Plant Tissue Cculture

Plant Tissue Culture:

Plant Tissue Culture Dr. Vijay Gulkari

Plant biotechnology and its products –including transgenic plants is the most assimilated technology for both farmers and companies. . :

Plant biotechnology and its products –including transgenic plants is the most assimilated technology for both farmers and companies. .

In the last few decades plant biotechnology applications have been widely developed and incorporated in many countries worldwide:

In the last few decades plant biotechnology applications have been widely developed and incorporated in many countries worldwide .

HISTORY OF PLANT TISSUE CULTURE:

HISTORY OF PLANT TISSUE CULTURE 1838-39 cellular theory (Cell is autonom and totipotent ) Schleiden-Schwann 1902 First attempt of plant tissue culture Harberlandt 1939 Continuously growing callus culture White 1946 Whole plant developed from shoot tip Ball 1950 Organs regenerated on callus Ball 1954 Plant from single cell Muir 1960 Protoplast isolation Cocking

HISTORY OF PLANT TISSUE CULTURE ……continue:

HISTORY OF PLANT TISSUE CULTURE ……continue 1962 MS media Murashige - Skoog 1964 Clonal propagation of orchids Morel 1964 Haploids from pollen Guha 1970 Fusion of protoplasts Power 1971 Plants from protoplasts Takebe 1981 Somaclonal variation Larkin

The potential of plant tissue culture tools is the key factor of support:

The potential of plant tissue culture tools is the key factor of support Tissue culture may be defined as the aseptic culture of cells, tissues, organs or whole plants under controlled nutritional and environmental conditions

Types of In Vitro Culture:

Types of In Vitro Culture Culture of intact plants (seed and seedling culture) Embryo culture (immature embryo culture) Organ culture 1. shoot tip culture 2. root culture 3. leaf culture 4. anther culture Callus culture Cell suspension culture Protoplast culture

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PLANT TISSUE CULTURE OR MICROPROPAGATION Applications : Conservation of endangered plants Large-scale propagation of horticultural plants Extraction of medicinally important compounds Improvement of agricultural crops Genetic engineering

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Two Hormones Affect Plant Differentiation: Auxin: Stimulates Root Development Cytokinin: Stimulates Shoot Development Generally , the ratio of these two hormones can determine plant development:  Auxin ↓Cytokinin = Root Development  Cytokinin ↓Auxin = Shoot Development Auxin = Cytokinin = Callus Development Basis for Plant Tissue Culture

Factors Affecting Plant Tissue Culture:

Factors Affecting Plant Tissue Culture Growth Media Minerals, Growth factors, Carbon source, Hormones Environmental Factors Light, Temperature, Photoperiod, Sterility, Media Explant Source Usually, the younger, less differentiated the explant, the better for tissue culture Genetics Different species show differences in amenability to tissue culture In many cases, different genotypes within a species will have variable responses to tissue culture; response to somatic embryogenesis has been transferred between melon cultivars through sexual hybridization

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The development of tissue culture techniques rest upon two properties of plant cells: cell totipotence and cell plasticity

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Cell totipotentiality is the genetically retained capacity that all living cells posses to originate a new genetically identical cell, and, after cellular division and differentiation processes, to be able to form tissues, organs, systems and complete individuals. It implies that all the information necessary for growth and reproduction of the organism is contained in the cell

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Cellular plasticity is the characteristic which marks the difference between plant and animal cells in their capacity of multiplication, division, differentiation and formation of a new individual

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The micropropagation of plants through in vitro culture can be broadly put under Five fundamental stages . Stage 0 Preparation of donor plant Stage I Introduction and establishment Stage II Propagation of plants Stage III Rooting and explant preparation for the ex vitro conditions Stage IV Ex vitro adaptation or plant acclimatization

Stage 0 Preparation of donor plant:

Stage 0 Preparation of donor plant In vitro introduction and establishment depends on the physiological and phytosanitary qualities of the plant,.

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….morph-physiological status and quality

Stage I Introduction and ……..:

Stage I Introduction and …….. Superficial disinfection by Fungicide ,bactericide and insecticide treatments

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Control of emission of phenolic compounds by the tissues. To minimize the production and their oxidation.

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Pretreatments with plant growth regulators

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..establishment.

....establishment:

....establishment

Stage II Propagation of plants :

Stage II Propagation of plants The aim of this phase is to increase the number of units in the tissue culture system until the desired number is obtained

Stage III Rooting and explant preparation for the ex vitro conditions: :

Stage III Rooting and explant preparation for the ex vitro conditions: The rooting stage may occur simultaneously to propagation in the same culture media used for multiplication of the explants.

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In some cases it is necessary to carry out media changes, including nutritional modification

Stage IV Ex vitro adaptation or plant acclimatization   :

Stage IV Ex vitro adaptation or plant acclimatization

plant acclimatization :

plant acclimatization