Pemanfaatan Teknik Kriopreservasi Dalam Penyimpanan Plasma Nutfah Tanaman

The two basic approaches for conservation of plant genetic resources are ex situ and in situ conservation. Cryopreservation is a potential method for long term preservation of plant germplasm. Cryopreservation technique could be used to preserve plant materials with unlimited time (until 20 years) because it bring the materials to metabolically inactive state and completely arrest the growth in liquid nitrogen at –196oC. This method is more efficient in terms of cost, time, space, and labour because it does not require frequenlty subculture. Cryopreservation techniques can be divided in to classical technique and new techniques. Classical technique can be applied to limited species but new techniques can be applied to wide range of species and many types of explant. The success of cryopreservation is not only indicated by the high rate of survival and regenerated culture, but also on the stability level of culture after cryopreservation

Regenerasi Tanaman Sedap Malam melalui Organogenesis dan Embriogenesis Somatik

Secara konvensional perbanyakan tanaman sedap malam dilakukan melalui umbi. Semakin kecil ukuran umbi semakin lama tanaman berbunga. Penerapan teknik kultur in vitro diharapkan dapat membantu perbanyakan tanaman secara masal. Hingga saat ini, teknik kultur in vitro tanaman sedap malam belum pernah dilaporkan di Indonesia. Penelitian ini bertujuan memperoleh formulasi media yang efektif menginduksi organogenesis dan embriogenesis kultur in vitro tanaman sedap malam serta memacu regenerasinya. Percobaan dibagi menjadi 4 tahap, yaitu (1) induksi tunas, (2) multiplikasi tunas, (3) induksi kalus embriogenik, dan (4) regenerasi kalus embriogenik. Media induksi tunas yang diuji adalah MS+BA 0 ppm, MS+BA 3 ppm, MS+BA 5 ppm, dan MS+BA 7 ppm. Pemacuan multiplikasi tunas lanjut dilakukan pada media subkultur MS+BA 7 ppm+glutamin 100 ppm, MS+BA 7 ppm, DKW+TDZ 7 ppm, dan DKW+TDZ 7 ppm+glutamin 100 ppm. Untuk induksi kalus embriogenik, media induksi kalus yang diujikan adalah MS+2,4-D 2,5 ppm, MS +2,4-D 5 ppm, dan MS+2,4-D 10 ppm. Untuk meregenerasikan kalus embriogenik, media yang diujikan MS+BA 2 ppm+TDZ 0,2 ppm, MS+BA 3 ppm+TDZ 0,4 ppm, MS+zeatin 1ppm+kinetin 1ppm, dan MS+zeatin 0,5 ppm+kinetin 2 ppm. Hasil percobaan menunjukkan bahwa pembentukan tunas terbanyak diperoleh dari media BA 3 ppm (80%) namun inisiasi tunas tercepat dihasilkan pada media BA 0 ppm. Formula media MS+BA 7 ppm+glutamin 100 ppm menghasilkan jumlah tunas dan akar terbanyak. Penggunaan MS+2,4-D 5 ppm dapat menginduksi kalus embriogenik dengan persentase pembentukan nodul sebesar 18,75% dan jumlah nodul yang terbentuk sebanyak 3,6 dengan visual kalus yang paling baik. Setelah disubkultur, calon tunas terbanyak (17) dihasilkan dari perlakuan MS+BA 2 ppm+TDZ 0,4 ppm. Kalus embriogenik pada media MS+zeatin 0,5 ppm+kinetin 2 ppm dapat berkembang membentuk benih somatik.Regeneration of tuberose through organogenesis and embryogenesis. Tuberose is normally propagated by the tuber. The smaller size of tuber the longer time plant to flower. The application of in vitro culture technique might be used for mass propagation. Up to know, the research of in vitro culture of tuberose in Indonesia has not been reported. The objective of the study was to find out media formulation for organogenesis and embryogenesis. The experiments consisted of 4 steps of (1) shoot induction, (2) shoot multiplication, (3) induction of embryogenic callus, and (4) regeneration of embryogenic callus. The treatments for shoot induction were MS+BA 0 ppm, MS+BA 3 ppm, MS+BA 5 ppm, and MS+BA 7 ppm. The shoots were multiplied on media MS+BA 7ppm+glutamine 100ppm, MS+BA 7 ppm, DKW+TDZ 7 ppm, and DKW+TDZ 7 ppm+glutamin 100 ppm. For induction of embryogenic callus, the treatments were MS+2.4-D 2.5 ppm, MS+2,4-D 5 ppm, and MS+2.4-D 10 ppm. For regeneration of embryogenic callus, the treatments were MS+BA 2 ppm+TDZ 0.2 ppm, MS+BA 3 ppm +TDZ 0.4 ppm, MS+zeatin 1ppm+kinetin 1ppm, and MS+zeatin 0.5 ppm+kinetin 2 ppm. The results showed that the highest shoot formation was obtained from media MS+BA 3 ppm but the earliest shoot initiation was obtained from media MS+BA 0 ppm. The media formulation of MS+BA 7 ppm+glutamine 100 ppm gave the highest number of shoot and root. The application of media MS+2.4-D 5 ppm could induce embryogenic callus with high percentage of nodul formation (18.75%) and high number of nodul (3.6) with the best visual calli. After subculturing, the highest number of nodul (17) was obtained from media MS+BA 2 ppm+TDZ 0.4 ppm. The embryogenic callus from media MS+zeatin 0.5 ppm+kinetin 2 ppm could develop to form somatic seed

