Micropropagation
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Micropropagation is a tissue culture technique used for the rapid asexual propagation of plants. It involves culturing small pieces of plant tissues or organs on growth media in controlled conditions. The process includes initiation of cultures from explants, multiplication through shoot proliferation or somatic embryogenesis, rooting of shoots, and transplantation of plantlets to soil. Micropropagation allows for large-scale production of genetically identical clones in a short period of time. It has applications in producing disease-free plants, conserving rare species, and commercializing new plant varieties.
![Stages of Micropropagation
Stage 0: Preparation of the mother plant
Stage 1: Initiation and establishment of cultures
Stage 2: Multiplication [shoot or rapid embryo; and rooting of shoot]
Stage 3: Germination of somatic embryo for rooting of in-vitro formed
shoots
Stage 4: Transfer of plantlets -- sterilized soil ---- Hardening
to greenhouse/ field conditions (transplantation)](https://image.slidesharecdn.com/micropropagation-200902054900/85/Micropropagation-10-320.jpg)
Micropropagation
- 1. Micropropagation Dr. Divya Sharma Assistant Professor A Biodiction (A Unit of Dr. Divya Sharma)
- 2. Micropropagation Tissue culture technique: Increasingly popular alternative means -- Plant vegetative propagation Plant tissue culture involves – o Asexual method of propagation o Primary goal ----- crop improvement In-vitro selection, genetic manipulation techniques ------- Higher plant
- 3. Clonal Propagation Clonal propagation ------- In-vitro selection Micropropagation Greek word --- Clone --- Twig ---- Identical copies Multiplication of genetically identical copies by asexual reproduction is called Clonal Propagation Process -- Asexual reproduction --- Multiplication occur ----- Individual plants genetically identical copies Multiplication of genetically, identically individual by asexual reproduction Rapid process -------- Commeralization of improvement plants like apple, pear etc. Micropropagation Apomixes (seed development without meiosis and fertilization) and vegetative propagation (regeneration of new plants from vegetative parts) Short time
- 4. Micropropagation Aseptic method of clonal propagation is called as Micropropagation Synonyms -- tissue culture Rapid multiplying stock plant material to produce a large number of progeny plants, using modern plant tissue culture methods Widely used for Orchids, Ferns, many interior foliage plants etc.
- 5. Features of Micropropagation Relatively short time Easy to manipulate production cycles Disease free plants can be produced In-vitro production can be planned Recycled many times to produce an unlimited number of clones Micropropagated plants may acquire new desirable traits, like bushy habit of ornamental plants and increased number of runners in strawberry Commercially viable method for horticultural crops Eg.: Orchids, Ferns, many interior foliage plants etc.
- 6. Rapid Clonal in-vitro Propagation of Plants From cells, tissue or organs Cultured aseptically on defined media Contained in culture vessels Maintained under controlled conditions of light and temperature
- 7. Commercialization of Micropropagation Broad commercial applications ------ 1970s and 1980s Murashige (1974)
- 8. Selection of plant material Part of plant Genotype Physiological condition Season Position on plant Size of explants
- 9. Explants used in Micropropagation Different kinds of explants used in micropropagation: Eg.: In case of orchids, shoot tips (Anacamptis pyramidalis, Aranthera, Calanthe, Dendrobium); Axillary bud (Aranda, Brassocattleya, Cattleys Laelia); Inflorescence segment (Aranda, Ascofinetia, Neostylis, Vascostylis); Lateral bud (Cattleya, Rhynocostylis gigantean); Leaf base (Cattleya); Nodal segment (Dendrobium; Flower stalk segment (Dendrobium, Phalaenopsis); and Root tips (Neottis, Vanilla)
- 10. Stages of Micropropagation Stage 0: Preparation of the mother plant Stage 1: Initiation and establishment of cultures Stage 2: Multiplication [shoot or rapid embryo; and rooting of shoot] Stage 3: Germination of somatic embryo for rooting of in-vitro formed shoots Stage 4: Transfer of plantlets -- sterilized soil ---- Hardening to greenhouse/ field conditions (transplantation)
