Plant Tissue Culture Basics
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Questions and Answers

Match the following terms with their descriptions:

Apical shoot = Tip of the plant that is responsible for growth Callus = Undifferentiated plant tissue Explants = Tissue taken from a plant for culture Somatic embryos = Embryos that develop from somatic cells

Match the macronutrients with their chemical formulas:

Nitrogen (N) = KNO3 and NH4NO3 Phosphorus (P) = KH2PO4 Potassium (K) = K2SO4 Calcium (Ca) = CaCl2. 2H2O

Match the types of PGRs (Plant Growth Regulators) with their categories:

Auxins = 2,4-D, IAA, IBA, NAA Cytokinins = BAP, Kinetin, Zeatin Abscisic Acid = Inhibits growth and promotes dormancy Gibberellins = Promotes stem elongation and seed germination

Match the micronutrients with their chemical components:

<p>Iron (Fe) = FeSO4. 7H2O Manganese (Mn) = MnSO4. 4H2O Copper (Cu) = CuSO4. 5H2O Zinc (Zn) = ZnSO4. 7H2O</p> Signup and view all the answers

Match the stages of organogenesis in plant tissue culture:

<p>Dedifferentiation = Begins after isolation of explant Redifferentiation = Formation of new tissue types Organogenesis = Development of organs from tissue culture Callus formation = Development of an undifferentiated mass</p> Signup and view all the answers

Match the types of tissue cultures with their usage:

<p>Batch cultures = Maintained as friable callus Mass cultures = Used for large-scale propagation Cell suspension cultures = Used for secondary metabolite production Meristem cultures = Used for disease-free plant propagation</p> Signup and view all the answers

Match the terms related to plant tissue cultures:

<p>Maintained as aggregated = Refers to the grouping of cells Cell wall removal = Can be performed mechanically or enzymatically Tissue culture = Technique for growing plants in controlled environments Somatic breeding = Breeding techniques using somatic cells</p> Signup and view all the answers

Match the following concepts with their descriptions:

<p>Somaclonal variation = Genetic differences from the same culture process generating mutant phenotypes Epigenetic changes = Modifications like histone methylation affecting gene expression Reactive Oxygen Species (ROS) = Produced under stress conditions affecting cellular functionality Chloroplasts = Primary source of ROS under light conditions</p> Signup and view all the answers

Match the following stress responses with their outcomes:

<p>Excess ROS = Degradation of pigments and proteins NADPH-dependent-oxidases = Response mechanism to stress conditions Homeostasis disturbance = Loss of cellular functioning during stress Mitochondria = Source of ROS in dark conditions</p> Signup and view all the answers

Match the following processes with their examples:

<p>DNA sequence changes = SNPs and chromosomal alterations Gene amplification = Common genetic change observed in regenerants Transposition = Movement of genetic elements within the genome Fusarium wilt resistance = Application of somaclonal variation in banana</p> Signup and view all the answers

Match the following terms related to plant tissue culture with their definitions:

<p>Biochemical causes = Factors leading to mutations in culture Physiological causes = Stress responses that affect plant tissues Genetic causes = Intrinsic mutations in plant DNA during culture Plant regeneration = Process that can be impaired by ROS and epigenetic changes</p> Signup and view all the answers

Match the following terms with their definitions:

<p>Dedifferentiation = A process allowing mature cells to revert to an undifferentiated state Redifferentiated cells = Cells that have lost the ability to divide after differentiation Transdifferentiation = Developmental changes between different cell lineages Meristem = Tissue capable of continuous growth and cell division</p> Signup and view all the answers

Match the following stages of cell development:

<p>Callus formation = Initial stage of undifferentiated mass of cells Dedifferentiation = Cells regain the ability to divide Redifferentiation = Process where cells begin to specialize again Transdifferentiation = Direct conversion of one cell type to another</p> Signup and view all the answers

Match the following types of meristem with their functions:

<p>Vascular cambium = Increases the thickness of stems and roots Cork cambium = Forms protective outer layers in plants Wound meristem = Facilitates healing and regeneration Apical meristem = Promotes vertical growth of the plant</p> Signup and view all the answers

Match the following statements to the concepts of cell differentiation:

