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Did you ever have a plant that was so unique or so beautiful that you wished you had hundreds or thousands of them to enjoy or to sell? Plant tissue culture (micropropagation) is a technique which will do just that for us. It is a fascinating and useful tool which allows the rapid production of many genetically identical plants using relatively small amounts of space, supplies and time.
Basically the technique consists of taking a piece of a plant (such as a stem tip, node, meristem, embryo, or even a seed) and placing it in a sterile, (usually gel-based) nutrient medium where it multiplies. The formulation of the growth medium is changed depending upon whether you are trying to get the plant to produce undifferentiated callus tissue, multiply the number of plantlets, grow roots, or multiply embryos for "artificial seed". Plant tissue culture research has become a thrust area during the last decade due to the renewed emphasis it has received in all areas of crop improvement programmes. In several publications, including books, and in syllabi prescribed for biotechnology courses of our universities, plant tissue culture along with plant genetic engineering have become synonymous with plant biotechnology.

BASIC REQUIREMENT OF A PLANT TISSUE CULTURE MEDIA

Plant Tissue Culture refers to the technique of growing plant cells, tissues, organs, seeds or other plant parts in a sterile environment on a nutrient medium. Culture media used for in vitro cultivation of plant cells are composed of following basic components: 1. Complex mixture of salts: Essential elements, or mineral ions. 2. Organic supplement: vitamins and/or amino acids. 3. Carbon source: usually sugar sucrose. 4. Gelling agents 5. Plant Growth Regulators 6. Antibiotics
Complex mixture of salts:

These include essential elements or mineral ions important for plant nutrition and their physiological function. The essential elements can further be divided into the following categories: a. Macroelements (or macronutrients) b. Microelements (or micronutrients) c. Iron Source
Macroelements: These elements are required in large amounts for plant growth and development. Nitrogen, phosphorus, potassium, magnesium, calcium and sulphur

(and carbon, which is added separately) are regarded as macroelements. These elements comprise at least 0.1% of the dry weight of plan
Microelements: These elements are required in trace amounts for plant growth and development. Manganese, iodine, copper, cobalt, boron, molybdenum, iron and

zinc are regarded as microelements, although other elements like aluminium and nickel are frequently found in some formulations.
Iron Source: Iron is usually added in the medium as iron sulphate, although iron citrate can also be used. Ethylenediaminetetraacetic acid (EDTA) is usually used in conjunction with the iron sulphate. The EDTA complexes with the iron so as to allow the slow and continuous release of iron into the medium. Uncomplexed iron can precipitate out of the medium as ferric oxide. Organic supplements:

These include vitamins and amino acids. Two vitamins, i.e., thiamine (vitamin B1) and myoinositol (a vitamin B) are essential for the culture of plant cells in vitro. However, other vitamins are often added to for historical reasons. The most commonly used amino acid is glycine. However, arginine, asparagine, aspartic acid, alanine, glutamic acid, glutamine and proline are also used. Amino acids provide a source of reduced nitrogen and, like ammonium ions, uptake causes acidification of the medium. Casein hydrolysate can be used as a source of a mixture of amino acids.

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The most commonly used carbon source is sucrose. It is readily assimilated and relatively stable. Other carbohydrates like glucose, maltose, galactose and sorbitol can also be used and may prove better than sucrose in specialized circumstances.
Gelling agents:

Plant tissue culture media can be used in either liquid or 'solid' forms, depending on the type of culture being grown. Agar, produced from seaweed, is the most common type of gelling agent, and is ideal for routine applications. For more demanding applications, a range of purer gelling agents are available. Purified agar or agarose can be used, as can a variety of gellan gums. Agar is a gelatinous substance derived from the seaweeds. It is an unbranched high molecular weight polysacchride obtained from the cell walls of some species of the red algae, primarily from the genra Gelidium and Gracilaria. Agar is insoluble in cold water but forms relatively inert gel that melts at about 1000C and solidifies at around 450C. Using Agar as a gelling agent main advantage is that agar does not react with any components of the medium and is not digested by enzymes from the plant tissue. If necessary, agar can be washed to remove inhibitory impurities. Agarose is a purified extract of agar without the agaropectin fraction and sulphate groups. It is required in lesser quantities since it has a higher gelling strength. Agarose is used for more demanding procedures like culturing protoplasts. It consists of -D(1-3) galactopyranose and 3,6-anhydro--L(1-4) galactopyranose linked into polymer chains of 20-160 monosaccharides units. Gellan gum, also known commercially as Phytagel or Gelrite or Gelwell, is used primarily as a gelling agent, alternative to agar, in microbiological culture. It is able to withstand 120 C heat, making it especially useful in culturing thermophilic organisms. One needs only approximately half the amount of gellan gum as agar to reach an equivalent gel strength, though the exact texture and quality depends on the concentration of divalent cations present. Gellan gum is used as gelling agent in plant cell culture on petri-dishes, as it provides a very clear gel, facilitating light microscopical analyses of the cells and tissue.
Plant growth regulators:

