Sains Malaysiana 47(6)(2018): 1093–1099

http://dx.doi.org/10.17576/jsm-2018-4706-03

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

(Proliferasi dan Penjanaan Semula yang Cekap ke atas Jasad seperti Protokorm (PLBS) Orkid

Terancam, Phalaenopsis bellina)

YEE-CHERN CHEW¹, MUHAMMAD HUSNI ABD HALIM¹, WAN MUHAMAD ASRUL NIZAM WAN ABDULLAH², JANNA ONG ABDULLAH² & KOK-SONG LAI^2*

¹Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

²Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences

Universiti Putra Malaysia, 43400 Serdang, Selangor Darul Ehsan, Malaysia

Received: 6 November 2017/Accepted: 1 February 2018

ABSTRACT

Phalaenopsis bellina is an important indigenous fragrant orchid threatened with extinction. In this study, we evaluated the effect of medium strength, sucrose, nitrogen (NH₄NO₃) and potato extract on proliferation of P. bellina protocorm-like bodies (PLBs) to improve micropropagation in this species. Optimal treatment for PLBs proliferation rate with an average fresh weight (FW) of 0.97±0.16 g was obtained through culturing on half strength (½) MS medium containing 20 g/L sucrose, 15 mM NH₄NO₃ and 20% w/v potato extract supplemented with 0.8 μM 2,4 dichlorophenoxyacetic acid (2,4-D). The optimal treatment produced large, healthy and greenish PLBs with reduction in the occurrence of culture browning. In contrast, treatments with high potato extract (>20% w/v) or NH₄NO₃ (>30 mM) concentrations tend to have inhibitory effect and resulted in low PLBs proliferation rate, with an average FW of 0.77±0.15 g and 0.69±0.15 g, respectively. Plant regeneration of PLBs was achieved on plant growth regulator (PGR)-free ½ MS medium. In total, 60 healthy PLBs from the optimal treatment were successfully regenerated, acclimatized with 100% survival percentage and grew well in a mixture of soil, sand and vermicompost (8:4:2 (w/w/w)). With the optimal treatment, PLBs proliferation rate was enhanced by 27.63%. Our findings offer an improved micropropagation protocol of the endangered P. bellina for conservation and commercial production.

Keywords: Indigenous orchid; Phalaenopsis bellina; proliferation; protocorm-like bodies; regeneration

ABSTRAK

Phalaenopsis bellina merupakan orkid harum asli yang diancam kepupusan. Menerusi kajian ini, kami menilai kesan sukrosa, nitrogen (NH4NO3) dan ekstrak kentang terhadap proliferasi jasad seperti protocorm (PLBs) P. bellina untuk meningkatkan perambatan spesies ini. Rawatan optimum untuk kadar proliferasi PLBs dengan purata berat segar 0.97±0.16 g diperoleh dengan pengkulturan pada separuh kekuatan (½) media MS yang mengandungi 20 g/L sukrosa, 15 mM NH₄NO₃ dan 20% w/v ekstrak kentang ditambah dengan 0.8 μM 2,4 asid diklorofenoksiasetik (2,4-D). Rawatan optimum menghasilkan PLBs yang besar, sihat dan hijau dengan pengurangan keperangan kultur. Sebaliknya, rawatan menggunakan kepekatan ekstrak kentang (>20% w/v) atau NH4NO3 (>30 mM) yang tinggi cenderung memberikan kesan rencatan dan menyebabkan kadar proliferasi PLBs yang rendah dengan purata berat segar masing-masing 0.77±0.15 g dan 0.69±0.15 g. Regenerasi tumbuhan daripada PLBs dihasilkan dengan ½ media MS tanpa zat pengatur tumbuhan (ZPT). Secara keseluruhan, 60 PLBs yang sihat berjaya dijana semula dan PLBs dapat menyesuaikan diri dengan 100% kelangsungan hidup dan tumbuh dengan baik di dalam campuran tanah, pasir dan vermikompos (8:4:2 (w/w/w)). Kadar proliferasi PLBs meningkat 27.63% dengan rawatan optimum. Penemuan kami mengusul satu penambahbaikan protokol pembiakan P. bellina yang terancam untuk tujuan pemeliharaan dan penghasilan komersial.

Kata kunci: Jasad seperti protocorm; orkid asli; penjanaan semula; Phalaenopsis bellina; proliferasi

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*Corresponding author; email: laikoksong@upm.edu.my