Protoplast
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Transcript of Protoplast
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INTRODUCTION
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Introduction
Solution to sexual incompatibility barriers – distant species;
Fusion of cells – full hybrids – crop improvement programmes;
EC Cocking (1960) – enzymatic degradation of plant cell wall - viable naked cells
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Freshly isolated mesophyll protoplasts
JB Power, UK
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Introduction
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ISOLATION OF PROTOPLASTS
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ISOLATION OF PROTOPLASTS FROM PLANTLETS OF Solanum phureja
Freshly isolated
First mitotic division
Multicellular colonies (7-d-old)
Macroscopic p-calli (several wks)
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Factors that influence yield and viability of protoplasts
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Factors that influence yield and viability of protoplasts
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Factors that influence yield and viability of protoplasts
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Factors that influence yield and viability of protoplasts
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Commercial Enzymes
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Factors that influence yield and viability of protoplasts
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Factors that influence yield and viability of protoplasts
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Factors that influence yield and viability of protoplasts
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Viability of the Protoplasts
Detection methods:
1)Observation of cyclosis or cytoplasmic streaming;
2)Oxygen uptake
3)Photosynthetic activity;
4)Staining with fluroescein diacetate - common
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Nutritional Requirements
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Plant regeneration
- Plant regeneration from hypocotyl protoplasts of Brassica napus
Stage
Event
AFreshly isolated
protoplasts
B 1-d-old protoplasts
C Callus (14-d culture)
D Shoot buds differentiated
ECalli with regenerated shoots (Zeatin + IAA)
F Plant regenerated
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