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Application of plant tissue culture/ micro-propagation
Tissue culture is the process of growing cells or tissues in sterile conditions. It allows for rapid cloning of plant materials. Plant tissue culture involves excising plant parts and growing them on nutrient media. This allows for mass multiplication of plant materials irrespective of season. Some key developments include Haberlandt's proposal of plant cell culture in 1902, and Murashige and Skoog's nutrient medium in 1962. Micropropagation is now used for conservation of rare species, producing disease-free plants, mutation breeding, and more. The future of this technique remains promising.
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Introduction to tissue culture which involves plant and animal cells, producing clones with same genotype. Historical insights including pioneers like Haberlandt, White, Murashige, and Skoog.
Explanation of various explants in plant tissue culture including shoots, leaves, roots, and pollen. Illustrates different plant parts used for micropropagation.
Detailed steps of micro-propagation, stressing on advantages such as high multiplication rates, ‘true to type’ clones, disease-free plants, and year-round supply.
Explores broad applications of micro-propagation including somaclonal variation, germplasm conservation, mutation breeding, embryo culture, disease-free plants, and genetic engineering.
Discusses challenges faced in micro-propagation such as cost and acclimatization issues, with successful commercialization cases like Taiwan's orchid industry highlighted.
Speculates on future developments in micro-propagation, encouraging readiness for upcoming discoveries.
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Application of plant tissue culture/ micro-propagation
- 1. Sushil Nyaupane Department ofNatural Resources and Environmental Design North Carolina A&T State University
- 2. What is tissueculture? Tissue culture is the term used for “the process of growing cells artificially in the laboratory”. Tissue culture involves both plant and animal cells. Tissue culture produces clones in which all product cells have the same genotype.
- 3. Plant Tissue Culture Plant tissue culture refers to the techniques of growing plant cells, tissues, organs, seeds or other plant parts in a sterile environment on a nutrient medium.
- 4. Who they are? Haberlandt T. Murashige F. Skoog
- 5. A glance inHistory In 1902, Haberlandt; father of plant tissue culture ( He proposed that plant cells could be cultured. In 1930- White cultured tomato root tip and subculutred to fresh medium containing salts, yeast extract and sucrose and vit B During this period, some plant growth regulators, additives and vitamins was discovered for the plant micro- propagation. discovery of first PGR ---IndoleAcetic Acid , called IAA, in 1937 In 1962, Murashige and Skoog published a recipe for MS media. In 1972, protoplast fusion has been done in toabacco.
- 7. Micropropagation israpid clonal in vitro propagation of plants from cells, tissues or organs cultured aseptically on defined media contained in culture vessels maintained under controlled conditions of light and temperature. Briefly, it is the art and science of multiplying plants in- vitro.
- 8. Explants Explant is anexcised piece of tissue or organ taken from the plant to initiate a culture. They can be : • shoot meristem, tip, bud • leaf or stem (internode) • root • anther / microspore • ovule • embryo associated seed parts
- 9. Shoot Explants Meristem TipShoot Tip
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- 15. Plant tissue culturetypes for micro-propagation
- 19. Yes, this isfrom lab.
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- 21. Steps of micro-propagation Stage 0- Selection and preparation of the mother plant(Sterilization of the explant tissues) Stage I- Initiation of culture ( Explant placed into growth media) Stage II- Multiplication ( Explant transferred to shoot media and shoot can be constantly divided) Stage III- Rooting ( Explant transferred to rooting media) Stage IV- Transfer to soil ( Hardening off)
- 23. Why do micro-propagation A single explant can be multiplied into several thousand plants in less than a year. Once established, it can give a continuous supply of young plants throughout the year i.e. irrespective of season. Taking an explant does not usually destroy the mother plant, so rare and endangered plants can be saved somehow. Clones through micro-propagation are ‘true to type’ as compared with seedlings, which show greater variability
- 24. Why do micro-propagation? This allows fast selection for crop improvement - explants are chosen from superior plants, then cloned. Plant ‘tissue banks’ can be frozen, then regenerated through micro-propagation. Disease and virus free plants can be produced through this technique. Greater credibility in international market as plantlets are produced through micro-propagation( Phyto-sanitary perspective)
- 25. How possible isthis ?? Micro-propagation of almost all vegetables and fruit crops is possible. Some examples,mustard, corn, soybean, azalea, dwarfing sweet cherry, strawberry, mango, banana, rose, orchid, nutraceutical plants, rhododendron, citrus, potato, tomato,legumes etc.
- 26. Applications of micro-propagation Somaclonal Variation Germplasm Conservation Mutation Breeding Inducing mutation Embryo culture Haploid and Dihaploid production In Vitro Hybridization- protoplast fusion Production of Disease free plants Molecular farming Genetic engineering Production of secondary metabolites
- 27. Applications .. Somaclonal variation Somaclonalvariation is a general phenomenon of all plant regeneration systems that involve a callus phase. There are two types of Somaclonal variation: Heritable genetic variation Non- Heritable genetic variation
- 28. Applications.. Advantages of somaclonalvariation It helps in crop improvement Creates additional genetic variants Plants with resistant and tolerant to toxins, herbicides, high salt and even mineral toxicity Suitable for breeding purposes Increased and improved production of secondary metabolites
- 29. Applications.. Germplasm conservation Micropropgationis utilized in conserving genetic resources. Depending upon the crop species and method of preservation, tissue culture can help in the preservation of genetic resources from 1 to 15 years. Cyopreservation – the preservation of germplasm in a dormant state at ultra-low temperatures, usually in liquid nitrogen (-196 °C) – is a type of tissue culture which can be used to preserve seeds.
