Tissue Culture: Definition, Types, Steps, Examples, Diagram, Techniques
Tissue culture is defined as a technique used to grow cells, tissues, or organs in a controlled, sterile environment. It allows scientists to produce thousands of identical plants from a single parent plant, making it a powerful tool in plant propagation. This method is especially useful in agriculture as part of strategies for enhancement in food production, helping in the rapid multiplication of high-yielding and disease-resistant crops.
- What is Tissue Culture?
- Types of Tissue Culture
- Steps of Tissue Culture
- Importance of Plant Tissue Culture
- Applications of Tissue Culture
- Advantages and Disadvantages of Tissue Culture
- MCQs on Tissue Culture
- Frequently Asked Questions (FAQs)
This technique has wide applications in biotechnology, where it supports genetic engineering, conservation of rare species, and production of improved plant varieties. Tissue culture also plays a vital role in research and commercial plant production, offering uniformity and quality assurance. Its precision and efficiency make it a valuable tool in modern scientific and agricultural practices.
What is Tissue Culture?
It is a refined biological technique used in growing and maintaining cells, tissues, or organs outside their natural environment, usually in a controlled laboratory atmosphere. In this method, a small portion of tissue, known as an explant, is taken from a plant, animal, or microbial source. The explant is transferred into a sterile state in a nutrient rich medium. The culture medium contains all the necessary nutrients, growth factors, and hormones to be used in conditions that promote optimum growth and development.
It is done by creating a controlled environment in tissue culture, which would provide the optimum setting for cell proliferation and differentiation. The researchers thereby carry on studies of cellular behaviour, conduct genetic manipulations, and grow large amounts of uniform cells or tissues that are used in various applications. Tissue culture assumes a central role in joining many advances in biotechnological, medical, agricultural, and environmental conservation domains through studies relating to complex biological activities or improvements in agriculture and therapeutic new remedies.
Types of Tissue Culture
Tissue culture techniques can vary depending on the part of the plant used and the purpose of the culture. These types allow scientists to grow and maintain cells in controlled environments. Each method has specific conditions and applications. The types of tissue culture can be broadly categorised into:
Organ Culture
Organ culture is when intact organs or parts of organs are grown under controlled conditions outside the body of an organism. The technique helps in maintaining the structural and functional properties of organs so that researchers can further elucidate growth, development, and responses to different stimuli. Organ cultures are used by investigators dealing with plants and animals. Plant organ cultures are applied to study development of the root system, flowering patterns, and responses to various environmental stresses. Organ cultures in animals contribute to biomedical research by studying functions of organs, mechanisms of various diseases, and treatments.
Cell Culture
Cell culture is concerned with the isolation and in vitro growth of single cells or small groups of cells. It is one of the most frequently employed techniques not only throughout biomedical sciences but also in the development of pharmaceuticals and genetic engineering. Cell cultures may be derived from animal tissues, plant tissues, or microbial cultures. They are used to study cellular behaviours like proliferation, differentiation, metabolism, response to drugs or toxins, etc. Tissue culture techniques have become an essential tool in the generation of cell lines, assessment of toxicity tests, and development of therapies against various diseases.
Plant Tissue Culture
Plant tissue culture has a variety of techniques which are oriented towards the propagation and manipulation of plant cells, tissues, or organs in an aseptic environment. It finds extensive applications in agriculture and horticulture for the mass production of disease-free plants, effective multiplication of elite cultivars, and conservation of endangered species. Applications in this field are mostly attributed to micropropagation, somatic embryogenesis, and genetic transformation. Through the tissue culture techniques, such as the development of genetically modified crops that have increased resistance to several pests, diseases and environmental stresses, plant tissue culture has completely revolutionised agricultural practice.
Animal Tissue Culture
Animal tissue culture is cultivated in a laboratory for research study purposes, testing of pharmaceutical compounds, and medical uses. This forms a very prominent area in understanding cellular physiology, mechanisms of various diseases, and drug responses in the controlled environment of a laboratory. Animal tissue cultures have applications in the development of cell-based assays, the production of vaccines and therapeutic proteins, and the modelling of human diseases in drug discovery and biomedical research. Primary isolation of cells directly from tissues and immortalised cells, otherwise cell lines, are some of the techniques applied in studying a complex biological process and developing novel treatments.
Microbial Culture
Microbial culture is the process of growing and maintaining microorganisms such as bacteria, fungi, algae, etc., in laboratory conditions. It is very important in studies on microbial physiology, genetics, and biochemical pathways. Cultures find application in aspects connected with biotechnology, the production of food, and the control of the environment. They also turn out to be very useful tools in producing antibiotics, enzymes, and biofuels through microbial fermentation processes.
