12 Botany | Chapter 5 Plant Tissue Culture

T. Selvan 50
Arputharaj | G. Muthu | P. Jesintha Mary | S. Kaja Sulthan | A. Moses Packiaraj………

Ghlm; 1

Chapter 5
Plant Tissue Culture
One Mark Question and Answer
1. Find wrong statement about (IPR)
a) In biotechnology, the transformed microorganisms and plants and technologies for the
production of commercial products are exclusively the property of the discoverer
b) The discoverer has the full rights on his property. It should be neglected by the others
without legal permission.
c) The right of discoverer must be protected and it does by certain laws framed by a country.
d) The IPR is protected by different ways like patents, copyrights, trade secrets and
trademarks, designs and geographical indications

2. Find the wrong statement about Patents

a) It is a special right to the discoverer/inventor that has been granted by the government through
legislation for trading new articles
b) A patent is a personal property which can't be licensed or sold by the person or organisation
just like any other property
c) Patent terms give the inventor the rights to exclude others from making, using or selling his
invention
d) It is difficult to keep secret certain inventions and therefore, guidance should be obtained from
a qualified patent attorney.

3. The grant is filled at the............ office which is not published It is a signed document,actually the
agreement that grants patent right to the inventor
a) Incom tax b) Municipal c) Collector d) Patent

4. "Pathogenicity of living organisms and viruses"find wrong example

a) Infect humans b) Animals c) Plants d) Micro Organisms

5. Find the wrong statement about Potential risks and consideration for safety aspects
a) Toxicity of allergy associated with microbial production
b) Decreasing number of antibiotic resistant pathogenic microorganisms
c) Safety aspects associated with contamination, infection or mutation of process strains.
d) Safety aspects associated with the industrial use of microorganisms containing in vitro
recombinants

6. "Bio safety guidelines are being implemented by"find the wrong department
a) The Institutional Bio-safety Committees
b) Ethical, Legal and Social Implications
c) The Review Committee on Genetic Manipulation
d) The Genetic Engineering Approval Committee

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7. ..............was established as an apexbody to accord approval of activities involving large scale use
of hazardous microorganisms and recombinants in research and industrial production
a) The Institutional Bio-safety Committees
b) Ethical, Legal and Social Implications
c) The Review Committee on Genetic Manipulation
d) The Genetic Engineering Approval Committee

8. The Ethical, Legal, and Social Implications(ELSI) program was founded in ....................
a) 1988 b) 1989 c) 1990 d) 1991

9. The concept Totipotency was proposed by

a) P.R.White b) Gott lieb Haberlandt c) Hildbrandt d) Carlson

10. ............................... developed root cultures, used Knop’s solutionalong with three vitamins like
pyridoxine,thiamine and nicotinic acid
a) P.R.White b) Gott lieb Haberlandt c) Hildbrandt d) Carlson

11. Who was used coconut water in plant tissue culture work
a) Murashige b) Kanta c) Chilton d) F.C. Steward

12. Produced transformed tobacco plants fromsingle cell transformation and gene insertion
a) Murashige b) Kanta c) Chilton d) F.C. Steward

13. The process of biochemical and structural changes by which cells become specialized inform and
function
a) Totipotency b) Differentiation c) Redifferentiation d) Dedifferentiation

14. The (HEPA) filter is used in ...........

a) autoclave b) pH meter c) laminar air-flow bench d) ovens

15. In culture facility growing the explant inoculated into culture tubes at
a) 22-28° C b) 22-30° C c) 28-30° C d) 28-32° C

16. In culture facility growing the explant inoculated with illumination of light
a) 2200 lux b) 2300 lux c) 2400 lux d) 2500 lux

17. Growing the explant inoculated with a photoperiod of

a) 8-16 hours b) 8-12 hours c) 6-12 hours d) 8-14 hours

18. Growing the explant inoculated with a relative humidity of

a) 40% b) 50% c) 60% d) 70%

19. Autoclaving at
a) 15 psi (120°C) for 15 to 30 minutes b) 15 psi (120°C) for 20 to 30 minutes
c) 15 psi (121°C) for 15 to 30 minutes d) 21 psi (121°C) for 15 to 30 minutes

20. Surface sterilization agents like ................ mercuric chloride, 70% ethanol under aseptic
condition inside the Laminar Air Flow Chamber
a) 1% b) 0.1% c) 0.01% d) 0.001%

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21. Gamborg.et.al using the .......................... medium

a) MS nutrient medium b) B5 medium c) White medium d) Nitsch’s medium

22. The pH of medium is normally adj usted between ............ for the best result
a) 2.6 to 3.0 b) 3.6 to 4.0 c) 4.6 to 5.0 d) 5.6 to 6.0

