ZOOLOGY

Additional Content for NEET-2024

Biotechnology and
Its Applications
Chapter Contents
z Introduction Introduction
Human population is rising steadily. The requirement of food is rising
z Tissue Culture
proportionally. Therefore, a continued enhancement of food production
has become a major necessity. New techniques like tissue culture are
of great importance to increase food production.

TISSUE CULTURE
As traditional breeding techniques failed to keep pace with demand and
to provide sufficiently fast and efficient systems for crop improvement,
therefore another technology called tissue culture got developed.

(1) What does tissue culture mean?

Plant tissue culture is the maintenance and growth of plant cells,
tissues and organs on a suitable culture medium in-vitro (in a
container e.g., test tube). The plant part which is cultured is called
explant, i.e., any part of plant taken out and grown in a test
tube, under sterile conditions in special nutrient media. This
capacity to generate a whole plant from any cell/explant is called
totipotency.
(2) Environmental conditions :
There are three important aspects of in-vitro culture namely
nutrient medium, aseptic conditions and aeration of the tissue.
(a) Nutrient medium : It will provide the nutrition that is
required for the desired development of the explants. Culture
medium contains carbon source (such as sucrose), inorganic
salts, vitamins, amino acids and growth regulators. The
growth regulators commonly used in plant tissue culture are
auxins like 2,4-D and cytokinins, such as BAP (benzyl
amino purine). Growth regulators are required for cell division
and organ regeneration from the cultures. Culture medium
may be liquid or solid. The optimum pH should be 5.7.

Corporate Office : Aakash Tower, 8, Pusa Road, New Delhi-110005. Phone : 011-47623456
2 Biotechnology and Its Applications Additional Content for NEET-2024

(b) Sterilisation : It is essential that the explants, culture vessels, media and the instruments used
for tissue culture be made free from microbes. For this purpose, explants are treated with specific
anti-microbial chemicals. This procedure is called surface sterilisation. The vessels, media and
instruments are suitably treated with steam (autoclave), dry heat or alcohol, or subjected to filtration
to make them free from microbes. This is called complete sterilisation.
(c) Aeration of the tissue : Proper aeration of the cultured tissue is also an important aspect of tissue
culture technique.
(3) Method of plant tissue cultures :
Plant tissue culture includes two major methods :
(a) Type of in-vitro growth – Callus and suspension cultures.
(b) Type of explant – Meristem culture, protoplast culture, embryo culture etc.
Callus and Suspension Cultures
(i) In callus culture, cell division in the explant forms an unorganised mass of cells called callus. It is
maintained on a medium gelled with agar. The medium usually contains the auxin 2,4-D, and a cytokinin
like BAP.
(ii) In suspension culture, explant is suspended into liquid medium containing auxin, 2,4-D and is
constantly agitated at the speed of 100–250 rpm (revolutions per minute).
(iii) In both the type of tissue cultures, with passage of time, cell/tissue dry matter increases and level of
nutrient decreases. To prevent the damage of newly formed cells, part of the cultures are regularly
transferred to new culture vessels containing fresh media. This process is termed as subculturing.
(iv) Shoot and root regenerations are generally controlled by auxin-cytokinin balance. Usually, an excess
of auxin promotes root regeneration and excess of cytokinin promotes shoot regeneration. Callus cultures
are first kept on a BAP-containing medium. After some time, shoots regenerate from callus cells. When
the shoots become 2-3 cm long, they are excised and transferred to an auxin-containing medium. Roots
regenerate from the lower ends of these shoots to yield complete plantlets.
(v) Plantlets are removed from culture vessels and established in the field. This transfer is done by specific
procedures called hardening. During hardening, plantlets are kept under reduced light and high
humidity. Hardening procedures make the plantlets capable of tolerating the relatively harsh environments
outside the culture vessels.
Meristem Culture : Pathogen free clones of plants can be obtained through meristem culture because
meristem (apical and axillary) is free of virus due to high concentration of auxins and rapid rate of
cell division. The apical meristem accompanied by 1-2 leaf primordia may be taken. For this the apical bud
is sterilised. The shoot tip is now placed over culture medium under aseptic conditions. Scientists have
succeeded in culturing meristems of banana, sugarcane and potato.
Protoplast fusion or Somatic hybridisation : It is fusion of protoplasts of two plants belonging to different
varieties, species and even genera. The cells are first treated with enzymes pectinase and cellulase. These
enzymes dissolve the cell wall and as a result naked protoplasts are produced.