Steps of Tissue Culture
The process involves a sequence of steps to ensure successful growth. These steps must be performed in sterile conditions to avoid contamination. Growth media and controlled conditions are essential at each stage. Proper care is required throughout the process.The steps involved in the process of tissue culture are:
Explants and Preparation
Type of Explants Used: Standard explants include apical meristem, embryos, and callus tissues. Each type is applied for certain purposes of tissue culture, like rapid proliferation or genetic transformation.
Techniques of Preparation: These involve surface sterilisation, cutting, and precise dissection so that the explants are properly rid of contaminants, and ready to set in for culture.
Culture Media
Media Composition: Culture media are composed of some of the basic requirements or nutrients, which involve macro and micronutrients, vitamins, amino acids, and growth regulators like auxins and cytokinins. All these components are added in a balanced way for the cells to grow and differentiate.
Preparation and Sterilisation Methods: Media is prepared by mixing ingredients under sterile conditions. Sterilisation consists of passing media through a filter or autoclaving. This helps to avoid any kind of microbial contamination.
Culture Conditions
Environmental Control: The temperature, humidity, pH, and CO2 are optimised for successful tissue culture. This supposedly provides the most analogous environment that cells require to proliferate and differentiate.
Incubation and Maintenance: Cultures are incubated in a controlled environment, like growth chambers or bioreactors, where the growth of the cells will proceed and their progress with time is to be traced.
Importance of Plant Tissue Culture
Tissue culture spans disciplines such as biology, agriculture, and medicine because of its ability to grow cells and tissues in a controlled environment outside their natural context. In agricultural practices, it provides large-scale production of disease-free plants and propagation of plants that are facing extinction or are genetically altered to have new characteristics. Hence, advancing agricultural productivity and sustainability initiatives.
The importance of plant tissue culture has been a large tool in medicine in studying the mechanisms of many diseases, responses to drugs, and tissue regeneration. This creates the ability needed for the production of cell lines and three-dimensional tissue models, advancing biomedical research into innovations in personalised medicine and pharmaceutical development. Apart from that, tissue culture plays a very vital role in improving food security, understanding human health, and driving forward scientific discovery.
Applications of Tissue Culture
Tissue culture is widely used in agriculture, horticulture, and scientific research. It helps in rapid plant multiplication and conservation of endangered biodiversity. It also supports genetic studies and the development of disease-free plants. Tissue culture and its applications are described below-
Micropropagation: It can produce numerous identical plants or clones rapidly for crops with desirable traits, for example resistance to certain diseases, high yield, etc. Tissue culture can also yield plants produced from disease-free meristematic tissues.
Conservation of Rare and Endangered Plants: Tissue culture conserves plant species, whether they are rare, endangered, or difficult to propagate with seeds, by manufacturing multiple plants in a controlled environment.
Genetic Modification and Research: One of the other uses of tissue culture is that it enables scientists to make genetic manipulations in the plant cells, thus developing genetically modified plants rich in certain desirable traits, like resistance to pests or increased nutrition.
Advantages and Disadvantages of Tissue Culture
This technique has many benefits, including fast propagation and uniform quality. However, it also comes with limitations like high cost and the need for technical expertise. Both strengths and challenges exist in this method.The advantages and disadvantages of tissue culture are listed below-
Merits of Tissue Culture
Tissue culture allows swift and efficient propagation of plants and cells from a limited population. Thus creating an opportunity for mass production of disease-free plants and contributing to the conservation of rare species. This can facilitate genetic variations to develop crops with desirable traits, thus enhancing agricultural productivity and fertility. It reduces the risk of contamination under aseptic conditions, hence guaranteeing predictable results for experiments.
Challenges and Limitations
It is expensive in terms of special apparatuses and artificial media, the mastery of which requires handling delicate tissues. Despite strict handling, there is a risk of contamination, which can be very frustrating and decrease experimental reliability. A lack of access to resources and technical skills to a large extent may limit its applications, especially in developing regions or smaller research facilities.
MCQs on Tissue Culture
Q1. What is the advantage of producing somaclones through tissue culture?
The ability to produce plants with desirable characteristics
The production of plants that are genetically identical to the original plant
The ability to recover healthy plants from diseased plants
The production of hybrid plants with desirable traits
Correct answer: 2) The production of plants that are genetically identical to the original plant
Explanation:
Somaclones produced through tissue culture are genetically identical to the original plant from which they were grown. This allows for the production of large numbers of plants with desirable characteristics, such as disease resistance or increased yield.
