Effects of different phytohormones on callus induction

of Pereksia sacharosa
Introduction
Origin: : 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.
Objective
To 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 Statement
Which 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 sacharosa
Medical use: Raw / brewed it
Cytotoxicity activity toward cancer cell lines:
1. Human colon carcinoma cell line
2. Human lung carcinoma cell line
3. Human cervical carcinoma cell line
4. Human hormone dependent breast carcinoma.
Anti-tumor, anti-inflammatory and anti-ulcer.
Overview of Materials & Methods
Surface
Sterilization

2 g/L Auxin &
cytokinin + MS medium
MS medium only
&
Incubation
Light
Condition
Dark
Condition
Transfer
(Negative Control)
(Positive Control)
Overview of Materials &
Methods
Surface 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 Culture
Medium 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 Results
1. 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.
References
Oakley, 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.

References
Edwards 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!