Antibiotics:

Antibiotics are substances produced by certain microorganisms that suppress the growth
of other microorganisms and eventually destroy them. Their applications include:

a. Suppresses bacterial infections in plant cell and tissue culture.

b. Suppresses mould and yeast infections in cell cultures.
c. Eliminates Agrobacterium species after the transformation of plant tissue.

These antibiotics can be divided into different classes on the basis of chemical structure
and their mechanism of action:

1. Inhibitors of Bacterial Cell Wall Synthesis.

e.g. β-lactam antibiotics, Penicillins and Cephalosporins.
2. Antibiotics that affect Cell Membrane permeability.
(a) Antibacterial e.g. Colistin Sulphate, Polymixin B Sulphate, Gramicidin
(b) Antifungal e.g. Amphotericin B, Nystatin, Pimaricin
3. Bacteriostatic Inhibitors of Protein
Antibiotics that affect the function of 30 S or 50 S ribosomal subunits to cause a
reversible inhibition of protein synthesis. e.g. Chloramphenicol, Chlortetracycline
HCl, Clindamycin HCl, Doxycycline HCl, Erythromycin, Lincomycin HCl,
Oxytetracycline HCl, Spectinomycin sulphate, Tetracycline HCl, Tylosin tartrate,
Lincomycin HCl
4. Bactericide Inhibitors of Protein Synthesis
Antibiotics that bind to the 30 S ribosomal subunit and alter protein synthesis
which eventually leads to cell death. This group includes:
(a) Aminoglycosides: Apramycin, Butirosine, Gentamicin, Kanamycin,
Neomycin, Streptomycin, Tobramycin.
(b) Inhibitors of Nucleic Acid Metabolism: e.g. Rifampicin, Mitomycin C and
Nalidixic acid.
(c) Antimetabolites: Antibiotics, which block specific metabolic steps that are
essential to microorganisms e.g. Metronidazole, Miconazole, Nitrofurantoin,
Trimethoprim and Sulphomethoxazole.
(d) Nucleic Acid Analogs, which inhibit enzymes essential for DNA synthesis.
e.g. 5-Fluorouracil, Mercaptopurine

PREPARATION OF PLANT TISSUE CULTURE MEDIUM

1. Measure approximately 90% of the required volume of the deionized-distilled water

in a flask/container of double the size of the required volume.
2. Add the dehydrated medium into the water and stir to dissolve the medium
completely. Gentle heating of the solution may be required to bring powder into
solution.
3. Add desired heat stable supplements to the medium solution.
4. Add additional deionized-distilled water to the medium solution to obtain the final
required volume.
5. Set the desired pH with NaOH or HCl.
6. Dispense the medium into culture vessels.
7. Sterilize the medium by autoclaving at 15 psi (121°C) for appropriate time period.
Higher temperature may result in poor cell growth.
8. Add heat labile supplements after autoclaving.