<p>Increased developmental potency = A characteristic of dedifferentiated cells Callus mass = Result of accelerated cell division Chromatin organization change = Accompanies dedifferentiation Loss of division ability = Describes redifferentiated cells</p> Signup and view all the answers

Match the following terms related to plant tissue culture (PTC):

<p>Budding = Another term for redifferentiation in PTC Callus = Mass of undifferentiated cells formed during tissue culture Meristematic activity = Characteristic of dedifferentiated tissues Developmental changes = Can occur through transdifferentiation</p> Signup and view all the answers

Match the following processes with their characteristics:

<p>Dedifferentiation = Cells revert to a state allowing for division Redifferentiation = Cells begin the process of specialization Transdifferentiation = Involves a shift between cell lineages Potency increase = A feature of dedifferentiated tissues</p> Signup and view all the answers

Match the following types of cells with their context in plant biology:

<p>Callus cells = Undifferentiated cells that enable tissue culture Dedifferentiated cells = Cells that can act like meristem Redifferentiated cells = Cells that have specialized after division Transdifferentiated cells = Cells that change lineage type without division</p> Signup and view all the answers

Match the following characteristics related to cellular responses:

<p>Undifferentiated state = Achieved through dedifferentiation Differential specialization = Described by transdifferentiation Callus cell formation = Initial response to tissue culture Meristematic features = Exhibited by dedifferentiated tissues</p> Signup and view all the answers

Match the agricultural biotechnology pros with their descriptions:

<p>Reduction of pesticides application = Lower environmental impact Transgenic plants with better nutritional quality = Enhanced food value Production of proteins for therapeutic use = Biopharmaceutical manufacturing Mass propagation of selected plants = Increased crop availability</p> Signup and view all the answers

Match the ethical considerations of agricultural biotechnology with their potential impacts:

<p>Risk of escapes = Gene transfer to wild species Increased genetic erosion = Loss of biodiversity Insect-resistant plants = Impact on beneficial insects Patent monopolies = Ethical concerns over gene ownership</p> Signup and view all the answers

Match the topics regarding GMOs with their focus areas:

<p>Biosafety regulations = Safety measures for GMOs Testing for commercialization = Assessment of GMO viability SPS laws = Trade regulations for agricultural products Diagnostic techniques = Methods for identifying GMOs</p> Signup and view all the answers

Match the benefits of transgenic crops with their effects:

<p>Insect-resistant traits = Reduce pest damage Herbicide-resistant traits = Simplify weed management Improved crop yield = Increase food production Diverse agricultural biodiversity = Enhanced ecosystem resilience</p> Signup and view all the answers

Match the authors with their contributions in the field of plant biotechnology:

<p>Slater, A. = Plant genetic manipulation Thorpe, T.A. = Plant tissue culture history Fowler, M.R. = Advancements in crop science Scott, N.W. = Transgenic plant studies</p> Signup and view all the answers

Match the following terms with their definitions:

<p>Dedifferentiation = Increased developmental potency Callus = Mass of undifferentiated parenchyma type cells Transdifferentiation = Overproliferation of differentiated cells Somatic embryogenesis = Development of embryos from somatic cells</p> Signup and view all the answers

Match the process with its description:

<p>Organogenesis = Initiating development of shoot or root primordia Callus formation = Result of dedifferentiation Direct regeneration = From the same organ type Indirect regeneration = Through callus phase</p> Signup and view all the answers

Match the following auxins with their specific types:

<p>2,4-D = Dichlorophenoxyacetic acid IAA = Indole acetic acid NAA = Napthylacetic acid BAP = 6-Benzylaminopurine</p> Signup and view all the answers

Match the plant growth regulators with their category:

<p>Kinetin = Cytokinin Zeatin = Cytokinin Auxins = Growth hormones Cytokinins = Cell division regulators</p> Signup and view all the answers

Match the plant tissue culture steps with their order:

<p>Initiation of culture = I Multiplication/sub-culturing = II Rooting = III Hardening = IV</p> Signup and view all the answers

Match the advantages of tissue culture with their explanations:

<p>Fast propagation = Large volumes of mature plants produced quickly Cloning endangered species = Safe propagation of vulnerable plants Genetically identical plants = Large quantities produced Improved agriculture = Disease- and pest-free plants in sterile vessels</p> Signup and view all the answers

