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Juanita Joseph, Chan Eng Heng, Tan Soon Guan and Liew Hock Chark

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Juanita Joseph, Chan Eng Heng, Tan Soon Guan and Liew Hock Chark FOR Category Fl0800: Biological Sciences STUDIES ON THE POPULATION GENETICS OF FitzSimmons et al. 1995; Kichler, internet pers. comm.) were GREEN TURTLE (CHELONIA MYDAS) AND used and amplifications were performed using the Perkin- HAWKSBILL TURTLE (ERETMOCHELYS Elmer 2400 PCR system. Aliquots of PCR products will be IMBRICATA) IN MALAYSIA USING run on 4% Nusieve agarose 3:1 gels, stained using ethidium MICROSATELLITE bromide and visualized on the UV transiluminator. Non- radioactive confmnation tests for the heterozygots will also Juanita Joseph, Chan Eng Heng, Tan Soon Guan and be done using 6% sequencing acrylarnide gels. Liew Hock Chark Results and Discussion Faculty of Applied Science and Technology Blood samples were collected from various nesting popula- Universiti Putra Malaysia Terengganu, Mengabang Telipot, tions in Malaysia, viz., Sabah (Sabah Turtle Islands Park and 21030 Kuala Terengganu, Malaysia Pulau Sipadan), Melaka, Johor, Perak, Pahang and Tereng- ganu (Pulau Redang and Setiu). Further sample collection Keywords: population genetics, microsatellite, sea was also done from Sarawak Turtle Islands and Terengganu turtle, genetic variability, conservation. (Pulau Perhentian and Setiu). So far, we managed to screen all the green and hawksbill turtle's blood samples from Introduction Sabah Turtle Islands. Anyhow, we haven't done any statisti- This study applies modem molecular genetic techniques to cal analyses yet on the results obtained. Currently, we have address aspects of Malaysian Sea turtle biology that have standardised our molecular genetic techniques and expected remained unresolved using conventional methods. The en- to finish the lab analyses in May, 1999. We hope to finish dangered or threatened status of sea turtles in Malaysia dic- this project by the end of the year 1999 and submit a com- tates aggressive and comprehensive management plans to plete genetic appraisal of diversity and stock structure of expedite population recoveries. In light of that, specific Malaysian sea turtles. studies were undertaken to determine the genetic variability Conclusions within and among the various nesting populations, assessing the extent of differentiation among these populations and to Our preliminary results demonstrate that microsatellite tech- determine their conservation status. Our ultimate aim is to nique can be used as a rapid and sensitive genetic marker for provide the first complete genetic appraisal of diversity and the detection of polymorphism and to study the population stock structure for Malaysian green and hawksbill turtles in genetics of sea turtles in Malaysia. response to the need to identify conservation priorities. We hoped to establish conditions that permit nesting populations References to increase in numbers to some level at which a species is no Dutton, P.H. 1995. Molecular evolution of sea turtles with special longer at appreciable risk of extinction. reference to the leartherback, Dermochelys coriacea. PhD dis- sertation. p. 137. Materials and Methods Dutton, P.H. 1996. Methods for collection and preservation of sam- Blood samples from hatchlings will be collected from vari- ples for sea turtle genetic studies. pp. 17-24. In: Bowen, B.W. ous nesting populations in Malaysia. A standard bleeding and W.N. Witzell (eds.). Proceedings of the International Sympo- sium on Sea Turtle Genetics. NOAA Tech. Mem. NMFS- technique will be used as described by Owens and Ruiz SEFSC-396. p. 173. (1980), where blood is drawn from the dorsal cervical sinus FitzSimmons, N.N., Moritz, C. and Moore, S.S. 1995. Conservation using Y2 CC insulin syringe. Samples are preserved in a lysis and dynamics of microsatellite loci over 300 million years of ma- buffer (Dutton, 1996) and stored at -20°C. Genomic DNA rine turtle evolution. Mol. Bioi. Evol. 12: 432-440. from blood samples are then extracted using the Qiagen tis- Owens, D.W. and Ruiz, GJ. 1980. New methods of obtaining blood sue kit protocols. All microsatellite amplifications will be and cerebrospinal fluid from marine turtles. Herpetologica 38: performed in a 20 III mixtures containing template DNA, 124-135. PCR buffer, MgCl2, dNTP's, taq polymerase and microsatel- lite primers. 17 microsatellite primers (Dutton, 1995; Supported by IRPA Grant 09-02-04-0044 199.
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