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Induction, multiplication, and acclimatization of rodent tuber (Typhonium flagelliforme Lodd.) plant from Indonesia by in vitro organogenesis

Rodent tuber (Typhonium flagelliforme Lodd.) is a native Indonesian medicinal herb with high anticancer activity. Propagation of rodent tuber is hard to be achieved by using conventional methods. The purpose of this research is to optimize method and media for propagating rodent tuber through initiation, induction, multiplication, and acclimatization. Rodent tuber was obtained from Pekalongan, Indonesia. In vitro rodent tuber plantlets were induced through direct single node culture of tuber. Shoot induction was achieved on MS 1 mgL-1 2,4-D combined with 0.3 mgL-1 BAP and 1 mgL-1 NAA combined with 0.5 mgL-1. Induced shoots were initiated on MS media supplemented with (1, 0.5, 1.5 mgL-1) NAA and (1, 0.5, 0.5 mgL-1) BAP. Shoots were then multiplicated on MS media supplemented with (0.25, 0.5, 1, 1.5 mgL-1) NAA and 0.5 mgL-1 BAP to investigate the effect of NAA concentrations on shoot proliferation. Plantlets were acclimatizated on husk and compost (1:1) media and post-acclimatizated on husk, soil, and compost (1:1:1) media. The highest percentage of viable induced explants (66.67%) was obtained on MS 1 mgL-1 2,4-D and 0.3 mgL-1 BAP. The maximum number of shoots obtained in the initiation stage (14 shoots) was achieved on MS media supplemented with 0.5 mgL-1 NAA and 0.5 mgL-1 BAP. The maximum number of shoots obtained in the multiplication stage (8.2 ± 3.19) was achieved on MS media supplemented with 0.5 mgL-1 NAA and 0.5 mgL-1 BAP. The survival rate of rodent tuber during acclimatization period was 100% and during post-acclimatization period was 58%. Rodent tuber has been successfully multiplicated through direct organogenesis in vitro.

Bioactive compounds of fourth generation gamma-irradiated Typhoniumflagelliforme Lodd. mutants based on gas chromatography-mass spectrometry

Rodent tuber (Typhonium flagelliforme Lodd.) is an Indonesian anticancer medicinal plant. The natural genetic diversity of rodent tuber is low due to vegetative propagation. Plants genetic diversity has to be increased for obtaining clones which contain a high amount of anticancer compounds. In vitro calli were irradiated with 6 Gy of gamma ray to produce in vitro mutant plantlets. Mutant plantlets were acclimated and propagated in a greenhouse. This research was aimed to identify the chemical compounds in the leaves and tubers ofthe fourth generation of rodent tubers vegetative mutant clones (MV4) and control plantsby using GC- MS method. Leaves and tubers of MV4 each contained 2 and 5 anticancer compounds which quantities were higher compared to control plants. MV4 leaves contained 5 new anticancer compounds while its tubers contained 3 new anticancer compounds which were not found in control. The new anticancer compounds in leaves were hexadecanoic acid, stigmast-5-en-3-ol, ergost-5-en-3-ol, farnesol isomer a, and oleic acid while the new anticancer compounds in tubers were alpha tocopherol, ergost-5-en-3-ol, and beta-elemene. Rodent tuber mutant clones are very potential to be developed into anticancer drugs.

PENGEMBANGAN TANAMAN KELADI TIKUS (TYPHONIUM FLAGELLIFORME LODD.) ASAL INDONESIA SEBAGAI OBAT ANTIKANKER

Keladi tikus merupakan tanaman obat yang bermanfaat sebagai obat kanker. Keladi tikus memiliki kandungan kimia diantaranya adalah alkaloid, flavonoid, saponin, steroid dan glikosida. Keladi tikus memiliki keragaman genetik yang rendah. Untuk meningkatkan keragaman telah dilakukan mutasi secara fisik melalui iradiasi sinar gamma pada kultur in vitro keladi tikus. Media MS dengan penambahan 0.5 mg/l 2.4D dan 0.1 mg/l kinetin menghasilkan jumlah tunas terbanyak yaitu 14.38 plantlet. Kalus embriogenik mutan dihasilkan melalui induksi dengan sinar gamma 6 gray. Kalus embriogenik diregenerasikan menjadi 59 tunas mutan. Plantlet mutan yang dihasilkan telah dideteksi dengan marka molekuler RAPD dan menunjukan perubahan genetik. Beberapa galur mutan yang telah diperoleh memiliki keragaman tinggi secara morfologi dan genetik. Keragaman genetik klon mutan generasi pertama (MV1) sampai generasi keempat (MV4) yang diperoleh telah dideteksi secara genetik molekuler dengan RAPD. Beberapa klon MV5 (mutan stabil) memiliki kandungan senyawa bioaktif antikanker tinggi berdasarkan GC-MS. Analisis GC-MS menunjukkan bahwa tanaman mutan mengalami peningkatan kandungan senyawa antikanker dibanding kontrol serta senyawa baru yang tidak ditemukan di kontrol. Beberapa klon unggul yang dihasilkan perlu dikembangkan sebagai bahan baku obat antikanker. Keladi tikus unggul bermanfaat untuk mencegah dan mengobati penyakit kanker pada masyarakat Indonesia.