Hence, the correct answer is option 2) The production of plants that are genetically identical to the original plant.
Q2. What is the primary advantage of micropropagation in tissue culture?
It allows for the production of genetically modified plants.
It enables the rapid propagation of a large number of plants.
It facilitates the introduction of foreign DNA into plant cells.
It improves the nutritional quality of the cultured plants.
Correct answer: 2) It enables the rapid propagation of a large number of plants.
Explanation:
The primary benefit of employing micropropagation via tissue culture in the botanical context is its capability to generate a substantial quantity of clonal, genetically homogeneous plants within a brief temporal frame, originating from a minute segment of plant tissue. This technique's salient advantages include:
1. Mass Propagation: It enables the swift and extensive production of flora on a grand scale, which is particularly beneficial in agriculture and horticulture.
2. Disease-Free Plants: The utilization of meristematic tissue allows for the creation of pathogen-free specimens, which is crucial for maintaining plant health and productivity.
3. Uniformity: As all the plants produced are genetic replicas of the parent, this ensures a consistent expression of desirable traits among them.
4. Conservation: Micropropagation plays a vital role in the preservation and propagation of rare or threatened species, contributing significantly to biodiversity conservation efforts.
5. Year-Round Production: It circumvents the limitations imposed by seasonal and environmental factors, thus facilitating continuous cultivation throughout the year.
These distinctive features render micropropagation an excellent choice for applications in agriculture, horticulture, and environmental conservation domains.
Hence, the correct answer is option 2) It enables the rapid propagation of a large number of plants.
Q3. To obtain virus-free healthy plants from a diseased one by tissue culture technique, which part/parts of the diseased plant will be taken?
Apical meristem only
Palisade parenchyma
Both apical and axillary meristems
Epidermis only
Correct answer: 3) Both apical and axillary meristems.
Explanation:
Both apical and axillary meristems are free of virus for healthy plant cultivation because of strong interferon activity in this region. These tissues form a protective impermeable covering around themselves, which is non-penetrable by any pathogen. Therefore, these tissues are used in the production of disease-free plants by tissue culture.
Hence, the correct answer is option 3) Both apical and axillary meristems.
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Frequently Asked Questions (FAQs)
Q1. What is tissue culture?
Tissue culture is the technique of growing plant or animal cells in a nutrient medium under sterile conditions.
Q2. What are the three types of tissue culture?
The three main types are organ culture, cell culture, and callus culture.
Q3. What are the applications of tissue culture?
It is used in cloning, crop improvement, conservation of rare plants, and production of disease-free plants.
Q4. Who is the father of tissue culture?
Gottlieb Haberlandt is known as the father of tissue culture.
Q5. What are the principles of tissue culture?
It is based on the principle of totipotency, aseptic conditions, and the use of suitable growth media.
Frequently Asked Questions (FAQs)
In medicine, tissue culture helps in studying a disease, vaccine preparation, and the production of many therapeutic proteins and drugs through cell-based systems.
The advantages are rapid propagation and genetic manipulation. The disadvantages are high cost, technical complexity of work, and vulnerability to contamination.
Through tissue culture techniques, variations in genes can be studied, genes manipulated, and organisms that are identical in genes can be used for experiments and commercial purposes.
Tissue culture aids conservation by allowing the propagation of rare or endangered species, preserving genetic diversity, and producing large numbers of plants for reintroduction to the wild or for sustainable use, reducing pressure on wild populations.
Synthetic seeds are artificially encapsulated somatic embryos or other vegetative propagules. They combine the advantages of tissue culture (rapid multiplication, disease-free plants) with the ease of handling and storage of traditional seeds. Synthetic seeds can be used for propagation, conservation, and easy transport of valuable plant material.
Somaclonal variation, which occurs during tissue culture, can be both beneficial and problematic in plant breeding. It can generate new genetic variability, potentially leading to improved traits. However, it can also cause unwanted changes in clonally propagated plants. Plant breeders can exploit this variation to develop new cultivars or use techniques to minimize it when clonal fidelity is desired.
Tissue culture facilitates the production of transgenic plants by providing a controlled environment for introducing foreign DNA into plant cells and regenerating whole plants from transformed cells. It allows for the selection of successfully transformed cells and their subsequent development into complete transgenic plants.
