Plant tissue culture techniques

Plant tissue culture techniques

1. Topic: PLANT TISSUE CULTURE TECHNIQUE AND THEIR APPLICATIONS Applied Biotechnology (For II- M.Sc .,Zoology students-2019-2020) Dr.S.Selvaraj Assistant professor Dept.of Zoology Vivekananda college Tiruvedakam - west

2. Synopsis: 1.0 Introduction 2.0 Structure and growth of a plant 3.0 Types of culture 4.0 Technique of plant tissue culture 4.1 callus culture 4.2 suspension culture 5.0 Applications 6 .0 Reference

3. 1.0 Introduction: In vitro cultivation of plants, seeds and various parts of the plants. The cultivation process is invariably carried out in a nutrient culture medium under aseptic conditions. Unlike animal cells, highly mature and differentiated plant cell retain the ability of totipotency. Advantages: It is largely helped in the green revolution by improving the crop yield and quality. It cultures has contributed to the understanding of metabolism, growth, differentiation and morphogenesis of plant cells. To produced several pathogen-free plants, besides the synthesis of many biologically important compounds including pharmaceuticals.

4. 2.0 Structure and growth of a plant: An adult plant – both root and stem are characterized by apical growth regions which are composed of meristematic cells. Primary sources of all type of plants. It can growth 2 ways namely, Determinate growth – Ceastion of growth as the plant parts attain certain size and shape. E.g , leaves, flowers, fruits. 2. Indeterminate growth – continuous growth of roots and stems under suitable conditions – meristem can proliferate continuously . Terms used in tissue culture: Explant – It initiated from multicellular tissue fragments, called explants, obtained from living plants. plant tissues, such as leaf, stem, root, petiole, hypocotyl, cotyledon, embryo, or meristem. The explant is selected it is either haploid or diploid explant .

5. The plant growth can be achieved in two ways: 1. Shoots directly by appropriate media, 2.By somatic embr y oge n e sis. Callus – unorganized and undifferentiated mass of plant cell. E.g., mass of parenchymatous cells. Dedifferentiation – mature cells reverting to meristematic state to produce callus. When suitable culture medium. Redifferentiation – ability of the callus cell to differentiate into a plant organ or whole plant.

6. 3.0 T ypes of Plant tissue culture: Callus culture – differentiated tissues from explant which dedifferentiates in vitro to form callus. Organ culture – it used may be embryo, seed, root, endosperm, anther, ovary, ovule, meristem. Organized organ culture - well organized structure of a plant is used, organ is maintained and original. b) Un organized organ culture – the isolation of cells or tissue of a part of the organ and their culture in vitro.(callus) Cell culture – It is isolated individual cells, obtained from an explant tissue or callus is regarded as cell culture. Carried out dispension medium and as cell suspension culture. Protoplast culture – also cultured in laboratory condition. Organ culture Callus culture Cell culture Protoplast culture PTC

7. 4.0 Basic technique of plant tissue culture: The collection of explant – trimmed and to sterilized in a detergent solution – after washing with sterile distilled water, it placed at suitable culture medium and incubated – results in establishment of culture. Mother culture can be subdivided, as frequently as needed, to give daughter culture. Most important aspect of in vitro technique is carried out all the process id aseptic condition. Bacteria and fungi is most contaminants in PTC and certain toxic compounds. They grow much faster in culture and often kill plant tissue – absolutely essential that aseptic condition and maintained throughout the tissue culture.

8. Selection of explant Trimming Surface Sterilization Washes in sterilized with distilled water Establishment of culture Incubation Subculture Regeneration Hardening Transfer of plant into the green house or open field Explant (Bud, Root, Seed Fig: A diagrammatic of the outline procedure in plant tissue culture

9. 5.0 Callus culture technique: Undifferentiated and unorganized mass of plant cells. Tumor tissue which usually forms on wounds of differentiated tissues or organs. Callus cells are parenchymatous in nature although not truly homogenous. Callus formation in vitro is frequently - cut edges of stems or roots. Invasion of microorganisms or damage by infect feeding usually occurs through callus. Explant for Callus culture: T he starting materials for callus culture may be differentiated tissue from any part of the plant . Explant may be different stages of cell division, cell proliferation and organization into distinct specialized structures. Explant possess meristematic cells, then cell division and multiplication will be rapid .

10. Fig: Steps involved by using Callus culture technique

11. Plant regeneration from tissue culture varies with the following parameters: plant species, genotype within the species, source of the cultured tissue, age and health of the donor plant, nutrient medium, other factor Factors Affecting Plant Tissue Culture Efficiency

12. 6 .0 Applications of Plant tissue culture: To study the respiration and metabolism of plants . For the evaluation of organ functions in plants. Various plant diseases and work out method for their elimination. Single cell clones are useful for genetic, morphological and pathological studies. Embryonic cell suspension can be used for large cell clonal propagation. The production of soma clonal variations . Production of haploids and Mutant cells can be selected from cultures and used for crop improvement. Immature embryos can be cultured in vitro to produce hybrids, a process referred to as embryo rescue.

13. Thank you 7 .0 Reference: 1. Biotechnology, U. Satyanarayana, 2010, Pgs – 497 to 503.