Specific media manipulations can be used to direct the development of plant cells in culture due to plasticity and totipotency. Plant growth regulators are the critical media components in determining the developmental pathway of the plant cells. There are five main classes of plant growth regulator used in plant cell culture, namely: a. Auxins b. Cytokinins c. Gibberellins d. Abscisic acid e. Ethylene
Auxins: Auxins promote both cell division and cell growth. IAA (indole-3-acetic acid) is the most important naturally occurring auxin but its use in plant tissue culture media is limited because it is unstable to both heat and light. 2,4-Dichlorophenoxyacetic acid (2,4-D) is the most commonly used auxin and is extremely effective in most circumstances. Cytokinins: Cytokinins promote cell division. Of the naturally occurring cytokinins, only zeatin and 2iP (2-isopentyl adenine have some use in plant tissue culture

media. The synthetic analogues, kinetin and BAP (benzylaminopurine), are used more frequently. Non-purine-based chemicals, such as substituted phenylureas, are also used as cytokinins in plant tissue culture media.
Gibberellins: Gibberellins are involved in regulating cell elongation, in determining plant height and fruit-set. Only a few of the gibberellins like GA3 are used in

plant tissue culture media.

Abscisic acid: It is used in plant tissue culture to promote distinct developmental pathways such as somatic embryogenesis. Abscisic acid (ABA) inhibits cell division.

Ethylene: Ethylene is associated with controlling fruit ripening in climacteric fruits, and its use in plant tissue culture is not widespread. Some plant cell cultures produce ethylene, which, if it builds up sufficiently, can inhibit the growth and development of the culture.

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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. d. As a selective agent in plant transformation experiments.
Antimicrobial Compound Mechanism of Action Nature

Amoxycillin Amphotericin B solubilized powder Amphicillin sodium salt Bacitracin Carbenicillin,disodium salt Cephotaxime, sodium salt Chloramphenicol Ciprofloxacin hydrochloride Erythromycin Gentamycin sulphate Geneticin disulphate Hygromycin B Kanamycin acid sulphate Mycostain Penicillin, benzyl sodium salt Vancomycin hydrochloride Rifampicin

Inhibits synthesis of the bacterial peptidoglycan cell wall Act by increasing membrane permeability by binding to sterol moiety, primarily ergosterol present in the cell membrane of sensitive fungi Inhibit synthesis of the bacterial peptidoglycan cell wall Inhibit synthesis of the bacterial peptidoglycan cell wall Inhibit synthesis of the bacterial peptidoglycan cell wall Inhibit bacterial cell wall synthesis Inhibits protein synthesis by acting on 50S ribosomes Interfere with DNA replication by inhibition of DNA gyrase Inhibit protein synthesis by acting on 50S ribosomes Inhibit protein synthesis by binding to 30S ribosomes and decreasing the fidelity of translation of m RNA Inhibit prokaryotic and eukaryotic protein synthesis Inhibit bacterial cell wall synthesis Inhibit protein synthesis by interaction with 30s, 50S ribosomes Bind to sterols in fungal cell membrane, changing the cell wall permeability allowing for leakage of cellular contents Inhibit bacterial cell wall synthesis Interferes bacterial cell wall synthesis for Gram+ only Rifampicin inhibits DNA-dependent RNA polymearase in bacterial cells by binding its beta subunit, thus preventing transcription of messenger RNA (mRNA) and subsequent translation to Proteins. Inhibits protein synthesis

Bactericidal Antifungal Bactericidal Bactericidal Bactericidal Bactericidal Bactericidal Bactericidal Bactericidal Bactericidal Bactericidal Bactericidal Bactericidal Antifungal Bactericidal Bactericidal Bactericidal

Neomycin sulphate

Bactericidal

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Plant Tissue Culture Media

? Banana Medium ? BM Medium ? CHU (N6) Medium ? Gamborg B5 Medium ? Medium (MS) Murashige & Skoog ? Musrashige & SkoogModified Medium (MS) ? Nitsch Medium ? Orchid Medium ? Medium (SH) Schenk & Hidebrandt

Banana Medium
Use

APM1001/APM5001
Formula Ingredients in Grams/Litre Macroelements

Banana medium is used for in vitro micropropagation of Musa (Family Musaceae).