- 30. Applications.. Mutation Breeding 1927:Muller produced mutations in fruit flies using x- rays 1928: Stadler produced mutations in barley Basically, there are three groups of breeders 1) Mutation breeding is useless, we can accomplish the same thing with conventional methods 2) Mutation breeding will produce a breakthrough given enough effort 3) Mutation breeding is a tool, useful to meet specific objectives
- 31. Applications.. Types of mutation Spontaneous(natural mutation) Some type of spontaneous mutation have played an outstanding role in development of valuable crop cultivars and hybrids. But limitation of this is that, it can not form the basis of modern plant breeding Induced mutation Any mutation found in nature can be induced by mutation breeding.
- 32. Applications.. Inducing Mutations Physical mutagens (Irradiation) These mutagens helps in the chromosome aberrations and point mutations. Neutrons, Alpha rays Gamma, Beta, X-rays Chemical mutagens By using different carcinogenic chemicals which are highly toxic in nature and eventually result in point mutations.
- 33. Applications.. Embryo Culture Embryoculture is usually done from the need to rescue embryo from wide crosses where fertilization occurred, but not the embryo development. Objectives: Rescue F1 Hybrid from wide crosses Overcome seed dormancy by addition of hormone to media. For ex. GA To overcome immaturity in seed To rescue valuable genotype from dead or dying plant To speed up the generations in a breeding program
- 34. Applications.. Embryo culture asa source of genetic variation Hybridization Can introduce new genetic combinations through inter-specific crosses Can transfer mutant alleles between species Polyploidy It combines embryo culture with chromosome doubling to create new polyploid species
- 35. Application… Haploid and dihaploidproduction Plants produced through the anther culture are the haploids. Doubling the chromosomes without going into series of backcrossing produce homozygous plants. This technique shortens the time of breeding by half.
- 36. Application… In Vitro Hybridization(Protoplast fusion) Created by degrading the cell wall using enzymes. Very fragile in nature. Protoplasts can be induced to fuse with one another. Methods: Electrofusion Poly Ethylene Glycol(PEG) Addition of calcium ions at high PH Values
- 37. Applications.. Production of Diseasefree plant Heat treatment Plants grow faster than viruses at higher temperature. Meristemming Viruses are transported from cell to cell through plasmodesmata and vascular tissue. Apical meristem are virus free in nature. So, micro- propagation of these cells gives virus free plantlets. Even, not all the cells in the plant are infected. For example, adventitious shoots formed from single cells can give virus free shoots.
- 38. Applications.. Molecular farming Whereplants are treated as bioreactors for the production of specific compounds. Range from simple peptides to a thermoplastic. Two types of products: 1)High value compounds with small scale production requirements such as pharmaceutical products. For ex. Malaria epitope. 2)Compounds needed on a bulk scale with low production costs (plant biotechnology has greatest potential in this area) For ex. α-Amylase (food + detergent industries)-starch manipulation
- 39. Applications… Genetic Engineering Geneticengineering would not be possible without the development of plant tissue culture. Genetic engineering requires the regeneration of whole plants from single cells. Efficient regeneration systems are required for commercial success of genetically engineered products.
- 40. Application.. Protoplast fusionbetween male sterile cabbage and normal cabbage was done and cybrids were selected that contained the radish mitochondria and cabbage chloroplast.
- 41. Applications.. Production of secondarymetabolites Secondary metabolites are those cell constituents which are not essential for survival. For example, alkaloids, glycosides, terpenoids,latex,tannins etc. In vitro production of secondary metabolites is much higher from differentiated tissues compared to non- differentiated tissues. Azadirachtin from Azadirachta indica Insecticidal Berberine from Coptis japonica Antibacterial, anti inflammatory Capsaicin from Capsicum annum Cures Rheumatic pain
- 42. Problems of micro-propagation Expensive laboratory equipment and service No possibility of using mechanization Plants are not autotrophic Poor Acclimatization to the field is a common problem (hyperhydricity) Risk of genetic changes if 'de novo' regeneration is used Mass propagation cannot be done with all crops to date. In cereals much less success is achieved Regeneration is often not possible, especially with adult woody plant material.
- 43. Do you haveany idea about this orchid?
- 44. Micro-propagation a toolfor commercialization (Taiwan story) According to Taiwan Today, Jan 1, 2016 The nation’s orchids are exported to 36 countries in Northern America, Northern Europe and South Africa. This country began the export with US $ 23 million in 2004 but in 2015 they have exported with worth US $ 130 million. They are selling a Phalaenopsis or moth orchid in Dubai at the price of US $ 1,000. Won the bid to host the 23rd World Orchid Conference in 2020.
- 45. What may bethe reason for this achievement ? According to John Feng, CEO of SOGO TEAM CO LTD., they are exploiting micro-propagation as a technique in mass production as well as in variety improvement. In I-Hsin Biotechnology Corp’s Tissue Culture Lab only, they have conducted 12,000 breeding experiments which have yielded 2,300 Phalaenopsis varieties. This all is being possible through tissue culture.
- 46. ALBIFLORA ORCHID OFNEPAL In Nepal a total of 377 species of orchids belonging to 100 genera reported.
- 47. Future of micro-propagation …………………………………………………………………………………… Perhaps some of the greatest discoveries of plant tissue culture are yet to come. SO BE READY !
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