Regenerasi Kultur Lengkeng Dataran Rendah CV. Diamond River melalui Embriogenesis Somatik

. Beberapa kultivar lengkeng toleran dataran rendah telah diintroduksi ke Indonesia termasuk lengkengcv. Diamond River. Kultivar tersebut telah dibudidayakan secara komersial di daerah Kalimantan Barat. Namun,pengembangannya menghadapi kendala dalam hal penyediaan bibit. Dalam rangka memperoleh bibit lengkengdalam jumlah yang berlimpah, perlu penerapan teknik kultur in vitro. Penelitian ini bertujuan untuk menginduksidan meregenerasikan kalus embriogenik lengkeng cv. Diamond River. Induksi kalus dilakukan menggunakan daunmuda sebagai eksplan. Regenerasi kalus embriogenik dilakukan dalam 4 tahap. Pada tahap pertama digunakan airkelapa pada konsentrasi 5 dan 10%. Pada tahap kedua, diuji pengaruh auksin (IBA dan NAA) serta sitokinin (BAdan kinetin) masing-masing pada taraf 0,5 ppm. Pada tahap ketiga, diuji pengaruh auksin IBA dan NAA pada taraf0,1; 0,5; dan 1 ppm. Pada tahap keempat diuji perlakuan sukrosa pada taraf 2 dan 3% dengan atau tanpa auksin(IBA dan NAA) masing-masing pada taraf 0,5 dan 1 ppm. Hasil penelitian menunjukkan bahwa regenerasi melaluiembriogenesis somatik berpeluang diterapkan pada tanaman lengkeng cv. Diamond River. Respons kalus embriogeniklebih dominan ke arah pembentukan akar daripada tunas. Penggunaan media yang mengandung NAA 1 ppm mampumeningkatkan pembentukan tunas hingga mencapai lebih dari 30%, sedangkan penggunaan sukrosa 3% tanpa auksinmampu meningkatkan pembentukan planlet hingga mencapai 12%. Persentase keberhasilan aklimatisasi adalahsebesar 14%

Regenerasi Kultur Lengkeng Dataran Rendah cv. Diamond River melalui Embriogenesis Somatik

ABSTRAK. Beberapa kultivar lengkeng toleran dataran rendah telah diintroduksi ke Indonesia termasuk lengkengcv. Diamond River. Kultivar tersebut telah dibudidayakan secara komersial di daerah Kalimantan Barat. Namun,pengembangannya menghadapi kendala dalam hal penyediaan bibit. Dalam rangka memperoleh bibit lengkengdalam jumlah yang berlimpah, perlu penerapan teknik kultur in vitro. Penelitian ini bertujuan untuk menginduksidan meregenerasikan kalus embriogenik lengkeng cv. Diamond River. Induksi kalus dilakukan menggunakan daunmuda sebagai eksplan. Regenerasi kalus embriogenik dilakukan dalam 4 tahap. Pada tahap pertama digunakan airkelapa pada konsentrasi 5 dan 10%. Pada tahap kedua, diuji pengaruh auksin (IBA dan NAA) serta sitokinin (BAdan kinetin) masing-masing pada taraf 0,5 ppm. Pada tahap ketiga, diuji pengaruh auksin IBA dan NAA pada taraf0,1; 0,5; dan 1 ppm. Pada tahap keempat diuji perlakuan sukrosa pada taraf 2 dan 3% dengan atau tanpa auksin(IBA dan NAA) masing-masing pada taraf 0,5 dan 1 ppm. Hasil penelitian menunjukkan bahwa regenerasi melaluiembriogenesis somatik berpeluang diterapkan pada tanaman lengkeng cv. Diamond River. Respons kalus embriogeniklebih dominan ke arah pembentukan akar daripada tunas. Penggunaan media yang mengandung NAA 1 ppm mampumeningkatkan pembentukan tunas hingga mencapai lebih dari 30%, sedangkan penggunaan sukrosa 3% tanpa auksinmampu meningkatkan pembentukan planlet hingga mencapai 12%. Persentase keberhasilan aklimatisasi adalahsebesar 14%.ABSTRACT. Roostika, I., V.N. Arief, and N. Sunarlim. 2009. Regeneration of Lowland Longan cv. DiamondRiver through Somatic Embryogenesis. Several low-land longan cultivars have been introduced to Indonesia,including cultivar of Diamond River. This cultivar has been planted commercially and produced well in WestKalimantan. Unfortunately, the development of this cultivar was facing a problem on the availability of plantingmaterials. In order to provide large number of Diamond River seedlings, tissue culture technique was used. Theaim of the study was to induce and regenerate embryogenic calli of longan cv. Diamond River. A research on callusinduction was conducted using young leaves as explants source. Regeneration of embryogenic calli was conductedin 4 steps. The first, coconut water at the rate of 5 and 10% were used. The second, the auxin (IBA and NAA) andcytokinin (BA and kinetin) at the level of 0.5 ppm, respectively were tested. The third, the IBA and NAA at thelevel of 0.1, 0.5, and 1 ppm were used. The fourth, the sucrose at the level of 2 and 3% with or without addition ofIBA and NAA at the level of 0.5 and 1 ppm were used respectively. The results showed that somatic embryogenesisregeneration was potentially applied to longan cv. Diamond River. The root formation was more dominant than theshoot formation. The use of 1 ppm NAA could increase the shoot formation up to more than 30% whereas the useof 3% sucrose without auxin could increase the plantlet formation up to 12%. The 14% of plantlet produced by thistechnique grew well during acclimatization period