Vacuole Nucleus

Cellulase
+
Pectinase

Cell wall
Plant cell Cell wall
Plasmalemma Protoplast

Fig.: Protoplast Preparation

Corporate Office : Aakash Tower, 8, Pusa Road, New Delhi-110005. Phone : 011-47623456
Additional Content for NEET-2024 Biotechnology and Its Applications 3
The naked protoplasts are fused by electrofusion (high frequency alternating electric field with short current
pulses) or chemofusion (through sodium nitrate or PEG = polyethylene glycol). It results in hybrid protoplasts.
Hybrid protoplasts can be further grown to from a new plant. These hybrids are called somatic hybrid while
the process is called somatic hybridisation. The somatic hybrid may have a synkaryon (single fused
nucleus) or heterokaryon (having two unfused nuclei). The hybrid protoplast is called cytoplasmic hybrid
or cybrid if one of the two nuclei of this get degenerated. The first somatic hybrid was obtained by Carlson
et. al. (1972) between Nicotiana glauca and N. langsdorfi (species of Tobacco). The intergeneric somatic hybrid
is Pomato (Potato × tomato) but unfortunately, Pomato did not have all the desired combination of
characteristics for its commercial utilisation.

PEG
Parent A Parent B
protoplast protoplast

Cell fusion

Nuclear fusion

Somatic hybrid cell

Fig.: Protoplast fusion technique

(4) Applications of plant tissue culture :
(i) Rapid clonal propagation : By application of tissue culture it is possible to achieve propagation
of a large number of plants in very short durations. This method of producing thousands of plants
through tissue culture is called micropropagation. Each of these plants will be genetically
identical to the parental plant i.e., they are somaclones. Many important food plants like tomato,
banana, apple, etc. have been produced on commercial scale using this method.
(ii) Pathogen-free plants : Recovery of healthy plants from the diseased plants is possible by
meristem culture. e.g.,Resistance to tungro virus and leaf hopper in rice is developed by tissue
culture.

Example 1 : Indicate whether the following statements are true or false. Rectify each false statement to make
it true.
(i) One of the alternate sources of protein for animal and human nutrition is single cell protein.
(ii) Techniques of tissue culture and somatic hybridization offer vast potential for manipulation of
plants in-vivo to produce new varieties.
Solution : (i) True
(ii) False, in-vitro

Corporate Office : Aakash Tower, 8, Pusa Road, New Delhi-110005. Phone : 011-47623456
 4 Biotechnology and Its Applications Additional Content for NEET-2024

Example 2 : Fill in the blanks with suitable words in following statements:

(i) Method of producing thousands of plants through tissue culture is called ________.
(ii) Recovery of healthy plants from diseased plant is possible by using ________ as explant in
tissue culture.
Solution : (i) Micro-propagation

(ii) Apical or axillary meristem

ASSIGNMENT

SECTION - A
Objective Type Questions
1. Plant part used for tissue culture is sterilized by
(1) Chemicals (2) Heat
(3) UV-rays (4) Nutrient media
2. The ability of a plant cell to give rise to a complete plant is called
(1) Totipotency (2) Pluripotency
(3) Hardening (4) Protoplast fusion
3. To prevent the damage of newly formed cells, part of the cultures are regularly transferred to new culture vessels
containing fresh media. This process is termed as
(1) Callus culture (2) Suspension culture
(3) Subculturing (4) Hardening
4. During the somatic hybridisation, the cells are treated with enzymes
(1) PEG (2) Cellulase, pectinase
(3) Electrofusion (4) All of these
5. Pomato is an intergeneric somatic hybrid of
(1) Potato and Brinjal (2) Potato and Tomato
(3) Tomato and Brinjal (4) Potato and Tobacco
6. In tissue culture, plant genetically identical to the parental plant is called
(1) Synkaryon (2) Explant
(3) Sexual hybrid (4) Somaclone
7. Find the odd one w.r.t. somaclonal variations
(1) Produced during tissue culture
(2) Some may be useful and stable
(3) All are useful and very stable
(4) Some of the significant variations have been taken up in plant breeding
8. Choose correct option w.r.t. tissue culture
(1) The whole plant can be regenerated from explant
(2) Only apical meristem part of plant can grow in test tube under sterile conditions
(3) It is not related to totipotency
(4) Sterilisation is not a pre-requisite