Match the following concepts with their respective usages:

<p>Auxin ratio for shoots = High auxin and high cytokinin Auxin ratio for roots = High auxin and low cytokinin Cytokinin activity = Promotes cell division Auxin activity = Stimulates root formation</p> Signup and view all the answers

Match the following explant types with their roles:

<p>Leaf = Direct use for embryo production Meristem = Source of regenerative cells Callus = Intermediate form in indirect regeneration Somatic cells = Source for somatic embryogenesis</p> Signup and view all the answers

Match the following terms with their related concepts:

<p>Auxins = Promoters of cell elongation Cytokinins = Inhibitors of senescence Callus = Proliferating mass of dedifferentiated cells Wound responses = Initiation of dedifferentiation</p> Signup and view all the answers

Match the following types of regeneration:

<p>Direct regeneration = From existing organs Indirect regeneration = From callus formation Embryonic regeneration = Through somatic embryogenesis Tissue culture = In vitro plant propagation</p> Signup and view all the answers

Match the following cultivation techniques with their characteristics:

<p>Tissue culture = Propagation in sterile conditions Callus culture = Development of undifferentiated cells Organ culture = Culture of specific organ parts Protoplast culture = Direct regeneration from cells</p> Signup and view all the answers

Match the following concepts with their respective phases:

<p>Proliferation = Rapid growth of callus Differentiation = Formation of specialized cells Meristem culture = Source of new shoots Somatic tissue culture = Production of whole plants</p> Signup and view all the answers

Match the following plant growth regulator roles:

<p>Auxins = Stimulate root growth Cytokinins = Promote shoot development Gibberellins = Initiate seed germination Ethylene = Regulate fruit ripening</p> Signup and view all the answers

Match the following tissue culture steps with their functions:

<p>Initiation = Setting up the culture environment Multiplication = Increasing plant material Rooting = Encouraging root formation Hardening = Preparing plant for soil transfer</p> Signup and view all the answers

What happens to homeostasis under stress?

<p>It is not preserved.</p> Signup and view all the answers

Which organelles are primary sources of ROS under light conditions?

<p>Peroxisomes</p> Signup and view all the answers

What is somaclonal variation?

<p>Genetic differences from the same culture process which generates mutant phenotypes.</p> Signup and view all the answers

What is a common example of genetic change affecting somaclonal variants?

<p>SNPs, gene amplification, transposition, and chromosomal alterations.</p> Signup and view all the answers

What disease resistance is being studied in bananas using somaclonal variation?

<p>Fusarium wilt resistance.</p> Signup and view all the answers

What was one of the benefits of agricultural biotechnology as of 2008?

<p>Mass propagation of selected and healthy plants</p> Signup and view all the answers

Which of the following is a potential ethical concern of transgenic plants?

<p>Risk of escapes of new genes</p> Signup and view all the answers

How many hectares of transgenic crops were reported in 2008?

<p>125 million ha</p> Signup and view all the answers

What is plant tissue culture (PTC)?

<p>A system for propagating plants vegetatively using small parts of living tissues on defined artificial growth media under sterile conditions.</p> Signup and view all the answers

What are the main requirements for cells in plant tissue culture, as defined by White in 1939?

<p>Cells must remain undifferentiated yet capable of unlimited growth.</p> Signup and view all the answers

Who developed the concept of in vitro cell culture?

<p>Gottlieb Haberlandt.</p> Signup and view all the answers

What does micropropagation involve?

<p>Producing whole plants from different tissues</p> Signup and view all the answers

The ability of a cell to grow and develop into a fully differentiated organism is known as ______.

<p>totipotency</p> Signup and view all the answers

Match the following components of tissue culture media with their descriptions:

<p>Macronutrients = Essential mineral elements for growth Plant Growth Regulators (PGRs) = Substances that regulate plant growth Organic Base = Supplies amino acids and vitamins Fixed Carbon = Source of energy for plant cells</p> Signup and view all the answers

What is callus culture associated with?

<p>Solid tissue culture media</p> Signup and view all the answers

Plant tissue culture can be performed in the absence of seeds.