Summary

Banana medium is the modification of Murashige and Skoog medium(1962), which provides all essential elements to grow Banana in vitro.
Principle

Banana medium provides all essential Macroelements, Microelements, Vitamins, Amino acid & Plant growth regulators for the growth of Banana in vitro.
Macroelements :

Microelements

Potassium nitrate Ammonium nitrate Calcium chloride anhydrous Magnesium phosphate anhydrous Potassium phosphate monobasic Sodium phosphate monobasic

1900.00 1650.00 332.16 180.69 170.00 221.00 16.90 6.20 0.83 0.25 8.60 0.03 0.03 27.80 37.26 100.00 0.10 30000.00 10.00 0.18 8000.00

Potassium nitrate, ammonium nitrate provide nitrogen source to this media. This mixture of cation and anion balances the pH of the medium. Also this ratio play a very important role in plant growth.
Microelements:

Manganese, molybdenum, zinc, boron, copper and iron play an important role in plant metabolism while manganese plays a vital catalytic role in nitrogen metabolism. Iron used as a FeEDTA to overcome the problem of precipitation in the medium.
Vitamins:

Manganese sulphate.H2O Boric Acid Potassium iodide Molybdic acid (sodium salt).2H2O Zinc sulphate.7H2O Copper sulphate.5H2O Cobalt chloride.6H2O Ferrous sulphate.7H2O Na2.EDTA
Vitamins

Myo inositol concentartion was increased as it shows stimulatory effect in tobacco callus. Increased concentration of thiamine, pyridoxine and nicotinic acid have a stimulatory effect. Thiamine in the medium is a key elements of carbohydrate metabolism and biosynthesis of some amino acids. Ascorbic acid is provided to control phenol production in the medium.
Amino acid:

Carbohydrate Organic supplement

Myo-Inositol Thiamine HCL

Sucrose

Casein enzymic hydrolysate

Gelling agent

Plant Growth Regulators

Indole-3-acetic acid
Storage

The medium contains increased concentration of glycine (2mg/L).

Plant growth regulators:

Agar

Indole-3-acetic acid (IAA) is used in the medium forimproved growth and quanlity of banana plantlets. 6-Benzyl amino purine(BAP) induces shoot proliferation.

Shelf Life

Store at 2-80C and prepared medium at 2-80C.

Use before expiry date as mentioned on the label

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BM Medium
USE Plant growth regulators:

APM1002/APM5002
6-Benzyl amino purine(BAP) induces shoot proliferation.
Formula Ingredients in Grams/Litre Macroelements

BM Media is used for seed culture and micropropagation of orchids.

Summary

Van waes, (1986) has developed BM medium for in vitro cultivation of Protocorms from orchid seeds.
Principle

BM medium provides all essential Macroelements, Microelements, Vitamins, Amino acid & Plant growth regulators for the growth of Orchid in vitro. This medium is especially suitable for terrestrial orchids.
Macroelements :

Potassium nitrate Ammonium sulphate Calcium chloride anhydrous Magnesium sulphate Potassium phosphate monobasic
Microelements

2830.00 463.00 125.33 90.37 400.00 3.33 1.60 0.80 1.50 27.80 37.26 1.00 0.50 0.50 2.00

Potassium dihydrogen phosphate serves as a source of phosphate. This medium lacks in inorganic nitrogen.
Microelements:

Zinc and boron content in the medium is increased to provide proper nourishment to developing protocomes.
Vitamins:

Vitamins

Manganese sulphate.H2O Boric Acid Potassium iodide Zinc sulphate.7H2O Ferrous sulphate.7H2O Na2.EDTA

Thiamine content had been increased (0.5mg/l) in the medium. It is a most important element in carbohydrte metabolism and some amino acids biosynthesis. Biotin and folic acid along with other vitamins facilitates in vitro development of Protocorm.
Amino acid:

Thiamine HCL Pyridoxine HCL Nicotinic acid (Free acid)

Amino acid Storage

Glycine (free base)

Shelf Life

Glycine and glutamine serve as reduced nitrogen source.

Carbohydrte:

Store at 2-80C and prepared medium at 2-80C.

