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Molecular Ecology Comparison of Blue Leg Hermit Crab (Calcinus Elegans) Based on Spatial Factor in South Coast of Java Island

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Molecular Ecology Comparison of Blue Leg Hermit Crab (Calcinus Elegans) Based on Spatial Factor in South Coast of Java Island Available online at IJMARCC, Website: http://ejournal.undip.ac.id/index.php/ijmarcc International Journal o Marine and Aquatic Resource Conservation and Co-existence Research Article, 1 (1): 12-18, October 2014 Molecular Ecology Comparison o .lue /eg 0ermit Crab (Calcinus elegans) based on 1patial 2actor in 1outh Coast o Java Island Mulia3ati 0andayani1), 1utrisno Anggoro2), Ita Wido3ati2) and Imai 0ideyu4i3) ) Faculty of Fisheries and Marine Science, Brawi(aya )niversity, Indonesia 2) Faculty of Fisheries and Marine Science, Diponegoro )niversity, Indonesia 3) ,enetic -aboratories, Depart.ent Marine and Environ.ental Science, )niversity of the Ryu0yus E.ail1 .uliawati2fpi03ub.ac.id Received1 February 25, 20 5, Accepted1 April 7, 20 5 A.1TRACT The study was conducted to deter.ine the genetic diversity of Calcinus elegans based on sequences of the C8I .itochondrial DNA. Successfully 55 sa.ples of ,arut; 53 of Yogya0arta and 57 of Banyuwangi populations were a.plified by PCR using universal pri.ers -C8 590 and HC8 2 98 successfully a.plified C8I gene in A25 bp, with an overall haplotype totaled A5 at poly.orphic sites. All population showed high genetic diversity within population interpreted by the value of gene diversity (H)1 ,arut is 0.92AA C 0.025 ; Yogya0arta is 0.9AA8 C 0.0 5A and Banyuwangi is 0.9288 C 0.0257 and the value of nucleotide diversity (D)1 ,arut is 0.007 55 C 0.003972; Yogya0arta is 0.0079AA C 0.005387 and Banyuwangi is 0.00723 C 0.00502 . ,ene diversity and nucleotide diversity did not differ significantly between populations. Ehile genetic diversity a.ong populations interpreted fro. haplotypes si.ilarity or shared haplotype. Nu.ber of shared haplotypes a.ong three populations is 2 haplotypes with 90 sa.ples included. Banyuwangi population showed s.allest intensity interaction with other populations indicated by the s.allest value of haplotypes; poly.orphic sites; hetoroFigosity and distribution of larvae are influenced by spatial factor. However, three populations co.e fro. one ancestors and part of a large population. Thus the current and other environ.ental factors in South Coast of Java Island are effected on the genetic diversity a.ong populations of C. elegans with a correlation coefficient is r G 0.98 (genetic diversity with current) and r G H0.90 (genetic diversity with te.perature, salinity, pH, depth and distance). 7ey 3ords1 Calcinus elegans, genetic diversity, C8I, .itochondrial DNA, haplotype I8TROD:CTIO8 about 5,000 individuals per .onth in the period fro. April to August (not the season). Her.it crab is one of o.nivores ani.al since it plays an This study is co.ple.entary of her.it crab studies i.portant role in ecosyste. that is as the cleaner ani.al and previous conducted by about dyna.ic of tropical her.it crab plays an i.portant role in co..unity structure (Eric et al. asse.blage (Bertness, 98 ); reproduction of her.it crab 20 0). They are eaten by .any ani.als higher up in the food (Brian, 989); behavior of her.it crab (Coenobitidae) chain. They also as scavengers and help quic0ly recycle dead (Vanessa, 2008); .olecular analysis of the taxono.ic and .atter on the shores. So.e her.it crabs beco.e the i.portant distributional status for the genera Loxopagurus and Isocheles export co..odity that is being hunted .arine aquariu. (Mantello et al., 200A); and paripatric speciation drives enthusiasts in the world because of the beauty and uniqueness deversification and distributional pattern of her.it crab (Calado et al., 2003). 8ne of the. is the C. elegans which is (Decapoda1 Diogenidae1 Calcinus) (Malay and Paulay, 2009), usually called by the Blue leg hermit crab (Ielo.ang 0a0i .icrohabitat distribution of the her.it crabs Calcinus haigae biru) by supplier of aquariu. orna.ent in Indonesia. and Calcinus Hazletti (Brian and Bach, 20 0); .ultigene According to Haig and Ball ( 988), C.elegans was .olecular syste.atic confir. species status of .orphologically distributed along the East Coast of Africa to the Hawaiian convergent Pagurus her.it crab (Silva et al., 20 ) and Islands, Tua.otu Islands and Malu0u Islands. But not only in revisiting Pagurus pilosipes (Io.ai et al., 20 ). the Malu0u, had the ani.al also distributed in so.e roc0y 8ne of the protein-coding genes that located on the .tDNA beaches and tidal flats along Indian 8cean (Morgan, 99 ). is gene cytochro.e C oxidase subunit I (C8I). These coding Exportir supplier of C. elegans in Java co.es fro. ,arut genes have been widely used as a .ar0er of .olecular in a beach (Peungpeu0 beach), Yogya0arta (Baron beach) and variety of genetic studies of various species and genetic Banyuwangi (Bla.bangan beach). All of these place located in .ar0ers in population studies. Hebert et al. (2003) suggests South Coast of Java (Ahyan, personal co..unication). that the DNA coding syste. for the ani.als can be based on a The lac0 of infor.ation about abundance of her.it crab in sequence diversity of C8I gene .itochondrial DNA that quite Indonesia. It is esti.ated that approxi.ate 30,000 individuals fulfilling to be used as .olecular .ar0ers. That is because the per .onth in the period fro. Septe.ber to March (season) and C8I gene has regions sustainable and high variability between species (-ee, 2005). © Copyright by The International Journal of Marine and Aquatic Resources Conservation and Co-existence 2 3 International Journal of Marine and Aquatic Resource Conservation and Co-existence ( )1 2- 8, 20 5 Muliawati Handayani, Sutrisno Anggoro, Ita Eidowati and I.ai Hideyu0i ,enetic diversity of C. elegans can be affected by the ,,TCAACAAATCATAAA,ATATT,,-3 ) and HC8 2 98 discrepancy of spatial factor (Row, 20 ). The study of so.e (5 -TAAACTTCA,,,T,ACCAAAAAATCA-3 ) (Fol.er et population of aquatic organis. are deter.ined by spatial al., 995). This pri.er has been widely used to study C8I gene factors, they are population siFe, strength of diffusion, for. of .ithochondrial DNA at invertebrates as in Sotelo et al. (2009a) aquatic and behavioral characteristics of larvae and the .odel and Sotelo et al. (2009b) which uses two pri.ers above at the of current .ove.ents (-argier, 2003). According to Dahlhoff species of European Ma(a Spider Crab and Necora Puber and and Ran0 (2000), genetic diversity related to the latitude and Io.ai et al. (20 ) also uses it on Pagurus pilosipes. PCR longitude or the gradient of tide and te.perature. A.ong those conditions were as follows1 pre-denaturation 95 KC for 2 .in, factors, the latitude and longitude describe the spatial location followed by denaturation 95 KC for 30 seconds, annealing at a of the habitat that beco.es the selection factor to .aintain the te.perature of 55 KC for 30 seconds, and extension at a life and deter.ine biologic interaction li0e gen flow, .utation, te.perature of 72 KC for .in, cycle 30 ti.es, then ends with a and gen drift. Therefore, spatial factor has an i.portant role in final extension at 72 KC for 7 .in. Chec0ing the results of PCR deter.ining the genetic diversity of a population by identifying was perfor.ed by electrophoresis on L gel agarose. the quantity of poly.orphis.s appearance. ,enetic diversity 2) Sequencing C8I gene of C. elegans and fenotipic co.es fro. DNA variation and gene regulation. Sequencing was perfor.ed by the sequencing principle of Further.ore Stansfield and Elrod (2007), states the a.ount of dideoxy Sanger .ethod using Auto.atic DNA sequence CEM genetic diversity within a species depends on the nu.ber of 8800 with star 0it (Bec0.an Coulter). Sequencing C8I gene individuals, the range of geographical spread, the rate of .itochondrial DNA of C. elegans had been deposited in ,en population isolation and the genetic syste.s. An i.portant role ban0 with accession nu.ber AB 820 38 N AB82028 ( 55 is also perfor.ed by the processes of natural selection and the sa.ples). factors that affect the spatial and te.poral changes in the 3) Analysis of .olecular data genetic co.position of a species or population. Therefore, the The sequences of C8I DNA .itochondrial were analyFed ai. of this study was to deter.ine how genetic diversity within by using Clustal O (version 2. ) to align the sequences results population and a.ong population in C. elegans based on (Tho.pson et al., 997); Arlequin to deter.ine haplotipe and sequence analyFes of C8I gene .itochondrial DNA and the nucleotide diversity in the population (Ta(i.a, 983); AM8VA relationship the genetic diversity a.ong population with spatial (Analysis Moleculer of Variance) to analyFe population factor in ,arut, Yogya0arta and Banyuwangi. structure (Excoffier et al., 992) and ME,A version 5.0 (Molecular Evolutionary ,enetic Analysis) to create a MATERIA/ A8D MET0OD phylogeny tree by neighbor oining .ethod, where the .atrix calculation of genetic distance with .odel of Ii.ura-2 Sampling of crab para.eters were i.ple.ented on the pair wise distance calculation (Ta.ura et al., 2007). Ehile to 0now the ,enetic .aterials used in this study were about 55 sa.ples relationship between genetic diversity with the environ.ent, fro. ,arut; 53 sa.ples fro. Yogya0arta and 57 sa.ples fro. then tested the correlation and regression between genetic Banyuwangi that were collected by purposive sa.pling. -eg diversity with spatial factors, so 0nown the degree of .uscle tissue sa.ples of C. elegans were ta0en by the correlation of r value. autoto.i .ethod on the legs (Ao0i et al., 2008).
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