Commercial exploitation of micropropagation& its tech in fruit crops
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Micropropagation is a process where a whole plant can be regenerated from small plant tissues or cells in a suitable culture medium. It allows for rapid multiplication of plants in a relatively short time and small space. The process involves 5 main steps - selection of explant, initiation and establishment of aseptic culture, multiplication of shoots, rooting, and acclimatization of plantlets. Some key advantages of micropropagation are that it enables fast commercial propagation of new varieties, provides a continuous supply of young pathogen-free plants, and allows for storage and exchange of germplasm. Micropropagation has been developed for many horticultural crops in India like banana, citrus, grapes, and strawberries.
Commercial exploitation of micropropagation& its tech in fruit crops
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- 2. Presentation on Commercial Exploitation of Micro-propagation in fruit crops & its Techniques Presentation on Commercial Exploitation of Micro-propagation in fruit crops & its Techniques presented by pawan Kumar nagar m.sc. (Horti) pomology reg. 04-2690-2015 baca, aau, anand 2
- 4. wHat iswHat is micropropagationmicropropagationA whole plant can be regenerated from a small tissue or plant cells in a suitable culture medium under controlled environment. The plantlets so produced are called tissue-culture raised plants. The advantage is that in a relatively short time and space a large number of plants are obtained. The advantage is that in a relatively short time and space a large number of plants are obtained. type of micropropagationtype of micropropagation 1.Direct micropropagation 2.Indirect micropropagation type of micropropagationtype of micropropagation 1.Direct micropropagation 2.Indirect micropropagation 4
- 5. wHy dowHy do micropropagationmicropropagation wHy dowHy do micropropagationmicropropagation A single explant can be multiplied into several thousand plants in less than a year - this allows fast commercial propagation of new cultivars. Once established, a plant tissue culture line can give a continuous supply of young plants throughout the year. In plants prone to virus diseases, virus free explants (new meristem tissue is usually virus free) can be cultivated to provide virus free plants. Plant ‘tissue banks’ can be frozen, then regenerated through tissue culture. Plant cultures in approved media are easier to export than are soil-grown plants, as they are pathogen free and take up little space (most current plant export is now done in this manner). A single explant can be multiplied into several thousand plants in less than a year - this allows fast commercial propagation of new cultivars. Once established, a plant tissue culture line can give a continuous supply of young plants throughout the year. In plants prone to virus diseases, virus free explants (new meristem tissue is usually virus free) can be cultivated to provide virus free plants. Plant ‘tissue banks’ can be frozen, then regenerated through tissue culture. Plant cultures in approved media are easier to export than are soil-grown plants, as they are pathogen free and take up little space (most current plant export is now done in this manner). 5
- 6. Advantages of micropropagationAdvantages of micropropagation From one to many propagules rapidly Multiplication in controlled lab conditions Continuous propagation year round Potential for disease-free propagules Inexpensive per plant once established Precise crop production scheduling Reduce stock plant space Long-term germplasm storage Production of difficult-to-propagate species DisadvantagesDisadvantagesDisadvantagesDisadvantages Specialized equipment/facilities required More technical expertise required Protocols not optimized for all species Plants produced may not fit industry standards Relatively expensive to set up 6
- 7. genetically modified plant breeding does not produce seeds does not respond well to vegetative reproduction7
- 8. Steps of MicropropagationSteps of Micropropagation 8
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- 10. Micropropagation involved in 5 steps: 10
- 11. Steps involved in theSteps involved in the in vitroin vitro MicropropagationMicropropagation Cleaning of glassware Preparation of nutrient medium Selection and sterilization of explant. Inoculation of aseptic explant in to nutrient medium. Proliferation of shoots on a multiplication medium. Transfer of shoots for sub-culturing. Rooting and hardening of plantlets Field trials. 11
- 12. 12Source : Cadila pharmaceuticals limited
