Effects of different phytohormones on callus induction of Pereksia sacharosa

IntroductionOrigin: : Argentina, Bolivia, Brazil and

Paraguay.Distribution: Low lands of Brazil, Bolivia,

Paraguay ; High land of Argentina ; Mediterranean, African, Asia and Australia.

Common names: Sacharosa, Guguchi, Guyapa and ‘Jarum Tujuh Bilah’ in Malay.

ObjectiveTo establish the suitable surface sterilisation

protocols for the leaf and stem explants of P. sacharosa

To study the effects of different type of auxin and cytokinin on callus indction from the leaf and stem of P. sacharosa

To identify the leaf or stem of P. sacharosa which much more suitable for callus induction.

Problem StatementWhich part of the stem (node or internode) is

better for callus induction ?Which part of leaf mid vein section is better

for callus induction ?Which phytohormones has best inductive

effect on callus induction of Pereksia sacharosa.

Does lighting condition affect the growth of the callus?

Literature Review

Family Cactaceae

Perennial (Annual)

Succulent

Areoles with Spines and glochid or both.

Flattened and photosynthetic leaves

Flower, Fruits and Seeds

Pereskia sacharosa

• Native: Argentina, Bolivia, Brazil and Paraguay.

•Habitat: Forest edge, clearings.

•6 to 8 meters height

•Lanceolate to oblanceolate with board lamina, 8 to 12 cm long

•Areole with spines, longest 5cm

•Flowers either in white or rose-colored about 8 cm in diameter during rainy season

•Fruits are harden with 2.5 cm to 4 cm in diameter.

The uses of Pereskia sacharosaMedical use: Raw / brewed itCytotoxicity activity toward cancer cell lines:1.Human colon carcinoma cell line2.Human lung carcinoma cell line3.Human cervical carcinoma cell line4.Human hormone dependent breast

carcinoma.Anti-tumor, anti-inflammatory and anti-ulcer.

Overview of Materials & Methods

Surface Sterilization

2 µg/L Auxin & cytokinin + MS medium

MS medium only

&Incubation

Light Condition

Dark Condition

Transfer

(Negative Control)

(Positive Control)

Overview of Materials & MethodsSurface Sterilization: Sodium Hypochlorite (0.5~1.0% for 10~20

mins) + Tween 20, Rinse with sterile water.

Types of Auxins used: 2,4-D , Picloram, Dicamba, NAA, IAA

Types of Cytokinins used : BAP and Kinetin

Protocols of Plant Tissue CultureMedium Preparation

•Compositions of MS medium•Addition of phytohormone•pH•Agar Powder•Autoclave

Surface Sterilization&

Transfer with Aseptic Technique

•Wipe working surface•No hand crossing•Reduce air flow

Incubation•Temperature

•Light Intensity

Observation, Data record & Analysis

Expected Results1. High concentration of 2,4-D is preferable for

callus induction of leaf explant.2. Cytokinin is preferable for callus induction

of the explant of stem with areole.

ReferencesOakley, L., Pin, A. & Lowry, M. 2013. Pereskia

sacharosa. In: IUCN 2014. IUCN Red List of Threatened Species. Version 2014.1. <www.iucnredlist.org>. Downloaded on 19 June 2014.

Anderson E.F., (2001), The Cactus Family, Oregon : Timber Press, Inc.

Britton N.L. & Rose J.N., (1919), The Cactaceae : Descriptions and Illustrations of Plants of the Cactus Family, Washington.

Chuah E.L. & Chan L.K., (2007), Induction of Somatic Embryogenic Callus from the Leaves of Pereskia grandifolia, Biotechnology 6(1) : 45-48 p.

ReferencesEdwards E.J. & Donoghue M.J., (2006),

Pereskia and the Origin of the Cactus Life-Form,

Rubluo A. et al., (2002), Auxin induced morphogenetic responses in long-term in vitro subcultured Mammillaria san-angelensis Sanchez-Mejorada (Cactaceae), Scientia Horticulturae 95, 341-349 p.

Thank You!