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Biotechnology in Horticulture Application of Plant Cell and Tissue Culture

Biotechnology in Horticulture Application of Plant Cell and Tissue Culture. Clonal Propagation Disease Elimination Secondary Products Germplasm Storage Genetic Improvement. www.Hortilover.net. 1. Clonal Propagation.

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Biotechnology in Horticulture Application of Plant Cell and Tissue Culture

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  1. Biotechnology in HorticultureApplication of Plant Cell and Tissue Culture • Clonal Propagation • Disease Elimination • Secondary Products • Germplasm Storage • Genetic Improvement www.Hortilover.net

  2. 1. Clonal Propagation Explant Source: Shoot tips, stem nodes, Leaf discs, flower buds Advantages: Rapid multiplication Problems: Mortality during soil establishment Somaclonal variation Application: Foliage plants Floricultural crops Landscape plants Some fruits Seed potatoes

  3. Apical Shoot Tip Culture

  4. Root Induction on Shootlets Root Induction on Desert Milkweed Left (A) - Roots are formed on callus due to excess auxin levels in the medium (undesirable) Right (B) - Root formation without callusing (desirable) A B

  5. Paper Bridge Method of In Vitro RootingSyngonium podophyllum (Arrow Head)

  6. Tissue Culture Propagation of River Birch Source: McCown

  7. Tissue culture grown birch plants (Right) are fuller with multiple branches at the base compared to cutting grown plants (Left).Source: B. McCown

  8. Cloning of Plants by Leaf Disc Culture

  9. Leaf disc culture of Salpiglossis sinuata

  10. Cytokinin and Auxin Influence on Differentiation of Callus TissuesSalpiglossis sinuata

  11. Use high cytokinin and low auxin concentrations in leaf disc culture in cloning of Salpiglossis sinuata (Velvet Flower)

  12. Clonal Propagation of Orchids A B C D A-obtain a meristem explant from a lead, B-an excised meristem, C- five protocorms formed from a meristem, D-multiplication of protocorms and shoot formation

  13. Commercial Orchid Cloning Labs

  14. Commercial Tissue Culture PropagationTwyford Lab, Santa Paula, California

  15. Clonal Propagation of Foliage PlantsTwyford Lab

  16. Clonal Propagation of Fruit Crops

  17. Commercial Production of Ferns by Tissue Culture

  18. Mechanical Transplanting of Tissue Culture Grown Plants

  19. Tissue Cultured Ferns Grown in Greenhouses

  20. Staghorn Ferns Grown from Tissue Culture

  21. 2. Disease Elimination Explant Sources: Apical meristems, shoot apex Pathogens: Viruses, Bacteria, Mycoplasma (MLO) Advantages: Cleaning of infected plants Problems: Plant regeneration Obtaining disease-free tissues (use of heat treatment) Application: Carnation Strawberry Potato Nursery crops Foliage plants

  22. Hydrangeas Clonally Propagated from Clean Stock Plants Mycoplasma infected plants are routinely cleaned by meristem cultures

  23. Clean Stock Program Used for Commercial Potato Procedures for cleaning virus infected clones and subsequent generation of nuclear seed potatoes for distribution

  24. Seed Potato ProductionND State Seed Department A B C D Shoots (A) from virus-free merstems multiplied in vitro (B) are transferred into soil medium and grown in a screened greenhouse (C, D) to ward off insect vectors

  25. Potato Mini-tuber Production in Vitro

  26. 3. Secondary Products Products: Essential Oils, Food Additives, Pharmaceuticals, Perfumes, Insecticides, Pigments Explant Sources: Callus, Cells, Embryos, Floral Tissues, Roots Advantages: “Pharming” in vitro Use of Bioreactors for Mass Production Problems: Energy Use Efficiency Scale-Up Production Slow Cell Cycles Examples: Chocolate, Shikonin, Perfumes, Taxol, Ginsengoside

  27. Chocolate Production Via Asexual EmbryogenesisTheobroma cacao

  28. Callus Cultures

  29. Examples of Secondary Products Produced in Tissue Culture *S=Suspension culture; C=callus culture

  30. Ginseng Root Culture

  31. In Vitro Production of Cotton Fiber

  32. 4. Germplasm Storage Explant Source: Meristems, Tissues, Cells, Pollen, Embryos, Seeds Methods: Hypobaric Storage (low atmospheric pressure) Cryogenic Storage (- 196 oC) Problems: Plant Regeneration After Storage Possible Chromosomal Damage Application: Long Term Preservation Germplasm Exchange Between Countries

  33. Cryogenic Storage of Plant Cell and Tissues • Liquid nitrogen (-196 oC, -320 oF) • Permanent storage of pollen, seed, meristems

  34. 5. Genetic Improvement Methods: 1) Haploid Production by Anther Culture 2) Embryo Rescue 3) Test Tube Pollination 4) Somatic Hybrids 5) Mutant Selection in Vitro 6) Genetic Engineering 7) Seed Encapsulation Problems: 1) Difficulty in Plant Regeneration 2) Lack of Marker Genes 3) Stability of Recombinant Traights 4) Use of Multigenic Traits Uses: 1) Resistance to Diseases, Insects, Herbicides 2) Stress (Cold, Heat, Salinity) Tolerance 3) Higher Yield 4) Nutritional Quality 5) Vaccine Production 6) New Horticultural Traits, etc.

  35. Plant Protoplast Culture

  36. Protoplast Isolation, Fusion, and Plant Regeneration

  37. Somatic Hybrid Between Tomato and Potato

  38. Somatic Hybrid in Genus Putunia First Row: Two parental lines 2nd Row: F1 hybrid (left) and somatic bybrid (right) 3rd and 4th Rows: Segregation of F2 progeny in somatic hybrids

  39. Plant Transformation Methods: 1) Use of Ti Plasmids as Vectors 2) Mostly Agrobacterium tumefesiens-mediated 3) Direct Transmission by Gene Gun 4) Pollen-mediated Gene Transformation Problems: 1) Lack of Desirable Genes to Transfer 2) Use of Selective and Marker Genes 3) Difficulty in Transforming Multi-genic Traits 4) Regeneration of Transgenic Plants 5) Public Perception and Acceptance

  40. Agrobacterium-Mediated Gene Transfer

  41. Designing Plasmid Vectors as Vehicles of Gene Transfer

  42. Genetic Transformation in Tobacco Bottom-Transgenic tobacco plant containing Round-Up resistance gene (left)

  43. Transgenic Plants A transgenic corn plant having additional copies of alcohol dehydrogenase gene Not all transgenic plants are useful.

  44. Transfer of Luciferus Gene from Firefly to Plants

  45. Examples of Transgenic PlantsSalpiglossis sinuata, Velvet Flower

  46. Transgenic Plants of Velvet Flower Bottom-rolC gene construct Right-Control (left) and transgenic plants (right) with rolC gene

  47. Genetic Improvement of TomatoIncreasing Solid Content

  48. Papaya Ring Spot Virus

  49. Artificial Seed Encapsulation Coating Materials 1. Sodium Alginate 2. Potassium Alginate 3. Sodium Alginate + Gelatin 4. Mixture of Carragreenan, Guar Gum, Locust Bean Gum, Agar, Tragacanth Gum, Sodium Pectate, Carboxymethyl Cellulose Uses: For Somatic Embryo Plantlets For Germinates Seeds Products Gel-CoatTM (U.S. Pat. 4,562,663, PGI)

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