- 13. Stage 0 : Selection and maintenance of stock plants for culture initiation EXPLANTEXPLANT SHOOT TIPS FROM YOUNG SUCKERSHOOT TIPS FROM YOUNG SUCKER APICAL MERISTEM (1-2 Cm3)APICAL MERISTEM (1-2 Cm3) SURFACED STERILIZATIONSURFACED STERILIZATION 13
- 14. ExplantExplant Cell, tissue or organ of a plant that is used to start in vitro cultures. 14
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- 16. Stage- I : Initiation and establishment of aseptic culture 16 Source: Jain Irrigation Systems Ltd
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- 19. Initiation 19
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- 21. Stage –II : Multiplication of shoot 21
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- 25. Stage –IV: Acclimatization / Hardening 25
- 26. Healthy/elite plantlets are exposed to the natural conditions in a step wise manner. It is a gradual acclimatization of in vitro grown plants to in vivo condition. The plantlets are transferred to the pots/polyghene bag and immediately irrigated with inorganic/nutrient solution. Plants are kept in the hardening room where controlled conditions of light, humidity and temperature are maintained. Plants are maintained under high humidity for 10-20 days and subsequently transferred in the field so as to grow under natural conditions. The success rate of micropropagation depends on the survival of the plantlets when transferred from culture to the soil (field). 26
- 27. Secondary Hardening 27 Plants transferred to nursery bagsPlants transferred to nursery bags Kept for 6 to 8 weeks under 50% shadeKept for 6 to 8 weeks under 50% shade Regular foliar spraysRegular foliar sprays Variation if observed is discardedVariation if observed is discarded Plant ready for sale (1 feet height)Plant ready for sale (1 feet height)
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- 30. Factors Affecting Plant micro-propagation 1. Growth Media 2. Environmental Factors 3. Explants Source 4. Genetics 30
- 31. Method of Micro-propagationMethod of Micro-propagation 31
- 32. Method of Micro-propagation A. Meristem tip cultureA. Meristem tip culture B. Callus cultureB. Callus culture C. Suspension cultureC. Suspension culture E. Protoplast cultureE. Protoplast culture D. Embryo cultureD. Embryo culture F. Ovule cultureF. Ovule culture G. Anther cultureG. Anther culture H. Cell cultureH. Cell culture 32
- 33. In addition to major use of tissue culture techniques for rapid clonal multiplication of plants, this technique is highly important for several purpose as under: 1. Production and maintenance of pathogen free stock plants; 2. Long term in vitro conservation of germplasm. 3. Selection and regeneration of transgenic plants. 4. Conservation of germplasm. Other Implications of Micro-propagationOther Implications of Micro-propagation 33
- 34. Micro-propagation Techniques & ProcedureMicro-propagation Techniques & Procedure A. Micro-Propagation by Nodes B. Micro-propagation by Node Cuttings in a Liquid Medium Procedure 1. Sterilize Petri dishes and prepare the laminar flow chamber by disinfecting the internal surfaces with alcohol. Sterilize the tools with an instrument sterilizer and place them on a sterile dish. 2. Open the tube, take off the plantlet and place it on a Petri dish with the help of forceps. 3. Remove the leaves and cut the nodes. 4. Open a tube containing fresh sterile medium and place a node inside, trying to plunge it slightly into the medium with the bud up. Close the tube. 5. Seal the tube with a gas-permeable plastic tape and label it correctly. It is recommended to place two explants in 16 x 125 mm tubes, three in 18 x 150 mm tubes, five in 25 x 150 mm tubes, and 20-30 in magenta vessels. 34
- 35. Applications of Tissue Culture 1. Embryo culture 2. Meristem culture 3. Micropropagation 4. Somatic embryogenesis and Organogenesis 5. Somaclonal variation and in vitro selection 6. Anther culture Haploid & Dihaploid Production 7. Protoplast culture (In vitro hybridization – Protoplast Fusion) 8. Germplasm preservation 35
- 36. Micro-propagation technology developed for some Horticultural Crops in India Sr. No. Crop Institute 1. Annona IIHR, Banglore; BARC, Mumbai; NCL, Pune 2. Banana NCL, Pune; IIHR, Banglore; TNAU, Coimbatore 3. Citrus NBRI, Lucknow; NRC- Citrus, Nagpur 4. Grape IARI, New Delhi 5. Guava GBPUAT, Pantnagar 6. Papaya IARI, New Delhi 7. Pineapple BARC, Mumbai 8. Strawberry TERI, New Delhi 36
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- 38. Tissue culture BananaTissue culture Banana 38
- 39. Tissue culTure DaTe palm 39
- 40. Tissue culTure pomegranaTeTissue culTure pomegranaTe 40
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