Plant Tissue Culture Lecture PDF

Applications of Plant Cell and Tissue Culture Agriculture is the backbone of many societies, providing the food, fiber, and resources essential for human survival and economic development. Na by Nazariyah Yahaya Principles of Plant Cell and Tissue Culture Cell Totipotency Con...

Applications of Plant Cell and Tissue Culture Agriculture is the backbone of many societies, providing the food, fiber, and resources essential for human survival and economic development. Na by Nazariyah Yahaya Principles of Plant Cell and Tissue Culture Cell Totipotency Controlled Environment Aseptic Technique Plant cells have the ability to Cultures are grown in a Maintaining sterile conditions regenerate into a whole plant carefully regulated is crucial to prevent from a single cell, known as environment with specific contamination and ensure totipotency. nutrients, lighting, and successful cultures. temperature to support growth. Techniques in Plant Cell and Tissue Culture Callus Culture 1 Inducing undifferentiated cell masses from plant explants. 2 Suspension Culture Growing plant cells in a liquid medium with agitation. Organogenesis 3 Regenerating whole plants from plant organs or tissues.  Products in the pipeline Agronomic benefits Oranges resistant to citrus canker Disease-resistant sweet potatoes Pest- and disease- resistant cassava Disease-resistant bananas  Products in the pipeline Enhanced nutritional qualities “I think in the long term we will have foods that are less hazardous because biotechnology will have eliminated or diminished their allergenicity.” — Steve Taylor, Ph.D. Department of Food Science and Technology, University of Nebraska  Products in the pipeline Functional foods Bananas to deliver a hepatitis vaccine Apples to protect against Respiratory Syncytial virus Potatoes to protect against cholera, E. coli and Norwalk virus  Better for the environment “The results clearly show that soil, air and water quality are enhanced through the responsible use of current biotechnology-derived soybean, corn and cotton crops.” — Teresa Gruber, executive director of the Council for Agricultural Science and Technology (CAST) Banana tissue culture  Pineapple tissue culture 2 3 1 4 7 6 5 Applications of Plant Cell and Tissue Culture Culture Micropropagation Secondary Metabolite Production Rapid multiplication of genetically identical Culturing cells to synthesize valuable plant plantlets. compounds. Genetic Engineering Germplasm Conservation Transforming plant cells to introduce Preserving plant genetic resources through in desirable traits. vitro storage. Micropropagation and Clonal Propagation Shoot Multiplication Root Induction Acclimatization Field Cultivation Multiplication Stimulating the Gradual adaptation of Transplanting and Rapid proliferation of growth of in vitro plantlets to ex growing shoots from adventitious roots on vitro conditions. micropropagated meristematic tissues. plantlets. plants in the field. Clonal Propagation Identical Replication Rapid Multiplication Clonal propagation is the process of This technique allows for the mass creating genetically identical copies of a production of desirable plant varieties, plant through asexual means, such as increasing the availability of high- from cuttings or tissue cultures. quality crops. Preservation of Traits Cloning ensures the consistent expression of valuable genetic traits, such as disease resistance or desirable flavors, across multiple generations of a plant. Advantages of Clonal Propagation Faster Cultivation Uniform Traits Genetic Stability Clonal propagation allows for Cloning ensures that all plants Clones are genetically the rapid multiplication of in a crop share the same identical to the parent plant, plants, accelerating the time it desirable characteristics, eliminating the risk of takes to produce a full crop. improving the consistency and undesirable genetic variations. quality of the final product.  Agricultural applications Clonal propagation and genetic engineering crops A wide variety of plant species can be clonally propagated in vitro from plant tissue It is useful where seed production is difficult Producing virus-eradicated plants  Clonal propagation-Advantage Speed of plant multiplication and quality Uniformity of plants produced within a controlled environment, independently of season and climate. Increases in the resistance of plants to chilling, fungal toxins, ions, salts, pests and disease.  