23. Protoplast Fusion done through the ...........................

a) Polyethylene Glycol b) Nicotinic acid c) Pyridoxin d) Thaiamine

24. In Protoplast culture the cell wall formation occurs within ...................
a) 0-24 hours b) 24-48 hours c) 48-72 hours d) 72-96 hours

25. Biosynthesis and isolation of indole alkaloids from ...................plant cell culture
a) Capsicum annum b) Papaver sominiferum
c) Catharanthus roseus d) Digitalis purpuria

26. Secondary metabolites Digoxin is produce by .............

a) Catharanthus roseus b) Digitalis purpuria
c) Papaver sominiferum d) Capsicum annum

27. Find the correct pair

a) Digoxin ------Analgesic b) Codeine ------Cardiac tonic
c) Capsaicin ------Antimalarial d) Vincristine ------ Anticarcinogenic

28. Find the incorrect pair

a) Digoxin ------- Cinchona officinalis b) Codeine ------ Papaver sominiferum
c) Capsaicin ----- Capsicum annum d) Vincristine ----- Catharanthus roseus

29......................................can be be used for the production of synthetic seeds

a) Callus b) Somatic embryoids c) embryoids d) Plant lets

30.Synthetic seeds are produced by encapsulation of embryoids in.............................

a) Agarose gel b) Benzylaminopurine c) Indol Acetic Acid d) NaCl

31. In banana micropropagation induction of shoots with in ............................

a) 160 days b) 162 days c) 165 days d) 168 days

32. In vitro icropropagation of banana suckers surface sterilized with 1% NaoCl for ........................
a) 30 seconds b) 30 minutes c) 30 hours d) 30 days

33. Find the wrong statement

a) Millions of artificial seeds can be produced at any time at low cost
b) It is easy to test the genotype of plants.
c) The period of dormancy of artificial seeds is increase
d) Artificial seeds produce identical plants

34. Cryopreservation by cooling to very low temperature of ........................

a) –190°C b) –196°C c) –197°C d) –198°C

35. In Cryopreservation we using ........................

a) Nitrogen b) oxygen c) liquid nitrogen d) Benzylaminopurine
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36. ................ induce shoot and root formation

a) cytokinins b) gibberellins c) ethylene d) abscisic acid

Two Mark Question and Answer

1, what is plant tissue culture?
Plant tissue culture is used to describe the in vitro and aseptic growth of any plant part on a tissue
culture medium.

2, Mention any two media

1. MS (Murashige and Skoog 1962)
2. B5 medium (Gamborg.et.al 1968),
3. White medium (white 1943)
4. Nitsch’s medium (Nitsch & Nitsch 1969).

3, Define agar
A complex mucilaginous polysaccharide obtained from marine algae (sea weeds) used as solidifying
agent in media preparation.

4, What is meant by sterilization?

Sterilization is the technique employed to get rid of microbes such as bacteria and fungi in the
culture medium, vessels and explants.

5, what is called embryoids?

The callus cells undergoes differentiation and produces somatic embryos, known as Embryoids.

6, Define Organ culture

The culture of embryos, anthers, ovaries, roots, shoots or other organs of plants on culture media.

7, What is meant by cell suspension culture?

The growing of cells including the culture of single cells or small aggregates of cells in vitro in
liquid medium is known as cell suspension culture.
8, What are the Basic concepts of Tissue culture?
Basic concepts of plant tissue culture are totipotency, differentiation, dedifferentiation and
redifferentiation.

9, Define Protoplasmic fusion

Protoplasts are cells without a cell wall, but bound by a cell membrane or plasma membrane. Using
protoplasts, it is possible to regenerate whole plants from single cells and also develop somatic
hybrids.

10, Mention the types of Plant tissue culture

 1.Organ culture
 2.Meristem culture
 3.Protoplast culture
 4.Cell suspension culture.

11, What is meant by hardening?

Hardening is the gradual exposure of in vitro developed plantlets in humid chambers in diffused
light for acclimatization so as to enable them to grow under normal field conditions.
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12, What is meant by somatic hybridization?

The fusion product of protoplasts without nucleus of different cells is called a cybrid. Following this
nuclear fusion take place. This process is called somatic hybridization

13, What is meant by explant?

The tissue taken from a selected plant transferred to a culture medium often to establish a new
plant.

Three Mark Question and Answer

1, What is meant by Cryopreservation?
Cryopreservation, also known as Cryo- conservation, is a process by which protoplasts, cells, tissues,
organelles, organs, extracellular matrix, enzymes or any other biological materials are subjected to
preservation by cooling to very low temperature of –196°C using liquid nitrogen.

