Skip to main content

Suspension culture and development - methodology, kinetics of growth and production formation, elicitation methods, hairy root culture. Detailed notes


Suspension culture and development - methodology, kinetics of growth and production formation, elicitation methods, hairy root culture. Detailed notes




1. Introduction

Suspension culture is a type of plant tissue culture in which single cells or small cell aggregates are grown in liquid nutrient medium under continuous agitation. It is mainly used for:
Large-scale biomass production
Secondary metabolite production
Cell physiology and biochemical studies
Genetic manipulation and selection.


2. Methodology of Suspension Culture


2.1 Source of Explant

Usually initiated from friable callus
Callus derived from:
Leaf
Stem
Root
Hypocotyl
Friable callus is preferred as it disintegrates easily into single cells.

2.2 Preparation of Cell Suspension


Friable callus is transferred into liquid MS medium
Medium contains:
Carbon source (usually sucrose)
Auxins (2,4-D commonly used)
Culture maintained in:
Conical flasks
Orbital shaker (100–150 rpm)


2.3 Culture Conditions


Parameter
Requirement
Temperature
25 ± 2°C
pH
5.6 – 5.8
Light
Dark or diffused light
Aeration
Provided by shaking
Agitation
Prevents cell clumping


2.4 Subculturing


Periodic transfer to fresh medium (7–14 days)
Maintains exponential growth
Prevents nutrient depletion


2.5 Types of Suspension Cultures

Batch culture
Continuous culture
Immobilized cell culture

3. Kinetics of Growth and Product Formation


3.1 Growth Curve of Suspension Culture
Suspension culture follows a sigmoid (S-shaped) growth curve:


1. Lag Phase
Cells adapt to new environment
Little or no division
Enzyme synthesis occurs


2. Exponential (Log) Phase
Rapid cell division
High metabolic activity
Maximum biomass accumulation


3. Linear Phase
Growth rate slows
Nutrients start declining


4. Stationary Phase
Cell division stops
Biomass remains constant
Secondary metabolite production is maximum

5. Decline Phase


Cell death occurs
Toxic metabolites accumulate


3.2 Measurement of Growth
Fresh weight
Dry weight
Packed cell volume (PCV)
Cell number (haemocytometer)


3.3 Product Formation Kinetics
Secondary metabolites are classified based on growth association:


1. Growth-associated products
Produced during exponential phase
Example: Proteins, enzymes


2. Non-growth-associated products
Produced during stationary phase
Example: Alkaloids, phenolics


3. Mixed growth-associated products
Produced throughout growth cycle


4. Elicitation Methods
4.1 Definition
Elicitation is the process of stimulating secondary metabolite production in plant cell cultures using elicitors.


4.2 Types of Elicitors
A. Biotic Elicitors
Derived from biological sources
Examples:
Fungal cell wall extracts
Yeast extract
Chitosan
Polysaccharides


B. Abiotic Elicitors


Non-biological
Examples:
Heavy metals (Ag⁺, Cd²⁺)
UV radiation
Temperature stress
Salicylic acid
Jasmonic acid


4.3 Mechanism of Elicitation


Elicitor binds to cell membrane receptor
Activation of signal transduction pathways
Increase in intracellular Ca²⁺
Activation of defense genes
Enhanced secondary metabolite synthesis


4.4 Advantages of Elicitation
Increased metabolite yield
Reduced culture time
Cost-effective production
5. Hairy Root Culture
5.1 Definition
Hairy root culture is a genetically transformed root culture induced by infection with Agrobacterium rhizogenes.


5.2 Mechanism of Hairy Root Induction


A. rhizogenes transfers Ri plasmid T-DNA
rol (root locus) genes integrate into plant genome
Causes:
Profuse root branching
Fast growth
Hormone-independent growth


5.3 Methodology


Plant explant infected with A. rhizogenes
Co-cultivation for 24–48 hours
Transfer to antibiotic medium to remove bacteria
Selection of transformed roots
Maintenance in hormone-free liquid medium


5.4 Characteristics of Hairy Root Cultures
Genetic stability
Rapid growth rate
High biosynthetic capacity
No need for growth regulators


5.5 Applications
Production of secondary metabolites:
Alkaloids (e.g., atropine)
Terpenoids
Phenolics
Study of root metabolism
Biotransformation reactions


6. Advantages of Suspension and Hairy Root Cultures
Uniform growth
Easy scale-up in bioreactors
Controlled conditions
Consistent metabolite yield


7. Limitations
Genetic instability in long-term cultures
Shear sensitivity of cells
High operational cost


8. Conclusion
Suspension culture and hairy root culture are powerful tools in plant biotechnology for large-scale production of biomass and valuable secondary metabolites. The use of growth kinetics studies and elicitation strategies significantly enhances productivity.



