STUDY ON THE PROTOPLAST FUSION BETWEEN MUNGBEAN [IRGNA RADIATA (L.) WILCZEK] AND ITS WILD RELATIVE [VIGNA SUBLOBATA L.]: ANATOMICAL STRUCTURE OF FUSION PRODUCT

• Main Author: Setiari, Nintya
• Format: Theses
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/5129
• Institution: Institut Teknologi Bandung

<b>Abstract :</b><p align=\"justify\"> <br /> A study of protoplasts fusion has been conducted between hypocotyls protoplast of mungbean and leaf protoplast of its wild type. The objective of this study were to investigate protoplast fusion and to observe anatomical structure of parental protoplasts and its fusion products. Protoplasts used in the fusion were isolated from hypocotyl of 5 days old mungbean seedling and the first leaves of 7 days old of its wild type seedling. Protoplasts were fused using PEG (Polyethylene Glycol) and the fusion product were then cultured in KM8p liquid medium. The viability and cell wall regeneration of parental protoplasts and fusion product were observed under fluorescence microscope, while cell division were observed under inverted microscope. The fusion. product and both parental protoplasts were fixed in 2.5% glutaraldehyde, postfixed in I %OsO4i dehydrated in alcohol series and then embedded in LR White resin. Semithin section at a thickness of 0.7 pm were made using Reichert ultramicrotoin, stained with toluidine blue and observed under light microscope. The result showed that viability of protoplasts of leaves, hypocotyl and fusion product at 2 h after culture were 87.48%, 92.1% and 86.95%, respectively. Cell wall regeneration in leaves and hypocotyl protoplasts were started 2 h after cultur, cell wall regeneration on leaves protoplasts, hypocotil protoplasts and fusion product after 8 h in culture were 64.86%, 72.51% and 75.06%, respectively. First cell division was occurred rapidly on hypocotyl protoplasts, i.e. 2d after culture. In leaves protoplasts, first division was observed after 4d in culture and in the fusion product was occurred after 3d in culture. Result on observation of fusion process and their anatomical structure of fusion product showed that there were 2 kinds of fusion product found, namely heterofusion which was occurred between leaves and hypocotyl protoplasts, and homofusion which was occurred between leaves protoplasts or hypocotyl protoplasts. From this result, it can be concluded that, the method used could induce protoplasts fusion and the fusion product can be easily distinguished from the parental protoplasts due to the concurrent existence of chloroplasts from leaves protoplasts and arryloplasts from hypocotyl protoplasts.