Use before expiry date as mentioned on the label.

Reference:

Sucrose serves as a carbon source.

Organic supplements:

Chu C.C., et. al., 1975. Scientia Sinic., 18, 659-668.

Casein hydrolysate used as a supplement, which is a sources of free amino acid.

CHU (N6) Medium

USE Macroelements :

APM1003/APM5003
Potassium nitrate serves as a source of nitrate. High concentration of ammonium ions has an inhibitory effect on the growth and quality of rice callus. The increased concentration of phosphate has a beneficial effect on the growth of rice callus.
Microelements:

CHU (N6) Medium is used to promote for organ culture and cell suspension culture.
Summary

Chu C.C. et al. (1975) has developed CHU (N6) Medium for in vitro anther culture of Oryza sativa (Family-Graminae). It is useful to generate new useful genetic varieties in gramineous plants by the initiation, growth, and differentiation of callus from rice pollen culture..
Principle

Boron, Managanese and zinc improves the quality of anther callus.

Vitamins:

CHU (N6) medium provides all essential Macroelements, Microelements, Vitamins & Amino acid for the growth of Orchid in vitro. This medium is not only to culture of rice anther but also for in vitro studies of other graminous plants.

Thiamine has been found to be beneficial for the growth and quality of rice callus. It is a most important elements of carbohydrate metabolism and biosynthesis of some amino acids.

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Ready to use... Formula Ingredients in Grams/Litre Macroelements Vitamins

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Thiamine HCl Pyridoxine HCl Nicotinic acid (Free acid)
Amino acid Storage

Potassium nitrate Ammonium sulphate Calcium chloride anhydrous Magnesium sulphate Potassium phosphate monobasic
Microelements

2830.00 463.00 125.33 90.37 400.00 3.33 1.60 0.80 1.50 27.80 37.26

1.00 0.50 0.50 2.00

Glycine (free base) Store at 2-80C and prepared medium at 2-80C.

Shelf Life

Manganese sulphate.H2O Boric Acid Potassium iodide Zinc sulphate.7H2O Ferrous sulphate.7H2O Na2..EDTA

Use before expiry date as mentioned on the label.

Reference:

Chu C.C., et. al., 1975. Scientia Sinic., 18, 659-668.

Gamborg B5 Medium
Use

APM1004/APM5004
Ammonium sulphate Calcium chloride anhydrous Magnesium sulphate Sodium phosphate monobasic
Microelements

Gamborg B5 Medium is used for callus culture and cell suspension culture.
Summary

Gamborg B5 Medium is established by Gamborg O.L. (1968) for callus and cell suspension culture of Glycine max (Family- Fabaceae). This medium is widely used for in vitro plant cell, tissue and organ culture.
Principle

134.00 113.24 122..09 130.42 10.00 3.00 0.75 0.21 2.00 0.025 0.025 27.80 37.26 100.00 10.00 1.00 1.00 8000.00

Gamborg B5 Medium provides all essential Macroelements, Microelements, & Vitamins for the growth of plant cell, tissue and organ culture in vitro.
Macroelements :

Increased nitrate content was found to beneficial for soyabean root callus. Potassium nitrate served as the sole source of nitrate in the medium. Ammonium sulphate fulfilled the ammonium requirement in the medium without altering cell growth. Sodium dihydrogen phosphate served as the source of phosphate in the medium.
Microelements:

Manganese sulphate.H2O Boric Acid Potassium iodide Molybdic acid (sodium salt).2H2O Zinc sulphate.7H2O Copper sulphate.5H2O Cobalt chloride.6H2O Ferrous sulphate.7H2O Na2.EDTA
Vitamins

Boron, Managanese and zinc provided for proper nourishment. Molybdenum, copper and cobalt were not included in the medium.
Vitamins:

Myo-Inositol Thiamine HCL Pyridoxine HCL Nicotinic acid (Free acid)

Gelling Agent

Agar

Thiamine content had been increased in the medium which supported the growth of soyabean cell suspension culture. The medium lacks glycine.
Formula Ingredients in mg per liter Macroelements

Storage

Shelf Life

Store at 2-80C and prepared medium at 2-80C.

Use before expiry date as mentioned on the label.

