GENETIC CHARACTERIZATION OF SEVERAL PROMISING ACCESSION OF Jatropha curcas L. BASED ON RAPD MARKER

MAFTUCHAH MAFTUCHAH, AGUS ZAINUDIN, RULLY DYAH PURWATI, HADI SUDARMO

Abstract


ABSTRACT

The objective of this research was to obtain genetic relationshipamong 13 Jatropha curcas L. accession plants based on Random Amplified Polymorphic DNA marker. This experiment used 13 accessionsof J. curcas L. potential to have higher seed productivity, including HS-49,SP-16, SP-38, SP-8, SM-33, SP-34, SM-35, IP-1A, IP-1M, IP-1P, IP-2A,IP-2M, and IP-2P. Polymerase Chain Reaction (PCR) was performedusing 10 selected primers of RAPD markers (OPA 2, OPA 9, OPA 13,OPA 15, OPA 18, OPA 19, OPA 20, OPF 8, OPF 10, and OPF 15). PCRproduct was used to determine genetic distance which implemented Un-weighted Pair-Group Method With Arithmetic (UPGMA) procedure andconstructed phylogeny trees using Numerical Taxonomy and MultivariateSystem (NTSYS) software version 1.8. The confidence level of UPGMAwas then tested by Boostrap using WinBoot program. Ten primers used inthis research were able to be applied in genomic DNA of J. curcas L. plantwhich had resulted about four (OPA 19) to ten band numbers (OPA 9)with the band size around 72-1,078 bp. However, OPA 13 primer was notable to give different band size. Genetic relationship analysis has foundtwo main groups, firstly accession plants consisted of HS-49, SP-16, SP-18, SP-8, SM-33, SM-35, and SP-34 (coefficient 0.8). In this group, SP-38clustered with SP-8, and SM-33 with SM-35 (coefficient 0.91). In thesecond group, the accessions consisted of IP-1A, IP-1M, IP-1P, IP-2A, IP-2M, and IP-2P (coefficient 0.78). In this group, accession of IP-1Aclustered with IP-1M (coefficient 0.85), IP-1P with IP-2M (coefficient0.87), and IP-2A with IP-2P (coefficient 0.90). Then, the first and secondgroups formed genetic relationship with coefficient 0.66.Key words: Genetic characterization, Jatropha curcas L., RAPD,molecular marker, promising accession.

ABSTRAK

Penelitian ini bertujuan untuk mendapatkan informasi keragamangenetik dan hubungan kekerabatan berbagai aksesi jarak pagar terpilihberdasarkan analisis molekuler Random Amplified Polymorphic DNA(RAPD). Penelitian menggunakan 13 aksesi Jatropha curcas yangmemiliki potensi produksi tinggi (HS-49, SP-16, SP-38, SP-8, SM-33, SP-34, SM-35, IP-1A, IP-1M, IP-1P, IP-2A, IP-2M, dan IP-2P). Isolasi DNAgenom J. curcas dilaksanakan dengan metode Zheng yang dimodifikasi.Polymerase Chain Reaction (PCR) dilakukan menggunakan 10 primersRAPD (primer OPA 2, OPA 9, OPA 13, OPA 15, OPA 18, OPA 19, OPA20, OPF 8, OPF 10, dan OPF 15). Produk PCR yang dihasilkan digunakanuntuk menentukan tingkat kekerabatan menggunakan Un-weighted Pair-Group Method With Arithmetic (UPGMA) dan diagram filogenetik denganprogram Numerical Taxonomy and Multivariate System (NTSYS) versi1.8. Kesepuluh primer yang digunakan mampu mengamplifikasi DNAjarak pagar dengan jumlah produk pita antara 4 (primer OPA 19) hingga10 pita DNA (OPA 9), dengan ukuran pita 72-1.078 bp. Primer OPA 13tidak dapat memberikan perbedaan pita DNA. Hasil analisis kekerabatanmenunjukkan adanya dua kelompok utama. Kelompok pertama terdiri atasaksesi HS-49, SP-16, SP-18, SP-8, SM-33, SM-35, dan SP-34 (koefisien0,80). Dalam kelompok pertama, SP-38 berkelompok dengan SP-8, danSM-33 dengan SM-35 (koefisien 0,91). Kelompok kedua terdiri atas aksesiIP-1A, IP-1M, IP-1P, IP-2A, IP-2M, dan IP-2P (koefisien 0,78). Dalamkelompok kedua, IP-1A berkelompok dengan IP-1M (koefisien 0,85), IP-1P dengan IP-2M (koefisien 0,87), dan IP-2A dengan IP-2P (koefisien0,90). Selanjutnya, kelompok pertama dan kelompok kedua membentukkekerabatan pada koefisien 0,66.

Kata kunci: Karakterisasi genetik, Jatropha curcas L., RAPD, markamolekuler, aksesi harapan.


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DOI: http://dx.doi.org/10.21082/jlittri.v17n2.2011.83-88

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