Plant tissue culture is the process of growing plant cells, tissues or organs in an artificial nutrient medium under sterile conditions. It allows for the rapid multiplication of plant cells and the regeneration of whole plants. The key steps involve selecting an explant, sterilizing it, inoculating it onto a nutrient medium, incubating to form callus tissue, subculturing the callus to induce regeneration of plantlets, hardening the plantlets, and transferring them to soil. Plant tissue culture has many applications including micropropagation, clonal propagation, breeding new plant varieties, and producing secondary metabolites. It is an important biotechnology tool in agriculture and horticulture.

1. PLANTTISSUE
CULTURE
PREPARED BY:MISS SMITA R. MANKAR.
ASSISTANT PROFESSOR,
P.R. PATIL INSTITUTE OF PHARMACY TALEGAON SP ,DIST-WARDHA

2. CONTENT
• INTRODUCTION
• HISTORY
• BASIC TERMS IN TISSUE CULTURE •
• DIFFERENT STEPS IN TISSUE CULTURE
• TYPES OF PLANT TISSUE CULTURE
• APPLICATIONS
• USES
• ADVANTAGES

4. INTRODUCTION
• Plant tissue cultureis an in vitro cultureor growth of cells. tissue or organs of
plant in a sterile condition and well formulated media to producean entire
plant.
• The controlledconditionsprovide the culturean environment conducive for
their growth and multiplication.
• These conditionsincludepropersupply of nutrientsmedium, adequate
temperatureand proper gaseou liquid environment.

5. HISTORY OF PLANT TISSUE CULTURE.
• 1902 - Haberlandt proposedconcept of in vitro cell culture•
• 1922 - Kolte and Robbins successfully culturedroot and stem tipsrespectively
• 1926 - Went discovered first plant growth hormone-Indoleacetic acid
• 1939 - Gautheret, White and Nobecourt established endlessproliferationof callus
cultures
• 1957 - Skoog and Miller gave concept of hormonal control (auxin: cytokinin) of
organ formation

6. • 1960 - Cocking was first to isolate protoplast by enzymatic degradation of cell wall.
• 1962-Murashige and Skoog developed MS medium with higher salt concentration.•
• 1964-Guha and Maheshwari producedfirst haploid plantsfrompollen grains of Datura
(Antherculture).
• 1970 - Power et al, successfully achieved protoplast fusion.
• 1971-Takebeet al.regenerated first plantsfrom protoplasts.
• 1981-Larkin and Scowcroft introducedthe term somacionalvariation.

7. BASIC TERMS USED IN TISSUE CULTURE.
• Explant
• An excised piece of differentiated tissue or organ is regarded as an explant. The explant
may be taken from any part of plant body e.g.,leaf, stem, root.
• Callus
• The unorganized and undifferentiated mass of plant cells is to as callus. Generally when
plant cells are culturedin a medium, they divide to form callus.

8. • De-differentiation
• The phenomenon of mature cells reverting to meristematic state toproduce callus is
dedifferentiation.
• Re-differentiation
• The ability of the callus cells to differentiate into a plant organ or awhole plant is regarded as
re-differentiation.
• Totipotency
• The ability of an individual cell to develop into a whole plant is referred to as totipotency.

9. TYPES OF PLANT TISSUE CULTURE
• Seed Culture.
• Embryo Culture.
• Callus Culture.
• Organ Culture.
• Protoplast Culture.
• Anther Culture.

12. STEPS INVOLVED IN PLANT TISSUE CULTURE.
• 1.Preparation of suitable nutrient medium:
• MS medium, White's medium, B5 medium are few culture media that can be obtained
from the market for the lab preparation of nutrient medium needed for tissue culture.
• The culture medium consists of: Inorganic nutrients:2 types of inorganic nutrients are
used in the culture medium.
• 6 macronutrients like nitrogen, phosphorus, sulphur, calcium, potassium and
magnesium and 6 micronutrients like iron, molybdenum, copper, zinc, boron and
manganese are also added in the medium.

