its a brief knowledge about Tissue Culture and Protoplast Culture.i want to share my work through slideshare.

1. Tissue Culture and
Protoplast Culture
Zahra Anwar
M.Phil scholar
Department of MMG

2. Learning Objectives
 Tissue Culture
 Plant Tissue Culture
 Requirements
 Types of Cultures
 Protoplast Culture
 Applications of Protoplast Cultures
 Factors Affecting protoplast culture
 Advances by Protoplast Cultures
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3. Tissue Culture
– A method of biological research in which fragments of tissue from an animal or
plant are grown in vitro in artificial medium under aseptic conditions and
continue to survive and function.
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4. Plant Tissue Culture
– The aseptic culture of plant protoplast, cells ,tissues or organ under aseptic
conditions which leads to cell multiplication or regeneration of organs or whole
plants.
– Basic Concept of Plant Tissue Culture
 Plasticity
 Totipotency
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5. Cont….
– So the tissue can be regenerated from explants such as leaf , ovary, protoplast,
petiole, root, anthers, etc.
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6. Requirements of Tissue Culture
– Tissue Cultures has several critical requirements :
 Appropriate tissue
 A suitable growth medium
 Aseptic conditions
 Laboratory & Media preparation room
 Transfer area
 Culturing facilities
 Hardening facilities
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7. Micro propagation
– The art and science of multiplying plants in
vitro..
– It implies
 Regeneration
 Multiplication
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8. Cont…
– Many different explants may use for
micro propagation
 Axillary buds
 Meristems are commonly used.
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9. Micro propagation Stages9

10. Types of Cultures
Root tip
culture
Shoot tip
culture
Complete
flower
culture
Anther
and Pollen
Culture
Protoplast
Culture
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12. Protoplast Culture
– Unit of biology composed of cell’s nucleus and the surroundings protoplasmic
materials.
– It is a plant cell that has its cell wall completely removed using either
 Mechanical
 Chemical means.
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14. Mechanical Method14

15. Enzymatic Method15

16. Purification of protoplast
– Protoplast are separated after enzymatic digestion and tranfer to suitable media
– Such as
 Mannitol
 Sorbitol
 Sucrose
 Cellular debris may can be separated by using nylon mesh.
 Viabilty can be checked by using fluorescent dyes visualize by microscope.
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19. Applications of Protoplast Culture
– Protoplast can be used to study:
 Metabolic studies including photosynthesis
 For DNA Transformation
 For somatic Hybridization
 Ingesting foreign material in to Cytoplasm
 Single cell system
 Wall synthesis & decomposition
 Virus uptake &replication
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20. Factors Affecting Protoplast Culture
 Plant species and varieties
 Plant age and Organ
 Pre culture conditions
 Pre treating to tissue before isolating protoplast
 Density
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21. Protoplast- up to now
– Transfer of useful genes from one species to another
– Techniques used in strain improvement for bringing genetic recombinants.
– Techniques used in microbial engineering: new strains produced for industrial
purposes.
– Protoplast fusion used by:
 Molecular Biologist
 Biotechnologists
 Industries
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22. Cont…..
 Best beer production(by S.uvarum)
 Palm oil Production
 Date Fruit Production
 Potato Production
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