Q.(a) Mention the applications of Bio-Technology. OR

(a) Biotechnology applies across agriculture, medicine, industry, environment, forensics and food; (b) protoplast culture isolates wall-less cells and regenerates them into whole plants.

(a) Applications of Biotechnology

Biotechnology, the use of living organisms or their products for practical purposes, has wide-ranging applications:

• Agriculture: development of genetically modified (Bt/GM) crops with pest, disease, or herbicide resistance; production of microbial biofertilizers and biopesticides to reduce chemical input use.
• Medicine: production of recombinant therapeutic proteins such as human insulin and growth hormone; development of recombinant vaccines; gene therapy for correcting genetic disorders; and molecular diagnostic kits for disease detection.
• Industry: production of industrial enzymes, biofuels (bioethanol/biodiesel), and biodegradable bioplastics using microbial or plant systems.
• Environment: bioremediation, using microorganisms to break down pollutants in soil or water, and phytoremediation, using plants to absorb and detoxify contaminants.
• Forensics: DNA fingerprinting for identification in criminal investigations and paternity testing.
• Food and nutrition: biofortified crops such as Golden Rice (enriched in beta-carotene), and improved fermentation/dairy processes.

(b) Steps in Protoplast Culture

1. Select healthy, actively growing leaf or other tissue as the source material and surface-sterilise it.
2. Digest the cell walls enzymatically, using a mixture of cellulase (which digests cellulose) and pectinase/macerozyme (which digests the middle lamella), carried out in an osmotically stabilised (isotonic) solution containing an osmoticum such as mannitol or sorbitol, to release naked protoplasts without bursting them.
3. Purify the released protoplasts by filtering the digest through a fine mesh to remove undigested tissue debris, followed by centrifugation to separate healthy, intact protoplasts from broken cells and other debris.
4. Check the viability of the isolated protoplasts, commonly using a stain such as fluorescein diacetate (FDA). …