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In Vitro Culture Techniques : The Biotechnological Principles

 
     
 
In vitro Androgenesis
In vitro androgenesis is the formation of sporophyte from the male gametophyte on artificial medium. It is most commonly found in family Solanaceae and Poaceae (Graminae). Success of in vitro androgenesis is based on adjustment of developmental stage of pollen, minerals of culture medium and growth regulators as well as thermal shock or other treatments. 

Methods of in vitro androgenesis is given in Fig. 8.10. Pollen grains are isolated from excised anther and extracted in a beaker. Well collected pollens are washed properly and centrifuged and decanted. They are inoculated in liquid medium, then subcultured in solid MS medium. In culture pollen grains can be induced to produce callus or embryo from which whole plants are regenerated in one month. This technique is most successful in Brassica, Datura, Petunia, etc. Mainly there are two ways of in vitro morphogenesis of pollen grains, direct and indirect (Fig. 8.10).

  Types of in vitro pollen morphogenesis in Datura (based on Sangwan, 1981)  
 

Fig. 8.10. Types of in vitro pollen morphogenesis in Datura (based on Sangwan, 1981)

 

(i)

Direct androgenesis. Direct androgenesis is also called pollen derived embryogenesis. Here pollen directly acts as a zygote and, therefore, passes through various embryogenic stages similar to zygotic embryogenesis. When pollen grains has reached globular stage of embryo, wall of pollen is broken and embryo is released. The released embryo develops cotyledons, then the plants. Direct androgenesis is very common in many plants of the family Solanaceae and Brassicaceae (Fig. 8.10) (Prakash and Giles, 1987).

(ii)

Indirect androgenesis. In indirect androgenesis the pollen grains, instead of normal embryogenesis, divide erratically to develop callus (Fig. 8.10). Indirect androgenesis has been found in barley, wheat, Vitis, coffea, etc. Possibility of pollen morphology from the dividing pollen varies. The haploid callus, embryo or plantlets may originate from (i) the continued division of vegetative cells of the pollen (the generative cell soon degenerate), (ii) multiple division of generative cells (non-vegetative cell), and (Hi) division products of both generative and vegetative cells (Sangwan, 1981).

 

Content

Totipotency

Historical background

Requirements for cell and Tissue Cultures

 

A tissues culture laboratory

 

Nutrient media

 

 

Inorganic chemicals

 

 

Growth hormones

 

 

Organic constitutents

 

 

Vitamins

 

 

Amino acids

Culture of plant materials

 

Explant culture

 

Callus formation and its culture

 

Organogenesis

 

Root culture

 

Shoot culture and micropropagation

 

Cell culture

 

 

Benefits from cell culture

 

Somatic embryogenesis

 

Somaclonal variation

 

Protoplast culture

 

 

Isolation

 

 

Regeneration

 

Protoplast fusion and somatic hybridization

 

 

Fusion products

 

 

Method of somatic hybridization

 

Anther and pollen Culture

 

 

Culturing techniques

 

In vitro androgenesis (direct and indirect androgenesis)

 

Mentor pollen technology

 

Embryo culture

 

Embryo rescue

 

Protoplast fusion in fungi

 
     
 
 
     



     
 
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