2, Digoxin, Codeine, capsaicin --Write it's uses and it’s Plant sources.
Secondary Plant source Uses
Metabolites
Digoxin Digitalis purpurea Cardiac tonic
Codeine Papaver somniferum Analgesic
Capsaicin Capsicum annuum pain treatment
Vincristine Catharanthus roseus Anticarcinogenic
Quinine Cinchona officinalis Antimalarial

3, Differentiate Somaclonal variations and Gametoclonal variation

Somaclonal variations Gametoclonal variation
Somatic variations found in plants Gametophytic variations found in plants
regenerated in vitro (i.e. variations found in regenerated in vitro gametic origin (i.e.
leaf, stem, root, tuber or propagule) variations found in gametes and gametophytes)
4, Write the applications of somatic embryogenesis
1. Somatic embryogenesis provides potential plantlets which after hardening period can establish
into plants.
2. Somatic embryoids can be used for the production of synthetic seeds.
3. Somatic embryogenesis is now reported in many plants such as Allium sativum, Hordeum
vulgare, Oryza sativa, Zea mays and this is possible in any plant.

5, Write a Short note on IPR

Intellectual property right (IPR) is a category of rights that includes intangible creation of the human
intellect, and primarily consists of copyrights, patents, and trademarks. It also includes other types
of rights, such as trade secrets, publicity rights, moral rights, and rights against unfair competition.
In biotechnology, the transformed microorganisms and plants and technologies for the production
of commercial products are exclusively the property of the discoverer.

6, Define artificial seeds

Artificial seeds or synthetic seeds (synseeds) are produced by using embryoids (somatic embryos)
obtained through in vitro culture. They may even be derived from single cells from any part of the
plant that later divide to form cell mass containing dense cytoplasm, large nuclceus, starch grains,
proteins, and oils etc., To prepare the artificial seeds different inert materials are used for coating
the somatic embryoids like agarose and sodium alginate.

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Five Mark Question and Answer

1, Describe Basic concepts of Tissue culture
Basic concepts of plant tissue culture are totipotency, differentiation, dedifferentiation and
redifferentiation.
Totipotency : The property of live plant cells that they have the genetic potential when cultured in
nutrient medium to give rise to a complete individual plant.
Differentiation : The process of biochemical and structural changes by which cells become
specialized in form and function Redifferentiation: The further differentiation of already
differentiated cell into another type of cell. For example, when the component cells of callus have
the ability to form a whole plant in a nutrient medium, the phenomenon is called redifferentiation.
Dedifferentiation: The phenomenon of the reversion of mature cells to the meristematic state
leading to the formation of callus is called dedifferentiation. These two phenomena of
redifferentiation and dedifferentiation are the inherent capacities of living plant cells or tissue. This
is described as totipotency.

2, list out the application of plant tissue culture

 Improved hybrids production through somatic hybridization.
 Somatic embryoids can be encapsulated into synthetic seeds (synseeds). These encapsulated
seeds or synthetic seeds help in conservation of plant biodiversity.
 Production of disease resistant plants through meristem and shoot tip culture.
 Production of stress resistant plants like herbicide tolerant, heat tolerant plants.
 Micropropagation technique to obtain large numbers of plantlets of both crop and tree species
useful in forestry within a short span of time and all through the year.
 Production of secondary metabolites from cell culture utilized in pharmaceutical, cosmetic and
food industries.

3, Explain the technique involved in sterilization

Sterilization is the technique employed to get rid of microbes such as bacteria and fungi in the
culture medium, vessels and explants.
i. Maintenance of Aseptic Environment: During in vitro tissue culture maintenance of aseptic
environmental condition should be followed, i.e., sterilization of glassware, forceps, scalpels, and
all accessories in wet steam sterilization by autoclaving at 15 psi (121°C) for 15 to 30 minutes or
dipping in 70% ethanol followed by flaming and cooling.
ii. Sterilization of culture room: Floor and walls are washed first with detergent and then with 2%
sodium hypochlorite or 95% ethanol. The cabinet of laminar airflow is sterilized by clearing the
work surface with 95% ethanol and then exposure of UV radiation for 15 minutes.
iii. Sterilization of Nutrient Media: Culture media are dispensed in glass containers, plugged with
non-absorbent cotton or sealed with plastic closures and then sterilized using autoclave at 15 psi
(121°C) for 15 to 30 minutes. The plant extracts, vitamins, amino acids and hormones are
sterilized by passing through Millipore filter with 0.2 mm pore diameter and then added to
sterilized culture medium inside Laminar Airflow Chamber under sterile condition.
iv. Sterilization of Explants: The plant materials to be used for tissue culture should be surface
sterilized by first exposing the material in running tap water and then treating it in surface
sterilization agents like 0.1% mercuric chloride, 70% ethanol under aseptic condition inside the
Laminar Air Flow Chamber.