<p>True</p> Signup and view all the answers

What are auxins and cytokinins used for in plant tissue culture?

<p>They are plant growth regulators that promote different types of growth and development in cultures.</p> Signup and view all the answers

Somaclonal variation can lead to undesirable traits in plants.

<p>True</p> Signup and view all the answers

What is the purpose of sterilization in tissue culture?

<p>To eliminate contaminants and ensure a pure and sterile environment for plant growth.</p> Signup and view all the answers

What is the main cause of undesirable traits in callus cultures?

<p>Genetic instability</p> Signup and view all the answers

Somaclonal variation can only be produced through environmental stresses.

<p>False</p> Signup and view all the answers

What type of genetic changes may affect regenerants in somaclonal variants?

<p>SNPs, gene amplification, transposition, and chromosomal alterations</p> Signup and view all the answers

Chloroplasts and peroxisomes are primary sources of __________ under light conditions.

<p>ROS</p> Signup and view all the answers

Match the following terms related to somaclonal variation with their descriptions:

<p>SNPs = Single nucleotide polymorphisms affecting DNA sequence Transposition = Movement of DNA segments within the genome Gene amplification = Increase in the number of copies of a gene Chromosomal alterations = Changes in the structure or number of chromosomes</p> Signup and view all the answers

Which of the following is a benefit of agricultural biotechnology?

<p>Production of proteins for industrial or therapeutic use</p> Signup and view all the answers

Herbicide-resistant transgenic plants can help in increasing biodiversity.

<p>False</p> Signup and view all the answers

What is one risk associated with the introduction of new genes into plants?

<p>Escape of new genes to other plant species.</p> Signup and view all the answers

In 2008, there were _______ million hectares of transgenic crops reported.

<p>125</p> Signup and view all the answers

Match the benefits of agricultural biotechnology with their descriptions:

<p>Reduction of pesticide application = Leads to lower environmental impact Improved nutritional quality = Enhances the health benefits of crops Mass propagation = Allows for the production of large quantities of plants Regulation of trade = Ensures safety and quality of agricultural products</p> Signup and view all the answers

What is an explant in the context of tissue culture?

<p>Any portion of leaf, shoot, stem, root, or living plant part used for regeneration</p> Signup and view all the answers

Younger and rapidly growing tissues are not suitable for explants in tissue culture.

<p>False</p> Signup and view all the answers

What are the three basic components of a tissue culture medium?

<p>Essential mineral elements, organic base, and fixed carbon</p> Signup and view all the answers

Tissue culture media contains approximately ______% water.

<p>95</p> Signup and view all the answers

What is the process that begins shortly after the isolation of explant tissues in plant tissue culture?

<p>Dedifferentiation</p> Signup and view all the answers

Succrose is included as a micronutrient in typical media composition for plant tissue culture.

<p>False</p> Signup and view all the answers

Match the types of tissue cultures with their descriptions:

<p>Callus culture = Used with solid TC media to initiate cell suspension Suspension culture = Associated with liquid TC conditions Protoplast culture = Cell walls removed for suspension Microspore culture = Uses haloid pollen or anthers as explants</p> Signup and view all the answers

Which of the following is a macronutrient needed for plant cell growth?

<p>Potassium</p> Signup and view all the answers

What two phases are involved in organogenesis during plant tissue culture?

<p>Dedifferentiation and redifferentiation</p> Signup and view all the answers

The primary growth regulators that promote cell division in plant tissue culture are called __________.

<p>Cytokinins</p> Signup and view all the answers

Fixed carbon in the tissue culture medium is commonly supplied as glucose.

<p>False</p> Signup and view all the answers

Name one type of culture that is usually associated with solid tissue culture media.

<p>Callus culture</p> Signup and view all the answers

Which plant growth regulator is often used for root formation in tissue culture?

<p>IAA</p> Signup and view all the answers

Match the following macronutrients with their chemical formulas:

<p>Nitrogen = KNO3 and NH4NO3 Phosphorus = KH2PO4 Potassium = K2SO4 Calcium = CaCl2. 2H2O</p> Signup and view all the answers

Callus can be maintained as friable in batch cultures.

<p>True</p> Signup and view all the answers

List one organic supplement that can be included in typical plant tissue culture media.