Clonal propagation-Advantage Increase nutritional quality of plants Transwitch technique (antisense DNA or RNA techniques) Identification and cloning a specific gene in the plant cells, the duplicate gene is inserted back into the chromosome by any transformation method. http://www.sciencemag.org/news/2017/05/h ow-transgenic-petunia-carnage-2017-began  Transwitch technique Classic example of RNA silencing. Chalcone synthase (CHS) (Pigmentation modification) Secondary Metabolite Production Cell Selection Identifying high-producing cell lines through screening. Elicitation Inducing stress to stimulate secondary metabolite synthesis. Bioreactor Cultivation Scaling up production using controlled bioreactor systems. Biochemicals and Foods This presentation explores the fascinating world of biochemicals and foods derived from native plants. We'll delve into the production, constituents, and biotransformation of these valuable natural resources, and examine their medicinal and nutritional applications. Biochemical Constituents of Native Plants 1 Phytochemicals 2 Essential Nutrients Native plants are rich in diverse Many native plants are excellent phytochemicals with potential sources of vitamins, minerals, health benefits, such as and other essential nutrients for antioxidants and anti- human and animal nutrition. inflammatory compounds. 3 Unique Compounds Specialized metabolites in native plants may possess novel therapeutic or functional properties for various applications. Extraction and Purification Techniques Solvent Extraction Chromatographic Separation Selective solvents are used to efficiently Advanced purification techniques, such as extract target biochemicals from native column chromatography, can isolate plant materials. individual compounds from complex plant extracts. Membrane Filtration Supercritical Fluid Extraction Membrane-based processes can remove This innovative technique uses carbon impurities and concentrate desired dioxide in a supercritical state to biochemicals, ensuring high purity and selectively extract target compounds. potency. Biotransformation of Native Plant Compounds Enzymatic Conversion Enzymes can be used to catalyze the transformation of native plant compounds into more potent or useful derivatives. Microbial Fermentation Microorganisms, such as bacteria and fungi, can metabolize plant compounds to produce novel biochemicals. Chemical Modification Strategic chemical reactions can alter the structure of native plant compounds to enhance their functionality or bioactivity. Medicinal and Nutritional Applications Cardiovascular Cognitive Function Immune Support Nutritional Value Health Certain native plant Many native plants Native plants are a Native plant-derived compounds possess contain bioactive rich source of biochemicals have neuroprotective compounds that can essential vitamins, been shown to properties and may modulate the minerals, and other support heart health enhance cognitive immune system and nutrients for optimal and reduce the risk performance and promote overall human and animal of cardiovascular brain health. wellness. nutrition. disease. Regulatory Considerations and Certifications Regulatory Compliance Ensuring adherence to local and international regulations for the safe and responsible use of native plant-derived biochemicals. Quality Assurance Implementing rigorous testing and certification processes to guarantee the purity, potency, and efficacy of native plant products. Sustainability Certifications Obtaining certifications that recognize sustainable and ethical cultivation practices for native plant production. Mass production of secondary metabolites using tissue culture Genetic Engineering and Transformation 1 Gene Insertion 2 Regeneration Introducing foreign genes into plant cells Regenerating whole transgenic plants using vectors. from transformed cells. 3 Molecular Analysis 4 Scale-up Verifying and characterizing the transgenic Propagating and cultivating the genetically plants. modified plants. Genetic Engineering in Crops 1 Targeted Modifications 2 Improved Resilience Genetic engineering allows scientists Genetically engineered crops can be to precisely insert, delete, or modify made more resistant to pests, specific genes within a plant's diseases, and environmental stresses, genome to improve desired traits. leading to higher yields and reduced losses. 3 Enhanced Nutritional Value Genetic engineering can be used to increase the production of beneficial nutrients, vitamins, and other compounds in crops. Techniques in Genetic Engineering Gene Insertion 1 Introducing new genes from other organisms to confer desirable traits. 