PENETAPAN KERAGAMAN GENETIK NILAM (Pogostemon sp.) HASIL FUSI PROTOPLAS DENGAN TEKNIK RAPD

Keragaman genetik dan kckerabatan tanaman nilam hasil fusi protoplas antara Nilam Jawa (Girilaya) dan Nilam Aceh (Sidikalang dan TT 75) dianalisis dengan menggunakan penanda RAPD. Bahan tanaman yang digunakan dalam penelitian ini adalah 9 genotipa yang tcrdiri dari 3 tetua dan 6 tanaman hibrida somatik (9 II 33, 9 II 21, 2 IV 8, 9 IV14, 9 II 7 dan 9 II 10). Primer yang digunakan dalam analisis tcrdiri atas 5 primer acak yaitu OPD 03, OPD 20, OPH 09, OPH 19 dan Abi 117.17. DNA dickstraksi dengan metode OROZCO-CASTJLLO et al. (1994) yang sudah dimodifikasi. Konsentrasi DNA ditetapkan dengan metode sambrook el al. (1989) dengan pcrbandingan kuantifikasi spektrofotometrik. Koefisien kemiripan dan kckerabatan antar genotipa dianalisis dengan menggunakan program NTsys ver. 1.80 dan UPGMA untuk menentukan sidik gerombol dan dendogram. Hasil analisis menunjukkan bahwa koefisien kemiipan dari amplifikasi DNA dengan 5 primer acak dari ke- 9 genotipa berkisar antara 0.48-1.0. Berdasarkan niatrik jarak genetik, kescmbilan genotipa tanaman yang diuji membentuk 2 kelompok besar yaitu kelompok I, tetua Girilaya (Nilam Jawa) dan kelompok II tcrdiri dari tetua Sidikalang dan TT 75 (Nilam Aceh) serta hibrida somatik. Kelompok II, tcrbagi menjadi dua sub kelompok yaitu sub kelompok I (9 II 33 dan 9 II 7) dan sub kelompok II yang tcrdii dari sub-sub kelompok II-I (9 II 21, S, TT 75) dan sub-sub kelompok II-II (2 IV 8, 9 IV 14, 9 II 10). Kata kunci: Pogostemon sp., fusi protoplas, keragaman genetik, RAPD ABSTRACTS Assessment of genetic variability of patchoulli (Pogostemon sp.,) derived from protoplast fussion using RAPD Somatic hybrids of Pogostemon heyneaneus (cv. Girilaya) X P. cablin (cv. Sidikalang and TT 75) were tested for their genetic variability and relationship. The somatic hybrids tested were 9 II 33, 9 II 21, 2 IV 8, 9 IV14, 9 II 7 and 9 II 10. DNA of the plant materials used were extracted by using the modified method of orozcocastulo et al. (1994) and quantified spectrophotometrically according to SAMBROOK el al. (1989). Five random primers, OPD 03. OPD 20, OPH 09. OPH 19 and Abi 117.17, were applied to amplify the extracted DNA. The genetic relationship among the somatic hybrids were estimated by using the index of similarity to perform genctical matrix and dendogram. Index of similarity among genotypes were calculated by using NTsys ver. 1.80 program. Then, cluster analyses to perform dendogram were achieved based on similarity estimates by using the Unweighted Pair-Group Method Arithmetic Average (UPGMA). Results showed that index of similarities of the amplified DNA from 5 random primers ranged from 0.48 to 1.0. The somatic hybrids and their parental plants subjected to RAPD analyses were classified into 2 major groups, first, the parental group of Java patchouli and second, others parental plants, Aceh patchouli (Sidikalang and TT 75), and the somatic hybrids. The second group was then classified into 2 minor groups. First group consisted of somatic hybrids nos. 9 II 33 and 9 II 7, while the second were classified into 2 groups which consisted nos. 9 II 21, S, TT 75 and nos. 2 IV 8, 9 IV 14 and 9 II 10. Key words : Pogostemon sp., protoplast fusion, genetic variability, RAPD