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CALLUS CULTURE Agustina Monalisa Tangapo
CALLUS CULTURE Agustina Monalisa Tangapo

CALLUS CULTURE

CALLUS CULTURE Agustina Monalisa Tangapo

Agustina Monalisa Tangapo

BRAINSTORMING

BRAINSTORMING

BRAINSTORMING
BRAINSTORMING
Characteristic of plant in vitro culture 1. Environmental condition optimized (nutrition, light, temperature). 2.

Characteristic of plant in

vitro culture

1.

Environmental condition optimized (nutrition, light, temperature).

2.

Ability to give rise to callus, embryos, adventitious roots and shoots.

3.

Ability to grow as single cells (protoplasts,

microspores, suspension cultures).

4.

Plant cells are totipotent, able to regenerate a

 

whole plant.

Three fundamental abilities of plants  Totipotency  Dedifferentiation  Competency

Three fundamental abilities of

plants

Totipotency

Dedifferentiation

Competency

Important factors  Growth Media  Minerals, growth factors, carbon source, hormones  Environmental Factors

Important factors

Growth Media

Minerals, growth factors, carbon source, hormones

Environmental Factors

Light, temperature, photoperiod, sterility, growth media

Explant Source

Usually, the younger, less differentiated explant, the better for

tissue culture

Different species show differences in amenability to tissue culture

In many cases, different genotypes within a species will have variable responses to tissue culture; response to somatic

embryogenesis has been transferred between melon cultivars

through sexual hybridization

Hormone

Auxins

Cytokinins

Gibberellins

Abscisic Acid

Polyamines

Product Name

Indole-3-Acetic Acid Indole-3-Butyric Acid Indole-3-Butyric Acid, Potassium Salt -Naphthaleneacetic Acid

2,4-Dichlorophenoxyacetic Acid

p-Chlorophenoxyacetic acid Picloram Dicamba

6-Benzylaminopurine

6-,-Dimethylallylaminopurine (2iP) Kinetin Thidiazuron (TDZ)

N-(2-chloro-4-pyridyl)-N’Phenylurea

Zeatin

Zeatin Riboside

Gibberellic Acid

Abscisic Acid

Putrescine

Spermidine

Function in Plant Tissue Culture

Adventitous root formation (high concen) Adventitious shoot formation (low concen) Induction of somatic embryos Cell Division Callus formation and growth Inhibition of axillary buds Inhibition of root elongation

Adventitious shoot formation Inhibition of root formation Promotes cell division Modulates callus initiation and growth Stimulation of axillary’s bud breaking and growth Inhibition of shoot elongation Inhibition of leaf senescence

Stimulates shoot elongation Release seeds, embryos, and apical buds from dormancy Inhibits adventitious root formation Paclobutrazol and ancymidol inhibit gibberellin synthesis thus resulting in shorter shoots, and promoting tuber, corm, and bulb formation.

Stimulates bulb and tuber formation Stimulates the maturation of embryos Promotes the start of dormancy

Promotes adventitious root formation Promotes somatic embryogenesis Promotes shoot formation

Callus  An un-organised mass of cells  A tissue that develops in response to
Callus  An un-organised mass of cells  A tissue that develops in response to

Callus

An un-organised mass of cells

A tissue that develops in response to injury caused by physical or chemical means

Most cells of which are differentiated although may be and are often highly unorganized within the tissue

It can be multiplied through solid or suspension culture

It can be multiplied through solid or suspension culture Callus (mass of parenchymatous cells) o r
It can be multiplied through solid or suspension culture Callus (mass of parenchymatous cells) o r
It can be multiplied through solid or suspension culture Callus (mass of parenchymatous cells) o r

Callus (mass of parenchymatous cells)

or suspension culture Callus (mass of parenchymatous cells) o r g a n o g e

organogenesis

Somatic embryogenesis

Plant regeneration

Kalus adalah suatu kumpulan sel amorphous yang terjadi dari sel-sel jaringan yang membelah

diri secara terus menerus

BAGAIMANA MENDAPATKAN KULTUR KALUS????

BAGAIMANA MENDAPATKAN KULTUR KALUS????

