Embriogenesis Somatik dari Eksplan Daun Anggrek Phalaenopsis sp L. (Somatic Embryogenesis from Leaf Eksplant of Phalaenopsis Orchids) (original) (raw)
Related papers
Embriogenesis somatik dari eksplan daun anggrek phalaenopsis sp L
2009
Somatic embryogenesis has been recoqnized as one of the process on plant micropropagation techniques. This process occured through regeneration by direct embryo formation and through an intermediary callus phase. This research was conducted through an intermediary callus phase. The experiment was initiated with callus induction from leaf explant on five modifications of MS medium i.e :1/2MS without plant hormone (MI-0); ½ MS containing 1mg/L BA + 0.5 mg/L 2.4-D + 1mg/L NAA (MI-1);1/3 MS containing 2 mg/L 2.4-D (MI-2); ½ MS supplemented with 0.5 mg/L 2.4-D + 0.5 mg/L BAP +0.2 mg/L thidiazuron (MI-3); ½ MS supplemented 2 mg/L thidiazuron and 1 mg/L BAP (MI-4). After the tissues were swollen, the explants were placed on callus proliferation medium ½ MS supplemented with 0.2 mg/L thidiazuron and 0.5 mg/L 2.4-D (MP). After two months, calli were regenerated in regeneration medium ½ MS supplemented with 0.4 mg/L BAP and 0.2 mg/L 2.4-D (MR). The results of this research showed that MI-1 and MI-3 were the best swelling explant mediums before the callus produced in both MP and MR medium. Callus produced was increased in every subculture. However, the level of callii production decreased on the following subculture. Plantlets were regenerated from somatic embryos derived from callii on MR medium. The results of this study may contribute to our advancement of scientific knowledge achievements tissue culture techniques to support inconventional plant improvement.
International Journal on Advanced Science, Engineering and Information Technology, 2016
Papilionanthe hookeriana Rchb.f. (Orchidaceae), popularly known as 'Pencil orchid' in Indonesia, is a perennial ephiphyte orchid, found only at Dendam Tak Sudah Lake in Bengkulu. The aims of this research were to find the best sterilization technique of P. hookeriana explant and to induce maximum formation of embryogenic calli. Rapid multiplication of this orchid was achieved through culture of shoot tips and young leaf segments of mature plants by in vitro cultured on Murashige and Skoog (MS) basal medium enriched with 50 g L-1 banana pulp. The experiment used Completely Randomized Design (CRD) with three replications. In the first stage, the explants were sterilized using three compositions of sterilizaton material. In the second stage, the explants were planted on the MS basal medium with addition of five levels of 2,4-Diclhorophenoxy Acetis Acid concentrations, namely 0, 0.25, 0.50, 0.75, and 1.00 mg L-1. The result showed that the best sterilization method to reduce explant contaminant was method 3, in which the explant was washed with detergent, rinsed with flowing water, soaked in 0.1% (v/v) HgCl 2 solution for 30 minutes, soaked again in the 10% (v/v) Sodium hypochloride solution for 20 minutes, rinsed three times with sterile water before planted and then soaked in sterile water added 10% (v/v) betadhine before planted on treated medium. This method was able to reduce contamination levels up to 70% from explants cultured for 5 months on MS medium. MS medium added with 1.00 mg L-1 2,4-D produced the highest number of embryogenic calli, and the biggest callus diameter (3.5 cm), characterized by transparent green color and friable callus structure.
Optimizing Tissue Culture Media for Efficient Callus Induction and Regeneration from Rice Seeds
2018
An efficient system was established for high frequency embryogenesis and regeneration of a local variety, NEH Megha Rice1 (Oryza sativa L.). In the present study Callus was induced on MS (Murashige and Skoog) media supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), 0.5 g/L L-proline, 0.3 g/L casein hydrolysate 30 g/L sucrose and 8 g/L agar. NEH Megha Rice1 showed maximum callus induction frequency on MS medium supplemented with 2.5 mg/L 2,4-D. The regeneration efficiency from healthy embryogenic calli was investigated on MS medium supplemented with various concentrations of BAP (6-Benzylaminopurine) with a fixed concentration of NAA (Naphthalene acetic acid). The regeneration efficiency of calli was also investigated on three different agar concentrations. NEH Megha Rice1 showed maximum regeneration frequency on MS medium supplemented with 5mg/L BAP and 0.05mg/L NAA in 1.2 % agar.
Somatic embryogenesis and plant regeneration from callus cultures of Cleome rosea Vahl
2010
This paper describes a protocol for the efficient vegetative propagation of Cleome rosea by somatic embryogenesis. Leaf and stem explants from nursery-grown seedlings of C. rosea were cultivated on Murashige and Skoog (MS) medium supplemented with indole-3-acetic acid (IAA), a -naphthaleneacetic acid (NAA), 4-amino-3,5,6-trichloropicolinic acid (picloram) or 2,4-dichlorophenoxyacetic acid (2,4-D). Nodular calli were produced from both explant types in the presence of 4.5 and 9.0 µM 2,4-D. Embryo development and maturation were achieved when calli from stem explants were transferred to media containing a ten-fold reduction of 2,4-D concentration initially used (0.45 and 0.90 µM). Leaf-derived calli did not form embryos with the same treatments. The highest frequency of embryogenic callus formation (85%) and number of embryo per callus (13.45 ± 2.8) were achieved during the first subculture on medium supplemented with 0.90 µM 2,4-D. Embryo conversion into plantlets was achieved follow...
Tissue Culture, In Vitro Organogenesis and Regeneration of
Introduction: Plantago lanceolata is one the most important species of Plantago genus and has valuable medicinal secondary metabolites. Materials and Methods: The effect of different factors on germination of P. lanceolata seeds was studied and leaf and root explants of in vitro growth seedling were cultured on Murashige and Skoog (MS) medium supplemented with combination of 6-benzylaminopurine (BAP) or thidiazuron (TDZ) (0, 0.5, 1, 1.5, 2 and 3 mg/L) and auxins: α-naphthaleneacetic acid (NAA) (0, 0.2, 0.5, 1 mg/L), indole-3-acetic acid (IAA) (0.1, 0.5, 1, 1.5 mg/L) or indole-3-butyric acid (0.5, 1, 1.5, 2 mg/L) (IBA) at different concentrations. Results: The results showed that cold pre-treatment, daylight and 1/8 MS salt concentration are more suitable for high germination. The best shoot organogenesis rate (95%) in leaf explants was observed in 1:1 mg/L, and 1.5:1 mg/L TDZ: IBA. The highest percentage of shoot organogenesis (100%) was observed in most of the plant growth regulator (PGR) treatments in root explants. About 58.67 and 60 shoot numbers obtained with 2 mg/L TDZ in leaf and 1:1.5 mg/L BAP: IBA in root explants, respectively. Conclusions: It can be suggested that the best shoot organogenesis and proliferation medium is MS basal medium containing cytokinin TDZ and auxin IBA in comparison with other hormone compounds on leaf and root explants. The result behind this fact is high callus induction and regeneration potential of explants in all concentrations.
In Vitro Cellular & Developmental Biology - Plant, 2001
Embryogenic calluses were induced from 73% of Phalaenopsis shoot-tip explants excised from flower stalk buds by culturing for 7 mo. on New Dogashima Medium (NDM) containing 0.5 mM a-naphthaleneacetic acid (NAA), 4.4 mM 6-benzylaminopurine and 29.2 mM sucrose. The sucrose concentration was increased to 58.4 mM 4 mo. after initiation of the callus culture. These calluses were successfully subcultured as cell suspension cultures in liquid NDM supplemented with 5.4 mM NAA and 58.4 mM sucrose. By simply reducing the sucrose concentration to 29.2 mM, the cells grew into plantlets through a developmental process similar to that of Phalaenopsis seedlings. The occurrence of somaclonal variants was less than 10% in six out of eight genotypes examined. These results suggest that the embryogenic callus and cell suspension culture could be utilized as the materials for micropropagation and breeding of Phalaenopsis orchids.
1986
Embryogenic callus was initiated from radicles of mature embryos removed from imbibed seeds (24 h). Embryogenic and other nonembryogenic types of callus proliferated on a modified half-strength Murashige-Skoog medium (MS) basal medium (BM) supplemented withmyo-inositol, casein hydrolysate (CH), L-glutamine (gln) and growth regulators kinetin (KN), N6-benzyladenine (BAP) each (20×10−6M), 2,4-dichlorophenoxyacetic acid (2,4-D) (50×10−6M) Embryogenic callus bearing suspensor-like cells in a mucilaginous gel matrix was isolated and maintained by subculture every 10 to 12 days on BM with KN, BAP each (2×10−6M) and 2,4-D (5×10−6M). Somatic embryos developed spontaneously from the callus on this medium at 23±1° C. Closer examination revealed that numerous polyembryonic clusters, comprised of elongated cells (suspensors) and small dense cells with large nuclei (somatic embryos), occurred in the viscous gel. When this enriched embryonal-suspensor mass was subcultured to low 2,4-D (1×10−6M), globular embryos developed by 40 to 60 days. Upon transfer to a liquid medium without growth regulators, the embryos elongated and developed cotyledons and shoots with needles. Plantlet development was completed by 30 days in a basal medium without CH, gln and growth regulators. The total culture time was 150 days. Approximately 40±10 embryos were formed from 500 mg of initial callus. Somatic embryogenesis became aberrant if embryos remained attached to the callus mass and were not subcultured within 10 to 12 days according to the described protocol. Somatic embryos were encapsulated in an alginate gel and stored at 4° C for nearly two months without visible adverse effects on viability.
Somatic embryogenesis and plant regeneration from callus cultures of Phlox paniculata Linn
Scientia Horticulturae, 2002
Callus was obtained from leaf explants of Phlox paniculata on Murashige and Skoog (MS) basal medium supplemented with 0.5 mg/l BA, 2.0 mg/l IAA and 3% (w/v) sucrose. Somatic embryogenesis was achieved when this callus was subcultured on MS medium supplemented with 1.0±2.0 mg/l BA, 0.25±0.5 mg/l IAA and 3% (w/v) sucrose. The frequency of somatic embryogenesis was greater in callus derived from immature than mature leaf explants. Somatic embryos proliferated rapidly by secondary somatic embryogenesis after transfer to MS medium supplemented with 1.5 mg/l BA, 0.5 mg/l IAA and 3% (w/v) sucrose. Somatic embryos germinated and developed into normal plantlets on half-strength MS medium supplemented with 2% (w/v) sucrose without growth regulator. Embryogenic calli maintained their regeneration capacity for 15 months. About 92% of the somatic embryo-derived plantlets were acclimatized in the greenhouse and successfully transferred to ®eld conditions.