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

Abstract

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.

Callus-mediated Somatic Embryogenesis and Plant Regeneration in Vanda tricolor Lindl. var. Pallida

“…Yet, it was not significantly different with half-strength MS supplemented with BAP at low concentration to 0.02 mg L -1 (Table2). In line with that, there was a same report on protocorms in Phalaenopsis(Winarto et al, 2016), regeneration of Phalaenopsis bellina (Rchb.f.) PLBs(Chew et al, 2018) and SE in Tolumnia Louise Elmore 'Elsa'(Shen et al, 2018) which regenerated into plantlets after being transferred to hormone-free half-strength MS medium.…”

Orchid Micropropagation Using Conventional Semi-Solid and Temporary Immersion Systems: A Review

“…The bisected shoot tips of the Phalaenopsis hybrid were most suited for producing PLBs and tissue multiplication on MS medium supplemented with varying concentrations of thidiazuron (TDZ) [84]. Winarto and Samijan [85] witnessed maximum axillary shoot proliferation (7.0 shoots per explant, 1.0 cm shoot height, and 9.8 leaves per explant) on MS appended with 1.5 mg L −1 BAP and 0.25 mg L −1 NAA, while root production was higher on medium containing 150 mL L −1 CW. Pant and Thapa [86], while using the shoot tips of Dendrobium primulinum, obtained the highest rootless healthy shoots (4.5 shoots per cul- Kerbauy [87] used root tips of Catasetum to generate PLBs directly without forming calli on MS medium.…”

Pengaruh Jenis Eksplan dan Asam Amino pada Inisiasi dan Proliferasi Kalus Embriogenik Phalaenopsis Var. ‘Raiza Agrihorti’

“…Pada penelitian ini juga diperoleh informasi bahwa penambahan 225 mg/l L-Cystein dan 150 mg/l Casein Hydrolisate menyebabkan pencokelatan yang cukup tinggi, yaitu mencapai 52%. Beberapa penelitian mengungkapkan aplikasi jenis asam amino tertentu dalam konsentrasi yang tepat memberikan pengaruh yang positif terhadap keberhasilan kultur, tetapi pada konsentrasi yang tidak sesuai dapat menimbulkan keracunan sel dan menghambat regenerasinya (Winarto et al 2016;Rachmawati et al 2016).…”