<p>Thiamine (Vitamin B1)</p> Signup and view all the answers

What is the primary result of dedifferentiation in plant tissue culture?

<p>Formation of callus</p> Signup and view all the answers

Callus formation can only occur indirectly through the use of explants.

<p>False</p> Signup and view all the answers

Name one advantage of using tissue culture in plant propagation.

<p>Fast propagation of new cultivars</p> Signup and view all the answers

The term used to describe the overproliferation of differentiated cells leading to callus formation is called __________.

<p>dedifferentiation</p> Signup and view all the answers

Match the type of plant growth regulator (PGR) with its example:

<p>Auxin = 2,4-D Cytokinin = BAP Gibberellin = GA3 Abscisic Acid = ABA</p> Signup and view all the answers

Which of the following is NOT a phase of somatic embryogenesis?

<p>Direct use of seeds</p> Signup and view all the answers

Cytokinins are used predominantly for root formation in tissue culture.

<p>False</p> Signup and view all the answers

What is the main purpose of meristem tip culture?

<p>To produce disease-free plants</p> Signup and view all the answers

In the presence of __________, callus can form when using specific media compositions in tissue culture.

<p>auxin and cytokinin</p> Signup and view all the answers

What is the role of auxins in plant tissue culture?

<p>Encourage root formation</p> Signup and view all the answers

Regeneration of whole plants from cultured cells can happen in a single step.

<p>True</p> Signup and view all the answers

What is one method of initiating organogenesis in vitro?

<p>Directly from the same organ type</p> Signup and view all the answers

The process of developing embryos from somatic cells is called __________.

<p>somatic embryogenesis</p> Signup and view all the answers

Which hormone is commonly associated with inducing shoot formation?

<p>Cytokinins</p> Signup and view all the answers

Plant tissue cultures can be established without sterilization.

<p>False</p> Signup and view all the answers

Study Notes

Plant Tissue Culture (PTC) Methods

  • Apical shoot tips are used as explants to generate callus, essential for somatic breeding.
  • Callus can be friable and maintained in batch cultures; cell wall removal is achieved mechanically with enzymes.

Typical Media Composition (MS)

  • Macronutrients: KNO3, NH4NO3, KH2PO4, MgSO4, CaCl2, etc.
  • Micronutrients: FeSO4, MnSO4, ZnSO4, CoCl2, CuSO4, etc.
  • Carbon source: Sucrose is commonly used.
  • Organic supplements: Thiamine (Vitamin B1), Myoinositol, amino acids.
  • Plant Growth Regulators (PGRs): Auxins (2,4-D, IAA, IBA, NAA) and Cytokinins (BAP, Kinetin, Zeatin).

Differentiation and Dedifferentiation

  • Organogenesis involves dedifferentiation (formation of callus from mature explants) and redifferentiation (development of primordia).
  • Dedifferentiated tissues can act as meristems (e.g., vascular cambium, cork cambium).
  • Transdifferentiation allows changes between cell lineages without altering cellular potency.

Callus Formation

  • Callus is a mass of undifferentiated parenchyma cells, formed often in the dark.
  • Overproliferation or transdifferentiation of cells results in callus; dedifferentiation increases developmental potency.

Organogenesis

  • Initiation of shoot or root primordia can occur directly from organs or indirectly via callus.
  • In direct organogenesis, growth arises from the same organ type, while indirect involves an intermediary callus phase.

Somatic Embryogenesis

  • Embryos are developed from somatic cells, either directly from leaf tissues or indirectly through callus formation.

Plant Growth Regulators

  • Auxins: 2,4-D, IAA, NAA play critical roles in growth regulation.
  • Cytokinins: BAP, Kinetin, and Zeatin also influence growth patterns.
  • Ratios of Auxins and Cytokinins determine growth direction (shoots or roots).

Typical Tissue Culture Steps

  • Initiation of culture in vitro
  • Multiplication via sub-culturing on shooting media
  • Rooting on rooting medium
  • Hardening for acclimatization to soil

Advantages of PTC

  • Rapid propagation of new cultivars or endangered species.
  • Production of genetically identical plants and eliminating seeds.
  • Cultivation of disease-free plants enhances agricultural practices.

