R.; Horn, G.T.; Mullis, K.B. and Erlich, H.A., 1988. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. Vol. 239, pp: 487-491.
  55. Saito, M.; Kunisaki, N. and Urano, N., 2002. Collagen as the major edible component of sea cucumber. Food Sci. Vol. 67, pp: 1319-1322.
  56. Salarzadeh, A.R.; Afkhami, M.; DarvishBastami, K.; Ehsanpour, M.; Khazaali, A. and Mokhleci, A., 2012. Proximate Composition of Two Sea Cucumber Species Holothuria pavra and Holothuria arenicola in Persian Gulf. Ann Biol Res. Vol. 3, No. 3, pp: 1305-1311.
  57. Samyn, Y.; Appeltans, W. and Kerr, A.M., 2005. Phylogeny of Labidodemas and the Holothuriidae (Holothuroidea: Aspidochirotida) as inferred from morphology. Zool J Linn Soc. Vol. 144, pp: 103-120.
  58. Siddiqui, Y.D.; Arief, E.M.; Yusoff, A.; Hamid, S.S.A.; Norani, T.Y. and Abdullah, M.Y.S., 2013. Extraction, Purification and Physical Characterization of Collagen from Body wall of Sea cucumber Bohadschia bivitatta. Health Envi J. Vol. 4, No. 2, pp: 53-65.
  59. Sikorski, Z.E.; Kolakowska, A. and Pan, B.S., 1990. The nutritive composition of the major groups of marine food organisms. In Seafood: Resources, Nutritional Composition and Preservation; Sikorski, Z.E., Ed.; CRC Press: Boca Raton, FL, USA. pp: 29-54.
  60. Skierka, E. and Sadowska, M., 2007. The influence of different acid and pepsin on the extractability of collagen from the skin of Baltic cod (Gadus morhua). Food Chem. Vol. 105, pp: 1302-1306.
  61. Tamura, K.; Stecher, G.; Peterson, D.; Filipski, A. and Kumar, S., 2013. MEGA6: Molecular Evolutionary Genetics Analysis Version 6. Mol Biol Evol. Vol. 30, pp: 2725-2729.
  62. Telahigue, K.; Hajji, T.; Imen, R.; Sahbi, O. and El Cafsi, M., 2014. Effects of drying methods on the chemical composition of the sea cucumber Holothuria forskali. The Open Food Sci J. Vol. 8, pp: 1-8.
  63. Titterton, R. and Whatley, R., 2006. Recent marine Ostracoda from the Solomon Islands: part 1- Cypridoidea, Platycopina & Cladocopina. Journal of Micropalaeontology. Vol. 25, pp: 73-94.
  64. Trotter, J.A.; Lyons-levy, G.; Thurmond, F.A. and Koob, T.J., 1995. Covalant composition of collagen fibrils from the dermis of sea cucumber, Cucumaria frondosa, a tissue with mutable mechanical properties. Comp Biochem Physiol. Part A: Physiology. Vol. 112, No. 3-4, pp: 463-478.
  65. Waldschmidt, A.Ma.; Salomao, T.M.F.; Barros, E.G.;Campos, L. and Antônio, O., 1997. Extraction of genomic DNA from Melipona quadrifasciata (Hymenoptera: Apidae, Meliponinae). Braz. J. Gen, [online]. Vol. 20, No. 3.
  66. Wen, J.; Hu, C. and Fan, S., 2010. Chemical composition and nutritional quality of sea cucumbers. J Sci Food Agr. Vol. 90, pp: 2469-2474.
  67. Zeugolis, D. and Raghunath, L., 2011. Collagen: Materials Analysis and Implant Uses. Compr Biomater J. pp: 261-278.
  68. Zeugolis, D.I.; Paul, R.G. and Attenburrow, G., 2008. Extruded collagen-polyethylene glycol fibers for tissue engineering applications. J Biomed Materi Res. Part B: Applied Biomaterials. Vol. 85, pp: 343-352.
  69. Zhang, M.; Liu, W. and Li, G., 2009. Isolation and characterisation of collagens from the skin of largefin longbarbel catfish (Mystus macropterus).  J Food Chem. Vol. 115, pp: 826-831.

 

فصلنامه محیط زیست جانوری
دوره 11، شماره 3
مهر 1398
صفحه 363-378

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