2 Gene Silencing Suppressing the expression of undesirable genes to improve crop Genome Editing 3 performance. Precisely modifying existing genes using tools like CRISPR to enhance crop characteristics. Benefits of Genetically Engineered Crops Higher Yields Reduced Pesticide Use Enhanced Nutrition Genetic modifications can Engineered crops with built-in Genetic engineering can boost crop productivity, leading resistance can minimize the increase the production of to greater food and resource need for harmful chemical vitamins, minerals, and other supplies. pesticides. beneficial compounds in crops. Challenges and Considerations Environmental Impact Public Perception Regulatory Framework Potential unintended Concerns over the safety and Implementing effective consequences on ethics of genetic policies and guidelines to ecosystems and biodiversity modifications have led to ensure the responsible require thorough risk ongoing public debates. development and use of assessment and regulation. these technologies. Challenges and Future Prospects Somaclonal Variation Genetic instability during long-term culture Recalcitrance Difficulty in regenerating some plant species Scalability Limitations in commercial-scale bioreactor systems Regulatory Hurdles Navigating policies for genetically modified organisms Despite these challenges, plant cell and tissue culture remains a powerful and evolving field with promising future applications in areas like precision agriculture, pharmaceutical production, and ecosystem restoration.  Plant breeding Conventional plant-breeding: aimed to increase yield & crop quality Selection of lines resistant to pest and diseases Development in plant breeding-uses rDNA technology and are already bringing new crops to the marketplace Plant Breeding Concept Plant breeding is the process by which humans change certain aspects of plants over time in order to introduce desired characteristics Increase crop productivity Plant Breeding Methods Conventional breeding Mutation or crossing to introduce variability Selection based on morphological characteres Growth of selected seeds Challenge: reduce the time needed to complete a breeding program Breeding the papaya-Carica papaya Female Hermaphrodite Male  Crossing between the three different sex forms Hermaphrodite (XYh) X Hermaphrodite (XYh) = 1 female: 2 hermaphrodites Female (XX) X male (XY)= 1 female: 1 male Hermaphrodite (XYh) X male (XY)=1 Hermaphrodite, 1 female and 1 male  Papaya breeding (SOLO) Unisexsual plants are difficult to improve by breeding because the males and females are separate The males do not show the characters that are inherent in them and which will appear in the fruit of their progeny. Hermaphrodites-one can be select as the sole parents of known quality (Solo-developed by breeding in Hawaii)  Solo group-Eksotika Small rounded-female fruits Larger and more uniform pear-shaped- hermaphrodites To produce uniform fruit-farmer should practice to cull the female plants Foot-long oblong-shaped (Sekaki) P1-XYh P2-XYh FI Eksotika 2 (Line 19 and Line 20) Frangi/Paiola (F1 Hybrid) Bacteria dieback disease Modern Breeding Tools In vitro culture Genomic tools Genomic engineering Increase of breeding effectiveness and efficiency  Plant breeding Nonrecombinant DNA techniques Somaclonal Protoplast variation fusion  Somaclonal Variation Genetic variability produced by plant tissue culture Generate useful genetic variation to exploit and improve characteristics of crop and ornamental plants – e.g: Corn : herbicide resistance Wheat – grain colour and height Barley: grain yeild Soybean – height, maturity and seed protein and oil contain Tomato – drawft growth habits, early flowering and orange fruit colour Carrot – higher carotene content Oats – increase seed weight, seed number and grain yeilds Potatoes – yeolds Sugarcane – sugar content, yeil and disease resistance Stress resistance such as salt tolerant, draughts tolerant, heavy metal tolerance, insect tolerant and an improved seed quality. https://www.researchgate.net/publication/229407749_Developing_stress_tolerant_p lants_through_in_vitro_selection-An_overview_of_the_recent_progress  How to induce the biotic/abiotic stress resistance crops plant using somaclonal variations? Plant tolerant to abiotic stress can be acquired by applying the selecting agents such as NaCI (for salt tolerance), PEG or mannitol (for drought tolerance). Only explants capable of sustaining such environments survive in the long run and are selected. The selection of somaclonal variations appearing in the regenerated plants may be genetically stable and useful in crop improvement. Abiotic stress-drought, salinity, low or high temperature. Biotic stress-damage by living organism such as insect, virus, fungus, bacteria How to induce the biotic/abiotic stress resistance crops plant using somaclonal variations?  