Somaclonal Variation

  • Callus culture may lead to undesired mutations due to biochemical and genetic factors.
  • Useful in plant breeding to develop resistant variants.

Epigenetic Factors in PTC

  • Epigenetic changes like histone methylation can affect tissue culture outcomes.
  • Stress during culture regeneration can lead to reactive oxygen species (ROS) accumulation, impacting cellular functions.

Somaclonal Variants

  • Variants arise from genetic differences during culture, leading to mutations such as SNPs and chromosomal alterations.

Case Study: Fusarium Wilt Resistance

  • Somaclonal variation used to create banana plants resistant to Fusarium wilt.

Agricultural Biotechnology Benefits

  • Transgenic crops enhance agricultural efficiency.
  • Reduction in pesticide use and improvement in crop nutritional quality.
  • Mass propagation of healthy plants and better regulation of GMO testing.

Ethical Considerations

  • Potential risks include gene escape, new virus development, and reduced biodiversity.
  • Concerns about patent monopolies on genetically modified organisms and possible ecological impacts on beneficial insect populations.

Learning Outcomes

  • Understand the history and evolution of plant tissue culture (PTC).
  • Distinguish between solid and liquid culture techniques.
  • Describe various methods used in plant tissue culture.
  • Analyze the roles of auxins and cytokinins in plant cultures.
  • Familiarize with key terminology such as plasticity, totipotency, explant, and callus.
  • Outline the essential steps in the tissue culture process.
  • Explore the diverse applications of plant tissue culture.

Importance of PTC

  • PTC is vital for regenerating whole plant clones from transformed isolated plant cells or tissues.
  • In vitro approach ensures high regeneration frequency, although not necessarily linked to transformation frequency.

Definition and Concept

  • PTC involves propagating plants vegetatively using small living tissue parts (explants) in sterile artificial media.
  • Micropropagation refers to producing whole plants from minimal plant parts like shoot/root tips and embryos.

Historical Development

  • Gottlieb Haberlandt pioneered in vitro cell culture in 1902, exploring isolated plant cells.
  • White established continuous callus culture and introduced vitamins in the 1930s.
  • Skoog and Miller proposed hormone-controlled organogenesis in 1957, demonstrating auxin's effect on rooting.
  • Murashige and Skoog developed the widely used MS medium in 1962, improving tobacco tissue growth by five times.
  • Advancements in somatic hybridization and protoplast fusion occurred in the 1980s.

Key Concepts in PTC

  • Totipotency: A cell's ability to develop into a fully differentiated organism under suitable conditions.
  • Plasticity: Adaptation of plant cells to environmental changes through metabolic adjustments.
  • Explant: Any portion of a plant (leaf, stem, root) used to initiate cultures, favoring rapidly growing tissues.

Components of Tissue Culture Media

  • Contains essential mineral salts, organic supplements, and carbon sources like sucrose.
  • Standard media includes macronutrients (N, P, K), micronutrients (Fe, Mn, Zn), and plant growth regulators (PGRs).

Types of Culture

  • Callus Culture: Associated with solid media used to initiate cell suspension cultures.
  • Suspension Culture: Involves liquid media and is utilized for friable aggregated callus.

Process of Differentiation and Dedifferentiation

  • Involves transitioning phases:
    • Dedifferentiation: Explant isolation leads to accelerated cell division forming callus.
    • Redifferentiation: Callus cells specialize into various tissues, either directly or indirectly.

Organogenesis

  • The process of initiating shoot or root primordia can occur through direct or indirect pathways from the explant or callus.
  • Somatic Embryogenesis: Formation of embryos from somatic cells, potentially through direct or indirect methods (using a callus phase).

Plant Growth Regulators

  • Auxins: Key compounds include 2,4-D, IAA, NAA promoting root development.
  • Cytokinins: Include BAP, kinetin, zeatin, influencing shoot growth.

Tissue Culture Steps

  • Sterilize and select tissues, initiate culture in vitro, multiply explants, root cultures, and finally harden plants for soil transfer.

Advantages of PTC

  • Rapid production of mature plants facilitating breeding and conservation of endangered species.
  • Generation of genetically identical plants and improvement of desirable traits.
  • Enables disease and pest-free plant distribution enhancing agricultural outcomes.