13. • Organic nutrients:
• Carbon and nitrogen containing compounds are used. •
• Sucrose, fructose and other carbohydrates are the carbon source. •
• Vitamins and amino acids are the nitrogen source.
• plant growth hormones :Auxins, Gibberellins, Cytokinins,Abscisic acid, Ethylene etc. are some
examples of plant hormones.Auxins, cytokinins and Gibberellins are the most commonly used growth
hormones in culture medium prepared for tissue culture.
• Agar: It solidifies the medium and acts as a substratum. It doesn't have any nutritive importance. It is
obtained from sea weeds and is used to provide solid surface for growth. •All these important nutrients are
incorporated in the culture medium and transferred into suitable containers and then sterilized.

14. • 2.Selection of explant:
• Explants are any excised part of the plant to be used in tissue culturelike axillary buds, leaf and stem segments,
root tip, shoot tip, anther,Ovary and endosperm.. •Always young and healthy parts of plants are selected as
explants.
• 3.Sterilization of the explant:
• Explants are sterilized by using different types of disinfectants like sodium hypochlorite (2%), silver nitrate
(1%) calcium hypochlorite (9- 10% ), bromine water (1-2 %), mercuric chloride (0.1-1%), hydrogen peroxide
etc. and different types of antibiotics,
• 4.Inoculation of explant:
• The sterile explant is inoculated on the surface of the solidified nutrient medium under aseptic (free from
contamination) condition. •Laminar air flow can be used for this inoculation process.

15. • 5.Incubation:
• Incubation is done at 25+/-2 °C, with a relative humidity of 50-60%with 16 hours of photoperiod. •
• After defined period of incubation, an unorganized and undifferentiated (no root and shoot) mass of cells
called callus is obtained from each explant. In some cases, root and shoot directly develop from the
explant after incubation.
• 6. Transfer of callus (subculture) and regeneration:
• The callus obtained from incubation of explant is transferred intoanother medium and reincubated.
•Plantlet regenerates from the callus with the induction of roots and shoot directly from the callus.

16. • 7.Hardeningor acclimatization:
• Hardening is the gradual exposure of the plantletsfor acclimatization or adjustment to the
environmental conditions.
• 8.Transferof plantletsto the field:
• After the hardeningprocess, plantletsare transferred to either the green house or the field
conditionsoutside. •These plantletswill grow into independent and healthyplant

17. STERILIZATION OF EXPLANT

18. PREPARATION OF CULTURE MEDIA

19. PLANTTISSUE CULTURETECHNIQUE

20. FACTORS AFFECTING TISSUE
CULTURE
• Growth media
• • Minerals, carbon source, vitamins , hormones.
• • Environmental factors
• • Light, temperature, photoperiod, humidity.
• • Explant source
• • Usually younger ,less differentiated, explant is used for tissue culture

21. BENEFITS OF PLANTTISSUE CULTURE
• Plant tissue culture is one of the most rapidly growing areas of biotechnology because of its
high potential to develop improved crops and ornamental plants. With the advances made in
the tissue culture technology, it is now possible to regenerate species of any plant in the
laboratory.
• To achieve the target of creating a new plant or a plant with desired characteristics, tissue
culture is often coupled with recombinant DNA technology. The techniques of plant tissue
culture have largely helped in the green revolution by improving the crop yield andquality.

22. • The knowledge obtained from plant tissue cultures has contributed to our understanding of
metabolism, growth, differentiation and morphogenesis of plant cells. Further, developments
in tissue culture have helped to produce several pathogen-free plants, besides the synthesis
of many biologically important compounds, including pharmaceuticals. Because of the wide
range of applications, plant tissue culture attracts the attention of molecular biologists, plant
breeders and industrialists

23. APPLICATIONS OF PLANT TISSUE CULTURE
• 1. As a Source of Micro Propagation
• 2. As a Source of Clonal Propagation:
• 3. Productionof GeneticallyVariable Plants with New Traits:
• 4. Used for Plant Breeding, Plant Improvement and Plant Tissue Culture:
• 5. As Source of SecondaryMetabolite: