MICROPROPAGATION OF PHALAENOPSIS SPP. BY SOMATIC EMBRYOGENESIS TECHNIQUE

Phalaenopsis spp. was regularly produced through micropropagation by protocorm like bodies (PLBs); micropropagation takes a lot of labor, and has high cost of seedlings, energy and material. The purpose of this paper was to study the new technique of using in vitro embryogenesis culturing for microprogation. The method involved using protocorm like bodies as planting materials. PLBs were cut into slices and placed on the medium for callus initiation. The callus was initiated on the medium MS + BA (0.1 mg/l) supplemented with NAA (1 mg/l) or 2,4D (1 mg/l) and was proliferated on the medium MS + BA (0.1 mg/l) supplemented with NAA (1 mg/l). Somatic cell suspensions were initiated and proliferated on the medium MS + BA (0.1 mg/l) supplemented with NAA (0.5, 1 mg/l). Somatic cell suspensions were differentiated to embryonic cell suspensions on the MS medium supplemented with NAA (0.1 mg/l) + BA (0.5 mg/l). Embryonic cell suspensions were plated and regenerated on the medium: 1/2MS supplemented with NAA (0.1 mg/l) + BA (0.5 mg/l). Micropropagation of Phalaenopsis sp. via the embryogenesis technique was set up to produce 5,800 plantlets per one liter of somatic embryogenesis suspension.

MICROPROPAGATION OF CYMBIDIUM SPP. BY SOMATIC EMBRYOGENESIS TECHNIQUE

Cymbidium spp. was regularly produced through micropropagation by protocorm like bodies (PLBs) and multiple shoots; micropropagation takes a lot of labor, and has high cost of seedlings, energy and material. The purpose of this paper was to study the new technique of using in vitro embryogenesis culturing for microprogation to resolve the above difficulties. The method was to use PLBs as planting materials to produce somatic callus cell and embryos. Results were followed as: PLBs were cut into slices and placed on the medium for callus initiated and used as materials for embryo formation study. A fresh weight of callus were used for the experiment of about 100 mg. The medium for the initiation of embryonic callus was composed of: MS + BA (0.1 mg/l) + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1mg/l) or 2.4D (1 mg/l) and was proliferated on the medium MS + pepton (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) or 2.4D (1 mg/l). Somatic cell suspensions were initiated and proliferated on the medium consisting of MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (1 mg/l) + BA (0.1 mg/l). Somatic cell suspensions were differentiated to embryonic cell suspensions on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with BA (1 mg/l). Embryonic cell suspensions were plating and regeneration on the medium MS + peptone (1 g/l) + activated charcoal (1 g/l) supplemented with NAA (0.1 mg/l) + BA (1 mg/l). Micropropagation of Cymbidium spp. via embryogenesis technique was set up to produce 3,800 plantlets per one liter of somatic embryogenesis suspension.

In vitro Embryogenesis Derived from Shoot Tips in Mass Propagation of Two Selected-Clones of Phalaenopsis

Phalaenopsis is of high economic value and market demand in Indonesia; however, orchid products are mostly imported from other countries. ‘Kristina Dwi’ (KD) 69.274 and ‘Dedeh’ (D) 802.28 are two selected clones with high potential utilized and developed commercially. To support their commercialization, a reliable in vitro propagation protocol is essential. In the current study, an in vitro mass propagation protocol for KD 69.274 and D 802.28 clones was successfully established using shoot tips as explant sources. A high number of embryos, up to 8.2 embryos per explant, with 58.5% explant regeneration, and 3.5 regenerated-explants in average were regenerated from shoot tips of KD 69.274 clone cultured on half-strength Murashige and Skoog (MS) medium, with full strength micro, Fe-chellate and vitamin containing 0.5 mg/L thidiazuruon (TDZ) and 0.25 mg/L N6-benzyladenine (BA). The initial embryos were proliferated by culturing embryos individually on half-strength MS medium with 0.13 mg/L TDZ and 0.25 mg/L BA and resulted in high embryo regeneration up to 91.4%, with 10.2 embryos per explant and no embryo browning. The embryos were multiplied under periodical subcultures of 3 months each, resulting in gradual increasing number of embryos from the first subculture till the fifth subculture, with 23.6 embryos produced, then declined afterward. The embryos were easily germinated on half-strength MS medium with full strength of vitamin and hormone free, with 73.9% embryo germination and 14.9 germinated embryos. Healthy plantlets were stimulated on the same medium with 2 g/L activated charcoal (AC) and successfully acclimatized on Cycas rumphii bulk, with 88.3% survival plantlets. Finally, it can be summarized that a new in vitro mass propagation protocol, as new alternative choice for Phalaenopsis propagation, was successfully established.

Study of Oil Palm (Elaeis guineensis Jacq) In Vitro Embryogenesis using Young Leaves Explants

This study reported in vitro embryogenesis of oil palm using young leaves as explants. Explants were grown in solid modified MS or Eeuwens medium containing different concentrations of NAA and 2,4-D, i.e. media C1, C2, C3, C4 and C5, M1, M2, M3 and M4, to induce embryogenic calli. Compact and pearly-white, globular calli were obtained from the youngest leaf explants 28 weeks after culture.C1 media (MS medium + 107.41 µM of NAA + 100 mg.L-1 of asparagine + 100 mg.L of glutamine-1) produced the highest percentage of calli formation (30.56%), whereas C4 media (C1 supplemented with 67.86 µM of 2,-D ) was the optimal media for embryogenic callus induction. Direct embryoids were obtained from slightly older leaf explants on the C3 media containing NAA after 36 weeks of culture. However, four subcultures using the same medium with gradual reduction of auxin concentration were not successful to develop embryogenic callus and embryoid cells during the course of this study.