50 MCQs: Suspension Culture & Hairy Root Culture


1. Suspension culture is best initiated from
A. Hard callus
B. Compact callus
C. Friable callus
D. Meristem
Answer: C
2. Suspension cultures are grown in
A. Solid medium
B. Semi-solid medium
C. Liquid medium
D. Agar medium
Answer: C
3. Continuous shaking in suspension culture helps in
A. Cell differentiation
B. Aeration and uniform cell distribution
C. Callus formation
D. Organogenesis
Answer: B
4. Ideal agitation speed for suspension culture is
A. 10–20 rpm
B. 50–60 rpm
C. 100–150 rpm
D. 300 rpm
Answer: C
5. Commonly used auxin for suspension culture
A. IAA
B. NAA
C. 2,4-D
D. Kinetin
Answer: C
6. pH of suspension culture medium is usually
A. 4.0
B. 4.5
C. 5.6–5.8
D. 7.0
Answer: C
7. The growth curve of suspension culture is
A. Linear
B. Parabolic
C. Sigmoid
D. Exponential only
Answer: C
8. During lag phase
A. Rapid cell division occurs
B. Secondary metabolites form
C. Cells adapt to new environment
D. Cell death occurs
Answer: C
9. Maximum biomass accumulation occurs in
A. Lag phase
B. Exponential phase
C. Stationary phase
D. Decline phase
Answer: B
10. Secondary metabolites are mainly produced during
A. Lag phase
B. Log phase
C. Stationary phase
D. Decline phase
Answer: C
11. Packed cell volume (PCV) measures
A. DNA content
B. Protein level
C. Cell biomass
D. Metabolite concentration
Answer: C
12. Which is a growth-associated product?
A. Alkaloids
B. Phenolics
C. Terpenoids
D. Proteins
Answer: D
13. Non-growth associated products are produced in
A. Lag phase
B. Exponential phase
C. Stationary phase
D. Decline phase
Answer: C
14. Elicitation increases production of
A. Primary metabolites
B. Secondary metabolites
C. DNA
D. RNA
Answer: B
15. An elicitor is
A. Growth hormone
B. Nutrient
C. Stress-inducing agent
D. Antibiotic
Answer: C
16. Yeast extract is an example of
A. Abiotic elicitor
B. Chemical elicitor
C. Biotic elicitor
D. Physical elicitor
Answer: C
17. Salicylic acid acts as
A. Nutrient
B. Abiotic elicitor
C. Antibiotic
D. Growth inhibitor
Answer: B
18. Heavy metals used as elicitors include
A. Na⁺
B. K⁺
C. Ag⁺
D. Ca²⁺
Answer: C
19. Signal transduction during elicitation involves
A. DNA methylation
B. Protein degradation
C. Calcium influx
D. Cell lysis
Answer: C
20. Hairy root culture is induced by
A. Agrobacterium tumefaciens
B. Agrobacterium rhizogenes
C. Rhizobium
D. Bacillus
Answer: B
21. Hairy roots arise due to transfer of
A. Ti plasmid
B. Ri plasmid
C. pBR322
D. BAC
Answer: B
22. rol genes are responsible for
A. Shoot induction
B. Callus formation
C. Root proliferation
D. Flowering
Answer: C
23. Hairy root cultures grow
A. With high auxin
B. With cytokinin
C. Without growth regulators
D. Only on solid medium
Answer: C
24. A major advantage of hairy root culture is
A. Slow growth
B. Genetic instability
C. High metabolite production
D. Need for hormones
Answer: C
25. Hairy roots show
A. Poor branching
B. Profuse lateral branching
C. No growth
D. Callus formation
Answer: B
26. Hairy root cultures are mainly used for production of
A. Vitamins
B. Carbohydrates
C. Secondary metabolites
D. Lipids
Answer: C
27. Antibiotics are used in hairy root culture to
A. Induce roots
B. Eliminate Agrobacterium
C. Increase growth
D. Prevent differentiation
Answer: B
28. One example of alkaloid produced in hairy root culture is
A. Morphine
B. Atropine
C. Nicotine
D. All of the above
Answer: D
29. Immobilized cell culture involves
A. Free cells
B. Entrapment in a matrix
C. Solid callus
D. Root culture
Answer: B
30. Batch culture is characterized by
A. Continuous medium supply
B. Closed system
C. Constant nutrient addition
D. No growth phases
Answer: B
31. Continuous culture maintains cells in
A. Lag phase
B. Log phase
C. Stationary phase
D. Decline phase
Answer: B
32. Shear sensitivity is a problem in
A. Solid culture
B. Suspension culture
C. Seed culture
D. Root nodules
Answer: B
33. Bioreactors are used for
A. Small-scale culture
B. Large-scale suspension culture
C. Seed germination
D. Organ culture
Answer: B
34. Hairy root cultures are genetically
A. Unstable
B. Stable
C. Variable
D. Temporary
Answer: B
35. Chitosan is a
A. Growth regulator
B. Antibiotic
C. Biotic elicitor
D. Nutrient
Answer: C
36. UV radiation is classified as
A. Biotic elicitor
B. Abiotic elicitor
C. Chemical elicitor
D. Hormonal elicitor
Answer: B
37. Primary metabolites are mainly produced during
A. Stationary phase
B. Decline phase
C. Exponential phase
D. Lag phase
Answer: C
38. A major limitation of suspension culture is
A. Uniform growth
B. Easy scale-up
C. Genetic instability
D. Controlled conditions
Answer: C
39. Haemocytometer is used to measure
A. Metabolites
B. Protein
C. Cell number
D. DNA
Answer: C
40. Friable callus is preferred because it
A. Is hard
B. Grows slowly
C. Easily disperses into single cells
D. Forms shoots
Answer: C
41. Stationary phase is characterized by
A. Rapid cell division
B. Nutrient limitation
C. Cell elongation
D. DNA synthesis
Answer: B
42. Hairy roots differ from normal roots because they
A. Grow slowly
B. Need hormones
C. Grow rapidly and autonomously
D. Are genetically unstable
Answer: C
43. Jasmonic acid is used as
A. Nutrient
B. Growth hormone
C. Elicitor
D. Antibiotic
Answer: C
44. Agitation in suspension culture prevents
A. Cell division
B. Cell aggregation
C. DNA replication
D. Metabolite synthesis
Answer: B
45. The decline phase is due to
A. Nutrient abundance
B. Toxic metabolite accumulation
C. Increased oxygen
D. Hormone addition
Answer: B
46. Secondary metabolite production is part of
A. Growth metabolism
B. Defense response
C. Cell division
D. DNA replication
Answer: B
47. The most suitable medium for suspension culture is
A. White’s medium
B. Gamborg’s B5
C. Murashige and Skoog (MS)
D. N6 medium
Answer: C
48. Hairy root cultures are maintained in
A. Solid agar
B. Semi-solid medium
C. Liquid hormone-free medium
D. Soil
Answer: C
49. Biotransformation refers to
A. Cell division
B. Gene transfer
C. Conversion of substrates into products
D. Callus formation
Answer: C
50. Main advantage of suspension culture over callus culture
A. Slower growth
B. Uniform exposure to nutrients
C. Less oxygen
D. Genetic variation
Answer: B






