Induksi embriogenesis dan deteksi senyawa nikotina pada tahap awal perkembangan embrio dari mikrospora tembakau (Nicotiana tabacum "vorstenlanden")

SANTOSA, Djoko, Dr. Ari Indrianto, SU

2004 | Tesis | S2 Biologi

Penelitian ini bertujuan untuk menginduksi mikrospora Nicotiana tabacum “vorstenlanden” menjadi embriogenik dan mendeteksi senyawa nikotina secara kualitatif dan kuantitatif pada tahap awal perkembangan embrio dari mikrospora tersebut. Mikrospora pada stadium uninukleat akhir diberi praperlakuan stres berupa starvasi nitrogen dan karbohidrat pada suhu 34oC selama 3, 5 dan 7 hari. Mikrospora yang menjadi embriogenik disubkultur ke medium embriogenesis (androgenesis), diinkubasi dalam keadaan gelap, pada suhu 25oC selama 45 hari. Analisis nikotina dilakukan dengan metode kromatografi lapis tipis (KLT), dipakai silika gel GF 254 sebagai fase diam dan campuran metanol dan amonia (100:1,5, v/v) sebagai fase gerak. Kadar relatif nikotina diamati dengan metode kromatografi gas (KG). Parameter penelitian yang diamati meliputi penetapan stadium perkembangan mikrospora, persentase viabilitas mikrospora embriogenik, pembelahan dua inti sampai multinukleat, serta penetapan senyawa nikotina secara kualitatif dan kuantitatif. Hasil penelitian menunjukkan bahwa mikrospora embriogenik dapat dihasilkan dari semua praperlakuan stres, praperlakuan stres selama 7 hari menghasilkan embrioid multinukleat terbanyak (3,90 ± 0,21 %), nikotina tampak setelah 45 hari mikrospora di medium androgenesis, dan konsentrasi relatif nikotina berturut-turut pada praperlakuan 3 dan 7 hari adalah 1,1087% dan 1,3192%.

The aims of this study is to induce microspores of Nicotiana tabacum “vorstenlanden” become embriogenic and to detect nicotine compound on early stage of microspore derived-embryos development qualitatively and quantitatively. Microspores in late-uninucleate stage, subjected to stress pretreatment in nitrogen-carbohydrate starvation and heat shock (34o C) for 3, 5 and 7 days. The resulting embriogenic microspores were then subcultured to androgenesis medium incubated in dark room at 25o C for 45 days. Analysis of nicotine compound was carried out by thin layer chromatography (TLC) method. The TLC method absorbent silica gel GF254 as stationary phase and solvent system containing methanol-ammonia (100: 1,5; v/v) as mobile phase were used. The relative concentration of nicotine was observed by gas chromatography (GC) method. The parameters observed in this study include determination of microspore development stage, viability of the embriogenic microspores, percentage of binucleate to multinucleate stage of embryoid development, and nicotine compound analysis. The results showed that the embriogenic microspores were produced from all stress pretreatments toward normal microspores. Stress pretreatment for 7 days produced the best result that is 3,90 ± 0,21 % multinucleate embryoid, the presence of nicotine was observed at day 45, and stress pretreatment for 3 and 7 days produced 1.1087% nicotine and 1.3192% respectively.

Kata Kunci : Biologi Tanaman,Pemuliaan Tanaman,Mikrospora,Induksi Embriogenesis, Nicotiana tabacum “vorstenlanden”, androgenesis, microspores, nicotine