Regenerasi Tanaman Sedap Malam Melalui Organogenesis dan Embriogenesis Somatik - Ika Rostika; Ika Mariska; R Purnamaningsih (Indonesian Center for Horticulture Research and Development, 2005)
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.