Comments

Post a Comment

Popular Posts

Fourth Semester M.Sc. Degree Examination, March 2021Time: 3 HoursBotanyBO 241: BIOINFORMATICS AND BIOPHYSICS(2019 Admission)

Fourth Semester M.Sc. Degree Examination, March 2021 Time: 3 Hours Botany BO 241: BIOINFORMATICS AND BIOPHYSICS (2019 Admission) 1. Answer the following questions. 1. Expand EMBL and DDBJ. 2. What do bootstrap values indicate? 3. What is multiple sequence alignment? 4. What is SNP? 5. Define transcriptome. 6. What is Smith Waterman algorithm? 7. Comment on Phylip. 8. What are the factors that determine the electrophoretic mobility of a particle? 9. Differentiate between resolution and resolving power of the microscope. 10. Which are the factors that determine the sedimentation of a component during centrifugation? (10 x 1= 10 Marks) II. Answer the following questions in not more than 50 words. 11. (a) What is the difference between rooted and unrooted phylogenetic tree? OR (b) What is ORF? What is its significance in functional genomics? 12. (a) Explain the use of GENSCAN. OR (b) Explain the assumptions in molecular clock hypothesis. 13. (a) Write a brief explanation on KEGG. OR (b) Co...
Post a Comment
Read more

Secondary Databases (PROSITE, PRINTS, BLOCKS)

Secondary Databases (PROSITE, PRINTS, BLOCKS  Secondary Databases Introduction Biological databases are broadly classified into primary and secondary databases. Primary databases store raw experimental data (e.g., nucleotide or protein sequences), whereas secondary databases contain derived information obtained by analyzing primary sequence data. Secondary databases are mainly used to: Identify protein families Detect conserved motifs, patterns, and domains Predict protein function Study structure–function relationships Examples of secondary databases include PROSITE, PRINTS, BLOCKS, Pfam, etc. 1. PROSITE Database Definition PROSITE is a secondary database that documents protein domains, families, and functional sites in the form of patterns and profiles. Developed by Swiss Institute of Bioinformatics (SIB) Maintained along with UniProt Principle PROSITE is based on the idea that functionally important regions of proteins are conserved during evolution. These conserved regions can ...
Post a Comment
Read more

Intellectual Property Rights (IPR) – Detailed Notes

Intellectual Property Rights (IPR) – Detailed Notes 1. Introduction Intellectual Property Rights (IPR) are legal rights granted to creators and inventors over their creations or inventions. They protect innovation and creativity, providing the owner exclusive rights to use, sell, or license their creation. IPR encourages research, development, and economic growth by rewarding creativity. 2. Importance of IPR Protects inventions, designs, and creative work. Prevents unauthorized use, copying, or commercialization. Encourages innovation and research. Provides financial benefits to inventors through licensing or royalties. Supports economic growth and competitiveness. Safeguards traditional knowledge and biodiversity. 3. Types of Intellectual Property Rights A. Patents Definition: Exclusive right granted to an inventor for a new invention for a limited period (usually 20 years). Requirements: Novelty – must be new and not published. Inventive step – non-obvious to someone skilled in the f...
Post a Comment
Read more

Electroporation – Detailed Notes

Electroporation – Detailed Notes Definition : Electroporation is a physical method of gene transfer in which cells are exposed to a brief, high-voltage electric pulse, creating temporary pores in the cell membrane. This allows DNA, RNA, proteins, or other molecules to enter the cytoplasm. It is widely used in bacteria, yeast, plant protoplasts, and mammalian cells. Key Concept: The electric field destabilizes the membrane, making it permeable to macromolecules. 1. Principle Cells are suspended in a conductive medium. A brief electrical pulse induces transient pores in the plasma membrane. DNA or other molecules present in the medium enter the cell through these pores. Membrane reseals after the pulse, and the molecule is retained inside the cell. Advantages of Principle: Direct and rapid. Works in many cell types. Does not require chemical carriers or viral vectors. 2. Materials Required Cells – bacterial, yeast, plant protoplasts, mammalian cells. DNA/RNA/other macromolecule – purifie...
Post a Comment
Read more

Third Semester M.Sc. Degree Examination, January 2023 Botany BO 231 PLANT BREEDING, HORTICULTURE AND BIOSTATISTICS

Third Semester M.Sc. Degree Examination, January 2023 Botany BO 231 PLANT BREEDING, HORTICULTURE AND BIOSTATISTICS Time: Three Hours (2019 Admission Onwards) I. Answer the following questions. 1.What is green super rice? 2.What are the functions of ICAR-NBPGR? 3.Give the importance of floral biology in plant breeding. 4.How do you develop a synthetic variety? 5.Where can you find gene-for-gene relationships? 6.Describe the significance of biodiversity policy. 7.What is Olericulture? 8.Describe the advantages of in door garden. 9.What is Students's t-test? 10. Explain Ogive graph. (10 × 1 = 10 Marks) 11.Answer the following questions in not more than 50 words . 11. (a) Explain hybridization and mention it's procedure. OR (b) Write short notes on the concept of centers of origin proposed by Vavilov. 12. (a) Describe cytoplasmic male sterility and its uses. OR (b) Explain the role of interspecific and intergeneric hybridization. 13. (a) What is seed certification? How is it done? ...
Post a Comment
Read more

Third Semester M.Sc. Degree Examination, February 2024 231: PLANT BREEDING, HORTICULTURE AND BIOSTATISTICS

Third Semester M.Sc. Degree Examination, February 2024                 Botany BO 231: PLANT BREEDING, HORTICULTURE AND BIOSTATISTICS (2019 Admission onwards) Time: 3 Hours I.Answer the following questions. 1.What is atomic gardening? 2.Name the cardamom research institute in Kerala. 3.Explain advantages of distant hybridisation. 4.Describe plant variety rights. 5.Write short notes on arboriculture. 6.What is vermicomposting? 7.Give short notes on cut flower industry. 8.What is ANOVA? 9.Describe the properties of binomial distribution. 10. Explain the use of LSD. Max. Marks: 75 (10 x 1 = 10 Marks) II.Answer the following questions in not more that 50 words. 11. (a) What do you mean by genetic modification techniques? OR (b) What is center of diversity of a species? 12. (a) Compare auto and allopolyploidy. OR (b) What are requirements of back cross breeding? 13. (a) Describe ideotype breeding and its significance. OR (b) What is the role of seed cer...
Post a Comment
Read more

