Establishment of axenic culture and callus induction of peperomia pellucida (L.) Kunth by using Leaf and Nodal Explants

Tan, Eng Keat. (2016) Establishment of axenic culture and callus induction of peperomia pellucida (L.) Kunth by using Leaf and Nodal Explants. [Final Year Project Report] (Unpublished)

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Abstract

Peperomia pellucida Kunth. (shining bush) of family Piperaceae typically grow as annual herb with shallow-rooted and succulent stems. P. pellucida having medicinal properties which is used widely in South East Asia as breast cancer treatment, reduce blood glucose level, treat rheumatism and fatigue and renal treatment. The purpose of this study is to establish an effective surface sterilized protocol by using Clorox® and to induce callus from leaves and nodal explants of P. pellucida by using 2, 4-D. The concentrations of Clorox® solution include 5%, 10%, 15% and 20%, and time exposures for each concentrations of Clorox® solution were 5minutes, 10minutes and 15minutes. The explants were cultured onto PGR-free MS (Murashige and Skoog) medium. Results shown using 10% of Clorox® solution for 15 minutes for leaf explant was 72.50 ± 7.94% of axenic whereas for nodal explant using 10% of Clorox® solution for 15 minutes showed 80.88 ± 6.21% axenic were treatments generate highest percentage of axenic culture within treatments. For callus induction, highest percentage of axenic culture within treatments were applied and explants cultured onto full strength and half strength MS media incorporated with plant growth regulator which is 2, 4-D at concentration of 0 mg/l, 1.0 mg/l, and 2.0 mg/l. Result showed full strength MS media with 2, 4-D at concentration at concentration of 1.0 mg/l and half strength MS media incorporated with 2, 4-D at concentration of 1.0 mg/l and 2.0 mg/l produced 100% of percentage of leaf explants growing callus after 4 weeks. However, the highest percentage of nodal explant growing callus is 45.0 ± 5.0% which is treatment with full strength MS medium and 2, 4-D at concentration 2.0 mg/l after 4 weeks. There was additional experiment on direct organogenesis by using leaf explant of P. pellucida. The result shown the highest shoot and root growing percentage after one month include half strength of MS media with 0 g/l of sucrose. This project can be enhance with include ethanol and benomyl to increase axenic explant percentage. Besides, treatments and media could be modified to improve the efficiency of explant growing callus whereas to promote growing of shoot and root of leaf explant as P. pellucida needs low nutrient source to grow shoot and root.

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