Nonrecombinant DNA techniques Protoplast fusion – Protoplast to be transformed with foreign genes using methods such as microinjection and electroporation – Genetic characteristics of cells from 2 unrelated plant species or genera OR plants from same species but with different genetic characteristics can be combined by fusing 2 protoplast from different genetic backgrounds. – Fusion – using chemicals or electroporation – Genetic materials are being mixed – a hybride cells with new characteristics – Useful: sexually incompatible or cross with difficulty Plant Germ Plasm Banks Germ plasm banks, also known as gene banks, are repositories that store and preserve the genetic diversity of plants. These specialized facilities play a crucial role in safeguarding the invaluable genetic resources essential for global food security and sustainable agriculture. Na  Germplasm storage Genetic meterial of an organisme Plant breeder depends on germplasm of plants. Wild relatives and ancient crops are mostly found in developing countries The plant are tolerent to a variety of insect, pest, bacterial and fungal infection, harsh enviroment conditions, such as drought and other stresses. This is due to the years of genetic selection Plant breeders uses these ancient germplasm to introduce the desirable traits such as insect resistance into modern crop. Crossing susceptible plants with more resistant varieties. Enviromental degradation, urbanization, changing in farminf practices are the major reason for the shortage or ‘extinct’ Importance of Genetic Diversity in Plants Resilience to Pests Adaptability to Breeding for Improved and Diseases Climate Change Traits Diverse gene pools provide Genetic diversity allows Germ plasm banks serve as plants with the genetic plants to adapt to changing a reservoir of valuable adaptability to withstand environmental conditions, genes, enabling plant evolving threats, such as such as temperature breeders to develop new new pests and diseases, fluctuations, drought, and crop varieties with desirable ensuring the long-term extreme weather events, characteristics, such as sustainability of crop crucial for maintaining food higher yields, enhanced production. security in the face of a nutritional value, and changing climate. resistance to abiotic stresses. Objectives and Functions of Germ Plasm Banks Conservation Characterization Germ plasm banks aim to collect, They evaluate and document the unique document, and preserve the genetic traits and characteristics of the stored diversity of plants, safeguarding them for genetic resources, providing valuable current and future use. information for researchers and breeders. Distribution Research Germ plasm banks facilitate the exchange They support various research initiatives, and distribution of plant genetic resources from understanding plant evolution to to scientists, breeders, and other developing new crop varieties, by providing stakeholders, promoting the utilization of access to a diverse array of genetic these valuable resources. materials. Collection and Preservation of Plant Genetic Resources 1 Exploration Germ plasm collectors undertake expeditions to identify and gather plant genetic resources from diverse geographic regions and environments. 2 Documentation Detailed information about the collected samples, including their origin, characteristics, and potential uses, is meticulously recorded and cataloged. 3 Storage The collected plant materials are stored using various techniques, such as seed banking, in vitro culture, and cryopreservation, to ensure long-term preservation. Characterization and Evaluation of Germ Plasm Phenotypic Evaluation Genotypic Evaluation Data Management Germ plasm banks assess Advances in molecular Detailed information about the physical characteristics techniques allow germ the characteristics and of plant samples, such as plasm banks to analyze the performance of germ plasm plant height, leaf shape, and genetic makeup of plant samples is stored in flower color, to document samples, revealing their databases, enabling their diversity. underlying diversity and efficient retrieval and potential for breeding. utilization by researchers and breeders. Utilization of Germ Plasm for Plant Breeding Trait Identification Germ plasm banks provide a diverse array of genetic resources, allowing plant breeders to identify desirable traits, such