Kriopreservasi Tanaman Obat Langka Purwoceng Dengan Teknik Enkapsulasi-Vitrifikasi

Pruatjan (Pimpinella pruatjan Molk.) is an Indonesian endangered medicinal plant, so that it is highly protected. Cryopreservation can be applied to this plant for long-term preservation. The aim of this research was to obtain a method of encapsulation-vitrification by optimizing each step in cryopreservation protocol i.e. preculture, loading, dehydration with and without freezing in liquid nitrogen. The best treatment of each step would be applied in the following step. On preculture experiment, in vitro shoots were planted on the Driver and Kuniyaki (DKW) basal media containing 0.3 M sucrose and incubated for 1, 2, 3, 4, and 5 days. After those incubation period, shoot tips were encapsulated with 2.5% Na-alginate and soaking for 15 minutes in 100 ppm CaCl2 solution before planting. On loading experiment, precultured explants were loaded in DKW basal solution containing 2 M glycerol and 0.4 M sucrose for 0, 30, 60, and 90 minutes. On dehydration experiment, preculturead and loaded explants were dehydrated with PVS2 solution PVS2 (DKW + 30% glycerol + 15% DMSO + 15% ethyleneglicol + 0.4 M sucrose) for 0, 30, 60, 90, and 120 minutes. The parts of them were freezed in liquid nitrogen (-196oC). The result showed that cryopreservation through encapsulation-vitrification technique could be applied on pruatjan. The best preculture treatment was 5 days incubation period. The best loading treatment was 30 minutes. The best dehydration treatment was 90 minutes. The successful level of this research was still low (10%) so that it needs optimization method

Teknik Isolasi Dan Kultur Protoplas Tanaman Padi

Protoplastfusion or somatic hybridization technology is an alternativetechnology for production hybrids of plants that are difficultto be produced by conventional methods due to their sexualincompatibility. An experiment was conducted to developtechniques for isolation, purification, and culture of riceprotoplasts of cultivar IR64 and a wild rice species (Oryzaofficinalis). Optimization of protoplast isolation and purificationmethods from both rice genotypes were successfullydone. The highest protoplast density was obtained bydigesting embryonic callus or stems of young seedling in anenzyme solution containing of 2% cellulose, 0.1% pectolyase,0.5% macerozyme, 0.5% driselase, 5 mM ES, and 13% mannitolin CPW solution. The protoplast digestion was done forthree hours by soaking in the enzyme solution followed byshaking at 50 rpm under a room temperature. Purification ofthe protoplasts were done by separating them from plantdebris using a 25% sucrose solution. Protoplast regenerationwas not successful using although different media compositionsand conditions. Growth process from cell division tocell aggregate was only successful on IR64 protoplast cultureon a medium that contained AgNO3