INTRODUCING PLANT TISSUE CULTURE IN THE CLASSROOM CONCEPTS & HISTORICAL PERSPECTIVE Dr. Mike Kane University of Florida
Applications of Plant Tissue Culture Concepts & Terminology Micropropagation: A Historical Perspective Commercial Micropropagation: Present Status Sequential Micropropagation Stages
Plant Tissue Culture In vitro culture of plant cells, tissues, organs and whole plantlets
APPLICATIONS OF PLANT TISSUE CULTURE Eradication of plant pathogens Collection and storage of crops and native plants for breeding (germplasm storage) Production and extraction of valuable chemical products from cultured cells Clonal plant propagation (micropropagation)
Applications of Plant Tissue Culture For Plant Improvement Plant Biotechnology Introduction of novel genes into economically important plant species, resulting in crop improvement and production of novel products in plants Encompasses Plant Genetic Engineering Plant Tissue Culture
Plant Genetic Transformation
APPLICATIONS OF PLANT TISSUE CULTURE Eradication of plant pathogens Collection and storage of crops and native plants for breeding (germplasm storage) Production and extraction of valuable chemical products from cultured cells Plant micropropagation
MICROPROPAGATION Rapid clonal in vitro propagation of plants from cells, tissues or organs cultured aseptically on defined media contained in culture vessels maintained under controlled conditions of light and temperature
MICROPROPAGATION In vitro propagation Tissue culture propagation
SEXUAL REPRODUCTION 1N Mitosis Differentiation Parent #1 Parent #2 Meiosis Meiosis Gamete 1N Gamete (egg) 2N Zygote (1 cell) Cell division Embryo Whole Plant ASEXUAL REPRODUCTION Shoot cuttings Mitosis Differentiation Cell division Donor plant Shoot tips Lateral buds Genetically Identical (Clones)
Basic Micropropagation Steps Whole plant Explant Micropropagation Rooting Acclimatization Whole plant In vivo In vitro culture Ex vitro In vivo In vivo - ( in life ) Experimentation carried out within intact plants under natural conditions In vitro - ( in glass ) Experimentation carried out on plants or tissues maintained in clear glass or plastic vessels under artificial and usually sterile conditions Ex vitro - ( from glass ) Transfer of plants from in vitro conditions into soil or potting mixture
MICROPROPAGATION Small propagule Aseptic conditions Controlled environment Heterotrophic growth Rapid multiplication Greater initial costs MACROPROPAGATION Larger propagule Non-aseptic conditions Less environmental control Photoautotrophic growth Slower multiplication Nominal costs
Applications of Plant Tissue Culture for Clonal Propagation Concepts & Terminology Micropropagation: A Historical Perspective Commercial Micropropagation: Present Status Sequential Micropropagation Stages
Micropropagation: A Historical Perspective How did it all begin?
PLANT TISSUE CULTURE PREHISTORY Duhamel de Monceau 1756 Shoot regeneration from wound callus Callus Duhamel de Monceau Wound healing following debarking of elm
PLANT TISSUE CULTURE PREHISTORY Schleiden 1838 Schwann 1839 Cell Theory Cell basic unit of life Each living cell of a multicellular organism should be capable of independent development if provided with the proper external conditions Totipotency Concept Plant Cell
EARLY ATTEMPTS AT IN VITRO CULTURE Haberlandt 1902 Innate potential of cells Attempted culture of isolated leaf cells Formulated plant tissue culture principles Culture Medium: Mineral salts & glucose Unsuccessful results Haberlandt Eichhornia crassipes
EARLY ATTEMPTS AT IN VITRO CULTURE Knudson 1919 Root cell culture Knudson 1920s Asymbiotic orchid seed germination & culture Concept of in vitro plant production Knudson Orchids Orchid Seedlings Seedling Culture
ORGAN CULTURE WHITE 1934 Long-term tomato root culture Virus eradication following subculture of extreme root tip White Root culture
CALLUS CULTURE 1934 White Gautheret Nobecourt Callus Continuous growing callus cultures
TOWARD COMMERCIAL MICROPROPAGATION (1940s - 1970s) LOO 1945: Shoot-tip culture of Asparagus BALL 1946: Shoot regeneration from isolated shoot-tips Father of Micropropagation BALL Shoot-tip