Plant Tissue Culture Lecture Notes PDF

Lecture 1 a Introduction to plant cell tissue culture Prepared and presented by : Dr. Abdelaziz Mohamed Nasr Lesson Objectives Understand plant biotechnology and its effect on modern agriculture. Define plant tissue culture and identify its applications as...

Lecture 1 a Introduction to plant cell tissue culture Prepared and presented by : Dr. Abdelaziz Mohamed Nasr Lesson Objectives Understand plant biotechnology and its effect on modern agriculture. Define plant tissue culture and identify its applications as well as its disadvantages. Identify the different types of plant tissue cultures. Grades distribution Final theoretical: 40 Midterm: 10 Final practical: 20 Theoretical coursework: 20 (2 quizzes, 1 project) Practical coursework: 10 (2 quizzes) Total= 100 marks Plant biotechnology Application of science and technology to plants or their parts, to alter living or nonliving materials for the production of knowledge, goods, and services. Encompasses a multitude of scientific tools and techniques for screening and genetic manipulation of plants to develop beneficial or useful plant/plant products. Plant biotechnology New plant variety / trait has been developed. Past: Traditional breeding techniques was used (selection, hybridization/crossing) Present: Plant tissue culture techniques. Definition of plant tissue culture The use of isolated parts of plants, called explants, obtained from an intact plant body and kept on or in a suitable nutrient medium. This nutrient medium functions as a replacement for the cells, tissue, or conductive elements originally neighboring the explant. Such experimental systems are usually maintained under aseptic conditions. Some exceptions to this are experiments concerned with problems of phytopathology, or with the co-culture studies of cell material to study symbiosis. Definition of plant tissue culture The whole concept of plant tissue culture is based on Totipotency and Plasticity. Totipotency: the ability of any plant cell to convert into a whole plant if appropriate conditions of nourishment are provided. Plasticity: The ability of an organism to change its phenotype in response to different environments.  Brief history of plant tissue culture 1756 Henri Louis observed that 1964 when elm plants 1902 1926 Guha and are injured, an Gottileb Fritz W. Went 1946 Maheshwari unorganized Haberlandt Discovery of first Ernest Ball Production of mass of cells proposed the plant growth Plant production haploid plants forms at the principle of hormone, from shoot tip from anther wound site totipotency indole acetic acid culture culture 1878 1922 1939 1962 1970 Vöchting Kotte and Gautheret and Murashige and Power et. al observed Robbins Nobecourt; Skoog Fusion of that the upper Cultured root worked Optimized isolated plant part of the shoot and stem tips independently medium for the protoplasts produces buds Introduction of a growth of plants and the basal callus culture under in vitro part produces conditions with callus or roots higher salt concentrations Importance of plant tissue culture Improvem Production ent Micropropagated Embryo plants culture Secondary Anther and pollen metabolites culture Genetic Synthetic seeds transformation Protoplast fusion  Cryopreservation, Somaclonal variations, and breaking seed dormancy. Rules for explant selection Prerequisite parameters essential for establishing tissue culture of any plant species are the selection of superior-quality, disease-resistant/free plant, and explant type according to the need. For commercial multiplication of plants, shoot tips or nodal segments from the juvenile phase are used. For direct or indirect organogenesis or somatic embryogenesis, young leaves (especially in vitro leaves) are used. In vivo explants other than seed, shoot tip, or nodal segments are avoided for the culture establishment as the tissue becomes highly differentiated and forcing them into dedifferentiation and re-differentiation may impart physiological stress to the explant. Types of plant tissue cultures A. Organized Structure B. Unorganized Culture Structure Culture 1. Meristem culture 1. Callus culture 2. Shoot tip culture 2. Suspension culture 3. Node culture 3. Protoplast culture 4. Isolated root culture 4. Anther culture 5. Embryo culture Shoot and root apical meristems  A. Organized Structure Culture 1. Meristem Culture 2. Shoot tip Culture 3. Node Culture Excised shoot apices, each started from excised shoot tips, Separate lateral buds, each consisting of the apical or buds, larger than the shoot carried on a small piece of stem meristematic dome with or apices employed to establish tissue; stem pieces carrying without one or two leaf meristem cultures, having either single or multiple nodes primordia. several leaf primordia. may be cultured. A. Organized Structure Culture 4. Isolated Root Culture 5. Embryo Culture The growth of roots, unconnected Fertilized or unfertilized zygotic to shoots: a branched root (seed) embryos are dissected system may out of developing seeds or fruits be obtained. and cultured in vitro until they have grown into seedlings.  B. Unorganized Structure Culture 1. Callus Culture 2. Suspension cell Culture 3. Protoplast Culture The growth and maintenance of Populations of plant cells and The culture of plant cells that largely unorganized cell masses, small cell clumps, dispersed in have been isolated without a cell which arise from the disorganized an agitated, wall. growth of small plant organs, that is aerated, liquid medium pieces of plant tissue, or previously cultured cells. B. Unorganized Structure Culture 4. Anther Culture The culture of complete anthers containing immature pollen microspores. The objective is usually to obtain haploid plants by the formation of somatic embryos.  Advantages of plant tissue culture 1. Rapid Clonal Propagation: Tissue culture allows for the rapid production of numerous genetically identical plants from a single parent plant. This is particularly useful for propagating valuable or rare plant species. 2. Year-Round Propagation: Unlike traditional methods of plant propagation, tissue culture can be carried out year-round, regardless of the season or weather conditions. 3. Disease-Free Plants: Plants produced through tissue culture are typically free from diseases, pests, and pathogens because the culture starts from aseptic (sterile) explants. This is crucial for producing healthy and disease-resistant plants. 4. Preservation of Rare and Endangered Species: Tissue culture can be used to preserve and propagate rare and endangered plant species, helping in their conservation efforts. 5. Genetic Modification: Tissue culture provides a platform for genetic modification and transformation of plants, allowing for the introduction of desirable traits such as resistance to pests, diseases, or environmental stress. 6. Production of Secondary Metabolites: Tissue culture can be employed to produce valuable secondary metabolites, such as pharmaceuticals and essential oils, from plant cells. 7. Storage and Banking of Genetic Material: Plant tissue culture can be used for the long-term storage and banking of genetic material, including seeds, embryos, and meristems, ensuring genetic diversity is preserved.  Disadvantages of plant tissue culture 1. Labor-Intensive: Tissue culture requires a high level of technical expertise and labor, making it relatively expensive and less accessible to small-scale growers. 2. Genetic Uniformity: While genetic uniformity is an advantage in some cases, it can also be a disadvantage as it may lead to a lack of genetic diversity, making plants susceptible to new pests and diseases. 3. Risk of Somaclonal Variation: Somaclonal variation refers to genetic variations that can occur in tissue-cultured plants, leading to unexpected traits or mutations. This can be problematic when uniformity is desired. 4. Cost: Setting up and maintaining a tissue culture facility can be costly due to the need for specialized equipment, sterile conditions, and skilled personnel. 5. High Failure Rate: The success rate of tissue culture can vary depending on the plant species and the specific protocol used. Some plants are more difficult to culture than others, leading to a high failure rate in certain cases. 6. Environmental Impact: The energy and resource requirements of tissue culture facilities can have an environmental impact, particularly if they rely on artificial lighting and climate control. 7. Ethical Concerns: There can be ethical concerns related to the genetic modification of plants through tissue culture, especially when it involves transgenic or genetically modified organisms (GMOs). What is the take-home message from today’s lecture. Let’s discuss. Pre-reading for next class Plant cell components Plant growth, division and differentiation. Any more questions?