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Briefly describe the essential components of the nutrient medium used for the plant tissue culture technique. Also, write the names of any two plant tissue culture media frequently used in the laboratory .

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Nutrient Medium : Virtually all tissue culture media are synthetic or chemically defined; only a few of.them use complex organics, e.g., potato extract, as their normal constituents. A synthetic medium consists of only chemically defined compounds. A variety of recipes have been developed since none of them is suitable for either all plant species or for every purpose. Most of these recipes have been elaborated from those of White (itself evolved from a medium for algae) and Gautheret (based on Knop’s salt solution) The composition is as follow : 

Inorganic nutrients: In addition to C, H and O, all nutrient media provide the 12 elements essential for plant growth, viz., N, P, K, Ca, S, Mg (these six are called macronutrients, and are needed in concentrations >0.5 mmol L-1 or > 0.5 mM), Fe, Zn, Mn Cu, B and Mo (these six are known a micronutrients, and are required in concentrations <0.5 mmol L” ’ ) Generally, iron is provided as iron.EDTAcomplex to keep it available at higher (>5.8) pH. Nitrate is superior to ammonium as the sole N source, but use of NH+ checks the drift of pH towards alkalinity. 

Vitamins : For optimum callus growth, the following vitamins are required . inositol, thiamine, pyridoxine and nicotinic acid of which thiamine is essential and the rest are promotory. Pantothenic acid is also known to be promotory but is not included in most of the recipes. 

Carbon source : Sucrose (20-50 g L) is the most commonly used carbon source for all cultured plant materials, including even green shoots. In some systems, e.g., monocots, glucose may be superior to sucrose. Plant tissue can utilize other sugars like maltose, galactose, lactose, mannose and even starch but these are rarely used. 

Growth regulator: The following growth regulators (GRs) are used in plant tissue culture. Auxins, e.g., IAA (indole-3-acetic acid), IBA (indole-3-butyric acid), NAA (napthalene acetic acid), NOA (naphthoxy acetic acid), 2, 4-D (2,4-dichlorophenoxy acetic acid) etc., are commonly used to support cell division and callus growth (especially 2. 4-D), somatic embryo (SE) induction, rooting, etc. Cytokinins like kinetin (furfurylamino purine), BAP (benzylamino purine), zeatin, 2-ip (isopentenyl adenine), TDZ (thidiazuron, a compound having cytokinin activity) are employed to promote cell division, regeneration of shoots often SE induction and to enhance proliferation and growth of axillary buds. Abscisic acid (ABA) promotes SE and shoot bud regeneration in many species are markedly improves SE maturation. Of the over 20 gibberellins known, GA3 is almost exclusively used. It promotes shoot elongation and SE germination. 

Complex organic additives: In earlier studies, complex additives like yeast extract, coconut milk, casein hydrolysate, com milk, malt extract and tomato juice are used to support plant tissue growth. White’s medium, Murashige and Skoog (MS) are the two common media used for plant tissue culture.

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