Pengujian Nomor-nomor Harapan Padi Tahan Al Dan PH Rendah Hasil Seleksi in Vitro Dengan Kultur Hara

Rice productivity in acid soil is very low because of low pH,low availability of N, P, K, Ca, Mg, Mo, toxicity of Al and Mn.Development of Al tolerant varieties could increase riceproductivity in acid soil. Somaclonal variation and in vitroselection method can be used to develop new Al tolerancevarieties. A rapid screening method is needed to select alarge number of new genotypes or new inbred lines in plantbreeding, such as solution culture methods to evalu-ate Altolerantrice. This methods was used to know the responseto Al in the seedling stage, root development, and pHchanging. In this experiment solution culture method wasused to evaluate the new genotypes derived from somaclonalvariation and in vitro selection methods. These newgenotypes have been tested the tolerance characteristic byusing AlCl36H2O at 6 concentrations (0, 100, 200, 300, 400,and 500 ppm). Yoshida solution with two Al concentrationwere used to tested these genotypes. Measurement of Altolerance was based on root development by using RelativeRoot Length (RRL), the relativity of root length at 45 ppm and0 ppm. Almost all of the genotypes have RRLs higher than0.7, which means that there was a positive correlationbetween the in vitro method and solution culture method. Inthis experiment pH changes were not applicable to measurethe tolerance of the rice genotypes to Al and low pH

In Vitro Culture Manipulation on Pruatjan for Secondary Metabolite Production

Purwoceng (Pimpinella pruatjan Molk. atau Pimpinellaalpina KDS.) adalah tanaman obat langka yang dapat dimanfaatkansebagai bahan obat afrodisik, diuretik, dan tonik.Kultur in vitro tidak hanya dapat digunakan untuk konservasidan perbanyakan tanaman, melainkan dapat juga diterapkanuntuk produksi metabolit sekunder. Melalui teknik ini,produksi metabolit sekunder tidak bergantung kepada sumbertanaman di lapang. Penelitian ini dilakukan dengan tujuanuntuk meningkatkan kadar stigmasterol melalui kultur invitro dengan menggunakan prekursor asam mevalonat. Penelitiandibagi menjadi dua tahap, yaitu induksi kalus danmanipulasi kultur in vitro untuk meningkatkan kadar stigmasterol.Pada tahap induksi kalus, terdapat 16 perlakuan yangmerupakan kombinasi perlakuan 2,4-D dan piklorammasing-masing pada taraf 0,5; 1,0; 1,5; dan 2,0 ppm. Untukmeningkatkan kadar stigmasterol, digunakan asam mevalonatpada taraf 0, 250, 500, dan 750 ppm dengan masa inkubasiselama 4 dan 6 minggu. Kandungan stigmasterol dianalisismenggunakan GC-MS. Hasil penelitian menunjukkanbahwa media P2 (DKW + 2,4-D 0,5 ppm + pikloram 1,0ppm) adalah media terbaik untuk induksi kalus. Eksplan daunlebih baik daripada eksplan petiol. Hasil analisis GC-MSmenunjukkan bahwa kandungan stigmasterol tertinggi(0,0356 ppm) diperoleh dari kalus dengan masa inkubasi 4minggu pada media dengan penambahan asam mevalonat250 ppm. Peningkatan taraf asam mevalonat tidak mampumeningkatkan kandungan stigmasterol. Kadar tersebut miripdengan kandungan stigmasterol pada planlet dari GunungPutri (0,0365 ppm) dan Dieng (0,0414 ppm). Dibandingkandengan kadarnya dalam akar tanaman dari lapang, kandungantersebut sekitar 10-100 kali lipat lebih tinggi

