Culture Media in Plant Tissue Culture

By: Pharma Tips | Views: 9194 | Date: 30-Mar-2011

Plants in nature can synthesize their own food material. In contrast, plants growing in vitro are heterotrophic, Le., they cannot synthesize their own food material.

Culture media:

Media - Plants in nature can synthesize their own food material. In contrast, plants growing in vitro are heterotrophic, Le., they cannot synthesize their own food material.

-Plant tissue culture media therefore require all essential minerals plus a carbohydrate source usually added in the form of sucrose and also other growth hormones (regulators and vitamins).

-Growth and morphogenesis of plant tissues in vitro are largely governed by the composition of the culture media.
-Although the basic requirements of cultured plant tissues are similar to those of whole plants, in practice nutritional components promoting optimal growth of a tissue under laboratory conditions may vary with respect to the particular species.

-Media compositions are thus formulated considering specific requirements of a particular culture system. 'For example, some tissues show better response on a solid medium while others prefer a liquid medium.Considerable progress has been made during the past two decades on the development of media for growing plant cells, tissues and organs aseptically in culture.

-A significant contribution to formulation of a defined growth medium suitable for a wide range of applications was made by Murashige and Skoog (1962), In their work to adapt tobacco callus cultures for use as a hormone bioassay system, they evaluated many medium constituents to achieve optimal growth of calluses.

-In so doing, they improved upon existing types of plant tissue culture media to such an extent that their medium (the MS medium) has since proved to be one of the most widely used in plant tissue culture work gives the composition of different media.

Composition of Culture Media

1.    Inorganic Salts
2.    Carbon Sources
3.    Vitamins
4.    Phytohormones
5.    Organic supplements
6.    Trace elements


(1) Inorganic Salts:–
o     To induce a callus from an explant and to cultivate the callus and cells in suspension, various kinds of media (inorganic salt media) have
       been designed.
o    Agar or its substitutes is added into the media to prepare solid medium for callus induction.One of the most commonly used media for
     plant tissue cultures is that developed by Murashige and Skoog (MS) for tobacco tissue culture (28).

o    The significant feature of the MS medium is its very high concentration of nitrate, potassium and ammonia. The B5 medium established
       by Gamborg et al.

o    (29) is also being used by many researchers. The levels of inorganic nutrients in the B5 medium are lower than in MS medium.
o    Many other media have been developed and modified and nutrient compositions of some typical media will be described in Table 6.
o    However, it is not always necessary to test many kinds of basal media when a callus is induced.

o    It would be better to use only one or two kinds of basal media in combination of different kinds and concentrations of phytohormones.
o    The most suitable medium composition should be optimized afterwards in order to obtain higher level of products as well as higher
      growth rate.

(2) Carbon Sources:–
o    Sucrose or glucose at 2 to 4% are suitable carbon sources which are added to the basal medium.
o    Fructose, maltose and other sugars also support the growth of various plant cells.
o    However, the most suitable carbon source and its optimal concentration should be chosen to establish the efficient production process of
      useful metabolites.
o    These factors depend on plant species and products, therefore it is necessary to optimize the medium compositions including carbon
       sources in each case.
o    From an economical point of view, the use of more inexpensive carbon sources is appropriate in industry and crude sugars such as
       molasses have been examined.

(3) Vitamins: –
o    The basal media described above such as MS medium include myo-inositol, nicotinic acid, pyridoxine HCl and thiamine HCl. Among
       these vitamins, thiamine is an essential one for many plant cells and other vitamins stimulate the growth of the cells in some cases.
o    The level of myo-inositol in the medium is 100 mg/L which is very high although it is not clear whether such a high level of the vitamin is
       required.

(4) Phytohormones: –
o    Phytohormones or growth regulators are required to induce callus tissues and to promote the growth of many cell lines.
o    As an auxin, 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneaceic acid (NAA) is frequently used.
o    The concentration of auxins in the medium is generally between 0.1 to 50 µM. Kinetin or benzyladenine as a cytokinin is occasionally
      required together with auxins for callus induction at concentrations of 0.1 to 10 µM. Other derivatives of auxin and kinetin are also used
       in some cases.

o    Since each plant species requires different kinds and levels of phytohormones for callus induction, its growth and metabolites production,
       it is important to select the most appropriate growth regulators and to determine their optimal concentrations.
o    Gibberellic acid is also added to the medium if necessary.

(5) Organic supplements:–
o    In order to stimulate the growth of the cells, organic supplements are sometimes added to the medium.
o    These supplements include casamino acid, peptone, yeast extracts, malt extracts and coconut milk. Coconut milk is also known as a
       supplier of growth regulators.

(6) Trace element–

     Boron(B)                            BO

     Manganese(Mn)                 Mn++

     Zinc(Zn)                             Zn++

    Copper(Cu)                        Cu+,Cu++

    Molybdenum(Mo)              MoO4-

    Chlorine (Cl)                        Cl-

     Nickel(Ni)                                 -

     Aluminium(Al)                         -

     Cobalt(Co)                                -

     Iodine(I)                                    -

     Ferrous(Fe)                               -

    Sodium (Na)                             

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