The research was undertaken to obtain drought-tolerant patchouli putative mutant calli through gamma-irradiation and in-vitro selection using Polyethylene Glycol (PEG). Patchoulina 2 variety was used as the mother plant. Embryogenic calli were induced on three medium formulation (combination of 0.1; 0.3 and 0.5 mg/l 2.4D with 0.1 mg/l BAP). The induced embryogenic calli were then exposed to nine levels of Gamma irradiation (0, 5, 10, 15, 20, 25, 30, 35, and 40 Gray) in combination with two levels of PEG (0 and 20%). The results showed that MS media with 2.4-D 0.3 mg/l and BAP 0.1 mg/l was the best medium for callus induction. The ability of calli to survive decreased with increasing doses of gamma irradiation. The radiosensitivity of patchouli calli showed that LD20 at 14.06 Gray and LD50 at 26.98 Gray. The drought-tolerant patchouli putative mutant calli were obtained from gamma-ray irradiation doses 15, 20, and 25 Gray in selection media 20% PEG has better drought stress tolerance compared to parent Patchoulina 2 variety. Further selection in the glasshouse and field are needed to obtain a candidate variety of patchouli drought tolerance.
World patchouly oil need is almost 90% fulfilled by Indonesia as the main producer, however, their narrow genetic variation is a major obstacle in plant breeding. Chemical mutation such as colchine is believed to be able to wider patchouly genetic variation The aim of this study was to increase patchouly genetic variation using chemical mutagen. The variety of Patchoulina 2 was treated by colchicine on callus stage. Callus irradiated was grown into regeneration medium until formed plantlet and ready to acclimatization. The results showed that there were 15 different morphological putative mutants with different characters. The mutants varied in qualitative characters(leaf shape, leaf color, leaf base, and leaf edge)and quantitative characters(plant height, number of leaf stalks). Two distinguishable main groups of mutants shared 62.62% genetic similarity level based on morphology. These new mutants in this study would be potential to be developed as new varieties along with the morphological characters as their specific identities.
ABSTRAKNilam merupakan tanaman penghasil minyak atsiri, oleh sebab itu produksi, kadar dan mutu minyak merupakan faktor penting yang menetukan keunggulan suatu varietas. Tujuan dari penelitian ini adalah mengetahui stabilitas hasil dan mutu lima somaklon harapan nilam di tiga agroekologi. Penelitian dilaksanakan di Banten (100 m dpl), Kuningan (400 m dpl) dan Purwokerto (600 m dpl) selama dua tahun (2011 sampai 2012). Somaklon harapan A, B, C, D, E dan varietas Sidikalang sebagai pembanding ditanam dengan rancangan acak kelompok (RAK) empat ulangan. Data perbedaan lima somaklon dianalisa menggunakan uji Duncan pada taraf lima persen. Stabilitas hasil dianalisa berdasarkan metode Eberhart dan Russell. Parameter yang diamati adalah produksi terna (basah dan kering angin), produksi minyak, kadar minyak dan mutu minyak (kadar patchouli alkohol). Hasil penelitian diperoleh somaklon harapan nilam A dan E yang stabil pada hasil terna, kadar minyak dan mutu minyak serta mampu beradaptasi pada semua tempat pengujian. Somaklon harapan nilam A unggul pada produksi terna basah per tahun 36,62 t ha , kadar minyak 2,78%, kadar patchouli alkohol 32,31%. Perbedaan somaklon harapan A dan E terletak pada bentuk daunnya. Somaklon harapan A mempunyai bentuk pangkal daun tumpul (obtusus), ujung daun runcing-tumpul (acutus-obtusus) serta tepi daun bergerigi tumpul (crenatus). Somaklon harapan E mempunyai bentuk pangkal daun runcing (acutus), ujung daun runcing (acutus) serta tepi daun bergerigi tajam ganda (biserratus). Kata kunci: Pogostemon cablin, variasi somaklonal, hasil, mutu, adaptasi ABSTRACT Patchouli is an essential oil plant, therefore the production, content and quality of essensial oil is to be determined the benefits of a variety. The objective of the research was to evaluate morphological characters, yield potency and quality of five promising numbers at three agroecology. The research was conducted at three agroecology i.e Banten (100 m dpl), Kuningan (400 m dpl) and Purwokerto (600 m dpl) for two years (2011 until 2012). Somaclones A, B, C, D, E and F Sidikalang variety as a control planted in Randomized BlockDesign with four replications. Data were analized by using were DMRT at five percent. Analysis of the stability was performed using the method of Eberhart and Russell. Parameters observed were morphological characters, herbs production, oil content and oil quality (patchouli alcohol
ABSTRAKNilam banyak dibudidayakan di Indonesia, dan lebih dari 80% produksi minyak nilam dunia dipasok dari Indonesia. Masalah utama dalam budidaya nilam di Indonesia adalah penyakit, seperti penyakit layu bakteri, budok dan nematoda. Sampai saat ini varietas tahan terhadap penyakit, khususnya budok, belum diperoleh. Tanaman tahan dapat diperoleh dengan teknik transformasi gen. Transkripsi faktor WRKY telah diketahui dapat meregulasi serangan beberapa patogen penyebab penyakit tanaman. Gen OsWRKY76 terletak pada segmen kromosom 9 tanaman padi yang telah diidentifikasi terkait dengan ketahanan berspektrum luas. Penelitian bertujuan untuk mengintroduksikan konstruksi gen OsWRKY76 yang berasal dari padi ke dalam tanaman nilam melalui bantuan Agrobacterium tumefaciens. Pada percobaan pertama, tanaman nilam ditransformasi dengan A. tumefaciens strain EHA 105 yang mengandung gen OsWRKY76. Perlakuan terdiri atas waktu induksi eksplan yang akan ditransformasi (pre-kultur) di dalam medium MS, yaitu 5 dan 7 hari, dan waktu inokulasi A. tumefaciens yaitu 10 dan 20 menit. Pada percobaan kedua, analisis molekuler untuk mengkonfirmasi keberadaan gen OsWRKY76 dalam tanaman nilam menggunakan teknik PCR dengan primer hptII. Hasil penelitian menunjukkan waktu induksi eksplan terbaik sebelum transformasi adalah 5 hari, dengan perendaman di dalam suspensi A. tumefaciens selama 10 menit. Dari transformasi tersebut telah dihasilkan 187 kalus independen. Hasil analisis PCR terhadap galur-galur putatif transgenik independen, lima galur (T1, T8, T10, T11, T13) positif mengandung gen hptII, yaitu gen penanda ketahanan terhadap antibiotik higromisin yang berada satu konstruk dengan gen OsWRKY76. Hasil tersebut menunjukkan bahwa gen OsWRKY76 yang berasal dari padi dapat diintroduksikan pada tanaman nilam dan berpeluang sebagai kandidat tahan terhadap penyakit utama.
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