Plant tissue culture.rrrr sssss


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Plant tissue culture.rrrr sssss

  1. 1. Presented by Surya Pratap Singh M . Sc , Life Sciences Central University of Gujarat
  2. 2. INTRODUCTION  The aseptic culture of plant protoplasts, cells, tissues or organs under conditions which lead to cell multiplication or regeneration of organs or whole plants.
  3. 3. HISTORY OF PLANT TISSUE CULTURE  The idea of the totipotency of plant cell was given by Haberlandt in 1902.  White first time established successful root culture of tomato in 1937.  Vanoverbeek used coconut milk for growth and development of young Datura embryos in 1941.  Skoog and Miller demonstrated the role of auxin and cytokinin on root and shoot formation in tobacco – tissue in 1957.  Murashige and Skoog introduced the medium for tobacco culture in 1962.  Guha and Maheshwari obtained for the first time haploid Plants through anther culture of Datura.
  4. 4. TYPES OF PLANT TISSUE CULTURES Plant tissue culture Embryo culture Seed culture Meristem culture Protoplast cultureCell culture Callus culture Bud culture Organ culture
  5. 5. Callus – A tissue that develops in response to injury caused by physical or chemical means, most cells of which are differentiated although may be and are often highly unorganized within the tissue. In nature, this wound tissue produces a protective layer of cells to cover an injury, example. Differentiated Cells - products of cellular maturation, i.e. cell types with particular function, e.g. xylem tracheary elements; large cells that are highly vacuolated with relatively little cytoplasm Undifferentiated Cells - meristematic; progenitors of differentiated somatic cells, e.g. small, isodiametric, small vacuoles.
  6. 6. Tobacco Callus
  7. 7. CELL SUSPENSION CULTURE  Tissues and cells cultured in a agitated liquid medium produce a suspension of single cells and cells clumps of few to may cell, these are called suspension cultures.
  8. 8. 2nd Passage1st PassageFriable Callus
  9. 9. PROTOPLAST CULTURES  Isolated protoplasts have been described as "naked" cells because the cell wall has been removed by either a mechanical or an enzymatic process.  Protoplasts can be induced to reform a cell and divide if placed in a suitable nutrient medium than form callus.
  10. 10. Isolation of Protoplast (Separation of protoplasts from plant tissue) 1. Mechanical Method 2. Enzymatic Method
  11. 11. MECHANICAL METHOD Plant Tissue Collection of protoplasm Cells Plasmolysis Microscope Observation of cells Cutting cell wall with knife Release of protoplasm
  12. 12. ENZYMATIC METHOD Leaf sterlization, removal of epidermis Plasmolysed cells Plasmolysed cells Pectinase +cellulase Pectinase Protoplasm released Release of isolated cells cellulase Protoplasm released Isolated Protoplasm
  13. 13. MERISTEM CULTURE  Cultivation of auxiliary or apical shoot meristems, particularly of shoot apical meristem ,is known as meristem culture.
  14. 14.  Application  Production of virus free germplasm.  Mass production of desirable genotypes.
  15. 15. ANTHER CULTURE/POLLEN CULTURE  Anther culture is a technique by which the developing anthers from unopened flower bud are cultured on a nutrient medium where the microspores within the cultured anther develop into callus tissue or embryoids that give rise to haploid plant. Application  Production of haploid plant  Production of useful gametoclonal varition.
  16. 16. Figure Show the Anther culture and haploid plant
  17. 17. SOMATIC EMBRRYOGENESIS  In somatic embryogenesis the embryos regenerate from somatic cells, tissue or organs either de nove directly from the tissues, which is the opposite of zygotic or sexual embryogenesis.
  18. 18. TWO TYPES OF SOMATIC EMBRYOGENESIS  Direct embryogenesis  The embryos initiate directly from explants in the absence of callus formation.  Indirect embryogenesis  Callus from explants takes place from which embryos are developed.
  19. 19. APPLICATION  Production of artificial seeds.  As source materials for embryogenic protoplast.  For genetic transformation.  Production of primary metabolites specific to seeds such as lipids in oil seeds.
  20. 20. OVARY/OVULE CULTURE  Ovary or ovule culture involves development of haploid from unfertilized cells of embryosac present in ovary.
  21. 21. Application  Production of haploid plants.  Achievement of in vitro fertilization.