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Concluding remarks

Like all other plant parts, endosperm can also respond the same way under in vitro conditions irrespective of their genetic constitutions. Hence, it helped in changing the misconception that endosperm being a “dead tissue” has now been contradicted by several reports suggesting the full plant regeneration from endosperm. Despite the success of plant regeneration from endosperm cultures in a number of systems, this protocol for production of triploid plants remained unutilised mostly. It may be basically due to the difficulty in obtaining organogenic callus from the mature or immature endosperms. The ploidy variation exhibited by endosperm derived plantlets is another difficulty which limits this technique. More efforts should be focussed on endosperm regeneration from plants where seedlessness is employed to improve the quality of fruits and plants that has economically useful vegetative parts.

 

 

EXPERIMENT

AIM: Triploid plant production by in vitro endosperm cultures of Neem (Azadirachta indica A. Juss)

 

EQUIPMENTS: Autoclave, pH-meter, Magnetic stirrers, Magnetic beads, Weighing balance, Laminar-air-flow, Microsopes.

 

MATERIALS REQUIRED : Salts and vitamins of Murashige and Skoog's (MS; 1962), sucrose, agar, conical flasks, measuring cylinders and beakers of various sizes. Reagent glass bottles for storage, spatula, tissue rolls, distilled water. Cotton plugs, aluminium foils, muslin cloth, scissor, media stocks, 1N NaOH, 1N HCl, myo-inositol. Autoclavable polybags, rubber bands, borosil glass test-tubes (150mm x 25mm without rim). Black markers, micropippete, micropippete- tips, test-tube stands, autoclavable baskets, plastic trays, immature seeds of Azadirachta indica .

 

Plant Growth regulators (Sigma):

Benzyl amino purine (BAP)

6-furfuryl amino purine (Kinetin)

Thidiazuron (TDZ)

6-γ γ –Dimethylallylamino purine (2-iP)

Indole-3-acetic acid (IAA)

Naphthalene acetic acid (NAA)

Casein hydrolysate (CH)