Reference:

Gamborg O.L., Miler R.A. And Ojima K., 1968. Exp. Cell Res., 50, 151-158. 2500.00

Potassium nitrate

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Murashige & Skoog Medium (MS)

Use

APM1005/APM5005
Calcium chloride anhydrous Magnesium sulphate Potassium phosphate monobasic
Microelements

Murashige & Skoog Medium (MS) is used for micropropagation, organ culture, callus culture and cell suspension culture..
Summary

332.16 180..69 170.00 16.90 6.20 0.83 0.25 8.60 0.03 0.03 27.80 37.30 100.00 0.10 0 .50 0 .50 2.00 30000.00 500.00 5.00 8000.00

Murashige & Skoog Medium (MS) is established by Murashige & Skoog (1962) for in vitro callus culture of Nicotiana tabacum (family- Solanaceae).
Principle

Murashige & Skoog Medium (MS) provides all essential Macroelements, Microelements, & Vitamins for the growth of plant cell, tissue and organ culture in vitro. Medium with high concentration of salts is used for cultivating plant cell, tissue and organ culture.
Macroelements :

Manganese sulphate.H2O Boric Acid Potassium iodide Molybdic acid (sodium salt).2H2O Zinc sulphate.7H2O Copper sulphate.5H2O Cobalt chloride.6H2O Ferrous sulphate.7H2O Na2.EDTA
Vitamins

In this medium nitrogen is supplied as ammonium and nitrate ions. This mixture of cation and anion balances the pH of the medium. Also plays a important role in plant growth. Potassium dihydrogen phosphate serves as a source of phosphate in medium.
Microelements:

Amino Acid

Myo-Inositol Thiamine HCL Pyridoxine HCL Nicotinic acid (Free acid)

Carbohydrate

Boron, Managanese, molybdenum, copper, iron and zinc plays a vital catalytic role in plant metabolism. Boron plays a key role in carbohydrate metabolism in plant cells.
Vitamins:

Glycine (Free base) Sucrose

Buffering Agent

Thiamine, pyridoxin and nicotinic acid content had been increased in the medium which have a stimulatory effect.
Amino acid:

MES (Free acid) Plant Growth Regulators 6-Benzyl amino purine(BAP)

Gelling Agent

Agar

The medium contains increased concentration of glycine.

Formula Ingredients in mg per liter Macroelements

Storage

Shelf Life

Store at 2-80C and prepared medium at 2-80C.

Use before expiry date as mentioned on the label. 1900.00 1650.00

Reference:

Potassium nitrate Ammonium sulphate

Murashige T. and Skoog F., 1962. Physio. Plant., 15, 473-497

Murashige & Skoog Modified Medium

Use Principle

APM1006/APM5006
Murashige & Skoog Medium (MS) provides all essential Macroelements, Microelements, & Vitamins for the growth of plant cell, tissue and organ culture in vitro.
Macroelements :

Murashige & Skoog Medium (MS) is used for micropropagation, organ culture, callus culture and cell suspension culture..
Summary

Murashige & Skoog Medium (MS) is established by Murashige & Skoog (1962) for in vitro callus culture of Nicotiana tabacum (family- Solanaceae). This medium was modified by incorporating plant growth regulators for the development of micropropagated explants in vitro.

In this medium nitrogen is supplied as ammonium and nitrate ions. This mixture of cation and anion balances the pH of the medium. Also plays a important role in plant growth. Potassium dihydrogen phosphate serves as a source of phosphate in medium.
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Boric Acid Potassium iodide Molybdic acid (sodium salt).2H2O Zinc sulphate.7H2O Copper sulphate.5H2O Cobalt chloride.6H2O Ferrous sulphate.7H2O Na2.EDTA
Vitamins

Boron, Managanese, molybdenum, copper, iron and zinc plays a vital catalytic role in plant metabolism. Boron plays a key role in carbohydrate metabolism in plant cells. Manganese to iron ratio is very important for plant metabolism. Zinc is essential for synthesis of tryptophan which is a precursor of indole-3-acetic acid. Iron is supplemented as FeEDTA to overcome the problem of precipitation in the medium.
Vitamins:

6.20 0.83 0.25 8.60 0.025 0.025 27.80 37.30 100.00 0.40 0.50 0.50 2.00 30000.00 0.30 1.75 1.75 80.00 30.00

Thiamine, pyridoxin and nicotinic acid content had been increased in the medium which have a stimulatory effect.
Amino acid:

The medium contains increased concentration of glycine.