Micropropagation is a technique in plant tissue culture used to produce large numbers of genetically identical plants from a single parent plant. It involves growing plants from small pieces of tissue or single cells under sterile conditions.
Cryopreservation is a technique used to preserve tissue culture samples by storing them at extremely low temperatures (usually in liquid nitrogen). This allows long-term storage of valuable genetic material and reduces the need for continuous subculturing, which can lead to genetic changes over time.
In animal tissue culture, cellular totipotency is limited compared to plants. While some animal cells retain the ability to differentiate into multiple cell types (pluripotency), true totipotency is generally only observed in early embryonic cells. This limitation affects the types of tissues and organs that can be generated through animal tissue culture.
The explant source is crucial as it determines the genetic makeup and potential of the cultured tissue. Different plant parts (e.g., leaves, stems, or meristems) may have varying regeneration capacities and genetic stability, affecting the success and outcome of the culture process.
Aseptic technique is crucial in tissue culture to maintain a sterile environment free from microorganisms. It involves using sterilized equipment, working in a laminar flow hood, and following strict protocols to prevent contamination. Without proper aseptic technique, cultures can easily become contaminated, leading to failure of the experiment or loss of valuable plant material.
Somatic hybridization involves fusing protoplasts (cells without cell walls) from different plant species to create hybrid cells that can be regenerated into whole plants. Unlike traditional breeding, which relies on sexual reproduction, somatic hybridization can combine genomes of distantly related species that cannot naturally interbreed.
Agar serves as a solidifying agent in tissue culture media, providing physical support for the growing tissues. It also helps maintain the proper water potential and allows for the slow diffusion of nutrients, creating a stable environment for cell growth.
Sterilization is essential in tissue culture to prevent contamination by microorganisms like bacteria or fungi. These contaminants can outcompete the cultured cells for nutrients and space, potentially ruining the entire culture.
Growth regulators, such as auxins and cytokinins, are crucial in tissue culture for controlling cell division, differentiation, and organ formation. The balance of these hormones determines whether cells will form roots, shoots, or remain as undifferentiated callus.
Callus is an undifferentiated mass of cells that forms when plant tissues are cultured. It's often the starting point for regenerating whole plants or producing specific plant products. Callus formation is induced by manipulating growth hormones in the culture medium.
Explant selection is crucial as it affects the success and efficiency of tissue culture. Factors to consider include the age and health of the donor plant, the type of tissue used, and its developmental stage. Younger, actively growing tissues often yield better results than older, mature tissues.
The culture medium provides all the necessary nutrients, vitamins, minerals, and growth regulators for cell growth and development. Its composition is carefully tailored to the specific needs of the cultured tissue and the desired outcome of the culture process.
The main types of tissue culture are:
Primary culture refers to cells taken directly from living tissue and cultured for the first time. Secondary culture, or cell line, is derived from a primary culture after the first subculture. Secondary cultures can often be maintained for longer periods and may have different characteristics from the original tissue.
Organogenesis involves the formation of specific plant organs (like roots or shoots) from cultured tissue, while embryogenesis results in the formation of complete embryos that can develop into whole plants. Organogenesis typically produces plants identical to the parent, while embryogenesis can lead to more genetic variation.
Habituation in plant tissue culture refers to the ability of cultured plant cells to grow and divide without the addition of exogenous growth regulators. This phenomenon occurs when cells develop the capacity to produce their own growth hormones, often after prolonged culture periods. Habituation can affect the regeneration potential and genetic stability of cultured tissues.
Plant growth regulators, particularly auxins and cytokinins, play a crucial role in somatic embryogenesis. High auxin levels often induce embryogenic callus formation, while a reduction in auxin and increase in cytokinin can trigger the development of somatic embryos. The precise balance and timing of these regulators guide the embryogenic process.
Tissue culture is a technique where plant or animal cells, tissues, or organs are grown in a controlled artificial environment outside of their original organism. This method allows for the study, manipulation, and propagation of biological material under sterile conditions.
Cellular competence refers to the ability of plant cells to respond to developmental signals and undergo morphogenesis. In tissue culture, not all cells are equally competent to form new structures. Understanding and manipulating cellular competence is crucial for successful plant regeneration and can involve factors such as explant choice, culture conditions, and growth regulator treatments.
The choice of explant can significantly affect genetic stability in tissue culture. Explants from highly differentiated tissues (like leaves) may be more prone to genetic changes during dedifferentiation and redifferentiation processes. In contrast, explants from meristematic regions (like shoot tips) often maintain greater genetic stability due to their naturally dividing state.
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