BAGAIMANA MENDAPATKAN KULTUR KALUS????
 Two hormones affect plant differentiation:  Auxin: Stimulates root development  Cytokinin: Stimulates shoot

Two hormones affect plant differentiation:

Auxin: Stimulates root development

Cytokinin: Stimulates shoot development

Generally, the ratio of these two hormones can

determine plant development:

Auxin Cytokinin = Root development

Cytokinin Auxin = Shoot development

plant development:   Auxin ↓ Cytokinin = Root development   Cytokinin ↓ Auxin =

Control of in vitro culture

Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin

Leaf strip

AdventitiousControl of in vitro culture Leaf strip Shoot Root Callus Auxin Cytokinin

Shoot

RootControl of in vitro culture Leaf strip Adventitious Shoot Callus Auxin Cytokinin

Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin

Callus

Auxin

Cytokinin

Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
Control of in vitro culture Leaf strip Adventitious Shoot Root Callus Auxin Cytokinin
TUGAS!!!!!!! (^_^)

TUGAS!!!!!!! (^_^)

TUGAS!!!!!!! (^_^)
Callus culture

Callus culture

Callus culture

Callus formation

1. Meristems

2. Leaf sections

3. Bulb sections

4. Embryos

5. Anthers

6. Nucellus

De-differentiation

4. Embryos 5. Anthers 6. Nucellus De-differentiation Re-differentiation Explants Callus Protoplasts Development
Re-differentiation
Re-differentiation

Explants

Callus

De-differentiation Re-differentiation Explants Callus Protoplasts Development Suspension cells Organs (leaves,
De-differentiation Re-differentiation Explants Callus Protoplasts Development Suspension cells Organs (leaves,
De-differentiation Re-differentiation Explants Callus Protoplasts Development Suspension cells Organs (leaves,

Protoplasts

Development

Suspension cells

Organs

(leaves, roots, shoots, flowers,

)

Callus formation

Stimuli :

In vivo : wound, microorganisms, insect feeding

In vitro : Phytohormones

1. Auxin

2. Cytokinin

3. Auxin and cytokinin

4. Complex natural extracts

Callus  During callus formation there is some degree of dedifferentiation both in morphology and

Callus

During callus formation there is some degree of dedifferentiation both in morphology and metabolism, resulting in the lose the ability to photosynthesis.

Callus cultures may be compact or friable. Compact callus shows densely aggregated cells Friable callus shows loosely associated cells and the callus becomes soft and breaks apart easily. Habituation:

The lose of the requirement for auxin and/or cytokinin by the culture during long-term culture.

Genotypic effects on callus morphology

Genotypic effects on callus morphology

Genotypic effects on callus morphology
Three stages of callus culture  Induction : Cells in explant dedifferentiate and begin to

Three stages of callus culture

Induction: Cells in explant dedifferentiate and begin to divide

Proliferative Stage: Rapid cell division

Differentiation stage (sometimes): organogenesis or embryogenesis

Persiapan eksplan

Persiapan eksplan

Persiapan eksplan

Induction

Induction © 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS
Induction © 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS
Induction © 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS
Induction © 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS

© 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS

Division E. Sutton, UC Davis

Division

Division E. Sutton, UC Davis

E. Sutton, UC Davis

Callus © 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS

Callus

Callus © 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS
Callus © 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS

© 1998-2003, Branch of Shemyakin&Ovchinnikov IBCh RAS

Andrographis paniculata

Andrographis paniculata

Andrographis paniculata
Andrographis paniculata
Kalus yang tumbuh pada eksplan tunas pucuk manggis

Kalus yang tumbuh pada eksplan tunas pucuk manggis

Figure 1. Stages of callus growth in A. malaccensis Lam.a. Initiation of callus, b&c; Growth
Figure 1. Stages of callus growth in A. malaccensis Lam.a. Initiation of callus, b&c; Growth

Figure 1. Stages of callus growth in A.

malaccensis Lam.a. Initiation of callus, b&c;

Growth of the callus; d. Cell death

Differentiation  Organogenesis  Somatic embryogenesis

Differentiation

Organogenesis Somatic embryogenesis

Gambar. Induksi dan proliferasi kalus (1-2); Diferensiasi/(regenerasi tunas (3-4) dan Pembentukan plantlet induksi
Gambar. Induksi dan proliferasi kalus (1-2); Diferensiasi/(regenerasi tunas (3-4) dan Pembentukan plantlet induksi

Gambar. Induksi dan proliferasi kalus (1-2); Diferensiasi/(regenerasi tunas (3-4) dan Pembentukan plantlet induksi perakaran(5-6)

Lolium temulentum

Lolium temulentum

MANFAATNYA?????

MANFAATNYA?????

MANFAATNYA?????
Callus culture of some commercially important plants Podophyllum hexandrum Azadirachta indica Linum album

Callus culture of some

commercially important plants

Callus culture of some commercially important plants Podophyllum hexandrum Azadirachta indica Linum album

Podophyllum

hexandrum

Callus culture of some commercially important plants Podophyllum hexandrum Azadirachta indica Linum album

Azadirachta indica

Callus culture of some commercially important plants Podophyllum hexandrum Azadirachta indica Linum album

Linum album

THANKS FOR UR ATT

THANKS FOR UR ATT

THANKS FOR UR ATT