SCIENCE AND AGRO STUDIES

Objective:- To isolate protoplasts by enzymatic method. Protoplast:- Protoplast is the living material of the cell where as an isolated protoplast is the cell from which the cell wall is removed. In plant breeding programme many desirable combination of characters could not be transmitted through the conventional method of genetic manipulation. Principle:- Protoplasts are isolated by treating tissues with a mixture of cell wall degrading enzyme in solution, which contain osmotic stabilizer. A most suitable source of protoplasts is mesophyll tissue from fully expanded leaves of young plants or new shoots. The release of protoplast is very much dependent on the nature and composition of enzymes used to digest the cell wall. There are three primary components of the cell wall which have been identified as cellulose, hemicellulose and pectin substance. Pectinase (macrozyme) mainly degrades the middle lamella while cellulose and hemicellulase degrades the cellulose and hemicellulosic com...

Objective:- To isolate protoplast by mechanical method. Protoplast:- Protoplast is the living material of the cell where as an isolated protoplast is the cell from which the cell wall is removed. In plant breeding programme many desirable combination of characters could not be transmitted through the conventional method of genetic manipulation. Principle:- Protoplast can be isolated from almost all plant parts: roots, leaves, fruits, tuber, root nodules, pollen mother cell etc. Protoplast isolated by mechanical is a crude and tedious procedure. Cells are plasmolysed causing the protoplast to shrink from the cell wall. The protoplast obtained from this method is then cultured on suitable culture medium. The principle deficiency of this approach is that the protoplast released is few in number. Mechanical isolation was that of only historical event now. Materials Required:- i. Plant leaves ii. Mortar and pestle iii. Phosphate buffer pH-7.0 iv. 0.3 M sorbitol v. 0.3 M mannitol vi. Glass ...

Objective:- Protocol for somatic embryogenesis in carrot. Plant Material:- Hypocotyl of carrot seedling. Procedure:- 1. Wash seeds by submerging in water with a few drops of detergent in a beaker and shake by hand, or wrap seeds in two layers of cheese cloth/muslin cloth/nylon pouch and then wash with water. 2. Submerge the seeds in 70% alcohol for 30-60s. Decant the alcohol. 3. Transfer the seeds to a flask or beaker containing 20-40% commercial sodium hypochlorite for 15-20 min. Rinse 4x with sterile distilled water. 4. Place 2-3 seeds per culture vessel on the surface of MS agar medium. 5. Incubate the cultures at 25°C under 16 h photoperiod with -1000 lux light intensity for 1-2 weeks. 6. Collect the germinated seedlings when the cotyledons are fully expanded. Place each seedling on a sterile petri dish and excise the hypocotyl from each seedling and cut them transversely into two parts. 7. Place the hypocotyl sections on the following medium: MS + 1-2 mg/l 2,4-D. 8. Incubate the...

Auxotrophic mutants:-  > These are bacteria, yeast, protoplast or mammalian host cell strains that can’t produce a nutrient vital for growth due to genetic mutations. As a result, these strains are unable to survive in media lacking that specific nutrient unless it is provided externally. > The term auxotrophy refers to a nutritional dependency, where the mutant requires an auxiliary or supplemental source of the missing nutrient for its growth and survival. In contrast, the corresponding wild type strain can synthesize that specific nutrient and is not dependent on an exogeneous supply in the growth medium. > Auxotrophic mutants are often used in genetic studies, selection experiments, and research involving metabolic pathways and nutrient utilization. They are also very useful in gene cloning procedures. साइब्रिड्स ( Cybrids ):- जब कायिक संकरण के दौरान एक जाति से केन्द्रक व कोशिकाद्रव्य तथा दूसरी जाति से केवल कोशिकाद्रव्य आते हैं तो इस प्रकार बने ...