Highly Efficient Proliferation and Regeneration of Protocorm-like Bodies (PLBS) of the Threatened Orchid, Phalaenopsis bellina

Chew, Yee-Chern; Halim, Muhammad Husni Abd; Abdullah, Wan Muhamad Asrul Nizam Wan; Abdullah, Janna Ong; Lai, Kok-Song

doi:10.17576/jsm-2018-4706-03

Cited by 5 publications

(3 citation statements)

References 17 publications

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“…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. The addition of BAP in higher concentration from 0.02 mg L -1 evidently had an effect to slow down germination of SE to form plantlets and caused germination time to be delayed.…”

Section: Regeneration Of Se To Form Plantletsmentioning

confidence: 99%

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

2021

JJBS

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In this study, a protocol to induce indirect somatic embryogenesis from the basal leaf segments of Vanda tricolor Lindl. var. Pallida has been developed. 2 0 T The experiments consisted of two stages، i.e. induction of SEs from calli and regeneration of plantlet from SEs. 2 0 T Embryogenic calli obtained from previous experiment (0.05 mg LP -1 P NAA + 0.01 mg LP -1 P BAP) were used to induce somatic embryos (SEs) on half-strength Murashige and Skoog (MS) medium supplemented with (0.05 mg LP -1 P toP P 0.20 mg LP -1 P ) 6-benzylaminopurine (BAP) alone or in combination with 0.01 mg LP -1 P α-naphtalene acetic acidP P (NAA). Embryogenic calli, those cultured on 0.05 mg LP -1 P BAP and 0.01 mg LP -1 P NAA resulted in 90 % induction of SE structures at 30 d of culture period. Histological observation exhibited development of pro-embryo to form completed embryo. The pattern of SEs development started from embryogenic callus to form pro-embryo, followed by globular phase at 10 d of culture. Globular embryo elongated to form suspensor at 20 d of incubation period, and completed embryo. Regeneration of SEs into complete plantlets was attained on the half-strength MS medium without addition of any plant growth regulator (PGR). Based on the results of the present study, it can be concluded that 2 0 T half-strength MS medium supplemented with 2 0 T NAA 0.01 mg LP -1

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Section: Regeneration Of Se To Form Plantletsmentioning

confidence: 99%

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

2021

JJBS

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“…PLBs induction was performed according to Chew's methods (Chew et al 2018). Briefly, three months old in vitro seedlings of P. bellina were used for protocorm-likebodies (PLBs) induction (Figure 1(B)).…”

Section: Protocorm-like-bodies (Plbs) Inductionmentioning

confidence: 99%

“…Naturally, it could be found growing on trees branches in shady lowland forests where it experiences wet and dry cycles and receives abundant sunlight. Besides, P. bellina is also known as the most iconic orchid species as it has strong unique biological features such as versatile colours flower, cereal fruit loops fragrance, elegant appearance and high commercial value (Chew et al 2018). Furthermore, P. bellina is favoured amongst orchid farmer due to its lowcost of maintenance, ease of production and all year-round flowering habit.…”

Section: Introductionmentioning

confidence: 99%

Development of an Efficient Particle Bombardment Transformation System for the Endemic Orchid, Phalaenopsis bellina

Chew¹,

Abdullah²,

Kok³

et al. 2019

JSM

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Phalaenopsis bellina is an important indigenous orchid with high commercial value. In this study, we established an efficient particle bombardment transformation system for P. bellina using the protocorm-like-bodies (PLBs) as target tissues. Leaf derived PLBs were proliferated on ½ strength Murashige and Skoog (MS) medium supplemented with 0.8 µM 2,4 dichlorophenoxyacetic acid (2, 4-D). Both physical and biological parameters affecting the transformation system were optimised using the green-fluorescent protein (GFP) and β-glucuronidase (GUS) as reporter systems. Optimal bombardment conditions with 6 cm target tissues distance, 1100 psi acceleration pressure, 1.0 μm gold particle size, 27 mmHg chamber vacuum pressure, single bombardment time, spermidine as DNA precipitation agent, 72 h post bombardment incubation time, 2 μg plasmid DNA in 0.15:0.12 pmol ratio (pSMCHS:p35SGFP) were successfully determined. Surviving PLBs transformants were successfully recovered from the hygromycin selection medium and verified using genomic PCR analysis. The established system is not only useful for a simple and reliable transient gene analysis but as well as generating stable transformants for selective traits improvement in orchids. ABSTRAKPhalaenopsis bellina ialah salah suatu orkid yang mempunyai nilai komersial yang tinggi. Dalam kajian ini, kami telah membentuk sistem transformasi pembedilan zarah yang berkesan untuk P. bellina menggunakan jasad seperti protokorm (PLB) sebagai tisu sasaran. Daun daripada PLBs bercambah pada ½ kekuatan medium Murashige dan Skoog (MS) yang mengandungi 0.8 μM 2,4 asid d diklorofenok siasetik (2, 4-D). Parameter biologi dan fizikal yang mempengaruhi sistem transformasi telah dioptimumkan dengan menggunakan protein fluoresen hijau (GFP) dan β-glukuronidase (GUS) sebagai sistem pelapor. Keadaan pembedilan optimum dengan jarak tisu 6 cm, tekanan pecutan 1100 psi, saiz zarah 1.0 μm, tekanan vakum 27 mmHg, pembedilan tunggal, spermidine sebagai agen pemendapan DNA, 72 jam tempoh inkubasi selepas pembedilan, 2 μg DNA plasmid dalam 0.15:0.12 pmol nisbah (pSMCHS:p35SGFP) telah ditentukan. Transforman PLB telah berjaya dipulihkan selepas inkubasi di dalam medium pemilihan higromisin dan disahkan menggunakan analisis PCR genom. Sistem yang ditubuhkan ini tidak hanya berguna untuk analisis gen sementara yang boleh dipercayai dan mudah tetapi juga mampu menjana transformasi stabil bagi peningkatan ciri-ciri terpilih dalam orkid.Kata kunci: β-glukuronidase (GUS); jasad seperti protokorm (PLB); Phalaenopsis bellina; pembedilan zarah; protein fluoresen hijau (GFP)

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Protocorm-Like Body Induction and Development Using In Vitro Culture Techniques and Biotechnological Methods

Yeung,

Yeung

2024

Springer Protocols Handbooks

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Highly Efficient Proliferation and Regeneration of Protocorm-like Bodies (PLBS) of the Threatened Orchid, Phalaenopsis bellina

Cited by 5 publications

References 17 publications

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

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

Development of an Efficient Particle Bombardment Transformation System for the Endemic Orchid, Phalaenopsis bellina

Protocorm-Like Body Induction and Development Using In Vitro Culture Techniques and Biotechnological Methods

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