- 11. Stages of Micropropagation Maintained light, temperature and moisture regimes under which the mother plants Loosely covering growing branches with polythene Application of growth regulators Watering of plant from the base Stage 0: Preparatory stage Involves the preparation of mother plants to provide quality explants for better establishment of aseptic cultures
- 12. Stages of Micropropagation Explant selection Sterilization treatment Choice of growth medium Stage 1: Initiation of Cultures
- 13. Stage 2: Multiplication Through Callusing Initiation of culture on high Auxin media Development of unorganized mass of cells called callus Transfer small piece of callus on regeneration media Shoot bud regeneration Somatic embryo
- 14. Stage 2: Multiplication Adventitious Bud Formation Transfer to proliferation media Shoots can be constantly divided
- 15. Small bud present Apical dominance is removed Cytokinins Auxins Cluster formation Off types Stage 2: Multiplication Enhanced Axillary Branching
- 16. Somatic embryos carry a preformed radical and may develop directly into plantlet. Embryos sow very poor conversion into plantlets, especially under in-vitro conditions Require an additional step of maturation to acquire the capability for normal germination Adventitious and axillary shoots developed in cultures in the presence of a cytokinins generally lack roots Obtain full plants the shoots must be transferred to a rooting medium which is different from the shoot multiplication medium, particularly in its hormonal and salt compositions For rooting, individual shoots measuring 2 cm in length are excised and transferred to the rooting medium Stage 3: Rooting of shoots Rooting & Hardening
- 17. Tissue culture raised plants are characterized by: Low photosynthetic rates Non-functional stomata Low cuticular wax Abnormal leaf morphology Accilimitization Green house 2 weeks Temperature (±10º) Stage 4: Transplantation & Accilimitization
- 19. In-vitro regenerative protocol of V. reitzii Nodule cluster culture formation and shoot regeneration in V. reitzii; a). Donor plant (inflorescence of adult plant); b). Yellow nodular cultures induced in MS medium free of PGR; c). Nodule cluster subcultureed in MS culture medium supplemented with GA3 resulted in high proliferation rate and the subsequent development of adventitious microshoots; d). Elongation and growth of shoot in MS culture medium free of PGR; e). Acclimatization of plantlets; f). Growth in green house
- 22. Methods of Micropropagation Organogenesis Organogenesis via callus formation Direct adventitious organ formation Embryogenesis Direct embryogenesis Indirect embryogenesis Microcutting Meristem culture (Mericloning) Bud culture
- 23. Advantages of Clonal Propagation From one to many propagules rapidly Multiplication in controlled lab conditions Continuous propagation year round Potential for disease free propagules Inexpensive per plant once establishment
- 24. Disadvantages of Clonal Propagation Specialized equipment/ facilities required More technical expertise required Protocols not optimized for all species Plants produced may not fir industry standards Relatively expensive to set up
- 25. Applications of Clonal Propagation Produce disease and virus free plantlets Leads to simplification of international exchange of plants Increases the production of plants which are normally propagated very slowly such as Narcissus and other bulbous crops Introduction of disease free new cultivars is possible through micropropagation Large number of plants can be obtained in small spaces Rapid method for cloning of disease free trees Secondary metabolites In-vitro cultures can be stored for long time through cryopreservation Breeding cycle shortened Rare species ---- Conservation Production of identical sterile hybrid species. Eg.: Cabbage
- 26. Horticultural uses for plant tissue culture Clonal mass propagation Difficult or slow to propagate plants Introduction of new cultivars Vegetative propagation of sterile hybrids Pathology – eliminate viruses, bacteria, fungi etc Storage of germplasm
- 27. Thank you A Biodiction (A Unit of Dr. Divya Sharma)