Plant Growth Regulators:

Myo-Inositol Thiamine HCL Pyridoxine HCL Nicotinic acid (Free acid)

Amino Acid

Indole 3- butyric acid and a acetic acid are promote the - naphthalene elongation of cell or tissue and mainly roots. Adenine sulphate stimulates axillary bud growth and help for shoot growth. Indole -3- acetic acid is a naturally occuring plant growth regulators.
Formula IIngredients in mg per liter Macroelements

Glycine (Free base)

Carbohydrate

Sucrose

Plant Growth Regulators

Potassium nitrate Ammonium sulphate Calcium chloride anhydrous Magnesium sulphate Potassium phosphate monobasic Sodium phosphate monobasic
Microelements

1900.00 1650.00 332.16 180..69 170.00 148.00 16.90

Indole-3-acetic acid Indole-3-butyric acid a acetic acid - naphthalene Adenine hemisulphate 6-(g ,g -Dimethylallylamino) purine
Storage Shelf Life

Store at 2-80C. and prepared medium at 2-80C.

Use before expiry date as mentioned on the label

Reference:

Murashige T. and Skoog F., 1962. Physio. Plant., 15, 473-497

Manganese sulphate.H2O

Nitsch Medium
Use Microelements:

APM1007/APM5007
Boron, Managanese, molybdenum, copper, iron and zinc plays a vital catalytic role in plant metabolism. Boron plays a key role in carbohydrate metabolism in plant cells. Manganese to iron ratio is very important for plant metabolism. Zinc is essential for synthesis of tryptophan which is a precursor of indole-3-acetic acid. Iron is supplemented as FeEDTA to overcome the problem of precipitation in the medium.
Vitamins:

Nitsch Medium is used for Anther callus culture..

Summary

Nitsch Medium is established by Nitsch J. P. (1969) for in vitro anther culture of Nicotiana (Family-Solanaceae).
Principle

Nitsch Medium provides all essential Macroelements, Microelements, & Vitamins for anther callus culture in vitro.
Macroelements :

In this medium nitrogen is supplied as ammonium and nitrate ions. This mixture of cation and anion balances the pH of the medium. Also plays a important role in plant growth. Potassium dihydrogen phosphate which increases the growth rate of anther callus.
471

The medium contains essential vitamins for proper growth and development of anther callus. Thiamine content has been increased in the medium. Folic acid serves as a coenzyme. Biotin in the medium is an essential cofactor in fat, protein and carbohydrate metabolism.

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Myo-Inositol Thiamine HCL Pyridoxine HCL Nicotinic acid (Free acid) Folic acid D-Biotin 950.00 720.00 166.00 90.34 68.00 18.94 10.00 0.25 10.00 0.03 27.85 37.25
Amino Acid

The medium contains increased concentration of glycine.

Carbohydrate:

Sucrose serve as the source of carbohydrate.

Formula Ingredients in mg per liter Macroelements

100.00 0.50 0.50 5.00 0.50 0.05 2.00 20000.00

Potassium nitrate Ammonium sulphate Calcium chloride anhydrous Magnesium sulphate Potassium phosphate monobasic
Microelements

Glycine (Free base)

Carbohydrate Storage

Sucrose

Shelf Life

Store at 2-80C and prepared medium at 2-80C.

Manganese sulphate.H2O Boric Acid Molybdic acid (sodium salt).2H2O Zinc sulphate.7H2O Copper sulphate.5H2O Ferrous sulphate.7H2O Na2.EDTA

Use before expiry date as mentioned on the labe

Reference:

Nitsch J.P. And Nitsch C., 1969. Science, 163, 85-87 Indole 3- butyric acid and a acetic acid are promote the - naphthalene elongation of cell or tissue and mainly roots. Adenine sulphate stimulates axillary bud growth and help for shoot growth. Indole -3- acetic acid is a naturally occuring plant growth regulators.

Orchid Medium
Use Vitamins:

APM1008/APM5008
The medium contains essential vitamins for proper growth and development of Orchid culture. Thiamine content has been increased in the medium which is beneficial for the growth of orchid seedlings.
Carbohydrate:

Orchid Medium is used for Orchid culture..

Summary

Orchid Medium for in vitro cultivation of Orchids.