Corporate Office : Aakash Tower, 8, Pusa Road, New Delhi-110005. Phone : 011-47623456
Additional Content for NEET-2024 Biotechnology and Its Applications 5
9. Capacity of a plant cell to give rise to whole plant body is known as
(1) Reproduction (2) Budding
(3) Totipotency (4) Regeneration
10. Somaclones are obtained by
(1) Tissue culture (2) Plant breeding
(3) Genetic engineering (4) Irradiation
11. Culturing meristem of banana, sugarcane and potato is primarily significant to produce
(1) Somatic hybrids (2) Virus free plants
(3) Specific meristematic tissue (4) Hybrid vigour
12. An explant is
(1) Dead plant
(2) Part of the wood
(3) Part of the plant that expresses a specific gene
(4) Part of the plant used in tissue culture
13. Select correct statement (w.r.t. somaclones)
(1) Somaclones of tomato and banana have been produced on commercial scale
(2) They are genetically not identical
(3) They are produced by sexual reproduction
(4) Can be generated in field
14. Select the incorrect option (w.r.t. tissue culture)
(1) Genetically identical – Somaclones
(2) Capacity to generate a whole plant – Totipotency
(3) Plant cell without cell wall – Protoplast
(4) Virus free part of plant – Flower
15. Somatic hybridisation is accomplished by
(1) Grafting between two varieties
(2) Fusion of two protoplast of different varieties of plants
(3) Chromosome doubling in androgenic culture
(4) Recombinant DNA technology
16. From a virus infected plant, we can obtain healthy plants by culturing its
(1) Differentiated somatic tissue (2) Anther
(3) Meristem (4) Embryo
17. Match the column I with column II and choose the correct option.
Column I Column II
a. Totipotency (i) Spirulina
b. SCP (ii) Unorganised mass of cells
c. Explant (iii) Capacity to generate whole plant from any cell
d. Callus (iv) Plant part which is cultured
(1) a(i), b(ii), c(iii), d(iv) (2) a(iii), b(i), c(iv), d(ii)
(3) a(iii), b(iv), c(i), d(ii) (4) a(ii), b(i), c(iv), d(iii)

Corporate Office : Aakash Tower, 8, Pusa Road, New Delhi-110005. Phone : 011-47623456
 6 Biotechnology and Its Applications Additional Content for NEET-2024

18. Somaclones are

(1) Genetically similar to the original plant from which they were grown
(2) Genetically as well as morphologically dissimilar to the original plant
(3) Only genetically similar to each other
(4) Morphologically similar but genetically dissimilar
19. The capacity to generate a whole plant from any plant cell or explant is called
(1) Totipotency (2) Propagation
(3) Hybridisation (4) Heterosis
20. Thousands of plants produced through micro-propagation are genetically identical to the original plant, which
are called as
(1) Hybrids (2) Recombinants
(3) Somaclones (4) Somatic hybrids
21. To obtain virus-free plants, one can remove the meristem and grow it in vitro. This meristem can be
(1) Apical (2) Permanent tissues
(3) Axillary (4) Both (1) and (3)

SECTION - B
Previous Years Questions
1. Match List-I with List-II [NEET-2021]

List-I List-II

(a) Protoplast fusion (i) Totipotency

(b) Plant tissue culture (ii) Pomato

(c) Meristem culture (iii) Somaclones

(d) Micropropagation (iv) Virus free plants

Choose the correct answer from the options given below.

(a) (b) (c) (d)
(1) (iv) (iii) (ii) (i)
(2) (iii) (iv) (ii) (i)
(3) (ii) (i) (iv) (iii)
(4) (iii) (iv) (i) (ii)
2. A technique of micropropagation is [AIPMT-2015]
(1) Embryo rescue (2) Somatic hybridization
(3) Somatic embryogenesis (4) Protoplast fusion
3. To obtain virus-free healthy plants from a diseased one by tissue culture technique, which part/parts of the
diseased plant will be taken? [AIPMT-2014]
(1) Apical meristem only
(2) Palisade parenchyma
(3) Both apical and axillary meristems
(4) Epidermis only

Corporate Office : Aakash Tower, 8, Pusa Road, New Delhi-110005. Phone : 011-47623456
Additional Content for NEET-2024 Biotechnology and Its Applications 7
4. Which one of the following is a case of wrong matching? [AIPMT (Prelims)-2012]
(1) Micropropagation-In vitro production of plants in large numbers
(2) Callus–Unorganised mass of cells produced in tissue culture
(3) Somatic hybridization - Fusion of two diverse cells
(4) Vector DNA-Site for t-RNA synthesis
5. Which part would be most suitable for raising virus-free plants for micropropagation? [AIPMT (Prelims)-2012]
(1) Meristem (2) Node
(3) Bark (4) Vascular tissue
6. Somaclones are obtained by [AIPMT (Prelims)-2009]
(1) Plant breeding (2) Irradiation
(3) Genetic engineering (4) Tissue culture
7. In order to obtain virus-free plants through tissue culture the best method is [AIPMT (Prelims)-2006]
(1) Protoplast culture (2) Embryo rescue
(3) Anther culture (4) Meristem culture

‰ ‰ ‰

ANSWER
Section - A : Objective Type Questions
1. (1) 2. (1) 3. (3) 4. (2) 5. (2) 6. (4) 7. (3)
8. (1) 9. (3) 10. (1) 11. (2) 12. (4) 13. (1) 14. (4)
15. (2) 16. (3) 17. (2) 18. (1) 19. (1) 20. (3) 21. (4)
Section - B : Previous Years Questions

1. (3) 2. (3) 3. (3) 4. (4) 5. (1) 6. (4) 7. (4)

Corporate Office : Aakash Tower, 8, Pusa Road, New Delhi-110005. Phone : 011-47623456