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4, Give an account on Protoplast culture

Protoplasts are cells without a cell wall, but bound by a cell membrane or plasma membrane. Using
protoplasts, it is possible to regenerate whole plants from single cells and also develop somatic
hybrids. The steps involved in protoplast culture.
i. Isolation of protoplast: Small bits of plant tissue like leaf
tissue are used for isolation of protoplast. The leaf tissue is
immersed in 0.5% Macrozyme and 2% Onozuka cellulase
enzymes dissolved in 13% sorbitol or mannitol at pH 5.4. It
is then incubated over-night at 25°C. After a gentle teasing
of cells, protoplasts are obtained, and these are then
transferred to 20% sucrose solution to retain their viability.
They are then centrifuged to get pure protoplasts as
different from debris of cell walls.
ii. Fusion of protoplast: It is done through the use of a suitable
fusogen. This is normally PEG (Polyethylene Glycol). The
isolated protoplast are incubated in 25 to 30%
concentration of PEG with Ca++ ions and the protoplast
shows agglutination (the formation of clumps of cells) and
fusion.
iii. Culture of protoplast: MS liquid medium is used with some modification in droplet, plating or
micro-drop array techniques. Protoplast viability is tested with fluorescein diacetate before the
culture. The cultures are incubated in continuous light 1000-2000 lux at 25°C. The cell wall
formation occurs within 24- 48 hours and the first division of new cells occurs between 2-7 days
of culture.
iv. Selection of somatic hybrid cells: The fusion product of protoplasts without nucleus of different
cells is called a cybrid. Following this nuclear fusion take place. This process is called somatic
hybridization.

5, List out the advantages of artificial seeds

Artificial seeds have many advantages over the true seeds
• Millions of artificial seeds can be produced at any time at low cost.
• They provide an easy method to produce genetically engineered plants with desirable traits.
• It is easy to test the genotype of plants.
• They can potentially stored for long time under cryopreservation method.
• Artificial seeds produce identical plants
• The period of dormancy of artificial seeds is greatly reduced, hence growth is faster with a
shortened life cycle.

6, write a short note on Germplasm Conversion.

 Germplasm conservation refers to the conservation of living genetic resources like pollen, seeds
or tissue of plant material.
 It is maintained for the purpose of selective plant breeding, preservation in live condition
 It is used for many research works.
 It is a part of collection of seeds and pollen that are stored in seed or pollen banks, so as to
maintain their viability and fertility for any later use such as hybridization and crop improvement.
 It may also involve a gene bank, DNA bank of elite breeding lines of plant resources for the
maintenance of biological diversity and also for food security.

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7, write a short note on Bio Safety.

 Biosafety is the prevention of large-scale loss of biological integrity, focusing both on ecology
and human health.
 These prevention mechanisms include conduction of regular reviews of the biosafety in
laboratory settings, as well as strict guidelines to follow.
 Many laboratories handling biohazards employ an ongoing risk management assessment and
enforcement process for biosafety.
 Failures to follow such protocols can lead to increased risk of exposure to biohazards or
pathogens.

8, write a short note on Bioethics - Ethical, Legal and Social Implications (ELSI)
 Bioethics refers to the study of ethical issues emerging from advances in biology and medicine.
 It is also a moral discernment as it relates to medical policy and practice. Bioethicists are concerned with
the ethical questions that arise in the relationships among life sciences, biotechnology and medicine.
 It includes the study of values relating to primary care and other branches of medicine.
 The scope of bioethics is directly related to biotechnology, including cloning, gene therapy, life extension,
human genetic engineering, astroethics life in space, and manipulation of basic biology through altered
DNA, RNA and proteins.
 These developments in biotechnology will affect future evolution, and may require new principles, such
as biotic ethics, that values life and its basic biological characters and structures.
 The Ethical, Legal, and Social Implications (ELSI) program was founded in 1990 as an integral part of the
Human Genome Project.
 The mission of the ELSI program was to identify and address issues raised by genomic research that would
affect individuals, families, and society.
 A percentage of the Human Genome Project budget at the National Institutes of Health and the U.S.
Department of Energy was devoted to ELSI research.

Important Diagrams

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Madurai Distirct 12 Botany Special Guide 2020