Principle

Orchid Medium provides all essential Macroelements, Microelements, & Vitamins for Orchid culture in vitro.
Macroelements :

Sucrose serve as the source of carbohydrate. Banana powder serve as a complex source of carbohydrate in the medium.
Organic supplements:

The macroelements are half the concentration of the Murashige and Skoog medium. In this medium nitrogen is supplied as ammonium and nitrate ions. This mixture of cation and anion balances the pH of the medium. Also plays a important role in plant growth. Potassium dihydrogen phosphate which increases the growth rate of anther callus.
Microelements:

Meat peptone and tryptone are the forms of nitrogen, supplied to the medium.
Buffering Agent:

2-(N-morpholino) ethanesulfonic acid (MES) is commonly used as a buffering agent. This prevents acidification by buffering the medium.
Adsorbent Agent:

Boron, Managanese, molybdenum, copper, iron and zinc plays a vital catalytic role in plant metabolism. Boron plays a key role in carbohydrate metabolism in plant cells. Manganese to iron ratio is very important for plant metabolism. Zinc is essential for synthesis of tryptophan which is a precursor of indole-3-acetic acid. Iron is supplemented as FeEDTA to overcome the problem of precipitation in the medium.

Activated charcoal in the medium adsorbs toxic phenolics. Also used in rooting medium to adsorb root inhibiting agents.
Formula IIngredients in mg per liter Macroelements

Potassium nitrate Ammonium sulphate Calcium chloride anhydrous

950.00 825.00 166.08

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90.34 85.00 8.45 3.10 0.42 0.12 5.30 0.013 0.013 27.80 37.30 100.00 10.00 1.00 1.00 20000.00 Banana Powder
Organic Supplement

Magnesium sulphate Potassium phosphate monobasic

Microelements

30000.00 2000.00 2000.00 1000.00 2000.00 8000.00

Peptone from meat Tryptone

Buffering Agent:

Manganese sulphate.H2O Boric Acid Potassium iodide Molybdic acid (sodium salt).2H2O Zinc sulphate.7H2O Copper sulphate.5H2O Cobalt chloride.6H2O Ferrous sulphate.7H2O Na2.EDTA
Vitamins

MES (Free acid)

Adsorbing Agent:

Activated Charcoal
Gelling Agent:

Agar

Store at 2-80C and prepared medium at 2-80C.

Shelf Life

Storage

Use before expiry date as mentioned on the label.

Reference:

Nitsch J.P. And Nitsch C., 1969. Science, 163, 85-87 Indole 3- butyric acid and - naphthalene acetic acid are promote the elongation of cell or tissue and mainly roots. Adenine sulphate stimulates axillary bud growth and help for shoot growth. Indole -3- acetic acid is a naturally occuring plant growth regulators.

Myo-Inositol Thiamine HCL Pyridoxine HCL Nicotinic acid (Free acid)

Carbohydrate

Sucrose

Schenk & Hidebrandt Medium (SH)

Use

APM1009/APM5009
plant cells. Manganese to iron ratio is very important for plant metabolism. Zinc is essential for synthesis of tryptophan which is a precursor of indole-3-acetic acid. Iron is supplemented as FeEDTA to overcome the problem of precipitation in the medium. Copper content was increased as it shows stimulatory effect in monocotylenous callus. Iron is supplemented as ferric citrate to overcome the problem of precipitation in the medium. Ferric is converted to the active ferrous forms.
Vitamins:

Schenk & Hidebrandt Medium (SH) used for callus and cell suspension culture.
Summary

Schenk & Hidebrandt Medium (SH) was established by Schenk R.U & Hilderbrandt A.C. (1972) for in vitro callus culture of monocotyledonous and dicotyledonous plants.
Principle

Medium supports the growth of both monocotyledonous and dicotyledonous plant tissues by providing all essential nutrients .
Macroelements :

Myo inositol concentration was increased in the medium as it shows stimulatory effect in monocotyledonous and dicotyledonous callus.
Buffering Agent:

Nitrogen is supplied as ammonium and nitrate ions. This mixture of cation and anion balances the pH of the medium. Also plays a important role in plant growth. Potassium dihydrogen phosphate which increases the growth rate of anther callus.
Microelements:

2-(N-morpholino) ethanesulfonic acid (MES) is commonly used as a buffering agent. This prevents acidification by buffering the medium.
Adsorbent Agent:

Boron, Managanese, molybdenum, copper, iron and zinc plays a vital catalytic role in plant metabolism. Boron plays a key role in carbohydrate metabolism in
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Activated charcoal in the medium adsorbs toxic phenolics. Also used in rooting medium to adsorb root inhibiting agents. Indole 3- butyric acid and a acetic acid are promote the - naphthalene

