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Introduction

A phytohormones is need to ad into the basal medium in order for plant to growth and
develop. Medium for plant tissue culture must contain macronutrients, micronutrients,
vitamins, sugars and sugar alcohols. Auxin and cytokinin is the most famous plant growth
hormones that is added in the plant tissue culture. After all the nutrients added the medium
must be set into a suitable pH between 5.7 to 5.8 before autoclave.

Regeneration of plants by micropropagation of in vitro culture can be done by using


the existing shoot tips and the stems of the plant. Plants can be regenerated from unorganized
callus tissue derived from different from different explants. Callus culture can also be facilitated
the amplification of limiting plant material.

In plant tissue culture, organogenesis is a process of differentiation by which plant


organs like roots, shoots, buds etc. are formed from the unusual points of origin of an
organized explants where a preformed meristem is lacking. Plant development through
organogenesis is the formation of organs either de novo or adventitious in origin and plant
regeneration via organogenesis is a mono-polar structure.

In this experiment, our main goal is to culture both leave and stem of the Tobacco plant
in two different medium label by TP and TC. We also need to maintain the sterile environment
while conducting the experiment in order for us to avoid contamination of our media.
Method and Materials

1. Leaf Explants

The leaf cut into squares


In the petri dish, inside
using a scalpel and
laminar air flow, young
forceps. Before the
leaves were removed
cutting process, both are
from the tobacco plant
flame using a Bunsen
by cutting the petioles
burner.
from the stem.

The boxes and the tubes Leaves were culture in


were flamed and sealed the condition which is
with parafilm and stored upper surface down and
based on the label lower surface down
(light/dark) inside a TP and TC boxes
(4 explants for each
boxes)
2. Stems explants

In the laminar air flow, 3- The stems were cut using


4cm segments of stem scalpel and forceps into
were cut and the leaves is segment which in the
trim leaving few mm of internode part.
petiole attached to the
stem inside a sterile petri
dish.

The boxes and the tubes The stem is culture in the TP


were flamed and sealed and TC medium horizontally
with parafilm and stored and vertically.
based on the label
(light/dark)
Results

1. Organogenesis
Source of explant: Leaf and Stem
Period of culture: Week 3

Orientation of Formati % Photo


explant on of contamination
plant
organs
(yes/no)
Upper No 0%
surface Light
down

The leaves started to curl


cfkrmndfbojreg erg

Lower No 0%
surface Light
down

The leaves started to curl

Horizontal Yes 50% of


Light contamination on
one box.
There is formation of shoot at one box. While the
other are contaminated

Vertical No 0%
Light

The explants turn yellowish


Period of Culture: Week 4

Orientation of Formati % Photo


explant on of contamination
plant
organs
(yes/no)
Upper Yes 0%
surface Light
down

The leaves started forming new leaves around


the edges and curl up

Cfkr

Lower Yes 0%
surface Light
down

mndfbojregerg

The leaves started forming new leaves around


the edges and curl up

Horizontal Yes 100% of


Light contamination on
one box.
There is formation of shoot at one box. While the
other are contaminated

Vertical Yes 0%
Light

There is formation of shoots around the


stem.
2. Callogenesis
Source of explant: Leaf and Stem
Period of culture: Week 4

Orientation Condition % Photo and descriptions


of explant contamination
Upper 0%
surface Light
down

There is formation of callus at the edge of the


leaves

Dark 0%

There is formation of callus at the edge of the


leaves
Lower Light 0%
surface
down

There is formation of callus at the edge of the


leaves
Dark 0%

There is no formation of callus but the leaf


started to curl up
0%
Vertical Light

There is no formation of callus but the stem turn


yellowish
Dark 0%

There is no formation of callus


Horizontal Light

There is formation of callus at the end of the


stem cut
Dark

There is no formation of callus


Discussion

In this experiment, we were required to study the formation of organ and callus for the
explants of stem and leaf. This is called callogenesis and organogenesis.

Cell in plants are derived from the zygote through mitotic division containing the
complete genome. The formation of adventitious buds depending on the embryonic phase of
development. During this stage, the used of phytohormones on the media is really important
in order for the organ to develop. There are two types of organogenesis, in which
organogenesis that are direct and indirect. Plant generation from explants involve initiation of
basal callus formation and also root differentiation. Plants with mitotically active cells are good
in developing embryos through callus formation and can be used successfully to initiate
plantlets through organogenesis.

From the table above, we can see that the formation of the organ took 4 weeks to
actually develop nicely. One of the stem sample were contaminated. This is might due to the
crack that the cap of the box had. Differentiation of the cells is an outcome of the process of
dedifferentiation followed by the redifferentiation. Dedifferentiation favours unorganized cell
growth and the resultant developed callus has meristems randomly divided.