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STUDI KERAGAMAN GENETIK TANAMAN SIRSAK (Annona muricata L.) DI JAWA BERDASARKAN PENANDA MOLEKULER RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD)

Abstract

This study aimed to determine the genetic diversity and relationship of soursop plants in Java based on Random Amplified Polymorphic DNA (RAPD) molecular marker. The collection of soursop leaf samples was done by purposive random sampling with 10 individuals representing a population in Java include Central Java (consist of Sukoharjo and Karanganyar), East Java (consist of Ngawi and Pacitan), Yogyakarta (consist of Kulonprogo and Gunung Kidul) and West Java (only Bogor). RAPD analysis was done by using a Hexadecyltrimethylammonium Bromide (CTAB) DNA extraction method and then continued by the quality and quantity of DNA testing, primer selection, optimization DNA by PCR and separation PCR product with horizontal electhrophoresis. DNA band amplification results which were obtained then analyzed descriptively. The cluster analysis using Unweighted Pair Group with Arithmatic Average (UPGMA) method which available in POPGENE 1.32 program was used to calculate the genetic distance. Genetic relationship clustering of the soursop plant among population obtained a dendogram using the Numerical Taxonomy and Multivariate Analysis System (NTSYS) program version 2.00. The results showed that RAPD diversity of soursop populations in Java using 6 types of primer (A18, A20, P10, P11, P04 and P06) produced 152 polymorphic DNA bands (100%) of 152 DNA bands can be observed. Highest genetic diversity produced on Karanganyar population with genetic diversity average value (h) was 0,0525±0,1095 where as shortest genetic diversity on Pacitan population with genetic diversity average value (h) was 0,0418±0,1079. Dendogram of the relationship based RAPD marker showed that the population of Karanganyar and Pacitan have nearest relationship with genetic distance value was 0,0244 where as the population of Sukoharjo and Gunung Kidul have fartest relationship with genetic distance value was 0,0410. Keywords: genetic diversity, soursop, Java, RAPD Penelitian ini bertujuan untuk mengetahui keragaman genetik dan hubungan kekerabatan tanaman sirsak di Jawa berdasarkan penanda molekuler Random Amplified Polymorphic DNA (RAPD). Pengambilan sampel daun sirsak sejumlah 10 tanaman per lokasi dilakukan secara acak (purposive random sampling) di beberapa lokasi di Pulau Jawa yaitu Jawa Tengah (Sukoharjo dan Karanganyar), Jawa Timur (Ngawi dan Pacitan), Yogyakarta (Kulonprogo dan Gunung Kidul) dan Jawa Barat (Bogor). Analisis RAPD dilakukan dengan ekstraksi DNA menggunakan metode Hexadecyltrimethylammonium Bromide (CTAB), kemudian dilakukan uji kualitas dan kuantitas DNA, seleksi primer RAPD, optimasi DNA dengan PCR dan separasi DNA hasil PCR dengan elektroforesis horizontal. Pita DNA hasil amplifikasi yang diperoleh kemudian dianalisis keragamannya secara deskriptif. Untuk menghitung jarak genetik digunakan analisis gerombol (cluster analysis) menggunakan metode Unweighted Pair Group with Arithmatic Average (UPGMA) yang ada dalam program POPGENE 1.32. Pengelompokan ini akan menampilkan hubungan kekerabatan genetik antarpopulasi tanaman sirsak dalam bentuk dendogram yang dihasilkan dalam program Numerical Taxonomy and Multivariate Analysis System (NTSYS) versi 2.00. Hasil penelitian menunjukkan bahwa keragaman RAPD pada populasi tanaman sirsak di Jawa dengan menggunakan 6 jenis primer (A18, A20, P10, P11, P04 dan P06) menghasilkan 152 pita DNA polimorfik (100% dari 152 pita DNA yang dapat diamati. Keragaman genetik tertinggi diperoleh pada populasi Karanganyar dengan nilai rata-rata keragaman genetik (h) sebesar 0,0525±0,1095 sedangkan keragaman genetik terendah pada populasi Pacitan dengan nilai rata- rata keragaman genetik (h) sebesar 0,0418±0,1079. Dendogram hubungan kekerabatan berdasarkan penanda RAPD menunjukkan bahwa populasi Karanganyar dan Pacitan memiliki hubungan kekerabatan paling dekat dengan nilai jarak genetik 0,0244 sedangkan hubungan kekerabatan terjauh yaitu dari populasi Sukoharjo dan populasi Gunung Kidul dengan nilai jarak genetik sebesar 0,0410. Kata kunci: keragaman genetik, sirsak, Jawa, RAPD

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