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Zinc sulphate.7H2O Copper sulphate.5H2O Cobalt chloride.6H2O Ferrous sulphate.7H2O Na2.EDTA 2500.00 300.00 151.02 195.34 10.00 5.00 1.00 0.10
Vitamins

elongation of cell or tissue and mainly roots. Adenine sulphate stimulates axillary bud growth and help for shoot growth. Indole -3- acetic acid is a naturally occuring plant growth regulators.
Formula Ingredients in mg per liter Macroelements

1.00 0.20 0.10 15.00 20.00 1000.00 5.00 0.50 5.00

Potassium nitrate Ammonium phosphate monobasic Calcium chloride anhydrous Magnesium sulphate
Microelements

Myo-Inositol Thiamine HCL Pyridoxine HCL Nicotinic acid (Free acid)

Storage Shelf Life

Manganese sulphate.H2O Boric Acid Potassium iodide Molybdic acid (sodium salt).2H2O

Store at 2-80C and prepared medium at 2-80C.

Reference:

Use before expiry date as mentioned on the label. Schenk R.U. & Hilderbrandt a.C., 1972. Can. J. Bot., 50, 199-204.

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Plant Tissue Culture Chemicals Macro Elements

Ammonium nitrate

(Suitable for PTC*)

Calcium chloride

APCM10011 APCM50011-5Kg APCM10012 APCM10012-5Kg APCM10013 APCM50013-5Kg APCM10014

Potassium nitrate

(Suitable for PTC*)

Potassium phosphate monobasic

APCM10015 APCM50015-5Kg APCM10016 APCM10016-5Kg APCM10017 APCM10017-5Kg APCM10018 APCM10018-1Kg

(Suitable for PTC*)

Magnesium sulphate heptahydrate

(Suitable for PTC*)

Sodium phosphate monobasic

(Suitable for PTC*)

Potassium chloride

(Suitable for PTC*)

Sodium Nitrate

(Suitable for PTC*)

(Suitable for PTC*)

Micro Elements
Boric acid

APCM50101

Ferrous sulphate heptahydrate

(Suitable for PTC*)

Copper sulphate pentahydrate

(Suitable for PTC*) APCM50102

Manganese sulphate monohydrate

APCM50106 APCM50106-5Kg APCM50107

(Cupric sulphate pentahydrate) (Suitable for PTC*)

Cobalt chloride hexahydrate

(Suitable for PTC*)

Potassium iodide

APCM10108

APCM10103

(Suitable for PTC*)

Sodium Molybdate dihydrate

(Suitable for PTC*) APCM10109

EDTA Ferric Mono Sodium Salt

APCM50104

(Suitable for PTC*)

Zinc sulphate heptahydrate

(Suitable for PTC*)

EDTA Disodium salt dihydrate

APCM50105

(Suitable for PTC*)

APCM50111 APCM10111-1K

(Suitable for PTC*)

Vitamins
D-Biotin

APV0001

(Suitable for PTC*)

Folic acid

L-Ascorbic acid (Vitamin C)

APV0003

(Suitable for PTC*) APV0002 APV0002-10gm

Myo-Inosotol or (Meso-Inositol)

(Suitable for PTC*)

APV0004

(Suitable for PTC*)

475

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Ready to use... Nicotinic acid or Niacin(Free acid)

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APV0005
Thiamine Hydrochloride

(Suitable for PTC*)

Pyridoxine Hydrochloride

(Suitable for PTC*) APV0006 APV0006-25gm

APV0007 APV1007

(Suitable for PTC*)

Amino Acid
L-Glycine (Free base)

APA0001

L-Tyrosine

(Suitable for PTC*)

L-Glutamine

(Suitable for PTC*) APA0002 APA1002

APA0003 APA1003

(Suitable for PTC*)

Carbohydrate
D-Glucose

APC5001

Sucrose

APC5002

(Suitable for PTC*)

(Suitable for PTC*)

Plant Growth Regulators

Adenine Sulphate

APR0001

Indole-3-Acetic acid

APR0004

(Suitable for PTC*)

6-Benzy laminopurine (BAP)

(Suitable for PTC*) APR0002

Kinetin (6-Furfurylaminopurine)

(Suitable for PTC*)

Indole- 3-Butyric acid (IBA)

(Suitable for PTC*) APR0003

a- Naphthalene acetic acid

APR0005 APR0005-5gm APR0006

(Suitable for PTC*)

(Suitable for PTC*)

Adsorbent
Activated Charcoal

(Suitable for PTC*)

APG10001 APG50001

Sucrose

APC5002

(Suitable for PTC*)

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