Chan, Eng Heng

Joseph, Juanita

Liew, Hock Chark

Tan, Soon Guan

Universiti Putra Malaysia Institutional Repository

FOR Category Fl0800: Biological SciencesSTUDIES ON THE POPULATION GENETICS OFGREEN TURTLE (CHELONIA MYDAS) ANDHAWKSBILL TURTLE (ERETMOCHELYSIMBRICATA) IN MALAYSIA USINGMICROSATELLITEJuanita Joseph, Chan Eng Heng, Tan Soon Guan andLiew Hock CharkFaculty of Applied Science and TechnologyUniversiti Putra Malaysia Terengganu, Mengabang Telipot,21030 Kuala Terengganu, MalaysiaKeywords: population genetics, microsatellite, seaturtle, genetic variability, conservation.IntroductionThis study applies modem molecular genetic techniques toaddress aspects of Malaysian Sea turtle biology that haveremained unresolved using conventional methods. The en-dangered or threatened status of sea turtles in Malaysia dic-tates aggressive and comprehensive management plans toexpedite population recoveries. In light of that, specificstudies were undertaken to determine the genetic variabilitywithin and among the various nesting populations, assessingthe extent of differentiation among these populations and todetermine their conservation status. Our ultimate aim is toprovide the first complete genetic appraisal of diversity andstock structure for Malaysian green and hawksbill turtles inresponse to the need to identify conservation priorities. Wehoped to establish conditions that permit nesting populationsto increase in numbers to some level at which a species is nolonger at appreciable risk of extinction.Materials and MethodsBlood samples from hatchlings will be collected from vari-ous nesting populations in Malaysia. A standard bleedingtechnique will be used as described by Owens and Ruiz(1980), where blood is drawn from the dorsal cervical sinususing Y2 CC insulin syringe. Samples are preserved in a lysisbuffer (Dutton, 1996) and stored at -20°C. Genomic DNAfrom blood samples are then extracted using the Qiagen tis-sue kit protocols. All microsatellite amplifications will beperformed in a 20 III mixtures containing template DNA,PCR buffer, MgCl2, dNTP's, taq polymerase and microsatel-lite primers. 17 microsatellite primers (Dutton, 1995;FitzSimmons et al. 1995; Kichler, internet pers. comm.) wereused and amplifications were performed using the Perkin-Elmer 2400 PCR system. Aliquots of PCR products will berun on 4% Nusieve agarose 3:1 gels, stained using ethidiumbromide and visualized on the UV transiluminator. Non-radioactive confmnation tests for the heterozygots will alsobe done using 6% sequencing acrylarnide gels.Results and DiscussionBlood samples were collected from various nesting popula-tions in Malaysia, viz., Sabah (Sabah Turtle Islands Park andPulau Sipadan), Melaka, Johor, Perak, Pahang and Tereng-ganu (Pulau Redang and Setiu). Further sample collectionwas also done from Sarawak Turtle Islands and Terengganu(Pulau Perhentian and Setiu). So far, we managed to screenall the green and hawksbill turtle's blood samples fromSabah Turtle Islands. Anyhow, we haven't done any statisti-cal analyses yet on the results obtained. Currently, we havestandardised our molecular genetic techniques and expectedto finish the lab analyses in May, 1999. We hope to finishthis project by the end of the year 1999 and submit a com-plete genetic appraisal of diversity and stock structure ofMalaysian sea turtles.ConclusionsOur preliminary results demonstrate that microsatellite tech-nique can be used as a rapid and sensitive genetic marker forthe detection of polymorphism and to study the populationgenetics of sea turtles in Malaysia.ReferencesDutton, P.H. 1995. Molecular evolution of sea turtles with specialreference to the leartherback, Dermochelys coriacea. PhD dis-sertation. p. 137.Dutton, P.H. 1996. Methods for collection and preservation of sam-ples for sea turtle genetic studies. pp. 17-24. In: Bowen, B.W.and W.N. Witzell (eds.). Proceedings of the International Sympo-sium on Sea Turtle Genetics. NOAA Tech. Mem. NMFS-SEFSC-396. p. 173.FitzSimmons, N.N., Moritz, C. and Moore, S.S. 1995. Conservationand dynamics of microsatellite loci over 300 million years of ma-rine turtle evolution. Mol. Bioi. Evol. 12: 432-440.Owens, D.W. and Ruiz, GJ. 1980. New methods of obtaining bloodand cerebrospinal fluid from marine turtles. Herpetologica 38:124-135.Supported by IRPA Grant 09-02-04-0044199

Studies on the population genetics of green turtle (Chelonia mydas) and hawksbill turtle (Eretmochelys imbricata) in Malaysia using microsatellite

http://psasir.upm.edu.my/42524/1/Studies%20on%20the%20population%20genetics%20of%20green%20turtle%20%28Chelonia%20mydas%29%20and%20hawksbill%20turtle%20%28Eretmochelys%20imbricata%29%20in%20Malaysia%20using%20microsatellite.pdf

Studies on the population genetics of green turtle (Chelonia mydas) and hawksbill turtle (Eretmochelys imbricata) in Malaysia using microsatellite

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