as disease resistance or high yield potential. Hybridization Breeders can cross-pollinate diverse germ plasm samples to create new genetic combinations, leading to the development of improved crop varieties. Introgression Desirable traits from wild or underutilized germ plasm can be selectively introduced into elite breeding lines, expanding the genetic diversity of commercial crop cultivars. Challenges and Limitations of Germ Plasm Banks 1 Funding Constraints 2 Accessibility Barriers Adequate and sustained funding is Complex regulations and limited crucial for the long-term maintenance infrastructure can hinder the and expansion of germ plasm banks, accessibility and distribution of germ which face budgetary challenges in plasm resources to researchers and many countries. breeders worldwide. 3 Genetic Erosion 4 Technological Limitations The loss of natural habitats and the Advances in storage and displacement of traditional agricultural characterization techniques are needed practices threaten the genetic diversity to ensure the long-term viability and that germ plasm banks aim to preserve. utility of the germ plasm collections. Future Directions and Emerging Technologies Genomics Cryopreservation Bioinformatics Automation Genomic Advancements in Innovative data Automated systems technologies will cryogenic storage management and will streamline the enable more techniques will analysis tools will collection, comprehensive improve the long- enhance the processing, and characterization and term preservation organization, storage of plant utilization of germ of plant genetic accessibility, and genetic resources, plasm resources for resources. utilization of germ increasing crop improvement. plasm data. efficiency. Bioreactors in Plant Tissue Culture Bioreactors are controlled, sterile environments used to grow plant cells, tissues, or organs in a large-scale, efficient manner. They provide optimal conditions for rapid proliferation and differentiation of plant materials, enabling the production of valuable secondary metabolites, pharmaceuticals, and other high-value products. a Types of Bioreactors Stirred-Tank Airlift Bioreactors Packed-Bed Bioreactors Bioreactors Leverage buoyancy and gas Contain immobilized plant Utilize impellers to provide gas flow to circulate the cells or tissues on a solid provide mixing and aeration, culture medium, well-suited matrix, allowing for high cell aeration, supporting suited for plant cell and hairy densities and efficient suspension cultures of plant hairy root cultures. nutrient/waste exchange. plant cells or tissues. Bioreactor Design Considerations 1 Vessel Geometry 2 Impeller/Sparger Design Dimensions and shape impact mixing, mass Influences oxygen transfer, shear stress, transfer, and scalability. stress, and cell growth. 3 Material Selection 4 Automation & Control Biocompatibility, sterilizability, and optical Enables precise monitoring and regulation optical properties are crucial. regulation of key process variables. Monitoring and Control Systems pH Sensors Dissolved Oxygen Probes Maintain optimal pH levels for plant cell Ensure sufficient oxygen supply for aerobic growth and metabolism. plant cell cultures. Biomass Sensors Automated Sampling Track growth and productivity of plant cells or Enable frequent, sterile monitoring of culture cells or tissues in real-time. culture parameters. Oxygen and Mass Transfer Aeration 1 Efficient sparging or agitation to ensure adequate oxygen supply. 2 Oxygen Solubility Influenced by medium composition, composition, temperature, and Mass Transfer Kinetics 3 pressure. Modeling oxygen/nutrient transfer transfer rates to optimize productivity. productivity.  Nutrient Supply and Waste Removal Nutrient Feeding Waste Removal Medium Composition Replenish essential macro- and Eliminate metabolic byproducts and micro-nutrients. to maintain a healthy culture. Optimize formulations to support rapid plant cell growth. growth. Scaling Up Bioreactor Systems Lab-Scale Pilot-Scale Commercial-Scale Proof-of-concept and Increased volume for process optimization studies. process validation. Large-scale production for for industrial applications. applications. Challenges and Future Developments Developments Shear Sensitivity Designing bioreactors that minimize damaging damaging shear forces. Scalability Overcoming challenges in maintaining optimal optimal conditions at larger scales. Automation Advancing sensor technologies and control control systems for greater efficiency. Process Monitoring Developing robust, real-time monitoring techniques for key parameters.