The post Protoplast Culture and its Applications, MCQ for GPAT, GATE, DBT BET & CSIR NET appeared first on Gpatindia: Pharmacy Jobs, Admissions, Scholarships, Conference,Grants, Exam Alerts.
]]>Protoplast is the plant cell without a cell wall. This protoplast possesses other cellular components and a plasma membrane. Hence, this protoplast is a functional plant cell without a barrier i.e. cell wall.
1. Source of protoplast: Tissues and organs like leaves, roots, shoot apices, fruits, embryos and microspores are widely used as sources for protoplast. Callus and suspension cultures are also good sources for protoplast isolation.
2. Isolation of protoplast: It can be done by two methods
a) Mechanical method: In this method, the cells of the epidermis are subjected to plasmolysis, by which protoplast causes to shrink away from the cell wall. Then, the protoplast is released by tissue dissection. The main disadvantage of this method that, the yield and viability of protoplast is very low. This process is tedious and laborious.
b) Enzymatic method: This method has an advantage over the mechanical method, which has a high yield of protoplasts with viable cells. Hence, it is a widely used method. Enzymes like cellulases, hemicellulases, pectinases, are used to degrade cellulose, hemicellulose, and pectin in the plant cell wall.
One of the two methods is used to isolate protoplast by enzymatic method.
3. Purification of protoplast: It is done by filtration to remove cell clumps and undigested tissues, followed by centrifugation and washing of protoplast.
4. Viability test of protoplast: The following are the methods used to test the viability of protoplast
a) Testing photosynthetic activity of protoplast
b) Measurements of cell wall formation can be observed using Calcofluor white (CFW) stain, which bind to the newly formed cell wall and emits fluorescence.
c) Measurement of oxygen uptake by protoplast by using oxygen electrode
d) Phenosafranine stain: dead protoplast uptake this stain and turns red, whereas viable protoplast remains unstained.
e) Fluorescein diacetate (FDA) staining: viable cells get stained by the FDA and can be detected by fluorescence microscopy.
5. Culture of Protoplast
Cell wall development around the protoplast membrane is the prior step of protoplast culture. The development of the cell wall is followed by cell division to form small colonies. Protoplast is generally culture in semi-solid Agar medium or Liquid medium.
Advantages of liquid culture over Solid culture
Nutritional Requirements of Culture media
It is similar to those used in callus culture EXCEPT:
Protoplast culture methods
Regeneration of protoplast
Protoplast development can be done in two steps
The millions of single cells are formed by protoplast isolation and protoplast culture, which can be used for wide varieties of studies
1. Protoplasts are__________
a) Cell without a nucleus
b) Cell without a cell wall
c) Cell without plasma membrane
d) Cell without genetic material
2.__________ is the first step for protoplast culture
a) Selection of Explant
b) Viability testing for protoplast
c) Isolation of protoplast
d) Preparation of culture media
3. ___________ is the advantage of mechanical method for isolation of protoplast
a) The process is very tedious
b) Very less viable protoplast are isolated
c) Both
d) Only B
4. Cellulase enzyme in isolation of protoplast is used ________
a) To degrade proteins
b) To degrade cellulose
c) To degrade pectin
d) To degrade hemicellulose
5. ____________ is used to check viability of protoplast
a) Phenosafranine staining
b) Fluorescein diacetate (FDA) staining
c) Both
d) Only A
6. The following are the advantages of liquid protoplast culture EXCEPT:
a) It is easy to transfer
b) Dilutions can be easily done in liquid culture
c) Cell density can be easily manipulated
d) Less time consuming
7. Co-culture of protoplast is appropriate for ___________
a) Protoplast from two different organs of the same plant
b) Protoplast from the same organ of different plant
c) Protoplast from two different plant species
d) Protoplast from two same plant species
8. ___________media is used for protoplast culture
a) Synthetic media
b) Natural media
c) Nutritional media
d) None of the above
9. High Auxin/kinetin ratio in nutritional media for protoplast culture is preferred ______
a) To induce cell regeneration
b) To induce cell growth
c) To induce cell division
d) All
10. Advantage of Micro-drop protoplast culture technique is _____________
a) It requires Large numbers of protoplasts
b) It requires a Small number of protoplasts
c) It requires a large amount of culture media
d) It requires less amount of water
Reference
The post Protoplast Culture and its Applications, MCQ for GPAT, GATE, DBT BET & CSIR NET appeared first on Gpatindia: Pharmacy Jobs, Admissions, Scholarships, Conference,Grants, Exam Alerts.
]]>