❃LC-MS (LIQUID CHROMATOGRAPHY – MASS SPECTROMETRY)

LC-MS (LIQUID CHROMATOGRAPHY – MASS SPECTROMETRY)  ┏━━━━━ •❃°•°❀°•°❃•━━━━•━━━┓ 1. INTRODUCTION LC-MS is a hyphenated analytical technique combining Liquid Chromatography (LC) and Mass Spectrometry (MS). It is used for separation, identification, and quantification of compounds in complex mixtures. LC separates analytes based on polarity, size, or charge, while MS detects molecules based on mass-to-charge ratio (m/z). Developed in the 1970s–1980s, LC-MS is now widely used in pharmaceutical, clinical, environmental, and food analysis. Importance : Detects trace levels of compounds (ng–pg range) Analyzes non-volatile, thermally labile compounds that cannot be analyzed by GC-MS Provides structural information through mass fragmentation Example: Detection of drugs in plasma, protein identification in proteomics, pesticide residue analysis in food. 2. COMPONENTS OF LC-MS The LC-MS system has three main parts: A. Liquid Chromatograph (LC) Function: Separates components of a mixture befor...
Post a Comment
Read more

Fourth Semester M.Sc. Degree Examination, June 2022BotanySpecial Paper II - ElectiveBO 242 a: BIOTECHNOLOGY

Reg. No.: Name: N-6273 Fourth Semester M.Sc. Degree Examination, June 2022 Botany Special Paper II - Elective BO 242 a: BIOTECHNOLOGY Time: 3 Hours (2019 Admission Onwards) Max. Marks: 75 1. Instruction: Draw diagrams and illustrate with examples wherever necessary. Answer the following questions. 1. What are the desirable features of a cloning vehicle? 2. What is a palindrome? 3. What is the significance of Ori C site? 4. What is the actual function of restriction enzymes in a bacterial system?  5.Name any two bacteria and fungi used for alcohol fermentation. 6. What is a starter culture? 7. What are adapters? 8. What are probes? 9. What is biopiracy? 10. Define cybrids. (10 x 1 = 10 Marks) II. Answer the following questions in not more than 50 words .  11. (a) Why is callus culture a prerequisite for somaclonal variations? OR (b) How is virus elimination done via plant tissue culture? 12. (a) How is aeration maintained in a bioreactor? OR (b) What are the methods available f...
Post a Comment
Read more

••CLASSIFICATION OF ALGAE - FRITSCH

      MODULE -1       PHYCOLOGY  CLASSIFICATION OF ALGAE - FRITSCH  ❖F.E. Fritsch (1935, 1945) in his book“The Structure and  Reproduction of the Algae”proposed a system of classification of  algae. He treated algae giving rank of division and divided it into 11  classes. His classification of algae is mainly based upon characters of  pigments, flagella and reserve food material.     Classification of Fritsch was based on the following criteria o Pigmentation. o Types of flagella  o Assimilatory products  o Thallus structure  o Method of reproduction          Fritsch divided algae into the following 11 classes  1. Chlorophyceae  2. Xanthophyceae  3. Chrysophyceae  4. Bacillariophyceae  5. Cryptophyceae  6. Dinophyceae  7. Chloromonadineae  8. Euglenineae    9. Phaeophyceae  10. Rhodophyceae  11. Myxophyce...
Post a Comment
Read more

Protein Sequence DatabasesPIR, SWISS-PROT and TREMBEL

Protein Sequence Databases PIR, SWISS-PROT and TREMBEL 1. Introduction Protein sequence databases are biological databases that store information about amino acid sequences of proteins, along with their functional, structural, and biochemical characteristics. Since proteins are the functional molecules of the cell, protein databases are essential for understanding gene expression, metabolism, enzymatic activity, signaling pathways, and evolution. Protein sequence databases mainly contain data derived from translated nucleotide sequences and experimental protein studies. 2. Types of Protein Sequence Databases Protein sequence databases are broadly classified into: A. Primary Protein Databases Contain original protein sequence data Minimal or no manual annotation B. Secondary Protein Databases Derived from primary databases Provide curated functional and structural information C. Composite Protein Databases Combine protein data from multiple sources Reduce redundancy 3. Protein Informati...
Post a Comment
Read more