Indirect Organogenesis and Somatic Embryogenesis of Pineapple Induced by Dichlorophenoxy Acetic Acid

This research aimed to study the effect of 2,4-D,AdS, and basal media to the regeneration of pineapplethrough indirect organogenesis and somatic embryogenesis,and to study the complete event of somatic embryogenesis.Callus formation was induced by 21, 41, and 62 μM 2,4-Dwith addition of 9 μM TDZ. The non embryogenic calli weretransferred onto 4.65 μM Kn containing medium.Embryogenic callus formation was induced on MS or Bacbasal media consisted of N-organic compounds withaddition of AdS (0, 0.05 and 0.1 μM). The embryogenic calliwere regenerated on modified MS medium with addition of0.9 μM IBA, 1.1 μM BA, 0.09 μM GA3 or MS mediumsupplemented with 0.018 mM BA. The result proved that thesingle auxin of 2,4-D was not enough to induce embryogeniccells. Therefore the non embryogenic calli were regeneratedthrough organogenesis. The 21 μM 2,4-D yielded high level ofcallus formation (80%), higher fresh weight (0.2 g/explant)and higher number of shoot (25 shoots/explant in twomonths). Embryogenic calli were produced on N-organiccompounds enriched media. The regeneration mediumsignificantly affected the level of browning, where the MSmedium with addition of 0.018 mM BA yielded lower level ofbrowning. There was an interaction of embryogenic callusinduction medium and regeneration medium to the numberof mature somatic embryos. The embryogenic callusinduction on MS medium enriched with N-organiccompounds and 0.05 μM AdS followed by the regenerationof somatic embryos on MS medium with addition of 0.018mM BA was the best treatment which yielded 17 maturesomatic embryos/explant

Multiplikasi Tunas Belimbing Dewi (Averrhoa Carambola) Melalui Kultur in Vitro

Star fruit (Averrhoa carambola) is one of tropical fruits which had a high content of vitamin C, which was higher than that in apple and grape. As fresh consumption, star fruit had a good role in decreasing human blood pressure. Main constraints of star fruit development weather for conservation purpose and or for cultivation were still limited due to lack of seedlings availability. In vitro culture technique was one of the altrnative technologies capable of producing seedlings in a large quantity, uniform growth and relatively in a short period. One of the important keys in micropropagation work was the step of shoot initiation and multiplication. In this study we used two kind of explant, namely shoot with single node and shoot from germinated embrio. Experiment I, shoot with single node and shoot from germinated embrio were planted at WPM media + citric acid 100 mg/l. The next activities was focused on single node shoots which was subcultured at WPM + BAP 0.5 mg/l. In experiment II in vitro shoots from previous experiment was subcultured at WPM + BA (1 and 2 mg/l) + thidiazuron (0.1 and 0.2 mg/l). To stimulate shoot multiplication rate, shoot was subcultured at WPM or MS media in combination with IAA 0.5 mg/l and zeatin 2 mg/l. To improve vigourity of the plant, in vitro shoots resulted from multiplication media was planted at WPM or MS media containing paclobutrazol (0.4 and 0.8 mg/l) + BA 2 mg/l + thidiazuron 0.2 mg/l. Result showed that the use of single node shoot as an explant better than shoot comes from germinated embrio. Sub culture of star fruit shoot on WPM basal media containing BAP of 0.5 mg/l produce the shoot number about 4, and the shoot number could be increased until 18 by using IAA 0.5 mg and zeatin 2 mg/l. The treatment of schock temperature at 4-5oC during 4 days before planting could fasten shoot initiation time from 3 months to 1 months. An addition of 0.4 mg/l paclobutrazol on MS or WPM media containing 2 mg/l BA and 0.2 mg/l thidiazuron could improve vigourity of plantlet