Somaclonal Variation

  • Genetic mutations in long-term callus cultures can lead to beneficial variations.
  • Screened for unique traits, aiding in developing resistant or tolerant plants.

Epigenetics in PTC

  • Epigenetic changes influenced by methylation/demethylation in plant tissues during culture can affect growth and regeneration.

Ethical Considerations

  • Risks include gene escape to wild species, creation of new viruses, reduction in biodiversity, and pest resistance impacting beneficial insects.

Applications in Agriculture

  • Transgenic crops can reduce pesticide use while enhancing nutritional quality.
  • Mass propagation of disease-resistant varieties helps improve agricultural sustainability and biodiversity.

Somaclonal Variation

  • Callus cultures can lead to undesirable traits through mutation, affected by biochemical, genetic, and physiological factors.
  • Somaclonal variation facilitates the screening of unique biotypes useful in plant breeding and improving crop traits.
  • Mutation plant breeding with tissue culture accelerates development of resistant and herbicide-tolerant plants, reducing traditional breeding cycles from 10-15 years.

Epigenetics in Plant Tissue Culture (PTC)

  • Epigenetic changes, such as histone methylation/demethylation, influence plant traits.
  • Stress during tissue culture regeneration disrupts organelle function, leading to reactive oxygen species (ROS) accumulation which can degrade essential cell components like proteins and DNA.

Genetic Changes in Somaclonal Variants

  • Somaclonal variants display genetic differences within plants from the same culture process, resulting in mutant phenotypes.
  • Common genetic alterations include DNA sequence changes like SNPs, gene amplification, transposition, and chromosomal alterations.

Case Study: Banana and Fusarium Wilt

  • Somaclonal variation is utilized to develop banana plants that are resistant to Fusarium wilt, enhancing sustainability in banana cultivation.

Tissue Culture Explants

  • Explants can come from various plant parts such as leaves, shoots, stems, and roots, with younger tissues preferred for regeneration.
  • Regeneration involves transferring explants to artificial media enriched with hormones that promote differentiation.

Components of Tissue Culture Medium

  • Essential elements include macronutrients (like nitrogen, phosphorus, potassium) and micronutrients (like iron, manganese) in complex salt mixtures.
  • Organic bases provide amino acids and vitamins, while sucrose is a common carbon source, with tissue culture media containing approximately 95% water.

Culture Types

  • Callus culture typically uses solid media; suspension culture utilizes liquid conditions.
  • Various tissue types can be cultured, including protoplast, embryo, microspore, root, and shoot tip cultures.

Dedifferentiation and Differentiation

  • Organogenesis consists of dedifferentiation of explant cells followed by redifferentiation into specific plant structures.
  • Callus formation, characterized by undifferentiated cell mass, occurs as a result of overproliferation, often when auxin and cytokinin are presented together.

Regeneration Processes

  • Two regeneration pathways: direct (from the same organ type) and indirect (via callus formation).
  • Somatic embryogenesis allows development of embryos directly from somatic (non-reproductive) cells.

Plant Growth Regulators

  • Key auxins include 2,4-D, IAA, and NAA; important cytokinins include BAP, kinetin, and zeatin.
  • Auxin and cytokinin ratios are crucial for determining whether root or shoot formation occurs.

Advantages of Tissue Culture

  • Rapid production of large volumes of genetically uniform plants can support fast cultivar propagation and cloning of endangered species.
  • Enables regeneration of disease-free plants, creation of genetically modified organisms (GMOs), and improvement of plant traits.

Agricultural Biotechnology Benefits

  • Significant area (125 million ha) of transgenic crops reported in 2008, leading to reduced pesticide use and improved nutritional quality in crops.
  • Mass propagation of healthy plants contributes to agricultural productivity in both developed and developing countries.

Ethical Considerations in Biotechnology

  • Risks include gene escape, potential development of new viruses, genetic erosion, and unintentional insect resistance.
  • Patenting of genetic materials may lead to monopolies and associated moral dilemmas.

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Description

This quiz covers fundamental concepts related to plant tissue culture, including apical shoot explants, callus formation, and the maintenance of friable callus in batch cultures. It emphasizes the significance of these techniques in somatic breeding and the manipulation of plant cell walls.

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